Lung tissue extract useful for treating hyaline-membrane disease and method for producing the extract

Claim 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 μg 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 2 and 45.6 cm 2 respectively.

Claim 14

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 2 and 45.6 cm 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%.

Claim 15

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 2 and 45.6 cm 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%.