Osteogenic devices

• US 5,011,691 A
• Filed: 02/23/1989
• Issued: 04/30/1991
• Est. Priority Date: 08/15/1988
• Status: Expired due to Term

• Current Assignee: Stryker Corporation
• Original Assignee: Stryker Corporation
• Inventors: Rueger, David C., Oppermann, Hermann, Ozkaynak, Engin, Kuberasampath, Thangavel
• Primary Examiner(s): Nutter, Nathan M.
• Application Number:
  US07/315,342
• Time in Patent Office: 796 Days
• Field of Search: 530/350, 530/840, 530/395, 514/2, 514/21, 424/423, 424/424, 424/426
• US Class Current: 424/423
• CPC Class Codes: A61C 8/0006 Periodontal tissue or bone regeneration support means for endoprosthetic bone substitute or bone graft implants A61F2/2846
• CPC Class Codes: A61F 2310/00365 Proteins; Polypeptides; Degradation products thereof
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 9/0024 Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue compositions for intravenous administration, normal injectable solutions or dispersions for, e.g. subcutaneous administration A61K9/0019; brain implants A61K9/0085; (coated) prostheses, catheters or stents A61L
• CPC Class Codes: A61L 2430/02 for reconstruction of bones; weight-bearing implants
• CPC Class Codes: A61L 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• CPC Class Codes: A61L 27/56 Porous materials, e.g. foams or sponges
• CPC Class Codes: C07K 14/51 Bone morphogenetic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
• CPC Class Codes: Y10S 530/84 Bones; tendons; teeth; cartilage

Abstract

Disclosed are (1) osteogenic devices comprising a matrix containing osteogenic protein and methods of inducing endochondral bone growth in mammals using the devices; (2) amino acid sequence data, amino acid composition, solubility properties, structural features, homologies and various other data characterizing osteogenic proteins, (3) methods of producing osteogenic proteins using recombinant DNA technology, and (4) osteogenically and chondrogenically active synthetic protein constructs.

Claims

1. An osteogenic device for implantation in a mammal comprising osteogenic protein dispersed within a biocompatible, in vivo biodegradable matrix defining pores of a dimension sufficient to permit influx, proliferation and differentiation of migratory progenitor cells from the body of said mammal,

the improvement wherein said osteogenic protein is produced by expression of recombinant DNA in a host cell, and comprises a pair of polypeptide chains, each of which has an amino acid sequence sufficiently duplicative of the sequence of COP-5 or COP-7 such that said pair of polypeptide chains, when disulfide bonded to produce a dimeric species, has a conformation capable of inducing endochondral bone formation in association with said matrix when implanted in a mammal.

3. The device of claim 1 or 2 wherein the sequence comprises:

##STR25##

4. The device of claim 1 or 2 wherein the sequence comprises:

##STR26##

5. The device of claim 1 or 2 wherein the sequence comprises:

##STR27##

6. The device of claim 1 or 2 wherein the sequence comprises:

##STR28##

7. The device of claim 1 or 2 wherein the sequence comprises:

##STR29##

8. The device of claim 1 or 2 wherein the sequence comprises:

##STR30##

9. The device of claim 1 or 2 wherein the sequence comprises:

##STR31##

10. The device of claim 1 or 2 wherein the sequence comprises:

##STR32##

11. The device of claim 1 or 2 wherein the sequence comprises:

##STR33##

12. The device of claim 1 or 2 wherein the sequence comprises:

##STR34##

13. The device of claim 1 or 2 wherein the sequence comprises:

##STR35##

14. The device of claim 1 or 2 wherein the sequence comprises:

##STR36##

16. The device of claim 1, 2, or 15 wherein said matrix comprises close-packed particulate matter having a particle size within the range of 70-850 μ

m.

17. The device of claim 1, 2, or 15 wherein said particulate matter has a particle size within the range of 70-420 μ

m.

18. The device of claim 1, 2, or 15 wherein said matrix comprises demineralized, protein-extracted, particulate, allogenic bone.

19. The device of claim 1, 2, or 15 wherein said matrix comprises a material selected from the group consisting of collagen, hydroxyapalite, tricalcium phosphate, polymers comprising lactic acid monomer units, polymers comprising glycolic acid monomer units, demineralized, guanidine-extracted allogenic bone, and mixtures thereof.

20. The device of claim 1, 2, or 15 wherein said matrix is shaped to span a non-union fracture in said mammal.

21. The device of claim 1, 2, or 15 disposed within the marrow cavity of allogenic bone.

22. The device of claim 1, 2, or 15 wherein said matrix comprises demineralized, protein extracted, particulate, deglycosylated xenogeneic bone.

23. The device of claim 22 wherein said matrix is treated with a protease.

32. A method of inducing local cartilage or bone formation in a mammal comprising the step of implanting the device of claim 1, 2, or 15 in said mammal at a locus accessible to migratory progenitor cells of said mammal.

33. A method of inducing endochondral bone formation in a mammal comprising the step of implanting the device of claim 1 or 15 in said mammal at a locus accessible to migratory progenitor cells of said mammal.

34. A method of inducing endochondral bone formation in a non-union fracture in a mammal comprising the step of implanting in the fracture in said mammal the device of claim 20.

35. The device of claim 1 or 2 wherein the sequence comprises:

##STR37## wherein X₁ =(K or R);

X₂ =(H, R, or K);

X₃ =(P, S, E or Q);

X₄ =(Y, K or F);

X₅ =(D, S or E);

X₆ =(R, S, K or A);

X₇ =(V, L or I);

X₈ =(N, Q, D or S);

X₉ =(D, E or N);

X₁₀ =(I or V);

X₁₁ =(I or V);

X₁₂ =(A or S);

X₁₃ =(P, E, L or K);

X₁₄ =(Y or F);

X₁₅ =(H or D);

X₁₆ =(F, Y or N);

X₁₇ =(H, E or S);

X₁₈ =(E or A);

X₁₉ =(P, A or Q);

X₂₀ =(F or Y);

X₂₁ =(L, M or I);

X₂₂ =(A, P or T);

X₂₃ =(D, E or K);

X₂₄ =(H or S)p;

X₂₅ =(L, M or F);

X₂₆ =(N or K);

X₂₇ =(S, A or P);

X₂₈ =(T or A);

X₂₉ =(I, V, or T);

X.sub. 30 =(V, I or L);

X₃₁ =(T or S);

X₃₂ =(L or I);

X₃₃ =(N, H or R);

X₃₄ =(S, A, F or N);

X₃₅ =(V or I);

X₃₆ =(P or S);

X₃₇ =(G or E);

X₃₈ =(K, Q, T or S);

X₃₉ =(I or V);

X₄₀ =(K or E);

X₄₁ =(A, P or S);

X₄₂ =(V or A);

X₄₃ =(T or E);

X₄₄ =(E, Q or K);

X₄₅ =(L or M);

X₄₆ =(S N or D);

X₄₇ =(A, S or P);

X₄₈ =(I, L or V);

X₄₉ =(S or A);

X₅₀ =(M, I or V);

X₅₁ =(Y or F);

X₅₂ =(L, F or Y);

X₅₃ =(D or N);

X₅₄ =(E, D or N);

X₅₅ =(N or Q);

X⁵⁶ =(E, D, S or K);

X₅₇ =(N or K);

X₅₈ =(V or I);

X₅₉ =(K or R);

X₆₀ =(N, K or H);

X₆₁ =(Q, E, R or P);

X₆₂ =(D, E or N);

X₆₃ =(V or T);

X₆₄ =(E, D or R);

X₆₅ =(G, A, S or E);

X₆₆ =(G or H); and

X₆₇ =(R or H).

36. The device of claim 1 or 2 wherein the sequence comprises:

##STR38## wherein X₁ =(Y, K or F);

X₂ =(D, S, or E);

X₃ =(R, S, K or A);

X₄ =(Y, L or I);

X₅ =(N, Q, D or S);

X₆ =(D, E or N);

X₇ l =(I or V);

X₈ =(I or V);

X₉ =(A or S);

X₁₀ =(P, E, L or K);

X₁₁ =(Y or F);

X₁₂ =(H or D);

X₁₃ =(F, Y or N);

X₁₄ =(H, E or S);

X₁₅ =(E or A);

X₁₆ =(P, A or Q);

X₁₇ =(F or Y);

X₁₈ =(L, M or I);

X₁₉ =(A, P or T);

X₂₀ =(D, E or K);

X₂₁ =(H or S);

X₂₂ =(L, M or F);

X₂₂ =(N or K);

X₂₇ =(S, A or P);

X₂₅ =(T or A);

X₂₆ =(I, V, or T);

X₂₇ =(V, I or L);

X₂₈ =(T or S);

X₂₉ =(L or I);

X₃₀ =(N, H or R);

X₃₁ =(S, A, F or N);

X₃₂ =(V or I);

X₃₃ =(P or S);

X₃₄ =(G or E);

X₃₅ =(K, Q, T or S);

X₃₆ =(I or V);

X₃₇ =(K or E);

X₃₈ =(A, P or S);

X₃₉ =(V or A);

X₄₀ =(T or E);

X₄₁ =(E, Q or K);

X₄₂ =(L or M);

X₄₃ =(S N or D);

X₄₄ =(A, S or P);

X₄₅ =(I, L or V);

X₄₆ =(S or A);

X₄₇ =(M, I or V);

X₄₈ =(Y or F);

X₄₉ =(L, F or Y);

X₅₀ =(D or N);

X₅₁ =(E, D or N);

X₅₂ =(N or Q);

X⁵³ =(E, D, S or K);

X₅₄ =(N or K);

X₅₅ =(V or I);

X₅₆ =(K or R);

X₅₇ =(N, K or H);

X₅₈ =(Q, E, R or P);

X₅₉ =(D, E or N);

X₆₀ =(V or T);

X₆₁ =(E, D or R);

X₆₂ =(G, A, S or E);

X₆₂ =(G or H); and

X₆₄ =(R or H).

37. The device of claim 1 or 2 wherein the sequence comprises:

##STR39##

2. A device for implantation in a mammal, said device comprising chondrogenic protein dispersed within a biocompatible, in vivo biodegradable matrix defining pores of a dimension sufficient to permit influx, proliferation and differentiation of migratory progenitor cells from the body of said mammal,

the improvement wherein said protein is produced by expression of recombinant DNA in a host cell, and comprises a pair of polypeptide chains, each of which has less than about 200 amino acids, in a sequence sufficiently duplicative of the sequence of COP-5 or COP-7 such that said pair of polypeptide chains, when disulfide bonded to produce a dimeric species, has a conformation that is capable of inducing cartilage formation in association with said matrix when implanted in a mammal.

15. An osteogenic device for implantation in a mammal, said device comprising osteogenic protein dispersed within a biocompatible, in vivo biodegradable matrix defining pores of a dimension sufficient to permit influx, proliferation and differentiation of migratory progenitor cells from the body of said mammal,

the improvement wherein said osteogenic protein comprises substantially pure protein capable of inducing endochondral bone formation in said mammal, said osteogenic protein having a half maximum bone inducing activity of at least about 0.8 to 1.0 ng per mg of said matrix and comprising the amino acid sequence:

V--P--K--P--C--C--A--P--T.

24. The device of claim 15 wherein said osteogenic protein is unglycosylated.

25. The device of claim 24 wherein said osteogenic protein has a apparent molecular weight of about 27 kD when oxidized as determined by comparison to molecular weight standards in SDS-polyacrylamide gel electrophoresis.

26. The device of claim 15 wherein said osteogenic protein is glycosylated.

27. The device of claim 26 wherein said osteogenic protein has a molecular weight of about 30 kD when oxidized as determined by comparison to molecular weight standards in SDS-polyacrylamide gel electrophoresis.

28. The device of claim 15 wherein said osteogenic protein comprises a pair of polypeptide chains.

29. The device of claim 28 wherein one chain of said pair of polypeptide chains has a molecular weight of about 14 kD and the other has an apparent molecular weight of about 16 kD, both as determined after reduction by comparison to molecular weight standards in SDS-polyacrylamide gel electrophoresis.

30. The device of claim 28 wherein one chain of said pair of polypeptide chains has a molecular weight of about 16 kD and the other has an apparent molecular weight of about 18 kD, both as determined after reduction by comparison to molecular weight standards in SDS-polyacrylamide gel electrophoresis.

31. The device of claim 15 wherein said osteogenic protein has the approximate amino acid composition set forth below:

space="preserve" listing-type="tabular">______________________________________ Amino acid Rel. no. Amino acid Rel. no. residue res./molec. residue res./molec. ______________________________________ Aspartic acid/ 22 Tyrosine 11 Asparagine Valine 14 Glutamic acid/ 24 Methionine 3 Glutamine Cysteine 16 Serine 24 Isoleucine 15 Glycine 29 Leucine 15 Histidine 5 Proline 14 Arginine 13 Phenylalanine 7 Threonine 11 Tryptophan ND Alanine 18 Lysine 12 ______________________________________