Glycoprotein Hormone Analogs

a glycoprotein and hormone technology, applied in the field of glycoprotein hormone analogs, can solve the problems of affecting the development of glycoprotein hormone agonists and antagonists, the -subunit has a tendency to fold completely prior to heterodimer assembly, and the extracellular domain is more difficult to model, etc., to achieve the effect of preventing the formation of heterodimer

Inactive Publication Date: 2009-08-20
UNIV OF MEDICINE & DENTISTRY OF NEW JERSEY
View PDF0 Cites 15 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this process is required for assembly of salmon FSH and other piscine follitropins in which the seatbelt is latched to a cysteine in the aminoterminal region of the β-subunit (Xing et al., 2004c), it is inefficient for at least two reasons.
First, the β-subunit has a tendency to fold completely prior to heterodimer assembly unless it is prevented from doing so by the composition of the seatbelt or by a chaperone that interferes with seatbelt latching.
The second impediment to assembly by the wraparound pathway stems from the fact that the unlatched seatbelt destabilizes the transient complex composed of the α-subunit and the unlatched β-subunit (Xing et al., 2004d).
This portion of the extracellular domain has been more difficult to model, however, since its amino acid sequence is not similar to proteins of known structure.
The lack of structural knowledge has hampered development of glycoprotein hormone agonists and antagonists.
Although methods for producing glycoprotein hormones were developed in 1985 (Reddy et al., 1985), these are not applicable to all ligands.
For example, it has been particularly difficult to produce hormones and hormone analogs in which the seatbelt is latched to a cysteine in the N-terminal region of the β-subunit.
This has limited the development of analogs that can be used to stimulate fertility in not only mammalian systems but also those of salmon, trout, and other fish that express follitropins (in which the seatbelt is latched to a cysteine in the aminoterminal region of the β-subunit).

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Glycoprotein Hormone Analogs
  • Glycoprotein Hormone Analogs
  • Glycoprotein Hormone Analogs

Examples

Experimental program
Comparison scheme
Effect test

example 1

Development of a Lutropin Antagonist

[0111]The wraparound pathway can be used to prepare hCG analogs in which the seatbelt is latched to the α-subunit rather than to the β-subunit (Xing et al., 2001a). The efficiency of this process depends on the location of the seatbelt latch site. Seatbelt latch sites that are located on α-subunit loop 2 are usually the most efficient and, when the seatbelt is latched to some of these, the heterodimer retains its biological activity. The hCG seatbelt can be forced to latch to a cysteine added to the α-subunit when the normal seatbelt latch site—i.e., Cys26 in β-subunit loop 1—is disrupted. Using this approach (Xing et al., 2001a) prepared heterodimers in which the seatbelt became latched to cysteines that had been substituted for several α-subunit residues. With the exception of hCG analogs in which the seatbelt was latched to cysteines nearby the normal seatbelt latch site—e.g., α-subunit residues Leu41 and Ser43—this approach to crosslinking the...

example 2

Abilities of Additional Disulfide Crosslinks to Stabilize the Heterodimer Containing a Truncated Seatbelt in a Functional Fashion

[0120]Crosslinks between α86-β103 that were produced by co-expressing pMB2472 and pMB2674, (FIG. 8), α86-β106 pMB2472 and pMB2672, (FIG. 8) and α86-β108 (pMB2472 and pMB2673, (FIG. 8) permitted the formation of heterodimers containing a truncated seatbelt. These were prepared by expressing a construct that encodes sequences pMB1244 or pMB2472 with constructs that encode sequences pMB2674, pMB2672, or pMB2673 transiently in COS7 cells. These had the ability to inhibit binding of 125I-hCG to rat LH receptors and had low efficacies in cAMP accumulation signal transduction assays (FIG. 13). Of the three types of crosslink, that between α86 and β102 led to an analog that had the lowest efficacy (FIG. 13). This analog blocked the binding of 125I-hCG to rat LH receptors (FIG. 14) and inhibited hCG stimulated cyclic AMP accumulation (FIG. 15). Its ability to compe...

example 3

The Antagonist can be made in a Single Chain Format

[0127]Single chain analogs of hCG and other glycoprotein hormones are often expressed better than the individual subunits. This is most likely because in the single chain format their subunit components are present at extremely high concentrations relative to one another. This phenomenon would be expected to facilitate heterodimer assembly in the endoplasmic reticulum, its normal site. Most hCG single chain constructs have a format in which the α-subunit component is linked to the carboxyterminal end of the β-subunit component. This is done to take advantage of the long disordered “tail” of the α-subunit, which facilitates assembly. It is also done because addition of residues to the α-subunit terminus can reduce receptor interactions (Furuhashi et al., 1995b). The antagonist analog described in Example 1 lacks much of the seatbelt and all of the α-subunit carboxyterminus. Therefore, it would not be expected that a single chain cons...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
pHaaaaaaaaaa
timeaaaaaaaaaa
pHaaaaaaaaaa
Login to view more

Abstract

This invention relates to the field of glycoprotein hormone analogs and their uses as agonists, antagonists, targeting vectors, and immunogens. In particular, this invention describes a method for stabilizing a heterodimer that permits the preparation of functional glycoprotein hormone analogs. The analogs of present invention comprise at least one alpha subunit polypeptide and at least one beta subunit polypeptide, wherein the seatbelt region of the beta subunit is linked to the alpha subunit. The invention also provides for a beta subunit polypeptide wherein the C-terminal amino acid is from residue 10 to residue 20 of the seatbelt region.

Description

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT[0001]This research has been funded by NIH Grant HD14907. The U.S. government has certain rights to the invention.FIELD OF THE INVENTION[0002]The present invention relates to the field of glycoprotein hormone analogs and their uses as agonists, antagonists, targeting vectors, and immunogens. In particular, this invention describes a method for stabilizing a heterodimer that permits the preparation of functional glycoprotein hormone analogs.[0003]Glycoprotein hormones control the functions of the gonads and the thyroid gland. These hormones are αβ heterodimers that are stabilized by a portion of the β-subunit known commonly as the “seatbelt.” The seatbelt contains a cysteine at its carboxyterminus that enables it to form a disulfide bond with another cysteine in the β-subunit. In most vertebrates the seatbelt is disulfide bridged to a cysteine in β-subunit loop 1. The seatbelt is bridged to a cysteine in the aminoterminal...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/16C07K14/00C07K17/00C07H21/04C12N15/63C12N5/00
CPCA61K38/00C07K14/59A61K47/48238A61K47/62A61P15/00
Inventor MOYLE, WILLIAM R.
Owner UNIV OF MEDICINE & DENTISTRY OF NEW JERSEY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap