DNA Sequences coding for PTH variants, PTH variants, expression

Chemistry: molecular biology and microbiology – Micro-organism – per se ; compositions thereof; proces of... – Bacteria or actinomycetales; media therefor

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435 694, 43525233, 4353201, 530324, 536 2351, C07K 14635, C12N 1516, C12N 1570, A61K 3829

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active

054570476

DESCRIPTION:

BRIEF SUMMARY
BACKGROUND OF THE INVENTION

1. Field of the Invention
The invention relates to analogs of parathyroid hormone.
2. Brief Description of Related Art
Parathyroid hormone (PTH) regulates the level of calcium in blood plasma by exerting, inter alia, anabolic and catabolic effects on the skeleton and also by promoting calcium re-absorption by the kidneys. There is reason to believe that the bone-building effect of PTH can be exploited for the clinical treatment of osteoporosis; see J. Bone Mineral Res., 1 (1987) 377-382.
There are also indications that the multiple functions of PTH are exerted by different regions of the hormone molecule. For example, it has been found that the N-terminal 34 amino acids mediate the binding of the hormone to its kidney cell receptor with subsequent stimulation of adenylate cyclase (see Adv. Prot. Chem., 32 (1982) 323-395) but that a mitogenic effect originates from the central region sequence 28-48 (see DE-A-37 25 319.0) which effect is not mediated by adenylate cyclase but possibly by protein kinase C. It would therefore seem attractive to modulate individual hormone effects by controlled variation of the amino acid sequence of PTH or of PTH fragments and thus possibly to reduce the catabolic activity of the hormone or increase its anabolic activity.
With regard to the prior art, reference should also be made to U.S. Pat. Nos. 4,086,196 and 4,423,037 and John et al., in Adv. Prot. Chem., 32 (1982) 323.


SUMMARY OF THE INVENTION

For the above purpose, according to the invention a DNA sequence is provided that codes for a variant of PTH(1-84), in which region of positions 1 to 27, and which acids and is optionally also shortened C-terminally (position +84) by one or more amino acids or acids, that occurs in bPTH, pPTH, rPTH, cPTH or hPTH.
According to a further embodiment, a DNA sequence is provided that codes for a variant of PTH(1-84), in which region of positions 28 to 31, and which acids and is optionally also shortened C-terminally (position +84) by one or more amino acids or (position +84) by one or more amino acids, that occurs in bPTH, pPTH, rPTH, cPTH or hPTH.
According to a further embodiment, a DNA sequence is provided that codes for a variant of PTH(1-84), in which region of positions 32 to 34, except that tyrosine may not appear at position 34, and, further, which acids and is optionally also shortened C-terminally (position +84) by one or more amino acids or (position +84) by one or more amino acids, that occurs in bPTH, pPTH, rPTH, cPTH or hPTH.
According to a further embodiment, a DNA sequence is provided that codes for a variant of PTH(1-84), in which region of positions 30 to 34, and which acids and is optionally also shortened C-terminally (position +84) by one or more amino acids or (position +84) by one or more amino acids, that occurs in bPTH, pPTH, rPTH, cPTH or hPTH.
According to a further embodiment, a DNA sequence is provided that codes for a variant of PTH(1-84), in which or region of positions 28 to 34 and especially 30 to 34 and, another amino acid outside the region of positions 28 to 34 or 30 to 34, and which acids and is optionally also shortened C-terminally (position +84) by one or more amino acids or (position +84) by one or more amino acids, that occurs in bPTH, pPTH, rPTH, cPTH or hPTH.
The DNA sequence preferably codes for a PTH variant whose N-terminus is lengthened by proline. Also preferred are hPTH, bPTH, pPTH, rPTH and cPTH variants. As regards the numbering of the DNA sequences, 1 denotes the first nucleotide of the first PTH codon so that the numbers to be found in the following Table 1 denote the position of the first nucleotide shown in relation to the first nucleotide of the first PTH codon.
According to a further embodiment, the invention relates also to PTH variants that are coded by one of the above described DNA sequences, and especially variants of human (h), bovine (b), porcine (p), rat (r) and canine (c) PTH.


DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT OF THE INVENTION

The PTH variants according to the invention can

REFERENCES:
patent: 4423037 (1983-12-01), Rosenblatt
Chorev, M. et al. Biochemistry 29:1580-1586 Feb. 13, 1990.
Hendy, G. N. et al. "Nuceotide sequence cloned cDNAS encoding human preproparathyroid hormon. " PNAS 78(12):7365-7369 (1981).

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