Generating peptide intermediate, incubating in dicloromethane, precipitating and recovering enfuvirtide; viricide; HIV fusion inhibitor
Inventors
Han, Yeun-Kwei • Khatri, Hiralal N. • Topping, Robert J.
Assignees
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Publication Number
US-7696311-B2
Publication Date
2010-04-13
Expiration Date
Abstract
The invention provides methods of obtaining a peptide that include steps of synthesizing a peptide intermediate having one or more side chain protecting groups; providing a solvent to the peptide intermediate to form a peptide intermediate composition; and providing a precipitating agent in an amount sufficient to precipitate the peptide intermediate from the peptide intermediate composition, wherein the precipitating agent is an alcohol having three or more carbon atoms. Also provided are methods for precipitating peptides, methods for concentration peptides, and methods for filtering peptides.
Core Innovation
The invention concerns methods of obtaining a peptide used for scalable synthesis. The methods include synthesizing a peptide intermediate having one or more side chain protecting groups, wherein the peptide intermediate is used for synthesis of enfuvirtide and is selected from a defined set of sequence-defined intermediates identified by SEQ ID NOs.
The methods form a peptide intermediate composition by providing a solvent to the peptide intermediate, and then precipitate the peptide intermediate from the composition. The precipitating agent comprises an alcohol having three or more carbon atoms, with secondary or tertiary alcohols highlighted in the description.
In the described implementations, the peptide intermediate composition can be formed as an emulsion, and the precipitating agent is provided in an amount sufficient to precipitate the peptide intermediate. The description emphasizes tuning filterability and avoiding undesired solid behavior during precipitation, and provides comparative examples for a 16-residue enfuvirtide intermediate.
Claims Coverage
The provided independent claims define two main inventive methods, each centered on precipitating a specified enfuvirtide peptide intermediate using an alcohol having three or more carbons. Across the two independent claims, the inventive core comprises the selection of the peptide intermediate sequence set and the precipitation-agent constraint, with one claim additionally specifying the intermediate composition as an emulsion format.
Enfuvirtide peptide intermediate precipitation with a carbon-three-or-more alcohol
A method of obtaining a peptide by synthesizing a peptide intermediate having one or more side chain protecting groups for synthesis of enfuvirtide, wherein the peptide intermediate is selected from the group consisting of the enumerated SEQ ID NOs; providing a solvent to form a peptide intermediate composition; and providing a precipitating agent in an amount sufficient to precipitate the peptide intermediate from the peptide intermediate composition, wherein the precipitating agent comprises an alcohol having three or more carbon atoms.
Enfuvirtide peptide intermediate emulsion precipitation with a carbon-three-or-more alcohol
A method of obtaining a peptide by synthesizing a peptide intermediate having one or more side chain protecting groups for synthesis of enfuvirtide, wherein the peptide intermediate is selected from the group consisting of the enumerated SEQ ID NOs; providing a solvent to form a peptide intermediate composition in the form of an emulsion; and providing a precipitating agent in an amount sufficient to precipitate the peptide intermediate from the peptide intermediate composition, wherein the precipitating agent comprises an alcohol having three or more carbons.
Both independent claims cover precipitation-based peptide recovery from a solvent-created peptide intermediate composition, where the precipitating agent comprises an alcohol having three or more carbon atoms and the peptide intermediate is restricted to the enumerated enfuvirtide-associated sequence set. One independent claim additionally requires the solvent-created composition to be in the form of an emulsion.
Stated Advantages
Improved purity with reduced impurities when using the alcohol precipitating agent compared with heptane, as described by HPLC impurity reduction.
Fewer or reduced processing steps for washing and distillation compared with heptane precipitation, as described in the summary of advantages.
Potential reagent and/or processing time savings, as described in the summary of advantages.
Improved downstream yields, as described in the summary of advantages.
Documented Applications
Scalable peptide synthesis for obtaining peptide intermediates used for synthesis of enfuvirtide, including recovery of an enfuvirtide intermediate.
Use of precipitation with an alcohol having three or more carbons to isolate peptide intermediates and support filtration and downstream processing performance, as demonstrated by comparative examples for a 16-residue enfuvirtide intermediate.
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