University of Massachusetts

Prime Bio, Inc.

Other

Bal Ram Singh

B.b. Scientific L.l.c.

Institute of Advanced Sciences, Inc.

Raj Kumar

Shashi Kant Sharma

Zhong Zhang

Tadanori Tomita

Yu Zhou

Andrew M Ress

Kruti Patel

Chandra K Mayanil

Weiping Yang

Methods of making the M or L complex of the botulinum neurotoxin E with its associated neurotoxin binding protein(s) (NBP), and methods of using to treat pain and wrinkles by injecting locally. Methods of making and isolating the L and M complex comprise a multi-step process of: providing a bacterial cultured media comprising the complex of botulinum neurotoxin E and neurotoxin binding proteins; centrifuging and pelletizing the cultured media; stirring the pelletized culture at 0 to 10° C. over a period of 4 to 24 hours; centrifuging the stirred solution to obtain a supernatant; precipitating the supernatant and filtering to obtain the precipitate; and dissolving the precipitate, centrifuging and filtering to obtain a solution comprising M complex and/or L complex of botulinum neurotoxin E and its NBP; and passing the solution through a functionalized sephadex ion exchange column to isolate the complex.

Complex of botulinum neurotoxin E and its binding protein as a formulation with enhanced potency

The present invention describes a second-generation tetanus toxoid vaccine and a process for the preparation thereof, comprising the steps of: inducing anculture OD=0.5 by adding 0.2 mM IPTG; growing the culture at 14-16° C. for 14 to 20 hours; suspending the culture in 25 mM phosphate buffer containing 200 mM sodium chloride; adding 1% of triton-X-100 to the phosphate buffer, and adding the buffer to the culture; sonicating the culture for a period of 3 minutes (at 5 sec on/off pulse) at 4° C. on cold beads; centrifuging the culture for 60 to 90 minutes; collecting and purifying a supernatant using Ni-NTA affinity column with an eluant; and combining the supernatant into a pool with contaminated bands and concentrating using Centriprep-30 centrifuge filters (30 kDa pores).

Composition for oral or nasal delivery of tetanus, diphtheria, and pertussis vaccine alone or in combination using neurotoxin associated proteins

A recombinant SARS-CoV2 protein-based vaccine, and method of immunizing, is developed by either embedding the epitopes/domains, or chemically attaching, to an already established vaccine candidate, such as a detoxified recombinant tetanus neurotoxin (DrTeNT), to develop a novel vaccine to immunize against SARS-CoV2. The developed vaccine will have three novel contributions compared to the present vaccine technology; a) providing a novel and very effective vaccine platform; b) priming with DrTeNT will prepare the host immune system for better response; and c) oral delivery of the vaccine candidate with a group of neurotoxin binding proteins (NAPs) fromsp. A Detoxified recombinant tetanus neurotoxin (DrTeNT) is prepared by mutation of the active site amino acid residues is an effective vaccine candidate, and is to be used for embedding epitopes of SARS-CoV-2 virus protein for vaccination against Covid-19. DrTeNT is a risk-free vaccine, free of formalin or any other chemical adjuvants.

Tetanus vaccine platform for embedding COVID-19 vaccine

Compositions comprising the L complex or the M complex of the botulinum neurotoxin E with its associated neurotoxin binding protein(s) (NBP), methods of making, and methods of use to treat pain and wrinkles by injecting locally. Methods of making and isolating the L and M complex comprise a multi-step process of: providing a bacterial cultured media comprising the complex of botulinum neurotoxin E and neurotoxin binding proteins; centrifuging and pelletizing the cultured media; stirring the pelletized culture at 0 to 10° C. over a period of 4 to 24 hours; centrifuging the stirred solution to obtain a supernatant; precipitating the supernatant and filtering to obtain the precipitate; and dissolving the precipitate, centrifuging and filtering to obtain a solution comprising M complex and/or L complex of botulinum neurotoxin E and its NBP; and passing the solution through a functionalized sephadex ion exchange column to isolate the complex.

The present invention describes a neurotoxin associated protein from botulinum neurotoxin complex used as an oral or nasal delivery system for a vaccine. The vaccine is selected from tetanus, diphtheria and pertussis alone or in combination. Further the oral or nasal delivery of tetanus vaccine in combination with other drug molecules.

Presently claimed invention related to a pharmaceutical composition comprising ascorbic acid caffeoylquinic acid, rosmarinic acid. and glycosyl sulfones obtained fromand a process for isolating said composition from. Presently claimed invention also provides a method of treating neural crest derived tumors such as malignant neurofibroma and melanoma.

Pharmaceutical composition for treating cancer

Present invention provides a stable pharmaceutical composition comprising botulinum neurotoxin, anesthetic in a buffer. Present invention also provides a process for the preparation of the stable pharmaceutical composition. Theneurotoxin is stabilized by the formation of complex between the toxin and anesthetic. The present invention further provides a method for treating wrinkles using the said pharmaceutical composition.

Botulinum neurotoxin compositions

An isolated, biologically active 33 kDa hemagglutinin purified from the type Aneurotoxin complex and its uses are described.

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This application is based, inter alia, on the discovery of a binding interaction between the Hn-33 hemagglutinin polypeptide of the type Aneurotoxin complex and synaptosomal proteins, including synaptotagmin II (Syt II). Methods of screening for compounds that modulate, e.g., increase or decrease, this interaction are provided. Also provided are compositions and methods for targeting compounds to neuronal and cancer cells by coupling the compounds to Hn-33 or biologically active Hn-33 variants.

Methods of identifying a compound that increases or decreases the interaction of Hn-33 with synaptotagmin II

The present invention relates to the isolation of polypeptides derived from theneurotoxin and the use thereof as immunogens for the production of vaccines and antitoxins, as well as research and drug development applications.is responsible for food bone botulism, a severe and often deadly disease. Botulinum neurotoxin binds to neural cells and are translocated into the cytosol. The toxin then prevents neurotransmitter release by cleaving a SNARE protein. A double mutant E224A/E262 full length botulinum neurotoxin Type A holo form was successfully cloned and purified, which lacks the endopeptidase activity involved in the toxic action of the BoNT, and thus leading to its detoxification (DR BoNT/A). This new molecule can be used as an antidote against botulism, and also as a vaccine candidate for botulism. Due to the poor availability and extreme toxicity of native holo-toxin, a nontoxic form of the holo-toxin is highly desired for research and vaccine development. The full length DR BoNT/A protein has been expressed inas a soluble form.

Detoxified recombinant botulinum neurotoxin

The invention features a polypeptide complex synthesized by bacteria of the genusthat contains the serotype E botulinum neurotoxin and five neurotoxin associated polypeptides having molecular weights of about 118, 80, 65, 40, and 18 kDa, respectively. The complex is useful in the treatment of diseases or conditions that are caused by excessive release of acetylcholine from presynaptic nerve terminals.

Proteins within the type E botulinum neurotoxin complex

A stable composition includingneurotoxin and a cyclodextrin and a method of preservingneurotoxin and for producing aneurotoxin composition with improved stability properties in an efficient and economically advantageous manner. The invention seeks to alleviate the problems associated with rapid degradation or denaturation ofneurotoxin by providing a novel composition that exhibits improved stability properties. Theneurotoxin is preferably stabilized by forming a cyclodextrin inclusion complex.

An isolated, biologically active 33 kDa Hemagglutinin purified from the type Aneurotoxin complex and its uses are described.

The invention features a polypeptide complex synthesized by bacteria of the genusthat contains the serotype Eneurotoxin and five neurotoxin associated polypeptides having molecular weights of about 118, 80, 65, 40, and 18 kDa. respectively. The complex is useful in the treatment of diseases or conditions that are caused by excessive release of acetylcholine from presynaptic nerve terminals.

Growing community of inventors

Dartmouth, MA, United States of America

Bal Ram Singh