Claims for Patent: 6,916,787

Summary for Patent: 6,916,787

Title:	Method for producing hemin proteins using plant cells, resulting proteins and products containing same
Abstract:	A method for producing haemin proteins by (i) inserting into plant cells one or more nucleic acid molecules that each comprise at least one sequence coding for a protein component of an animal haemin protein capable of reversibly binding oxygen, or for a variant or portion of said protein component, and optionally a sequence coding for a selection agent; (ii) selecting cells containing nucleic acid coding for the protein component of the haemin protein; (iii) optionally propagating the transformed cells either in a culture or by regenerating whole transgenic or chimeric plants; and (iv) recovering and optionally purifying a haemin protein that includes a complex consisting of the protein or proteins coded for by said nucleic acid and at least one iron-containing porphyritic nucleus, or a plurality of such complexes.
Inventor(s):	Merot; Bertrand (Volvic, FR), Dieryck; Wilfrid (Saint-Pathus, FR), Lenee; Philippe (Noumea, FR), Marden; Michael (Aulnay-sous-Bois, FR), Gruber; Veronique (Chamalieres, FR), Pagnier; Renee-Josee (Le Kremlin-Bicetre, FR), Baudino; Sylvie (Orcines, FR), Poyart; Claude (Paris, FR)
Assignee:	Institut National de la Sante et de Recherche Medicale (Paris Cedex, FR) Meristern Therapeutics (Clermont-Ferrand, FR)
Application Number:	10/085,853
Patent Claims:	1. A recombinant hemin protein having the capacity to reversibly bind oxygen, comprising at least one iron-containing porphyrin nucleus, of plant origin, and a protein component comprising at least one polypeptide chain selected from the group consisting on hemoglobin, myoglobin, cytochromes, peroxidases, and catalases of animal origin. 2. The recombinant protein according to claim 1, wherein the at least one iron-containing porphyrin nucleus is iron-containing protoporphyrin IX, or a protoporphyrin differing from protoporphyrin IX in the nature of the side chains, carried by the .beta. atoms of pyrrole rings. 3. The recombinant protein according to claim 1, wherein the protein component comprises at least one .alpha. and/or .beta.-globin polypeptide chain, or variants of said polypeptide chain, wherein the variant of the .alpha. chain has at least 90% homology with an .alpha. chain having an amino acid sequence of SEQ ID NO:31 and the variant of the .beta. chain has at least 90% homology with a .beta. chain having an amino acid sequence of SEQ ID NO:33, and the hemin protein is capable of binding oxygen reversibly. 4. The recombinant protein according to claim 3, wherein the .alpha. or .beta.-globin chain, or variants of the said polypeptide chain, further comprises a chloroplast targeting signal, a mitochondrial targeting signal, or a N-terminal signal peptide in combination with a signal responsible for retaining a protein in the endoplasmic reticulum or a N-terminal signal peptide in combination with a vacuolar targeting signal. 5. The recombinant protein according to claim 3, wherein each .alpha. and/or .beta.-globin polypeptide chain lacks an NH.sub.2 terminal methionine. 6. The recombinant hemin protein according to claim 1, wherein the protein component comprises at least four polypeptide chains of .alpha. and/or .beta.-globin or variants of the said polypeptide chain, each said polypeptide chain being bound to an iron-containing protoporphyrin nucleus, wherein the variant of the .alpha. chain has at least 90% homology with an .alpha. chain having an amino acid sequence of SEQ ID NO:31 and the variant of the .beta. chain has at least 90% homology with a .beta. chain having an amino acid sequence of SEQ ID NO:33, and the hemin protein is capable of binding oxygen reversibly. 7. The recombinant protein according to claim 6, wherein the protein component comprises 2 .alpha.-globin chains and 2 .beta. globin chains, or variants of the said polypeptide chain. 8. The recombinant protein according to claim 1, wherein said protein binds oxygen with an affinity of between 7 and 40 mm Hg. 9. A pharmaceutical product comprising one or more recombinant hemin protein(s) according to claim 1 in association with a physiologically acceptable excipient. 10. A recombinant hemin protein having the capacity to reversibly bind oxygen, comprising at least one iron-containing porphyrin nucleus of plant origin, and a protein component comprising at least one polypeptide chain selected from the group consisting of hemoglobin, myoglobin, and cytochromes of animal origin. 11. The recombinant protein according to claim 10, wherein the at least one iron-containing porphyrin nucleus is iron-containing protoporphyrin IX, or a protoporphyrin differing from protoporphyrin IX in the nature of the side chains, carried by the .beta. atoms of pyrrole rings. 12. The recombinant protein according to claim 10, wherein the protein component comprises at least one .alpha. and/or .beta.-globin polypeptide chain, or variants of said polypeptide chain, wherein the variant of the .alpha. chain has at least 90% homology with an .alpha. chain having an amino acid sequence of SEQ ID NO:31 and the variant of the .beta. chain has at least 90% homology with a .beta. chain having an amino acid sequence of SEQ ID NO:33, and the hemin protein is capable of binding oxygen reversibly. 13. The recombinant protein according to claim 12, wherein the .alpha. or .beta.-globin chain, or variants of the said polypeptide chain, further comprises a chloroplast targeting signal, a mitochondrial targeting signal, or a N-terminal signal peptide in combination with a signal responsible for retaining a protein in the endoplasmic reticulum or a N-terminal signal peptide in combination with a vacuolar targeting signal. 14. The recombinant protein according to claim 12, wherein each .alpha. and/or .beta.-globin polypeptide chain lacks an NH.sub.2 terminal methionine. 15. The recombinant hemin protein according to claim 10, wherein the protein component comprises at least four polypeptide chains of .alpha. and/or .beta.-globin or variants of said polypeptide chain, each said polypeptide chain being bound to an iron-containing protoporphyrin nucleus, wherein the variant of the .alpha. chain has at least 90% homology with an .alpha. chain having an amino acid sequence of SEQ ID NO:31 and the variant of the .beta. chain has at least 90% homology with a .beta. chain having an amino acid sequence of SEQ ID NO:33, and the hemin protein is capable of binding oxygen reversibly. 16. The recombinant protein according to claim 15, wherein the protein component comprises 2 .alpha.-globin chains and 2 .beta. chains, or variants of the said polypeptide chain. 17. The recombinant protein according to claim 10, wherein said protein binds oxygen with an affinity of between 7 and 40 mm Hg. 18. A pharmaceutical product comprising one or more recombinant hemin protein(s) according to claim 10 in association with a physiologically acceptable excipient. 19. The recombinant protein according to claim 10, wherein the protein component comprises at least one .alpha. and/or .beta.-globin polypeptide chain, or variants of said polypeptide chain, wherein the variant of the .alpha. chain has a heme binding domain and the variant of the .beta. chain has a heme binding domain and the hemin protein is capable of binding oxygen reversibly.

Details for Patent 6,916,787

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Recordati Rare Diseases, Inc.	PANHEMATIN	hemin for injection	For Injection	101246	07/20/1983		2015-07-17