Qualcomm Incorporated

Wisconsin Alumni Research Foundation

Harvard College

Biomadison, Inc.

Children's Medical Center Corporation

University of California

Harvard Corporation

Stenmark, Paul

Min Dong

Alexei Yurievitch Gorokhov

Edwin Raymond Chapman

Tingfang Ji

Robert D Fish

Dhananjay Ashok Gore

Liang Tao

Lisheng Peng

Arak Sutivong

Paul Stenmark

Pål Erik Gustav Stenmark

Sicai Zhang

Ronnie Per Arne Berntsson

Hemanth Sampath

Ayman Fawzy Naguib

Feifan Yu

Sony Akkarakaran

Aamod Dinkar Khandekar

Avneesh Agrawal

Edward Harrison Teague

Rongsheng Jin

Peng Chen

Sulyman Barkho

Aina He

Andrew Doxey

Sony John Akkarakaran

Ronnie Per-Arne Berntsson

Aamond Khandekar

Aamod c/o Qualcomm Incorporated Khandekar

Alexei c/o Qualcomm Incorporated Gorokhov

Provided herein are agents that bind to the lipid binding groove of Frizzled (FZD). In some embodiments, the agent is a fragment of TcdB corresponding to amino acids 1285-1804 of SEQ ID NO: 1. In some embodiments, binding of the agent to the lipid binding groove of FZD inhibits TcdB entry into cells, and/or inhibits Wnt signaling. Methods of treatinginfection (CDI) and methods of treating cancer are also provided.

Compositions and methods for inhibiting Wnt signaling

Provided herein is a novel homolog ofneurotoxin (BoNT) identified in the bacterial species(BoNT/EN). BoNT/EN variants and chimeric toxins comprising domains from BoNT/EN and other BoNTs are also provided. Further provided herein are methods of producing BoNT/EN and its use, e.g., in therapeutic applications.

neurotoxin (BoNT/EN) and derivatives thereof

Disclosed herein are means for the detection and characterization of neurotoxins such asneurotoxin (BoNT) or tetanus neurotoxin. The present disclosure provides methods for determining potency and activity of neurotoxins in vitro and in vivo. Also disclosed are polypeptides comprising N- and C-terminal fragments of a reporter protein that are split by a linker comprising a neurotoxin cleavage site. Cleavage of the linker by a neurotoxin decreases reporter protein activity, thereby indicating activity of the neurotoxin. Compositions and kits comprising the disclosed polypeptides, nucleic acids comprising nucleotide sequences encoding such polypeptides, and cells expressing such polypeptides are also disclosed.

In vitro and cell based assays for measuring the activity of botulinum neurotoxins

Disclosed herein are means for the detection and characterization of neurotoxins such as botulinum neurotoxin (BoNT) or tetanus neurotoxin. The present disclosure provides methods for determining potency and activity of neurotoxins in vitro and in vivo. Also disclosed are polypeptides comprising N- and C-terminal fragments of a reporter protein that are split by a linker comprising a neurotoxin cleavage site. Cleavage of the linker by a neurotoxin decreases reporter protein activity, thereby indicating activity of the neurotoxin. Compositions and kits comprising the disclosed polypeptides, nucleic acids comprising nucleotide sequences encoding such polypeptides, and cells expressing such polypeptides are also disclosed.

Provided herein are Clostridialneurotoxin (BoNT) polypeptides of a novel serotype (BoNT/X) and methods of making and using the BoNT polypeptides, e.g., in therapeutic applications.

Botulinum neurotoxin and its derivatives

Disclosed herein is a botulinum neurotoxin (BoNT) polypeptide with a modified receptor binding domain (HC) having one or more amino acid mutations that modify the binding of the BoNT to the receptor. Specific mutations and combinations of mutations are also disclosed. Isolated modified HC, polypeptides comprising the modified HC, chimeric molecules, pharmaceutical compositions, and methods of making and using the same are also disclosed. Methods of identifying additional such modified receptor binding domains, are further disclosed.

Engineered botulinum neurotoxin

Disclosed herein are methods for the isolation and purification of a botulinum neurotoxin (BoNT) protein, or a polypeptide comprising a receptor binding domain of BoNT, from a solution. The method comprises contacting the solution containing the protein or polypeptide to a matrix which has attached thereto a non-toxic non-hemagglutinin (NTNHA) under conditions appropriate for binding, washing the matrix to thereby remove unbound materials, and eluting the protein or polypeptide with a solution that dissociates the bound protein from the NTNHA. Conditions appropriate for binding are a pH of less than 7.5 (e.g, 6). Conditions appropriate for dissociation are a pH greater than or equal to 7.5 (e.g., 8). Compositions specific to the methods are also disclosed.

Method for purification and activation of botulinum neurotoxin

Disclosed herein are botulinum neurotoxin (BoNT) polypeptides with a modified BoNT/B4 receptor binding domain (B4-H) having amino acid mutations that modify the binding of the BoNT to the human SytII receptor. Specific mutations and combinations of mutations are disclosed. Isolated modified Hs, polypeptides comprising such modified Hs, chimeric molecules, pharmaceutical compositions, and methods of making and using the same are also disclosed. Methods of identifying additional such modified receptor binding domains, are further disclosed.

Disclosed herein are modifiedneurotoxin (BoNT) polypeptides with a modified receptor binding domain ofserotype B, comprising one or more substitution mutations at positions corresponding 1248 or 1249 in serotype B, strain 1. Specific substitution mutations include I1248F, I1248Y, I1248H, I1248W, V1249W, V1249F, V1249Y, V1249H, I1248W/V1249F, I1248W/V1249Y, I1248W/V1249H, I1248F/V1249Y, I1248F/V1249H, I1248Y/V1249H, I1248F/V1249W, I1248Y/V1249W, I1248H/V1249W, I1248Y/V1249F, I1248H/V1249F, or I1248H/V1249Y. Other substitution mutations are also disclosed. Isolated modified receptor binding domains, chimeric molecules, pharmaceutical compositions, and methods of using the same are also disclosed.

Engineered botulinum neurotoxins

The present disclosure relates to isolated polypeptides that inhibit Wnt signaling, pharmaceutical compositions comprising the isolated polypeptides, and methods of use thereof. Nucleic acids, cells, and methods of production related to the isolated polypeptides and compositions are also disclosed.

Disclosed herein are botulinum neurotoxin (BoNT) polypeptides with a modified receptor binding domain ofserotype B (B-H), comprising one or more substitution mutations corresponding to substitution mutations in serotype B, strain 1, V1118M; Y1183M; E1191M; E1191I; E1191Q; E1191T; S1199Y; S1199F; S1199L; S1201V; or combinations thereof. Specific combination mutations include E1191M and S1199L, E1191M and S1199Y, E1191M and S1199F, E1191Q and S1199L, E1191Q and S1199Y, or E1191Q and S1199F. Other substitution mutations are also disclosed. Isolated modified receptor binding domains, chimeric molecules, pharmaceutical compositions, and methods of using the same are also disclosed.

A signaling channel that punctures traffic channels is used to send signaling, e.g., acknowledgments (ACKs). To send signaling, resources for the signaling channel are determined, e.g., based on a frequency hopping pattern. Signaling is spread with a spreading code (e.g., a Walsh code) to generate spread signaling, which is mapped to the resources for the signaling channel. Each resource may be partitioned into multiple clusters. A signaling message may be mapped to different clusters to achieve diversity. Traffic data may also be mapped to other resources for a traffic channel assigned for use. Traffic data mapped to the other resources for the signaling channel is punctured. The mapped signaling and traffic data are further processed (e.g., for OFDM or SC-FDMA) and transmitted.

Puncturing signaling channel for a wireless communication system

Methods for improving the specificity of assays for botulinum neurotoxins are described. Reporting constructs are provided that include cleavable linker sequence with genetically modified recognition and/or cleavage sites. These linker sequences act as a substrate for a botulinum neurotoxin, with cleavage of the reporting construct providing a detectable signal. When first and second neurotoxins have activity with the same substrate protein, mutation and/or deletion of a recognition and/or cleavage site associated with the second neurotoxin improves the specificity of the detectable signal for the first neurotoxin.

Compositions and methods for improved neurotoxin assays

Methods for the identification of inhibitors of botulinum neurotoxins are described. Cells are provided that are genetically engineered to express peptides that act as a substrate for a botulinum neurotoxin and the provide reporting groups. Spacer sequences between the reporting groups serve to optimize energy transfer between the reporting groups. Characterization of the energy transfer-dependent signal prior to and following exposure to a botulinum neurotoxin in the presence of a candidate inhibitor provides a measure of the effectiveness of the candidate inhibitor.

Growing community of inventors

Madison, WI, United States of America

Min Dong