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US 5,013,720 AGranted
SAP-6-Val proteins and methods
AssigneeAbbott Laboratories
InventorJeffrey A. Whitsett
Life of this patent
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AssigneeAbbott Laboratories
InventorJeffrey A. Whitsett
Estimating the term…
A novel hydrophobic surfactant-associated protein mixture, i.e., a SAP-6 proteins, has been isolated from pulmonary animal tissue. A small, novel pulmonary hydrophobic surfactant-associated SAP-6-Val protein having a molecular weight of about 6,000 daltons as determined by SDS-PAGE and about 3,500-4,000 daltons as determined by tricine-SDS-PAGE has been further isolated from the SAP-6 protein mixture. The amino acid residue compositions of the SAP-6-Val protein for human and bovine have been determined and disclosed. When a SAP-6-Val protein is combined with phospholipids, it enhances the surfactant-like activity of the phospholipids in lungs of animals and, therefore, uniquely imparts to the mixture significant pulmonary biophysical activity. Such a mixture results in enhanced adsorption of the phospholipids with properties similar to that of natural pulmonary surfactant material. SAP-6-Val proteins in combination with phospholipids is highly useful for replacing or supplementing natural pulmonary surfactant material for reducing or maintaining normal surface tension in lungs, and especially in lungs of patients suffering from hyaline membrane disease, HMD, or other syndromes associated with the lack or insufficient amounts of natural pulmonary surfactant material. A mixture of a SAP-6-Val protein and phospholipids may be administered as an aerosol spray or in aqueous normal saline with or without calcium chloride for treating or preventing HMD and other surfactant deficiency states. Also disclosed are methods of isolating the noncanine SAP-6-Val proteins from the SAP-6 proteins isolated from animal tissue.
An isolated lung specific hydrophobic SAP-6-Val protein, said protein comprises a plurality of valine amino acid residues having a first stretch of about six adjacent valine amino acid residues and a second stretch of about three adjacent valine amino acid residues separated from the first stretch by about two hydrophobic amino acid residues, said protein enhances surfactant-like activity of phospholipids in lungs of an animal, has a molecular weight of about 3,500-4,000 daltons as determined by tricine-SDS-PAGE and a molecular weight of about 6,000 daltons as determined by SDS-PAGE, and is substantially resistant to protease, endoglycosidase F and collagenase enzymes.
An isolated lung specific hydrophobic surfactant-associated human protein having a molecular weight of about 3,500-4,000 daltons as determined by tricine-SDS-PAGE, said human protein enhancing surfactant-like activity of phospholipids in lungs of an animal and comprising the following amino acid sequence:
An isolated lung specific hydrophobic surfactant-associated human protein having a molecular weight of about 3,500-4,000 daltons as determined by tricine-SDS-PAGE, said human protein enhancing surfactant-like activity of phospholipids in lungs of an animal and comprising the following amino acid sequence:
An isolated lung specific hydrophobic surfactant-associated bovine protein having a molecular weight of about 3,500-4,000 daltons as determined by tricine-SDS-PAGE, said bovine protein enhancing surfactant-like activity of phospholipids in lungs of an animal and comprising the following amino acid sequence:
A method of isolating from animal pulmonary or amniotic tissue a lung specific hydrophobic surfactant-associated SAP-6-Val protein for enhancing surfactant-like activity of phospholipids in lungs of an animal, the SAP-6-Val protein having a molecular weight of about 6,000 daltons as determined in a polyacrylamide gel containing sodium dodecyl sulfate and being substantially resistant to protease, endoglycosidase F, and collagenase enzymes, said method comprises the steps of:
A substantially pure lung specific hydrophobic surfactant-associated SAP-6-Val protein isolated from animal pulmonary or amniotic tissue having a hydrophobic region of amino acids which includes a first stretch of about 6 adjacent valine amino acids and a second stretch of about 3 adjacent valine amino acids separated from the first stretch by about 2 hydrophobic amino acid residues for enhancing surfactant-like activity of phospholipids in lungs of an animal, said SAP-6-Val protein further having a molecular weight of about 6,000 as daltons determined in a polyacrylamide gel containing sodium dodecyl sulfate and a molecular weight of about 3,500-4,000 daltons as determined by tricine-SDS-PAGE and being substantially resistant to protease, endoglycosidase F, and collagenase enzymes, said SAP-6-Val protein being produced by a process comprising the steps of:
A substantially pure lung specific hydrophobic surfactant-associated human SAP-6-Val protein isolated from human pulmonary or amniotic tissue having a hydrophobic region of amino acids which includes a first stretch of about 6 adjacent valine residues and a second stretch of about 3 valine residues separated from the first stretch by about 2 hydrophobic amino acid residues for enhancing surfactant-like activity of phospholipids in lungs of an animal, said protein further having a simple molecular weight of about 6,000 daltons as determined in a polyacrylamide gel containing sodium dodecyl sulfate and a molecular weight of about 3,500-4,000 daltons as determined by tricine-SDS-PAGE and being substantially resistant to protease, endoglycosidase F, and collagenase enzymes, said human SAP-6-Val protein being produced by a process comprising the steps of:
An isolated hydrophobic surfactant-associated SAP-6-Val protein, said SAP-6-Val protein enhancing surfactant-like activity of phospholipids in lungs of an animal, having a simple molecular weight of about 6,000 daltons determined in a polyacrylamide gel containing sodium dodecyl sulfate and being substantially resistant to protease, endoglycosidase F and collagenase enzymes.
An isolated, human hydrophobic surfactant-associated protein, said human protein enhancing surfactant-like activity of phospholipids in lungs of an animal and having the following sequence:
An isolated lung specific hydrophobic surfactant-associated canine protein having a molecular weight of about 6,000 daltons as determined by SDS-PAGE and a molecular weight of about 3,500-4,000 daltons as determined by tricine-SDS-PAGE, said canine protein enhancing surfactant-like activity of phospholipids in lungs of an animal and comprising the following amino acid sequence:
Dependencies are inferred from the claim text.