RECOMBINANT TECHNIQUES FOR THE PRODUCTION OF NEW NATRIURETIC AND VESSEL-WIDING PEPTIDES.

摘要

The cDNA sequence encoding porcine brain natriuretic peptide and related genes encoding canine and human peptides with natriuretic activity are disclosed. The gene is shown to make accessible the DNAs encoding analogous natriuretic peptides in other vertebrate species. The genes encoding these NPs can be used to effect modifications of the sequence to produce alternate forms of the NPs and to provide practical amounts of these proteins. The NPs of the invention can also be synthesized chemically. The invention peptides have the formula: tables id="TABLE-US-00001" num="00001" table frame="none" colsep="0" rowsep="0" tabstyle="monospace" tgroup align="left" colsep="0" rowsep="0" cols="2" colspec colname="1" colwidth="217pt" align="left"/ colspec colname="2" colwidth="0pt" align="left"/ TBODY VALIGN="TOP" ROW ENTRYRSUP1/SUP-Cys-Phe-Gly-Arg- Arg/ - Leu/ -Asp-Arg-/ENTRY ENTRY/ /ROW ROW ENTRY Lys Met/ENTRY /ROW ROW ENTRY (1)/ENTRY /ROW ROW ENTRY Ile- Gly/ -Ser- Leu/ -Ser-Gly-Leu-Gly-Cys-R-SUP2/SUP/ENTRY /ROW ROW ENTRY Ser Ser/ENTRY /ROW /TBODY /TGROUP /TABLE /TABLES wherein RSUP1 /SUPis selected from the group consisting of: tables id="TABLE-US-00002" num="00002" table frame="none" colsep="0" rowsep="0" tabstyle="monospace" tgroup align="left" colsep="0" rowsep="0" cols="2" colspec colname="1" colwidth="217pt" align="left"/ colspec colname="2" colwidth="0pt" align="left"/ TBODY VALIGN="TOP" ROW ENTRY (H);/ENTRY ENTRY/ /ROW ROW ENTRY Gly-;/ENTRY /ROW ROW ENTRY Ser-Gly-;/ENTRY /ROW ROW ENTRY Asp//ENTRY /ROW ROW ENTRY Lys/ -Ser-Gly-;/ENTRY /ROW ROW ENTRY Gly/ENTRY /ROW ROW ENTRY Arg/ Asp//ENTRY /ROW ROW ENTRY His/ - Lys/ -Ser-Gly-;/ENTRY /ROW ROW ENTRY Gln Gly/ENTRY /ROW ROW ENTRY Arg/ Asp//ENTRY /ROW ROW ENTRY Met/ - His/ - Lys/ -Ser-Gly-;/ENTRY /ROW ROW ENTRY Val Gln Gly/ENTRY /ROW ROW ENTRY Arg/ Asp//ENTRY /ROW ROW ENTRY Thr/ - Met/ - His/ - Lys/ -Ser-Gly-;/ENTRY /ROW ROW ENTRY Met Val Gln Gly/ENTRY /ROW ROW ENTRY Arg/ Asp//ENTRY /ROW ROW ENTRY Lys- Thr/ - Met/ - His/ - Lys/ -Ser-Gly-;/ENTRY /ROW ROW ENTRY Met Val Gln Gly/ENTRY /ROW ROW ENTRY Arg/ Asp//ENTRY /ROW ROW ENTRY Pro-Lys- Thr/ - Met/ - His/ - Lys/ -Ser-Gly-;/ENTRY /ROW ROW ENTRY Met Val Gln Gly/ENTRY /ROW ROW ENTRY Arg/ Asp//ENTRY /ROW ROW ENTRYSer-Pro-Lys- Thr/ - Met/ - His/ - Lys/ -Ser-Gly-;/ENTRY /ROW ROW ENTRY Met Val Gln Gly/ENTRY /ROW /TBODY /TGROUP /TABLE /TABLES or a 10- to 109-amino acid sequence shown as the native upstream sequence for porcine, canine or human BNP in FIG. 8 , or a composite thereof; RSUP2 /SUPis (OH), NHSUB2/SUB, or NR'R'' wherein R' and R'' are independently lower alkyl (1-4C) or is tables id="TABLE-US-00003" num="00003" table frame="none" colsep="0" rowsep="0" tabstyle="monospace" tgroup align="left" colsep="0" rowsep="0" cols="3" colspec colname="offset" colwidth="42pt" align="left"/ colspec colname="1" colwidth="175pt" align="left"/ colspec colname="2" colwidth="0pt" align="left"/ TBODY VALIGN="TOP" ROW ENTRY/ ENTRYAsn//ENTRY ENTRY/ /ROW ROW ENTRY/ ENTRYLys/ENTRY /ROW ROW ENTRY/ ENTRYAsn/ -Val/ENTRY /ROW ROW ENTRY/ ENTRYLys/ENTRY /ROW ROW ENTRY/ ENTRYAsn/ -Val-Leu/ENTRY /ROW ROW ENTRY/ ENTRYLys/ENTRY /ROW ROW ENTRY/ ENTRYAsn/ -Val-Leu-Arg/ENTRY /ROW ROW ENTRY/ ENTRYLys/ENTRY /ROW ROW ENTRY/ ENTRYAsn/ -Val-Leu-Arg- Arg//ENTRY /ROW ROW ENTRY/ ENTRYLys Lys/ENTRY /ROW ROW ENTRY/ ENTRYAsn/ -Val-Leu-Arg- Arg/ - Tyr//ENTRY /ROW ROW ENTRY/ ENTRYLys Lys His/ENTRY /ROW /TBODY /TGROUP /TABLE /TABLES or the amides (NHSUB2 /SUBor NR'R'') thereof, with the proviso that if formula (1) is tables id="TABLE-US-00004" num="00004" table frame="none" colsep="0" rowsep="0" tabstyle="monospace" tgroup align="left" colsep="0" rowsep="0" cols="3" colspec colname="offset" colwidth="21pt" align="left"/ colspec colname="1" colwidth="196pt" align="left"/ colspec colname="2" colwidth="0pt" align="left"/ TBODY VALIGN="TOP" ROW ENTRY/ ENTRYRSUP1/SUP-Cys-Phe-Gly-Arg-Arg-Leu-Asp-Arg-/ENTRY ENTRY/ /ROW ROW ENTRY/ ENTRY Ile-Gly-Ser-Leu-Ser-Gly-Leu-Gly-Cys-RSUP2/SUP/ENTRY /ROW /TBODY /TGROUP /TABLE /TABLES and RSUP1 /SUPis Asp-Ser-Gly-, RSUP2 /SUPcannot be Asn-Val-Leu-Arg-Arg-Tyr. The peptides of the invention can be formulated into pharmaceutical compositions and used to treat conditions associated with high extracellular fluid levels, especially congestive heart failure.