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Modulation of ovulation

A technology of ovulation rate and structural domain, applied in the direction of peptide/protein composition, drug combination, peptide, etc., can solve the problem of not being associated with the effect of regulating ovulation

Inactive Publication Date: 2008-02-13
AGRESEARCH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the study did not correlate the regions identified as potentially functionally significant loci with effects in regulating ovulation in vivo

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0094] Use of peptides and antibodies to the N-terminal domains of GDF-9 and GDF-9B in regulating ovulation in sheep.

[0095] Four 12-15mer peptides were synthesized that were identical to the N-terminal domains of the GDF-9 and GDF-9B protein sequences and included additional residues to facilitate their conjugation to keyhole limpet hemocyanin (KLH ) to produce antigen. The peptide sequence is:

[0096] DQESASSELKKPLV(C) (SEQ ID NO: 3);

[0097] SEYFKQFLFPQNEC (SEQ ID NO: 4);

[0098] QAGSIASEVPGPSR(C) (SEQ ID NO: 5); and

[0099] SREHDGPESNQC (SEQ ID NO: 6);

[0100] In this study, groups of 10 nonestrous Romney ewes were injected with each peptide-KLH conjugated antigen dissolved in Freund's complete adjuvant at a dose of 0.4 mg / ewe, and 10 ewes were Nonestrous Romney ewes were injected with KLH antigen at a dose of 0.4mg / ewe as a control group. Animals were then boosted (subcutaneously) at 4 time points at monthly intervals with additional antigen (0.2 mg / ewe at ea...

Embodiment 2

[0112] Regulation of bovine ovulation by peptides and antibodies of the GDF-9 and GDF-9B N-terminal domains.

[0113] Two 15mer peptides were synthesized that were identical to the N-terminal domains of the bovine GDF-9 and GDF-9B protein sequences and included additional residues where required to facilitate their conjugation to KLH for antigen production. The peptide sequence is:

[0114] DQESVSSELKKPLV (C) (SEQ ID NO: 7); and

[0115] QAGSIASEVPGPSR (C) (SEQ ID NO: 5).

[0116] Note that SEQ ID NO:7 is a functional variant of SEQ ID NO:3 comprising a single amino acid change (A→V at amino acid position 5).

[0117]In this study, groups of 10 Friesian cross heifers were injected with each peptide-KLH conjugated antigen dissolved in 2 ml of Freund's complete adjuvant at a dose of 0.4 mg / heifer for 4 days. Injection of 0.5 ml each time; 10 heifers were injected with peptide-KLH conjugated antigen at 0.4 mg / heifer, and 10 heifers were injected with KLH antigen as a control g...

Embodiment 3

[0141] Antibody pairs raised against GDF-9 and GDF-9B peptide fragments 3 Effect of H thymidine incorporation into rat granulosa cells in vitro.

[0142] Using the method described previously (McNatty et al., 2005), various antibody preparations were tested for their ability to neutralize the effects of sheep or murine GDF-9 and sheep GDF-9B on rat granulosa cells when added directly to granulosa cell cultures. A total of 100 μg / ml IgG was added to each treatment and consisted of IgG purified from sheep immunized with GDF-9 or GDF-9B peptide fragments, compared with IgG purified from sheep immunized with KLH. Antibodies are able to neutralize the effects of the growth factors they are directed against (see Figures 5 and 6). It was also shown that the response was dose-dependent in the 2 tested antibody samples against sheep GDF-9 and GDF-9B (P<0.001, see Figures 7 and 8).

[0143] Sheep and mouse GDF-9 and sheep GDF-9B against peptide fragments of the N-terminal region of th...

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Abstract

The present invention is directed to the identification of a novel domain of functional significance on the GDF-9 and GDF-9B molecules and to peptides and antibodies which interact with the domain to modulate the biological activity of these molecules to alter mammalian ovarian function and ovulation rate.

Description

field of invention [0001] The present invention relates to the identification of novel functionally significant domains on the GDF-9 and GDF-9B molecules and to agonists and antagonists that interact therewith to modulate the biological activity of these molecules to alter ovarian function and ovulation rates in mammals. Background of the invention [0002] GDF-9 and GDF-9B (also known as BMP15) are expressed in oocytes in developing follicles and play a role in mammalian fecundity (Fitzpatrick et al., 1998). GDF9, a member of the transforming growth factor beta (TGFβ) superfamily (McPherron and Lee, 1993), is expressed in oocytes from the primary stages of follicular development until ovulation (McGrath et al., 1995; Laitinen et al., 1998). GDF9B is closely related to GDF9 (Dube et al., 1998; Laitinen et al., 1998) and is expressed contemporaneously with GDF9 in mouse oocytes but slightly later than GDF9 in human primary follicles. In the ovary, GDF9 and GDF9B have now bee...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K14/475C07K16/22C07K14/495A61P15/08A61K39/395A61K38/10
Inventor 珍妮弗·L·朱恩格尔肯尼思·P·马克纳蒂劳埃德·G·穆尔罗伯特·S·鲍尔
Owner AGRESEARCH LTD
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