Claims for Patent: 5,770,404

Summary for Patent: 5,770,404

Title:	Preparation of hemoproteins from apoproteins and heme-like porphyrins derived from chlorophyll
Abstract:	Hemin-like porphyrins can be prepared from chlorophylls which may be used as intermediates to form functionally active hemoproteins. The porphyrins can be reacted with apoproteins in vitro or added to a cell culture and combined intracellularly with apoproteins to form functionally active hemoproteins.
Inventor(s):	Camiener; Gerald W. (Solon, OH)
Assignee:
Application Number:	08/659,416
Patent Claims:	1. A method of producing a hemoprotein comprising an apoprotein and a heme-like porphyrin wherein said heme porphyrin is represented by structure II or III: ##STR4## wherein R.sup.1 through R.sup.9 independently represent hydrogen, methyl, ethyl, vinyl, propyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, formyl, ethanalyl or propionalyl and Me is a metal atom; wherein said heme-like porphyrin is obtained from chlorophyll; comprising the steps of a) contacting a composition comprising said heme-like porphyrin and a carrier with a fermentation or incubation medium containing naturally-occurring cells, genetically modified cells, organelles, organisms or combinations thereof, which produce an apoprotein; and b) collecting the hemoprotein produced in step a). 2. A method of producing a hemoprotein comprising an apoprotein and a heme-like porphyrin wherein said heme porphyrin is represented by structure II or III: ##STR5## wherein R.sup.1 through R.sup.9 independently represent hydrogen, methyl, ethyl, vinyl, propyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, formyl, ethanalyl or propionalyl and Me is a metal atom; wherein said heme-like porphyrin is obtained from chlorophyll; comprising the steps of a) contacting a composition comprising said heme-like porphyrin and a carrier with a medium containing an apoprotein; and b) collecting the hemoprotein produced in step a). 3. A method as claimed in claim 1 wherein said heme-like porphyrin is selected from the group consisting of hemin, deuteroheme, mesoheme, hematoheme, etioheme, rhodoheme, protoheme, phylloheme, coproheme, iron chlorophyllin, dibromoprotoheme, isoheme, hemin C, pyrroheme, siroheme, their non-metal-containing counterparts, and the methyl, ethyl, propyl, formyl, acetyl, cysteine, and proprionyl derivatives thereof. 4. A method as claimed in claim 2 wherein said heme-like porphyrin is selected from the group consisting of hemin, deuteroheme, mesoheme, hematoheme, etioheme, rhodoheme, protoheme, phylloheme, coproheme, iron chlorophyllin, dibromoprotoheme, isoheme, hemin C, pyrroheme, siroheme, their non-metal-containing counterparts, and the methyl, ethyl, propyl, formyl, acetyl, cysteine, and proprionyl derivatives thereof. 5. A method as claimed in claim 1 wherein the apoprotein is selected from the group consisting of the apoprotein for hemoglobin, myoglobin, catalase, peroxidase, cytochrome, tryptophane pyrrolase, and cytochrome P-450. 6. A method as claimed in claim 2 wherein the apoprotein is selected from the group consisting of the apoprotein for hemoglobin, myoglobin, catalase, peroxidase, cytochrome, tryptophane pyrrolase, and cytochrome P-450. 7. A method as claimed in claim 1 wherein the cells, organelles, and organisms are selected from the group consisting of bacteria, yeast, fungi, algae, plants, plant cells, protozoans, metazoans, animals, and animal cells. 8. A method as claimed in claim 1, wherein the porphyrin is represented by structure II and the process further comprises the step of adding a metal ion to the hemoprotein of step b. 9. A method as claimed in claim 2, wherein the porphyrin is represented by structure II and the process further comprises the step of adding a metal ion to the hemoprotein of step b. 10. The method as claimed in 1, wherein said heme-like porphyrin is produced by opening the cyclopentanone ring of a chlorophyll. 11. The method as claimed in claim 10, wherein the step of opening the cyclopentanone ring of the chlorophyll is effected by alkaline hydrolysis. 12. The method as claimed in claim 11, wherein the chlorophyll is represented by structure I: ##STR6## wherein the pyrrole ring II of structure I may be reduced; R.sup.1 represents vinyl, formyl, or hydrogen; R.sup.2 represents methyl, formyl, or hydrogen; R.sup.3 represents --(CH.sub.2).sub.2 COOH, or --(CH.sub.2).sub.2 COO-phytyl; or the chlorophyll is chlorophyll c. 13. The method as claimed in claim 12, wherein the heme-like porphyrin is pyrrohemin obtained from chlorophyll a. 14. The method as claimed in 2, wherein said heme-like porphyrin is produced by opening the cyclopentanone ring of a chlorophyll. 15. The method as claimed in claim 14, wherein the step of opening the cyclopentanone ring of the chlorophyll is effected by alkaline hydrolysis. 16. The method as claimed in claim 15, wherein the chlorophyll is a represented by structure I: ##STR7## wherein the pyrrole ring II of structure I may be reduced; R.sup.1 represents vinyl, formyl, or hydrogen; R.sup.2 represents methyl, formyl, or hydrogen; R.sup.3 represents --(CH.sup.2).sub.2 COOH, or --(CH.sub.2).sub.2 COO-phytyl; or the chlorophyll is chlorophyll c. 17. The method as claimed in claim 16, wherein the heme-like porphyrin is pyrrohemin obtained from chlorophyll a. 18. A method for producing a hemoprotein comprising the steps of a) opening the cyclopentanone ring of a chlorophyll to obtain a heme-like porphyrin; b) contacting the heme-like porphyrin of step a) and a carrier with an apoprotein; and c) collecting the hemoprotein produced in step b). 19. The method as claimed in claim 18, wherein the step of opening the cyclopentanone ring of the chlorophyll is effected by alkaline hydrolysis. 20. The method as claimed in claim 19, wherein the chlorophyll is represented by structure I: ##STR8## wherein the pyrrole ring II of structure I may be reduced; R.sup.1 represents vinyl, formyl, or hydrogen; R.sup.2 represents methyl, formyl, or hydrogen; R.sup.3 represents --(CH.sub.2).sub.2 COOH, or --(CH.sub.2).sub.2 COO-phytyl; or the chlorophyll is chlorophyll c. 21. The method as claimed in claim 20, wherein the heme-like porphyrin is pyrrohemin obtained from chlorophyll a. 22. A method for producing a hemoprotein comprising the steps of a) opening the cyclopentanone ring of a chlorophyll to obtain a heme-like porphyrin; b) contacting the heme-like porphyrin of step a) and a carrier with a fermentation or incubation medium containing naturally-occurring cells, genetically modified cells, organelles, organisms or combinations thereof; c) collecting the hemoprotein produced in step b). 23. The method as claimed in claim 22, wherein the step of opening the cyclopentanone ring of the chlorophyll is effected by alkaline hydrolysis. 24. The method as claimed in claim 23, wherein the chlorophyll is represented by structure I: ##STR9## wherein the pyrrole ring II of structure I may be reduced; R.sup.1 represents vinyl, formyl, or hydrogen; R.sup.2 represents methyl, formlyl, or hydrogen; R.sup.3 represents --(CH.sub.2).sub.2 COOH, or --(CH.sub.2).sub.2 COO-phytyl; or the chlorophyll is chlorophyll c. 25. The method as claimed in claim 24, wherein the heme-like porphyrin is pyrrohemin obtained from chlorophyll a. 26. A method for producing a hemoprotein, comprising the steps of a) extracting a porphyrin from photosynthetic cells, b) converting said porphyrin of step a) into a heme-like porphyrin; (c) either (i) contacting the heme-like porphyrin of step b) and a carrier with a fermentation or incubation medium containing naturally-occurring cells, genetically modified cells, organelles, organisms or combinations thereof; or (ii) contacting the heme-like porphyrin of step b) and a carrier with an apoprotein; and d) collecting the hemoprotein produced in step c). 27. The method of claim 26 wherein the porphyrin of step a) is selected from the group consisting of a chlorophyll and a plant-produced porphyrin. 28. The method of claim 27, wherein the plant-produced porphyrin is protoporphyrin IX. 29. The method of claim 26, wherein said porphyrin of step a) is a metalloporphyrin. 30. The method of claim 29, wherein the hemeprotein of step a) is hemoglobin.

Details for Patent 5,770,404

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		2040-01-28