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Parathyroid hormone-like polypeptides

A parathyroid hormone, sequence technology, applied in the direction of parathyroid hormone, hormone peptides, peptide sources, etc., can solve the problems of increased porosity, thinning of load-bearing bones, and lack of trabeculae.

Inactive Publication Date: 2005-10-12
TEELEOSTIN LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This results in thinning of the load-bearing bone, increased porosity, and trabecular depletion

Method used

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  • Parathyroid hormone-like polypeptides
  • Parathyroid hormone-like polypeptides
  • Parathyroid hormone-like polypeptides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1: Identification of parathyroid hormone in the fish redfin puffer (Fugu rubripes)

[0074] Materials and methods

[0075] Polymerase chain reaction and automated sequencing of DNA clones encoding Fugu PTH(1-80)

[0076] Primers for polymerase chain reaction (PCR) were designed using the known PTH amino acid sequence based on preliminary data obtained from the Joint Genome Institute (http: / / www.jgi.doe.gov / programs / fugu.htm). Preliminary nucleic acid sequences from databases were checked by PCR and some errors were identified. New PCR primers were designed for the corrected nucleotide sequence; forward primer-[5'-CAGTGAGTGAAGTCCAGCTCA-3'] (SEQ ID NO: 5), and reverse primer-[5'-CTTCACTCCTGTGATTTGAGCA-3'] (SEQ ID NO :6). PCR amplification was performed on approximately 100 ng of genomic DNA isolated from pufferfish. PCR products were purified using a commercially available kit (UltraClean PCR Clean-Up DNA Purification Kit, Geneworks, Adelaide, Australia) and D...

Embodiment 2

[0102] Example 2: The synthetic effect of Fugu PTH on rat bone

[0103] Substances and materials

[0104] Fugu PTH synthesis was assessed in the bones of 50-60 g male Sprague-Dawley rats (3-4 weeks old).

[0105] Rats were administered subcutaneously with low or high doses (3 or 10 μg) of Fugu PTH(1-34) per 100 g body weight per day for 30 days. Synthetic PTH peptide was dissolved in 0.01M acetic acid, and daily injections were prepared in saline with 2% rat serum (from male Sprague-Dawley rats). Rats were weighed twice a week and the PTH dose adjusted for each animal's weight gain.

[0106] There were 12 rats each in the following treatment groups: Control, Human PTH (low or high dose) and Fugu PTH (low or high dose). Euthanize the rat by asphyxiation, then remove the tibia leaving most of the muscle on the bone. Place the samples in freshly prepared 4% paraformaldehyde. Fixed for 24 hours and then transferred to 70% ethanol in preparation for histomorphometry.

[0107]...

Embodiment 3

[0112] Example 3: Detection of fPTH homologues in other fish

[0113] The purpose of this example is to identify the aforementioned gene encoding parathyroid hormone-like polypeptide in different species of fish.

[0114] Methods and materials

[0115] Genomic DNA was extracted from muscle samples of the fish listed in Table 2 following standard techniques. Total RNA extracted following standard techniques was reverse transcribed using random hexamers to generate cDNA.

[0116] Degenerate PCR primers were designed based on the amino acid and nucleotide sequences of the N-terminal (highly conserved) and C-terminal (not very conserved) regions of Fugu and zebrafish PTH (Table 3). This strategy cannot be designed to amplify PCR products from genomic DNA that may contain large introns. Therefore cDNA was also synthesized to detect PTH genes that may contain large intronic sequences.

[0117] In order to detect sequences with low homology to Fugu and zebrafish PTH genes, and also...

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PUM

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Abstract

Novel parathyroid hormone polypeptides and biologically active fragments thereof are disclosed along with nucleic acid molecules encoding same. In particular, parathyroid hormone polypeptides and biologically active fragments (and encoding nucleic acid molecules) derived from fish species (eg Japanese pufferfish (Fugu rubripes)) are disclosed. Such polypeptides and fragments may be used for treatment of diseases associated with abnormal calcium homeostasis (eg osteoporosis, osteopenia, Paget's disease, bone cancer, hyperparathyroidism, hypoparathyroidism, hypercalcemia, psoriasis and other skin-related conditions).

Description

field of invention [0001] The present invention relates to parathyroid hormone and fragments thereof. More particularly, the present invention relates to parathyroid hormone nucleic acid sequences and polypeptides from non-mammalian sources. Background of the invention [0002] Osteoporosis is a major disease in the elderly, affecting both men and women. Osteoporosis is a progressive disease that results in a loss of total bone mass and increased fragility. These changes in bone can lead to spontaneous fractures of load-bearing bones including the hip and vertebrae. These fractures, together with disease-induced heightened fragility, can significantly lead to physical and mental deterioration of the patient. Postmenopausal osteoporosis usually results from the drop in estrogen levels characteristic of menopause. Decreased estrogen levels lead to accelerated bone turnover and an increased imbalance between resorption of old bone and formation of new bone. This results in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/00A61P19/10C07K16/26
CPCA61K38/00C07K16/26A61P17/00A61P17/06A61P19/08A61P19/10A61P3/14A61P35/00A61P5/18A61P5/20
Inventor 杰弗里·戴维·扎亚克雅尼娜·阿塔利亚·丹克斯
Owner TEELEOSTIN LTD
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