Platelet aggregating material from equine arterial tissue

Claim 1

A method of preparing a hemostatic composition comprising the steps of: (a) contacting cleaned, homogenized equine arterial tissue with a balanced salt solution to provide an aqueous extract containing extracted collagen material and unextracted arterial tissue, and (b) separating unextracted arterial tissue from said aqueous mixture to provide an extract containing an arterial collagen material, said extract being capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent characterized by the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85° C., stable at 56° C. for up to 60 minutes, and when exposed to α-chymotrypsin, losing its platelet aggregating activity when exposed to collagenase at 37° C. or when heated at 100° C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: ______________________________________ Lysine 29.8 Glycine 206.5 Histidine 31.2 Alanine 133.9 Hydroxylysine 4.8 1/2 Cysteine 13.2 Arginine 43.2 Valine 57.6 Hydroxyproline 20.1 Methionine 14.2 Aspartic Acid 65.4 Isoleucine 26.7 Threonine 38.1 Leucine 52.5 Serine 40.3 Tyrosine 15.3 Glutamic Acid 81.9 Phenylalanine 27.1 Proline 98.2 ______________________________________

Claim 2

A method of preparing a hemostatic composition comprising the steps of: (a) contacting cleaned, homogenized equine arterial tissue with a balanced salt solution to provide an aqueous extract containing extracted collagen material and unextracted arterial tissue, and (b) separating unextracted arterial tissue from said aqueous mixture to provide an extract containing an arterial collagen material, said extract being capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent characterized by the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85° C., stable at 56° C. for up to 60 minutes, and when exposed to α-chymotrypsin, losing its platelet aggregating activity when exposed to collagenase at 37° C. or when heated at 100° C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: ______________________________________ Lysine 29.8 Glycine 206.5 Histidine 31.2 Alanine 133.9 Hydroxylysine 4.8 1/2 Cysteine 13.2 Arginine 43.2 Valine 57.6 Hydroxyproline 20.1 Methionine 14.2 Aspartic Acid 65.4 Isoleucine 26.7 Threonine 38.1 Leucine 52.5 Serine 40.3 Tyrosine 15.3 Glutamic Acid 81.9 Phenylalanine 27.1 Proline 98.2 ______________________________________

Claim 26

A method of stimulating platelet aggregation in mammalian plasma or whole blood which comprises contacting said plasma or whole blood with an amount which is effective to stimulate platelet aggregation of a hemostatic agent isolatable from equine arterial tissue by extraction with aqueous solutions which is an equine arterial fibrillar collagen capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent characterized by the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85° C., stable at 56° C. for up to 60 minutes, and when exposed to α-chymotrypsin, losing its platelet aggregating activity when exposed to collagenase at 37° C. or when heated at 100° C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: ______________________________________ Lysine 29.8 Glycine 206.5 Histidine 31.2 Alanine 133.9 Hydroxylysine 4.8 1/2 Cysteine 13.2 Arginine 43.2 Valine 57.6 Hydroxyproline 20.1 Methionine 14.2 Aspartic Acid 65.4 Isoleucine 26.7 Threonine 38.1 Leucine 52.5 Serine 40.3 Tyrosine 15.3 Glutamic Acid 81.9 Phenylalanine 27.1 Proline 98.2 ______________________________________

Claim 27

A method of stimulating platelet aggregation in mammalian plasma or whole blood which comprises contacting said plasma or whole blood with an amount which is effective to stimulate platelet aggregation of a hemostatic agent isolatable from equine arterial tissue by extraction with aqueous solutions which is an equine arterial fibrillar collagen capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent characterized by the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85° C., stable at 56° C. for up to 60 minutes, and when exposed to α-chymotrypsin, losing its platelet aggregating activity when exposed to collagenase at 37° C. or when heated at 100° C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: ______________________________________ Lysine 29.8 Glycine 206.5 Histidine 31.2 Alanine 133.9 Hydroxylysine 4.8 1/2 Cysteine 13.2 Arginine 43.2 Valine 57.6 Hydroxyproline 20.1 Methionine 14.2 Aspartic Acid 65.4 Isoleucine 26.7 Threonine 38.1 Leucine 52.5 Serine 40.3 Tyrosine 15.3 Glutamic Acid 81.9 Phenylalanine 27.1 Proline 98.2 ______________________________________

Claim 29

A method on controlling external bleeding from a wound or incision comprising contacting said wound or incision with an amount which is effective to control bleeding of a hemostatic agent which is capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent isolatable from equine arterial tissue by extraction with aqueous solutions as an equine arterial fibrillar collagen having the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85° C., stable at 56° C. for up to 60 minutes, and when exposed to α-chymotrypsin, losing its platelet aggregating activity when exposed to collagenase at 37° C. or when heated at 100° C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: ______________________________________ Lysine 29.8 Glycine 206.5 Histidine 31.2 Alanine 133.9 Hydroxylysine 4.8 1/2 Cysteine 13.2 Arginine 43.2 Valine 57.6 Hydroxyproline 20.1 Methionine 14.2 Aspartic Acid 65.4 Isoleucine 26.7 Threonine 38.1 Leucine 52.5 Serine 40.3 Tyrosine 15.3 Glutamic Acid 81.9 Phenylalanine 27.1 Proline 98.2 ______________________________________

Claim 30

A method on controlling external bleeding from a wound or incision comprising contacting said wound or incision with an amount which is effective to control bleeding of a hemostatic agent which is capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent isolatable from equine arterial tissue by extraction with aqueous solutions as an equine arterial fibrillar collagen having the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85° C., stable at 56° C. for up to 60 minutes, and when exposed to α-chymotrypsin, losing its platelet aggregating activity when exposed to collagenase at 37° C. or when heated at 100° C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: ______________________________________ Lysine 29.8 Glycine 206.5 Histidine 31.2 Alanine 133.9 Hydroxylysine 4.8 1/2 Cysteine 13.2 Arginine 43.2 Valine 57.6 Hydroxyproline 20.1 Methionine 14.2 Aspartic Acid 65.4 Isoleucine 26.7 Threonine 38.1 Leucine 52.5 Serine 40.3 Tyrosine 15.3 Glutamic Acid 81.9 Phenylalanine 27.1 Proline 98.2 ______________________________________

Claim 31

A method of detecting abnormal platelet function comprising contacting platelets which are suspected of abnormal function in plasma or whole blood under test with a hemostatic agent which is capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent isolatable from equine arterial tissue by extraction with aqueous solutions characterized as an equine arterial fibrillar collagen having the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85° C., stable at 56° C. for up to 60 minutes, and when exposed to α-chymotrypsin, losing its platelet aggregating activity when exposed to collagenase at 37° C. or when heated at 100° C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: ______________________________________ Lysine 29.8 Glycine 206.5 Histidine 31.2 Alanine 133.9 Hydroxylysine 4.8 1/2 Cysteine 13.2 Arginine 43.2 Valine 57.6 Hydroxyproline 20.1 Methionine 14.2 Aspartic Acid 65.4 Isoleucine 26.7 Threonine 38.1 Leucine 52.5 Serine 40.3 Tyrosine 15.3 Glutamic Acid 81.9 Phenylalanine 27.1 Proline 98.2 ______________________________________ and comparing the aggregation response to the plasma or whole blood under study to the response of normal platelets.

Claim 32

A method of detecting abnormal platelet function comprising contacting platelets which are suspected of abnormal function in plasma or whole blood under test with a hemostatic agent which is capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent isolatable from equine arterial tissue by extraction with aqueous solutions characterized as an equine arterial fibrillar collagen having the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85° C., stable at 56° C. for up to 60 minutes, and when exposed to α-chymotrypsin, losing its platelet aggregating activity when exposed to collagenase at 37° C. or when heated at 100° C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: ______________________________________ Lysine 29.8 Glycine 206.5 Histidine 31.2 Alanine 133.9 Hydroxylysine 4.8 1/2 Cysteine 13.2 Arginine 43.2 Valine 57.6 Hydroxyproline 20.1 Methionine 14.2 Aspartic Acid 65.4 Isoleucine 26.7 Threonine 38.1 Leucine 52.5 Serine 40.3 Tyrosine 15.3 Glutamic Acid 81.9 Phenylalanine 27.1 Proline 98.2 ______________________________________ and comparing the aggregation response to the plasma or whole blood under study to the response of normal platelets.