Claims for Patent: 4,338,301

Summary for Patent: 4,338,301

Title:	Lung tissue extract useful for treating hyaline-membrane disease and method for producing the extract
Abstract:	A surface active material may be produced from the lung tissue of mammal by means of employing an acetone-treatment procedure as well as an organic solvents mixture-treatment procedure other than the conventional procedures of differential centrifugation, density gradient centrifugation and dialysis. The chemical composition of the active material has for example, a phospholipid content of 75.0-95.5%, a neutral lipid content of 1.8-14.0%, total cholesterol content of 0.0-3.0%, carbohydrate content is 0.1-1.5% and protein content is 0.5-5.0%, all based on the dried weight of the material. The pharmaceutical composition containing the surface active material defined as above is usable for clinical treatment of human hyaline-membrane disease.
Inventor(s):	Tetsuro; Fujiwara (Akita, JP), Yuji; Tanaka (Toda, JP), Tsunetomo; Takei (Kuki, JP)
Assignee:	Tokyo Tanabe Co., Ltd. (JP)
Application Number:	06/152,048
Patent Claims:	1. Surface active material containing phospholipid, neutral lipid, total cholesterol, carbohydrate, protein and water, which material is obtained from lung tissue of a mammal with or without further phospholipid, characterized in that the phospholipid content is 75.0-95.5%, the neutral lipid content is 1.8-14.0%, the total cholesterol content is 0.0-3.0%, the carbohydrate content is 0.1-1.5%, the protein content is 0.5-5.0% and water content is 1.7-6.0%, all based on the dried weight of said material, the minimum and maximum surface tension ranges of the material estimated by Wilhelmy's method wherein the material is added dropwise to the surface of physiological saline in an amount of 0.3-0.8 .mu.g per square centimeter of surface area thereof being 2.1-8.6 dynes/cm and 48.2-58.0 dynes/cm when surface areas are 21.0 cm.sup.2 and 45.6 cm.sup.2 respectively. 2. Surface active material as claimed in claim 1, wherein the phospholipid content is estimated by multiplying its phosphorous content by 25; the individual contents of neutral lipid, total cholesterol carbohydrate and protein being expressed by calculating in respectively glycerol trioleate equivalent, cholesterol equivalent, glucose equivalent and bovine serum albumin equivalent. 3. Surface active material as claimed in claim 2, wherein the phospholipid content consists mainly of phosphatidylcholine and phosphatidylglycerol, the contents of which are 63.0-85.5% and 3.0-12.0% respectively, both based on the total weight of phospholipid content. 4. Surface active material as claimed in claim 3, wherein the content of phosphatidylcholine having two saturated fatty acid residues based on the total weight of phosphatidylcholine is 67.5-90.3%. 5. Surface active material as claimed in claim 4, wherein one of the saturated fatty acid residues is palmitic acid residue. 6. Surface active material as claimed in any one of claims 1, 2, 3, 4 or 5 wherein the mammal is selected from the group consisting of cattle, horses, sheep and pigs. 7. A pharmaceutical composition useable for the treatment of hyaline-membrane disease comprising an effective amount of surface-active material as set forth in claim 1 and a pharmaceutically acceptable non-toxic carrier thereof. 8. A pharmaceutical composition as claimed in claim 7, wherein the effective amount for a mammal is between about 50 and 400 milligrams per one treatment. 9. A pharmaceutical composition according to claim 7, wherein the carrier is selected from the group consisting of water and electrolyte solution. 10. A pharmaceutical composition according to claim 9, wherein the electrolyte solution is physiological saline. 11. A pharmaceutical composition according to claim 7, wherein the effective amount is administered directly into an airway of a mammal suffering from hyaline-membrane disease. 12. A pharmaceutical composition according to claim 11, wherein the effective amount is administered in a suspension form. 13. A method of producing a surface active material containing phospholipid, neutral lipid, total cholesterol, carbohydrate, protein and water, which material is obtained from lung tissue of a mammal with or without further phospholipid, characterized in that the phospholipid content is 75.0-95.5%, the neutral lipid content is 1.8-14.0%, the total cholesterol content is 0.0-3.0%, the carbohydrate content is 0.1-1.5%, the protein content is 0.5-5.0% and water content is 1.7-6.0%, all based on the dried weight of said material, the minimum and maximum surface tension ranges of the material estimated by Wilhelmy's method wherein the material is added dropwise to the surface of physiological saline in an amount of 0.3-0.8 g per square centimeter of surface area thereof being 2.1-8.6 dynes/cm and 48.2 to 58.0 dynes/cm when surface areas are 21.0 cm.sup.2 and 45.6 cm.sup.2 respectively, comprising the steps of: (a) mincing lung tissue of a mammal; (b) bringing the minced lung tissue into contact with an electrolyte solution and subjecting the resultant mixture to one of filtration under pressure and centrifugation at about 500-1,500 rpm to obtain an extract; (c) centrifuging the extract at about 12,000-16,000 rpm to obtain a crude sediment; (d) forming an aqueous suspension of the crude sediment and dissolving sodium chloride in the aqueous suspension of the crude sediment to obtain a density adjusted suspension and then centrifuging the adjusted suspension at about 4,000-10,000 rpm so as to divide it into three layers, the top layer being an emulsified scum layer which is then taken up; (e) forming an aqueous suspension of the top layer and dialyzing the aqueous suspension through a semi-permeable membrane to remove inorganic salts and water-soluble organic compounds of low molecular weight both having been remained in the top layer, whereby an aqueous suspension containing the non-dialyzing material is obtained; (f) centrifuging the resultant suspension at about 12,000-16,000 rpm to take up a pure sediment; (g) suspending one of the pure sediments and the aqueous suspension containing the non-dialyzing material, in an aqueous sucrose solution and centifuging the suspension of aqueous sucrose solution at about 18,000-40,000 rpm to obtain another pure sediment; (h) drying the pure sediment and suspending the resultant dried pure sediment in acetone to take up an acetone-insoluble material which is then dried and suspended further in organic solvent mixture followed by filtration of the resultant suspension so as to obtain a purified filtrate; and (i) mixing the purified filtrate with sterile water and subjecting the resultant mixture to filtration, the filtrate of which is then concentrated under reduced pressure to obtain a solid residue, followed by one of lyophilization of the said solid residue so that the phospholipid content is 75.0-95.5% or adding further phospholipid solid residue to bring the phospholipid content to 75.0-95.5%. 14. Method as claimed in claim 13, wherein the mincing is performed by using a meat-grinder. 15. Method as claimed in claim 13, wherein the minced lung tissue before being used is lyophilized and then preserved. 16. Method as claimed in claim 13, wherein the electrolyte solution is physiological saline. 17. Method as claimed in claim 13, wherein the density of the adjusted suspension of step (d) has a value of 1.07-1.20. 18. Method as claimed in claim 13, wherein the semipermeable membrane is selected from the group consisting of cellophane membrane, collodion membrane and bladder membrane. 19. Method as claimed in claim 13, wherein one part of the dried pure sediment of step (h) is suspended in 100-300 parts of acetone at -10.degree.-+10.degree. C. 20. Method as claimed in claim 13, wherein one part of the dried acetone-insoluble material of step (h) is suspended in 100-300 parts of organic solvent mixture. 21. Method as claimed in claim 20, wherein the organic solvent mixture is selected from the group consisting of chloroform-methanol with a 2:1 volume ratio, chloroform-ethanol with a 2:1 volume ratio, chloroform-isopropanol with a 1:1 volume ratio and ethyl ether-ethanol with a 3:1 volume ratio. 22. Method as claimed in claim 13, wherein the further phospholipid is selected from the group consisting of phosphatidylglycerol, phosphatidylcholine having two saturated fatty acid residues, and a mixture thereof. 23. Method as claimed in claim 22, wherein the saturated fatty acid residue is palmitic acid residue.

Details for Patent 4,338,301

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	ZOSTAVAX	zoster vaccine live	For Injection	125123	05/25/2006		1999-07-06