CA1066189A - Rapidly solubilized ahf composition and process for preparing same - Google Patents
Rapidly solubilized ahf composition and process for preparing sameInfo
- Publication number
- CA1066189A CA1066189A CA268,228A CA268228A CA1066189A CA 1066189 A CA1066189 A CA 1066189A CA 268228 A CA268228 A CA 268228A CA 1066189 A CA1066189 A CA 1066189A
- Authority
- CA
- Canada
- Prior art keywords
- composition
- ahf
- carbohydrate
- amount
- weight
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/36—Blood coagulation or fibrinolysis factors
- A61K38/37—Factors VIII
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Inorganic Chemistry (AREA)
- Biochemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Molecular Biology (AREA)
- Hematology (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
ABSTRACT OF THE DISCLOSURE
The rate of solubility of a freeze-dried solid composition containing therapeutic amounts of anti-hemophilic factor (AHF) is greatly increased by carrying out the solubilization in the presence of at least a critical threshold amount of a water soluble carbohydrate. The carbohydrate is incorporated into the AHF composition in sufficient quantity to provide at least 2 weight units of carbohydrate (expressed in grams) per 100 volume units of AHF solution (expressed in milliliters).
This enhanced rate of solubility permits rapid treatment of hemophilic patients.
The rate of solubility of a freeze-dried solid composition containing therapeutic amounts of anti-hemophilic factor (AHF) is greatly increased by carrying out the solubilization in the presence of at least a critical threshold amount of a water soluble carbohydrate. The carbohydrate is incorporated into the AHF composition in sufficient quantity to provide at least 2 weight units of carbohydrate (expressed in grams) per 100 volume units of AHF solution (expressed in milliliters).
This enhanced rate of solubility permits rapid treatment of hemophilic patients.
Description
~66~89 The present invention relates to a solid composition, its method of manufacture and its use for treatment of hemo-philiacs.
Time ia of considerable importance in the treatment of he~ophiliacs both to the person administering emergency injections and to the persons receiving such injections because loss of blood by the hemophiliac and/or injury to the joints is aggravated during the time required for solubilizing the nor~ally solid composition to permit injection. The problem therefore to which the present application is directed is to decrease the solubilization time of the solid composition to permit prompt injection into an injured person.
It has been the practice to freeze dry blood plasma fractions in form of a solid composition to remove water there-fr~m and, just prior to use, to dissolve the solid composition in a liquid aqueous medium to form a solution which is then injected into the patient. The blood plasma fractions are ob-tained by removing water from blood plasma (blood from which white and red cells and blood platelets have been removed and which contains about 90 percent water and 10 percent solids).
The solids remaining after removal of the water from the blood plasma are divided into blood plasma fractions, those most useful for the purpose of this invention being the anti-hemophilic factor (AHF, Factor VIII) fraction (which may contain fibrinogen) and, if desired, a separate fibrinogen factor.
Time ia of considerable importance in the treatment of he~ophiliacs both to the person administering emergency injections and to the persons receiving such injections because loss of blood by the hemophiliac and/or injury to the joints is aggravated during the time required for solubilizing the nor~ally solid composition to permit injection. The problem therefore to which the present application is directed is to decrease the solubilization time of the solid composition to permit prompt injection into an injured person.
It has been the practice to freeze dry blood plasma fractions in form of a solid composition to remove water there-fr~m and, just prior to use, to dissolve the solid composition in a liquid aqueous medium to form a solution which is then injected into the patient. The blood plasma fractions are ob-tained by removing water from blood plasma (blood from which white and red cells and blood platelets have been removed and which contains about 90 percent water and 10 percent solids).
The solids remaining after removal of the water from the blood plasma are divided into blood plasma fractions, those most useful for the purpose of this invention being the anti-hemophilic factor (AHF, Factor VIII) fraction (which may contain fibrinogen) and, if desired, a separate fibrinogen factor.
-2-Solubilizing agents have been used to solubilize blood plasma fractions but the time it takes to dissolve such fractions leaves much to be desired. For example, U.S. Patent 2,826,533 discloses the addition of dextrose to a fibrinogen, blood plasma, fraction and it is known that the time of solubilization of the fibrinogen fraction can be reduced by adding dextrose thereto. It is also known to add dextrose to A~IF as, for example, described in the "Journal of Thrombosis Research", Volume 1, pages 191-200, 1972, published by Pergamon Press, Inc. which reported addition of dextrose to AHF in order to facilitate the chromatography of AHF. The article concluded that the yield of bovine Factor VIII from chromato-graphy on anion exchange media can be greatly improved by the inclusion of a low-molecular weight carbohydrate, such as dextrose, in the solvents. In addition to the foregoing, Cutter Laboratories, Inc. has added sufficient dextrose to its com-mercial AHF preparation so that when the preparation is recon-stituted according to the instructions, the resulting AHF
solution contains about one gram of dextrose per 100 milliliters of solution.
Finally, U.S. Patent 3,057,781 discloses stabilizing plasma with invert sugar and levulinic acid.
While dextrose has been used with blood plas~la fractions for various purposes, as is seen from the fore~oing, it was not appreciated before the present invention that by critically controlling the amount of a water soluble carbohydrate (which is free of le w linic acid) in the solid composition, it is possible to solubilize the solid composition with an 1066~L89 aqueous medium within about 90 seconds.
More specifically, this is accomplished in accordance with the invention by providing a solid composition comprising AHF, characterized by including an amount of at least one water soluble carbohydrate sufficient to solubilize the compo-sition in an aqueous medium within about 90 seconds, the ~IF
being present in an amount sufficient to form a therapeutically effective AHF concentration upon solubilization of the composition.
We also provide in accordance with the invention a method of preparing an A~F-containing composition adapted to be rapidly solubilized upon reconstitution with a reconstituting liquid to produce a solution containing a therapeutically effective amount of AHF, characterized by adding to said composition a water soluble carbohydrate in an amount sufficient to solubilize the composition in the reconstituting liquid within about 90 seconds.
Objects and advantages of the invention will become apparent from the following detailed disclosure taken in conjunction with the accompanying drawing, in which:
Fig. 1 graphically depicts the unexpected dramatic improvement in the rate of solubilization of solid composi-tions containing AHF which is obtained when the composition is solubilized in the presence of from 0 to 5 grams of dextrose per 100 ml. of reconstituted solution. The various curves are obtained using different reconstitution temperatures and volumes.
~66~89 The problem of reducing dissolution time of solid compositions of the type above disclosed is solved in accordance with the invention by the inclusion in the solid composition of a critical amount of water soluble carbohydrate. When one desires to use the composition of the invention, one need only add water and because of the presence of the carbohydrate in the solid composition, the composition dissolves in a very short period of time, for example, within 90 seconds, and can then be injected into the patient.
The amount of carbohydrate to be added is, as noted, critical in the sense that mere addition of carbohydrate does not in itself result in significant solubilization time improvements. A threshold concentration of carbohydrate must be reached before useful improvements in solubilization time are achieved, after which addition of more carbohydrate again exerts no significant effect on solubilization time. This threshold concentration will vary within the ranges disclosed depending upon factors which are believed to include the amount and identity of protein and salts in the AHF preparations as well as the identity of the carbohydrate selected. The exact optimum quantity of carbohydrate will thus vary with the carbohydrate chosen, the method of AHF preparation and even with separate runs using thh same preparatory method. Thus the appropriate quantity should be determined by elementary and conventional solubilization time assays for each lot of AHF.
The amount of carbohydrate should be sufficient to bring the solid AHF preparation into solution within about 90 seconds, and preferably 65 seconds, the AHF being present in the solid composition in an amount sufficient to form a thera-peutically effective AHF concentration upon solubilization of '1066~39 the composition. A therapeutically effective concentrationof AHF in such solutions illustratively ranges about from 3 to 100 International Units of AHF per ml. with a preferred range of about from 3 to 40 International Units per ml. and more commonly about 24 to about 28 units per ml. The amount of carbohydrate present in the solid AHF composition typically provides, upon reconstitution with water, or other suitable reconstituting liquid, a therapeutically effectlve solution of AHF containing at least about 2 weight units of carbohydrate (computed in Grams) for every 100 volume units cf solution (computed as milliliters). The a unt of carbohydrate will generally vary about from 2 to 10 grams, preferably about 2 to 5 grams of carbohydrate per 100 ml. of solution, with about 3 grams appearing to be optimum.
To provide the desired concentration of carbohydrate in solution, the carbohydrate is illustratively present in the solid composition in an amount of about 1.6 to 7.5 times the a~ount of total protein in the solid AHF composition.
Preferably, the amount of carbohydrate is about 2.0 to 5.0 times th~ weight of total protein. The preferred embodiment is about 2.0 times the weight of total protein. This solid composition can contain anywhere from about 2 to 200 Inter-national Units of AHF/gm protein, and still produce as a pr~ctical matter a solution of A~F upon reconstitution which has a therapeutically significant effect.
The water soluble carbohydrates useful in the invention include any which are capable of hydrating the AHF-containing composition. This includes without limitation the monosaccharides such as the commonly available hexoses, inclu-ding dextrose (glucose), mannose, galactose and fructose;
~066~9 the disaccharides such as maltose, lactose and sucrose; the trisaccharides, such as raffinose; and the short chain dextrins, e g. dextrins having a chain length of less than about four monosaccharide units. Mixtures of suitable carbohydrates may also be employed. The preferred carbohydrates are dextrose, su~rose, c~mltose and lactose, with dextrose being an especially preferred material! The carbohydrate must be biologically acceptable when the AHF is marketed for human administration.
The carbohydrate can be admixed with the AHF-containing composi-tion at any point during or prior to preparation of the lyophilized composition.
The AHF compositions, whose rate of solubility is enhanced by the addition of carbohydrate in accordance with the present invention, may be prepared by numerous procedures known to those skilled in the art. In the preferred embodiment, the solid mixture comprising AHF and fibrinogen is obtained by starting with plasma frozen at about minus 25 degrees C. which is then thawed to 4 or 5 degrees C, to produce a cryoprecipitate which is collected by centrifugation.
The cryoprecipitate is suspended in heparinized, citrated saline to which is added 3.5% by weight, of polyethylene glycol. The resulting mixture is centrifuged and the resulting fibrinogen precipitate is discarded and the supernatant retained.
To the supernatant is added about 7.5 weight units of polyethylene glycol (expressed as grams) per 100 volume units of supernatant (expressed as milliliters). The resulting suspension is mixed for about 15 minutes at room temperature and is then centrifuged and the resulting precipitate collected. This precipitate or 10661~9 solid mixture comprises AHF and fibrinogen and can be used as such or can be further purified by glycine fractionation. In any event, the water soluble carbohydrate can be added to either of such mixtures. Preferably, the solid mixture of AHF and fibrinogen is dissolved in an aqueous medium, for example, a dextrose citrated saline aqueous solution containing about 0.72% sodium chloride, 0.02M sodium citrate and an appropriate amount of the water soluble carbohydrate to produce the desired effect of an enhanced rate of solubilization. It is neither necessary nor desirable at this stage to add water to the point where the solution contains about 2 weight units of dextrose per 100 volume units of solution since such a dilute solution may unnecessarily extend the time required for lyophilization.
The dissolved solid mixture containing the carbohydrate is further clarified by passing it through a coarse filter which removes some of the fibrinogen and other insoluble pro-teins. Thereafter, the sample is further diluted with citrated saline, as desired, to a potency of about 3 to 75 International Units/ml. or left as a concentrate which normally contains from 250 to 1000 International Units/ml. The dissolved product is then sterile filtered through a "Millipore" membrane filter having an average pore size of about 0.3 microns. The filtered solution is filled under aseptic conditions into 10 ml. to 30 ml.
capacity vials, as desired, rapidly frozen and freeze-dried.
To administer the AHF preparation to a patient, the normal procedure is to reconstitute the lyophilized product to a solution containing about 3 to about 100 International Units ~rale r)l~rK
of AHF per m~., and more commonly about 24 to about 28 Units per ml., about 2 to about 10 grams of dextrose per 100 ml., about 1.4 to about 1.6 grams of protein per 100 ml., about 0.6 to about 0.~ grams of fibrinogen per 100 ml., and about 0.7 gra~ to about 6 grams of salts such as NaCl, sodium citrate, glycine and unidentified residual solids per 100 ml.
Iypically a 10 ml. vial of reconstituted ~F solution will contain about 270 International Units of AHF, about 0.3 gram of dextrose, about 0.15 grams of protein including about 0.07 gram of fibrinogen, about 0.51 gram of residuals and sufficient water to 10 ml. volume.
The lyophilized product is readily soluble in sterile water at room temperature and after the addition of the water is almost immediately ready for administration to he~ophillc patients as a result of the dextrose levels in the solution.
In order to show the dramatic unexpected results obtained by the present invention the following tests were conducted using 10 ml. and 30 ml. capacity vials containing the lyophilized product of the invention, the 10 ml. and 30 ml. vials being respectively filled with 10 ml. and 30 ml.
of water at room temperature and 37C. The 30 ml. vials contained;approximately three times as much product as the 10 ml. vials. The time required for complete dissolution of such sample was recorded. The rcsults are presented below in Table 1 and in Fig. 1.
~066189 Table 1 grams dextrose/ Fig. 1 Curve: A B C D
100 ml. of recon- Reconstituting stituted composi- Volume: 10 ml. 10 ml. 30 ml 30 ml.
tion Diluent Temp.: 37C Room Temp 37C Room Temp.
1176 secs. 195 secs. 100 secs. 330secs
solution contains about one gram of dextrose per 100 milliliters of solution.
Finally, U.S. Patent 3,057,781 discloses stabilizing plasma with invert sugar and levulinic acid.
While dextrose has been used with blood plas~la fractions for various purposes, as is seen from the fore~oing, it was not appreciated before the present invention that by critically controlling the amount of a water soluble carbohydrate (which is free of le w linic acid) in the solid composition, it is possible to solubilize the solid composition with an 1066~L89 aqueous medium within about 90 seconds.
More specifically, this is accomplished in accordance with the invention by providing a solid composition comprising AHF, characterized by including an amount of at least one water soluble carbohydrate sufficient to solubilize the compo-sition in an aqueous medium within about 90 seconds, the ~IF
being present in an amount sufficient to form a therapeutically effective AHF concentration upon solubilization of the composition.
We also provide in accordance with the invention a method of preparing an A~F-containing composition adapted to be rapidly solubilized upon reconstitution with a reconstituting liquid to produce a solution containing a therapeutically effective amount of AHF, characterized by adding to said composition a water soluble carbohydrate in an amount sufficient to solubilize the composition in the reconstituting liquid within about 90 seconds.
Objects and advantages of the invention will become apparent from the following detailed disclosure taken in conjunction with the accompanying drawing, in which:
Fig. 1 graphically depicts the unexpected dramatic improvement in the rate of solubilization of solid composi-tions containing AHF which is obtained when the composition is solubilized in the presence of from 0 to 5 grams of dextrose per 100 ml. of reconstituted solution. The various curves are obtained using different reconstitution temperatures and volumes.
~66~89 The problem of reducing dissolution time of solid compositions of the type above disclosed is solved in accordance with the invention by the inclusion in the solid composition of a critical amount of water soluble carbohydrate. When one desires to use the composition of the invention, one need only add water and because of the presence of the carbohydrate in the solid composition, the composition dissolves in a very short period of time, for example, within 90 seconds, and can then be injected into the patient.
The amount of carbohydrate to be added is, as noted, critical in the sense that mere addition of carbohydrate does not in itself result in significant solubilization time improvements. A threshold concentration of carbohydrate must be reached before useful improvements in solubilization time are achieved, after which addition of more carbohydrate again exerts no significant effect on solubilization time. This threshold concentration will vary within the ranges disclosed depending upon factors which are believed to include the amount and identity of protein and salts in the AHF preparations as well as the identity of the carbohydrate selected. The exact optimum quantity of carbohydrate will thus vary with the carbohydrate chosen, the method of AHF preparation and even with separate runs using thh same preparatory method. Thus the appropriate quantity should be determined by elementary and conventional solubilization time assays for each lot of AHF.
The amount of carbohydrate should be sufficient to bring the solid AHF preparation into solution within about 90 seconds, and preferably 65 seconds, the AHF being present in the solid composition in an amount sufficient to form a thera-peutically effective AHF concentration upon solubilization of '1066~39 the composition. A therapeutically effective concentrationof AHF in such solutions illustratively ranges about from 3 to 100 International Units of AHF per ml. with a preferred range of about from 3 to 40 International Units per ml. and more commonly about 24 to about 28 units per ml. The amount of carbohydrate present in the solid AHF composition typically provides, upon reconstitution with water, or other suitable reconstituting liquid, a therapeutically effectlve solution of AHF containing at least about 2 weight units of carbohydrate (computed in Grams) for every 100 volume units cf solution (computed as milliliters). The a unt of carbohydrate will generally vary about from 2 to 10 grams, preferably about 2 to 5 grams of carbohydrate per 100 ml. of solution, with about 3 grams appearing to be optimum.
To provide the desired concentration of carbohydrate in solution, the carbohydrate is illustratively present in the solid composition in an amount of about 1.6 to 7.5 times the a~ount of total protein in the solid AHF composition.
Preferably, the amount of carbohydrate is about 2.0 to 5.0 times th~ weight of total protein. The preferred embodiment is about 2.0 times the weight of total protein. This solid composition can contain anywhere from about 2 to 200 Inter-national Units of AHF/gm protein, and still produce as a pr~ctical matter a solution of A~F upon reconstitution which has a therapeutically significant effect.
The water soluble carbohydrates useful in the invention include any which are capable of hydrating the AHF-containing composition. This includes without limitation the monosaccharides such as the commonly available hexoses, inclu-ding dextrose (glucose), mannose, galactose and fructose;
~066~9 the disaccharides such as maltose, lactose and sucrose; the trisaccharides, such as raffinose; and the short chain dextrins, e g. dextrins having a chain length of less than about four monosaccharide units. Mixtures of suitable carbohydrates may also be employed. The preferred carbohydrates are dextrose, su~rose, c~mltose and lactose, with dextrose being an especially preferred material! The carbohydrate must be biologically acceptable when the AHF is marketed for human administration.
The carbohydrate can be admixed with the AHF-containing composi-tion at any point during or prior to preparation of the lyophilized composition.
The AHF compositions, whose rate of solubility is enhanced by the addition of carbohydrate in accordance with the present invention, may be prepared by numerous procedures known to those skilled in the art. In the preferred embodiment, the solid mixture comprising AHF and fibrinogen is obtained by starting with plasma frozen at about minus 25 degrees C. which is then thawed to 4 or 5 degrees C, to produce a cryoprecipitate which is collected by centrifugation.
The cryoprecipitate is suspended in heparinized, citrated saline to which is added 3.5% by weight, of polyethylene glycol. The resulting mixture is centrifuged and the resulting fibrinogen precipitate is discarded and the supernatant retained.
To the supernatant is added about 7.5 weight units of polyethylene glycol (expressed as grams) per 100 volume units of supernatant (expressed as milliliters). The resulting suspension is mixed for about 15 minutes at room temperature and is then centrifuged and the resulting precipitate collected. This precipitate or 10661~9 solid mixture comprises AHF and fibrinogen and can be used as such or can be further purified by glycine fractionation. In any event, the water soluble carbohydrate can be added to either of such mixtures. Preferably, the solid mixture of AHF and fibrinogen is dissolved in an aqueous medium, for example, a dextrose citrated saline aqueous solution containing about 0.72% sodium chloride, 0.02M sodium citrate and an appropriate amount of the water soluble carbohydrate to produce the desired effect of an enhanced rate of solubilization. It is neither necessary nor desirable at this stage to add water to the point where the solution contains about 2 weight units of dextrose per 100 volume units of solution since such a dilute solution may unnecessarily extend the time required for lyophilization.
The dissolved solid mixture containing the carbohydrate is further clarified by passing it through a coarse filter which removes some of the fibrinogen and other insoluble pro-teins. Thereafter, the sample is further diluted with citrated saline, as desired, to a potency of about 3 to 75 International Units/ml. or left as a concentrate which normally contains from 250 to 1000 International Units/ml. The dissolved product is then sterile filtered through a "Millipore" membrane filter having an average pore size of about 0.3 microns. The filtered solution is filled under aseptic conditions into 10 ml. to 30 ml.
capacity vials, as desired, rapidly frozen and freeze-dried.
To administer the AHF preparation to a patient, the normal procedure is to reconstitute the lyophilized product to a solution containing about 3 to about 100 International Units ~rale r)l~rK
of AHF per m~., and more commonly about 24 to about 28 Units per ml., about 2 to about 10 grams of dextrose per 100 ml., about 1.4 to about 1.6 grams of protein per 100 ml., about 0.6 to about 0.~ grams of fibrinogen per 100 ml., and about 0.7 gra~ to about 6 grams of salts such as NaCl, sodium citrate, glycine and unidentified residual solids per 100 ml.
Iypically a 10 ml. vial of reconstituted ~F solution will contain about 270 International Units of AHF, about 0.3 gram of dextrose, about 0.15 grams of protein including about 0.07 gram of fibrinogen, about 0.51 gram of residuals and sufficient water to 10 ml. volume.
The lyophilized product is readily soluble in sterile water at room temperature and after the addition of the water is almost immediately ready for administration to he~ophillc patients as a result of the dextrose levels in the solution.
In order to show the dramatic unexpected results obtained by the present invention the following tests were conducted using 10 ml. and 30 ml. capacity vials containing the lyophilized product of the invention, the 10 ml. and 30 ml. vials being respectively filled with 10 ml. and 30 ml.
of water at room temperature and 37C. The 30 ml. vials contained;approximately three times as much product as the 10 ml. vials. The time required for complete dissolution of such sample was recorded. The rcsults are presented below in Table 1 and in Fig. 1.
~066189 Table 1 grams dextrose/ Fig. 1 Curve: A B C D
100 ml. of recon- Reconstituting stituted composi- Volume: 10 ml. 10 ml. 30 ml 30 ml.
tion Diluent Temp.: 37C Room Temp 37C Room Temp.
1176 secs. 195 secs. 100 secs. 330secs
3 35 " 85 " 50 " 65 "
48 " 55 " 52 " 75 "
None (control)185 " 210 " 105 " 210 "
The control in the above table was identical to the other samples in all respects except that the control contained no dextrose.
The results of Table 1 are plotted in Fig. 1. The plotted data clearly shows the marked improvement in rate of solubility once a dextrose concentration exceeding 2 grams per 100 mls. of reconstituted composition is obtained. Similar desirable results are obtained with the other carbohydrates discussed hereinabove.
In the preceding exa~ple, polyethylene glycol was used to fractionate the blood plas'-~. However, other compounds can be used such as ethylene oxide-propylene glycol conden~ation products, and other procedures for fractionation can be employed to produce a product which is rapidly soluble according to the teachings herein.
48 " 55 " 52 " 75 "
None (control)185 " 210 " 105 " 210 "
The control in the above table was identical to the other samples in all respects except that the control contained no dextrose.
The results of Table 1 are plotted in Fig. 1. The plotted data clearly shows the marked improvement in rate of solubility once a dextrose concentration exceeding 2 grams per 100 mls. of reconstituted composition is obtained. Similar desirable results are obtained with the other carbohydrates discussed hereinabove.
In the preceding exa~ple, polyethylene glycol was used to fractionate the blood plas'-~. However, other compounds can be used such as ethylene oxide-propylene glycol conden~ation products, and other procedures for fractionation can be employed to produce a product which is rapidly soluble according to the teachings herein.
Claims (17)
OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
1. A solid composition comprising AHF, characterized by including an amount of at least one water soluble carbohydrate sufficient to solubilize the composition in an aqueous medium within about 90 seconds, the AHF being present in an amount sufficient to form a therapeutically effective AHF concentration upon solubilization of the composition.
2. The composition according to claim 1, wherein it includes fibrinogen.
3. The composition according to claim 1, wherein it includes proteins, the amount of carbohydrate being about from 1.6 to 7.5 times the amount by weight of protein in the composition.
4. The composition according to claim 3, wherein said carbohydrate is dextrose and said AHF is present in an activity of about from 2 to 200 International Units/gm protein.
5. The composition according to claim 1, wherein said carbohydrate is dextrose, said composition including protein and containing about 31 parts by weight of dextrose, about 16 parts by weight of protein including AHF in an activity of about 270 International Units/gm of material, and about 53 parts by weight of salts.
6. The composition according to any one of claims 1, 2 or 3, wherein said carbohydrate comprises a monosaccharide or a disaccharide.
7. The composition according to any one of claims 1, 2 or 3, wherein said carbonydrate comprises dextrose, mannose, galactose or fructose.
8. The composition according to any one of claims 1, 2 or 3, wherein said carbohydrate comprises lactose, sucrose or maltose.
9. A method of preparing an AHF-containing composition adapted to be rapidly solubilized upon reconstitution with a reconstituting liquid to produce a solution containing a therapeutically effective amount of AHF, characterized by adding to said composition a water soluble carbohydrate in an amount sufficient to solubilize the composition in the reconstituting liquid within About 90 seconds.
10. The method according to claim 9, wherein the composition contains a mixture of fibrinogen and AHF and is produced from blood plasma or an AHF-containing fraction thereof by fractionating the plasma and dissolving the mixture of fibrinogen and AHF thus obtained in an aqueous medium, and freeze-drying the resulting solution to obtain a clinically useful freeze-dried solid composition, said carbohydrate being added to the composition during or prior to the freeze-drying step.
11. The method according to claim 10, wherein the amount of carbohydrate added is an amount sufficient to produce at least about 2 weight units (expressed in grams) per 100 volume units (expressed in milliliters) concentration of carbohydrate.
12. The method according to claim 9, wherein the composition contains a mixture of protein and AHF and is produced by fractionating blood plasma or an AHF-containing fraction thereof, dissolving the mixture of protein and AHF thus obtained in an aqueous medium and freeze-drying the resulting solution to obtain a clinically useful freeze-dried solid composition, said carbohydrate being added to the composition during or prior to the freeze-drying step in a proportion of about from 1.6 to 7.5 times the amount by weight of protein in the composi-tion.
13. The method according to any one of claims: 9, 10 or 11, wherein said carbohydrate comprises dextrose, maltose, lactose or sucrose.
14. A method of rapidly solubilizing a solid AHF-containing composition to produce a solution containing a therapeutically effective amount of AHF, characterized by mixing said composition with an amount of carbohydrate sufficient to solubilize the composition within 90 seconds.
15. The method according to claim 14, wherein the amount of carbohydrate mixed with the. AHF is at least about 2 weight units of carbohydrate, expressed in grams, per 100 volume units of solubilized composition, expressed in milliliters.
16. The method according to claim 14 or 15 wherein the carbohydrate is added as an aqueous solution.
17. A composition comprising AHF, water and at least about 2 weight units of carbohydrate, expressed in grams, per 100 volume units of said composition, expressed in milliliters.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US64317875A | 1975-12-22 | 1975-12-22 | |
| US05/729,758 US4089944A (en) | 1975-12-22 | 1976-10-05 | Rapidly solubilized AHF composition and process for preparing same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1066189A true CA1066189A (en) | 1979-11-13 |
Family
ID=27094202
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA268,228A Expired CA1066189A (en) | 1975-12-22 | 1976-12-20 | Rapidly solubilized ahf composition and process for preparing same |
Country Status (7)
| Country | Link |
|---|---|
| JP (1) | JPS5282716A (en) |
| AT (1) | AT351158B (en) |
| CA (1) | CA1066189A (en) |
| DE (1) | DE2653534C2 (en) |
| ES (1) | ES454453A1 (en) |
| GB (1) | GB1540610A (en) |
| MX (1) | MX4730E (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4186192A (en) * | 1978-12-18 | 1980-01-29 | Cutter Laboratories, Inc. | Stabilized immune serum globulin |
| DE2916711A1 (en) * | 1979-04-25 | 1980-11-06 | Behringwerke Ag | Blood coagulation factors and process for their manufacture |
| DE3432083A1 (en) * | 1984-08-31 | 1986-03-06 | Behringwerke Ag, 3550 Marburg | PASTEURIZED, ISOAGGLUTININ-FREE FACTOR VIII PREPARATION AND METHOD FOR THE PRODUCTION THEREOF |
| HUP9701554D0 (en) * | 1997-09-18 | 1997-11-28 | Human Oltoanyagtermeloe Gyogys | Pharmaceutical composition containing plazma proteins |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US2826533A (en) * | 1954-03-15 | 1958-03-11 | Cutter Lab | Stable fibrinogen solutions and method for producing same |
-
1976
- 1976-11-25 DE DE19762653534 patent/DE2653534C2/en not_active Expired
- 1976-12-07 MX MX519176U patent/MX4730E/en unknown
- 1976-12-09 GB GB5145376A patent/GB1540610A/en not_active Expired
- 1976-12-20 CA CA268,228A patent/CA1066189A/en not_active Expired
- 1976-12-21 ES ES454453A patent/ES454453A1/en not_active Expired
- 1976-12-21 JP JP15299476A patent/JPS5282716A/en active Pending
- 1976-12-22 AT AT956676A patent/AT351158B/en not_active IP Right Cessation
Also Published As
| Publication number | Publication date |
|---|---|
| GB1540610A (en) | 1979-02-14 |
| DE2653534C2 (en) | 1986-08-28 |
| DE2653534A1 (en) | 1977-06-30 |
| MX4730E (en) | 1982-08-24 |
| AT351158B (en) | 1979-07-10 |
| JPS5282716A (en) | 1977-07-11 |
| ATA956676A (en) | 1978-12-15 |
| ES454453A1 (en) | 1978-04-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US4089944A (en) | Rapidly solubilized AHF composition and process for preparing same | |
| DE69329795T2 (en) | COMPOSITION CONTAINING COAGULATION FACTOR VIII; METHOD FOR THE PRODUCTION THEREOF AND THE USE OF A SURFACE-ACTIVE SUBSTANCE AS A STABILIZER | |
| US4478829A (en) | Pharmaceutical preparation containing purified fibronectin | |
| DE3400413C2 (en) | ||
| DE69432179T3 (en) | A FORMULATION OF THE COOLING FACTOR VIII | |
| DE69531204T2 (en) | FACTOR IX FORMULATIONS | |
| DE69333928T2 (en) | IMPROVED SOLUBILIZATION AND STABILIZATION OF THE FACTOR VIII COMPLEX | |
| EP0796623B1 (en) | Pharmaceutical preparation for the treatment of blood coagulation disorders | |
| DE69430745T2 (en) | OXYGEN REDUCED AQUEOUS FACTOR VIII CONTAINING SOLUTION | |
| DE69729786T2 (en) | HIGH-CONCENTRATED, LYOPHILIZED AND LIQUID FACTOR IX FORMULATIONS | |
| CA1248453A (en) | Pharmaceutical compositions containing anthracycline glycosides | |
| DE69527966T2 (en) | PHARMACEUTICAL PREPARATION FOR SUBCUTANEOUS, INTRAMUSULAR OR INTRADERMAL ADMINISTRATION OF FACTOR VIII | |
| EP0302754B1 (en) | Stable aqueous thrombin solution | |
| US3631018A (en) | Production of stable high-potency human ahf using polyethylene glycol and glycine to fractionate a cryoprecipitate of ahf concentrate | |
| EP0733702B1 (en) | Stable transglutaminase preparations and process for their preparation | |
| JP2818898B2 (en) | Dual-chamber syringe with active substance sensitive drug consisting of human protein | |
| JPS597693B2 (en) | Antithrombin preparation and its manufacturing method | |
| DE69029765T2 (en) | Stabilization of highly purified proteins | |
| DE69630291T2 (en) | PROTEIN FORMULATION CONTAINING COAJULATION ERRORS VIII OR IX IN A SACCHAROSE AQUEOUS SOLUTION | |
| EP2258402A2 (en) | Dried composition | |
| US4168303A (en) | Lyophilized native gamma globulin preparation for intravenous administration | |
| US4085095A (en) | Clottable fibrinogen free Factor VIII product and process | |
| US3933996A (en) | Composition comprising radioactive labeled-fibrinogen and albumin | |
| CH665356A5 (en) | PHARMACEUTICAL FORMULATION. | |
| JP4925822B2 (en) | Method for stabilizing plasma protein cryoprecipitates for use in virus-inactivated heat treatment |