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Claims for Patent: 5,674,710

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Claims for Patent: 5,674,710

Title: Recombinant techniques for production of human brain natriuretic peptide
Abstract: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.
Inventor(s): Seilhamer; J. Jeffrey (Milpitas, CA), Lewicki; John (San Jose, CA), Scarborough; Robert M. (Hayward, CA), Porter; J. Gordon (Newark, CA)
Assignee: Scios, Inc. (Mountain View, CA)
Application Number:07/477,226
Patent Claims: 1. A recombinant DNA in isolated and purified form consisting essentially of a DNA which encodes a peptide having natriuretic activity which peptide has the formula:

wherein R.sup.1 is selected from the group consisting of:

wherein R.sup.3 is the 101 amino acid sequence shown for human BNP in FIG. 8 at positions numbered 1-99 immediately upstream of the Ser residue to which R.sup.3 is bound, or a C-terminal portion thereof; and

R.sup.2 is (OH) or R.sup.2 is selected from the group consisting of:

Lys;

Lys-Val;

Lys-Val-Leu;

Lys-Val-Leu-Arg;

Lys-Val-Leu-Arg-Lys;

Lys-Val-Leu-Arg-Lys-His.

2. A recombinant expression system capable, when contained in a recombinant host cell, of expressing a DNA which encodes a peptide having natriuretic activity wherein the peptide has the formula:

wherein R.sup.1 is selected from the group consisting of:

wherein R.sup.3 is the 101 amino acid sequence shown for human BNP in FIG. 8 at positions numbered 1-99 immediately upstream of the Ser residue to which R.sup.3 is bound, or a C-terminal portion thereof; and

R.sup.2 is (OH) or R.sup.2 is selected from the group consisting of:

Lys;

Lys-Val;

Lys-Val-Leu;

Lys-Val-Leu-Arg;

Lys-Val-Leu-Arg-Lys;

Lys-Val-Leu-Arg-Lys-His.

3. A recombinant host cell or cell culture which has been manipulated so as to contain the expression system of claim 2.

4. A method to produce a peptide having natriuretic activity, which method comprises:

culturing the cells of claim 3 under conditions which permit the expression of the DNA encoding said peptide; and

recovering the peptide from the culture.

5. The recombinant expression system of claim 2 wherein R.sup.1 is Ser-Pro-Lys-Met-Val-Gln-Gly-Ser-Gly- and R.sup.2 is Lys-Val-Leu-Arg-Lys-His.

6. A recombinant host cell or cell culture of claim 3 wherein R.sup.1 is Ser-Pro-Lys-Met-Val-Gln-Gly-Ser-Gly- and R.sup.2 is Lys-Val-Leu-Arg-Lys-His.

7. The method of claim 4 wherein R.sup.1 is Ser-Pro-Lys-Met-Val-Gln-Gly-Ser-Gly- and R.sup.2 is Lys-Val-Leu-Arg-Lys-His.

8. The recombinant DNA of claim 1 wherein R.sup.1 is Ser-Pro-Lys-Met-Val-Gln-Gly-Ser-Gly- and R.sup.2 is Lys-Val-Leu-Arg-Lys-His.
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