BMP-2 variants with improved properties

a technology of bmp-2 and variants, which is applied in the direction of osteogenic factors, peptide/protein ingredients, drug compositions, etc., can solve the problems of limiting the development and use of bmp therapeutics, hampering development and production, and poor bmp expression yields, etc., to achieve improved expression yield, increase solubility, and improve the effect of properties

Inactive Publication Date: 2006-10-19
XENCOR INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0013] The present invention is related to variants of human bone morphogenetic proteins and other cysteine knot cytokine proteins with improved properties, including increased expression yield, expression in the absence of a pro-domain, increased solubility, increased specific activity, altered receptor, co-receptor, and inhibitor specificity, and decreased immunogenicity.

Problems solved by technology

A number of unfavorable properties of naturally occurring BMPs limit the development and use of BMP therapeutics.
BMP expression yields are typically poor and suitable expression hosts are limited, hindering development and production.
BMPs often possess multiple biological effects, including unwanted side effects.
Many BMPs are poorly soluble, reducing storage stability and bioavailability.
Finally, BMPs may induce unwanted immune responses.

Method used

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  • BMP-2 variants with improved properties
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  • BMP-2 variants with improved properties

Examples

Experimental program
Comparison scheme
Effect test

example 1

Structural Modeling

[0165] Hexameric complexes comprising a BMP-7 dimer or a BMP-2 dimer bound to two ALK-3 receptors and two ActRIIa receptors was constructed using the structure of BMP-7 bound to ActRIIa (PDB code 1LX5) and the structure of BMP-2 bound to ALK-3 (PDB code 1ES7). Using InsightII (Accelrys), the BMP structures were superimposed as follows: BMP-2 residues 22-32 superimposed with BMP-7 residues 47-56, BMP-2 residues 49-71 superimposed with BMP-7 residues 73-95, and BMP-2 residues 101-106 superimposed with BMP-7 residues 126-131. This yielded a backbone atom RMSD of 0.77 Å. The superposition was repeated so that chain A in the BMP-7 structure was superimposed onto chains A and C of the BMP-2 structure. The sequence alignment between BMP-2 and BMP-7 is shown in FIG. 3 and the structure of the hexameric complex is shown in FIG. 2.

[0166] Homology modeling was used to generate structures of additional BMP receptors bound to BMP-2 and BMP-7. As shown in FIG. 5, the sequence...

example 2

Identification of Exposed Hydrophobic Residues in BMPs

[0167] Structures of BMP-7 dimer (“dimer”) and BMP-7 dimer bound to ALK-3 and ActRIIa “hexamer”) were analyzed to identify solvent-exposed hydrophobic residues. The absolute and fractional solvent-exposed hydrophobic surface area of each residue was calculated using the method of Lee and Richards (J. Mol. Biol. 55: 379-400 (1971)) using an add-on radius of 1.4 Å (Angstroms). Each residue was also classified as core, boundary, or surface (see Dahiyat and Mayo Science 278: 82-87 (1997)).

[0168] Solvent exposed hydrophobic residues in BMP-7 were defined to be hydrophobic residues with at least 50 Å2 (square Angstroms) exposed hydrophobic surface area in the BMP-7 dimer (PDB code 1LX5, chain A, plus symmetry-related BMP-7 molecule). Exposed hydrophobic surface area was also measured in the context of the BMP-7 / ALK-2 / ActRIIa hexamer and RESCLASS was run to categorize each position as core, boundary, or surface.

TABLE 1Exposed hydrop...

example 3

Identification of Receptor and Inhibitor Interface Residues in BMP-7

[0169] Potential sites of interactions between BMP-7 and ALK-3, BMP-7 and ActRIIa, and BMP-7 and noggin were identified by examining the structure of the hexameric structure described in Example 1 and the co-crystal structure of BMP-7 and noggin (PDB code 1M4U). Next, distance measurements were used to identify residues that may participate in intermolecular interactions. Residues in BMP-7 that are within 5 Å (Angstroms) of the ALK-3, ActRIIa, or noggin interfaces (as measured by CA-CA distances) are shown below, along with the receptor or inhibitor positions that are contacted. Next, the receptor sequence alignments used for homology modeling were analyzed for polymorphisms. Information about receptor polymorphisms was used to design receptor-specific variants, described below. If the receptor positions are polymorphic, it is noted in Table 2; “na” indicates that the receptor positions were not sufficiently well-a...

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Abstract

The invention relates to variants of BMP-2 with improved properties and methods for their use.

Description

[0001] This application claims benefit under 35 U.S.C. §119(e) to U.S. Ser. No. 60 / 558,189 filed on Mar. 31, 2004, entitled “Cysteine Knot Cytokine Variants with Improved Properties”; U.S. Ser. No. 60 / 570,520 filed on May 11, 2004 entitled “Cysteine Knot Cytokine Variants with Improved Properties”; U.S. Ser. No. 60 / 578,432 filed on Jun. 9, 2004 entitled “Cysteine Knot Cytokine Variants with Improved Properties”; and U.S. Ser. No. 60 / 587,464, filed on Jul. 13, 2004 entitled “Cysteine Knot Cytokine Variants with Improved Properties”, all of which are expressly incorporated by reference in their entirety.FIELD OF THE INVENTION [0002] The invention relates to variants of bone morphogenetic proteins and other cysteine knot growth factors with improved properties, and to methods of making and using compositions utilizing these variants. BACKGROUND OF THE INVENTION [0003] Bone morphogenetic proteins (BMPs) are a well-known family of growth factors that contribute to developmental processes...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/18C07K14/475C07K14/51
CPCC07K14/51A61K38/00A61P19/00A61P19/08
Inventor DESJARLAIS, JOHNMARSHALL, SHANNONZALEVSKY, JONATHAN
Owner XENCOR INC
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