WO2010117976A1 - Spray-dried blood products and methods of making same - Google Patents

Spray-dried blood products and methods of making same Download PDF

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Publication number
WO2010117976A1
WO2010117976A1 PCT/US2010/030031 US2010030031W WO2010117976A1 WO 2010117976 A1 WO2010117976 A1 WO 2010117976A1 US 2010030031 W US2010030031 W US 2010030031W WO 2010117976 A1 WO2010117976 A1 WO 2010117976A1
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WO
WIPO (PCT)
Prior art keywords
blood
spray
dehydrated
platelets
plasma
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PCT/US2010/030031
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English (en)
French (fr)
Inventor
Thomas H. Fischer
Joseph A. Dacorta
E. Stan Eskridge
Arthur P. Bode
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Entegrion Inc
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Entegrion Inc
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Publication date
Priority to US13/262,931 priority Critical patent/US9867782B2/en
Priority to CA2757961A priority patent/CA2757961C/en
Priority to AU2010234607A priority patent/AU2010234607B2/en
Priority to JP2012504757A priority patent/JP2012523414A/ja
Priority to CN2010800227674A priority patent/CN102448475A/zh
Priority to BRPI1006722A priority patent/BRPI1006722A2/pt
Priority to ES10762281.3T priority patent/ES2684130T3/es
Priority to MX2011010633A priority patent/MX2011010633A/es
Application filed by Entegrion Inc filed Critical Entegrion Inc
Priority to EP10762281.3A priority patent/EP2416790B1/en
Publication of WO2010117976A1 publication Critical patent/WO2010117976A1/en
Anticipated expiration legal-status Critical
Priority to US15/872,727 priority patent/US11213488B2/en
Priority to US17/566,226 priority patent/US12414920B2/en
Priority to US19/204,890 priority patent/US20250332106A1/en
Ceased legal-status Critical Current

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    • 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
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1682—Processes
    • A61K9/1688—Processes resulting in pure drug agglomerate optionally containing up to 5% of excipient
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F13/00—Bandages or dressings; Absorbent pads
    • A61F13/00051—Accessories for dressings
    • A61F13/00063—Accessories for dressings comprising medicaments or additives, e.g. odor control, PH control, debriding, antimicrobic
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/14—Blood; Artificial blood
    • A61K35/16—Blood plasma; Blood serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/14—Blood; Artificial blood
    • A61K35/19—Platelets; Megacaryocytes
    • 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
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A61K9/0026—Blood substitute; Oxygen transporting formulations; Plasma extender
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
    • A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
    • A61L15/40—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing ingredients of undetermined constitution or reaction products thereof, e.g. plant or animal extracts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
    • A61L26/0028—Polypeptides; Proteins; Degradation products thereof
    • A61L26/0047—Specific proteins or polypeptides not covered by groups A61L26/0033 - A61L26/0042
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0057—Ingredients of undetermined constitution or reaction products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0061—Use of materials characterised by their function or physical properties
    • A61L26/0076—Sprayable compositions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/08—Plasma substitutes; Perfusion solutions; Dialytics or haemodialytics; Drugs for electrolytic or acid-base disorders, e.g. hypovolemic shock
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F13/00—Bandages or dressings; Absorbent pads
    • A61F2013/00089—Wound bandages
    • A61F2013/00297—Wound bandages safety barrier for protection of the operator
    • A61F2013/00306—Wound bandages safety barrier for protection of the operator dressing for protection of the operator during surgical procedure
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F13/00—Bandages or dressings; Absorbent pads
    • A61F2013/00361—Plasters
    • A61F2013/00365—Plasters use
    • A61F2013/00463—Plasters use haemostatic
    • A61F2013/00472—Plasters use haemostatic with chemical means
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F13/00—Bandages or dressings; Absorbent pads
    • A61F2013/00361—Plasters
    • A61F2013/00902—Plasters containing means
    • A61F2013/00927—Plasters containing means with biological activity, e.g. enzymes for debriding wounds or others, collagen or growth factors

Definitions

  • the present invention is directed to methods of preparing dried blood products using spray-drying as an alternative to conventional lyophilization (freeze-drying), and products made by the method. Using the method of the invention, increased recovery rates of dried product are possible. The final product displays at least three-fold concentration over native plasma, as well as increased reconstitution rates when mixed with liquids.
  • Spray-drying is a technology in which a solution is atomized in a stream of flowing gas for rapid solvent vaporization (e.g., dehydration). The result is the formation on a sub- second timescale of microparticles composed of the residual solute.
  • Spray-drying has been used as a industrial process in the material, 4 food 5 and pharmaceutical 6 ' 7 industries for decades, (e.g., see Bergsoe 8 for an earlier review). More recently, spray-drying has facilitated the preparation of protein therapeutics as microparticles for inhalation, 9 the formulation of advanced carrier-therapeutic microstructures, 10"12 and new classes of micromaterials.
  • Dried blood products are known in the art, and the predominant technique for achieving the dried product is lyophilization (freeze-drying).
  • U.S. Patent Nos. 4,287,087 and 4,145,185 to Brinkhous et al. disclose dried blood platelets that have been fixed with a crosslinking reagent such as formaldehyde.
  • U.S. Patent Nos. 5,656,498, 5,651,966; 5,891,393; 5,902,608; and 5,993,804 disclose additional dried blood products.
  • Such products are useful for therapeutic purposes because they are stable, have long shelf life, and can be used potentially in powder form to arrest bleeding in patients undergoing severe trauma. However, such products must be manufactured under strict sterile conditions in order to avoid contamination.
  • the present invention is directed to a method of preparing dehydrated blood products, comprising the steps of: (a) providing a hydrated blood product; (b) spray-drying the hydrated blood product to produce a dehydrated blood product, as well as dehydrated blood products made by the method.
  • the present invention is directed to a method of treating a patient suffering from a blood-related disorder, comprising the steps of: (a) rehydrating a therapeutic amount of the dehydrated blood products to produce a rehydrated therapeutic composition; and (b) administering the rehydrated therapeutic composition to the patient.
  • the present invention is directed to a bandage or surgical aid comprising the dehydrated blood products described above.
  • the present invention is directed to a method of preparing dehydrated fixed blood platelets, comprising the steps of: (a) providing hydrated fixed blood platelets; and (b) spray-drying the hydrated fixed blood platelets to produce a dehydrated fixed blood platelets, as well as dehydrated fixed blood platelets made by the method.
  • the present invention is directed to a method of treating a patient suffering from a blood-related disorder, comprising the steps of: (a) rehydrating a therapeutic amount of the dehydrated fixed blood platelets to produce a rehydrated therapeutic composition; and (b) administering the rehydrated therapeutic composition to the patient.
  • the present invention is directed to a bandage or surgical aid comprising the dehydrated fixed blood platelets described above.
  • the present invention is directed to spray dried fixed blood platelets having spherical-dimpled geometry, wherein when said spray dried fixed blood platelets are rehydrated to form a rehydrated fixed blood platelet composition, the composition has a turbidity (A 500 ) value less than that of a comparable rehydrated lyophilized composition of fixed blood platelets.
  • Figure 1 is an electron micrograph of microspheres of spray-dried plasma produced according to the present invention.
  • Figure 2 is a graph showing coagulation factor levels in various samples
  • Figure 3 depicts graphs showing native coagulation pathway turnover with spray dried plasma produced according to the method of the invention
  • Figure 4 is an electron micrograph showing fibrin ultrastructure from spray dried plasma produced according to the method of the invention.
  • Figure 5 is a graph depicting the turbidity and rehydration rate of spray-dried vs. lyophilized plasma at several concentrations
  • Figure 6 is an electron micrograph of rehydrated spray-dried derivatized blood platelets
  • Figure 7 is another electron micrograph of rehydrated spray-dried derivatized blood platelets; and Figure 8 are electron micrographs illustrating ristocetin agglutination of spray- dried rehydrated platelets made according to the method of the invention.
  • the present invention is directed to methods of preparing dehydrated blood products, and dehydrated blood products made by the method.
  • Useful hydrated blood products that may be dehydrated by the method of the invention include, but are not limited to, whole blood, blood plasma, blood platelets, red blood cells, blood serum, plasma, and combinations of these.
  • One particularly useful blood product that is suitable for the method of the present invention is blood platelets that have been fixed with a fixative agent, such as formaldehyde or paraformaldehyde.
  • the blood products may be modified with additional diagnostic or therapeutic agents, such as imaging agents, concentration factors, performance enhancement drugs, antimicrobial and antiviral reagents, universal donor solutions, and the like, as well as combinations of these.
  • One example of a useful modified product is STASIX (derivatized dried blood platelets) available from Entegrion, Inc. (Research Triangle Park, NC).
  • Spray drying is a method of transforming material in a fluid state into a dried particulate form by spraying a feed of a material into a warm drying medium.
  • Spray drying involves evaporation of moisture from an atomized feed by mixing the spray and the drying medium in a controlled fashion.
  • the drying medium is typically air, although other gases such as nitrogen may also be used. The drying proceeds until the desired moisture content is reached in the sprayed particles and the product is then separated from the drying medium.
  • the complete process of spray drying basically consists of a sequence of four processes.
  • the dispersion can be achieved with a pressure nozzle, a two fluid nozzle, a rotary disk atomizer or an ultrasonic nozzle. Selection upon the atomizer type depends upon the nature and amount of feed and the desired characteristics of the dried product. The higher the energy for the dispersion, the smaller are the generated droplets.
  • the manner in which spray contacts the drying air is an important factor in spray dryer design, as this has great bearing on dried product properties by influencing droplet behavior during drying.
  • the material is sprayed in the same direction as the flow of hot air through the apparatus. The droplets come into contact with the hot drying gas when they are the most moist.
  • the material is sprayed in the opposite direction of the flow of hot gas.
  • the hot gas flows upwards and the product falls through increasingly hot air into the collection tray.
  • the residual moisture is eliminated, and the product becomes very hot.
  • This method is suitable only for thermally stabile products.
  • the advantages of both spraying methods are combined.
  • the product is sprayed upwards and only remains in the hot zone for a short time to eliminate the residual moisture. Gravity then pulls the product into the cooler zone. This embodiment is particularly advantageous because the product is only in the hot zone for a short time, and is less likely to be affected by heat.
  • air is mostly used as drying medium, but other gases such as nitrogen may also be used.
  • the gas stream is heated electrically or in a burner and after the process exhausted to atmosphere. If the heating medium is recycled and reused, typically an inert gas such as nitrogen, is used instead of air.
  • an inert gas such as nitrogen
  • Use of nitrogen is advantageous when flammable solvents, toxic products or oxygen sensitive products are processed.
  • evaporation takes place from the saturated vapor film which is quickly established at the droplet surface. Due to the high specific surface area and the existing temperature and moisture gradients, heat and mass transfer results in efficient drying. The evaporation leads to a cooling of the droplet and thus to a small thermal load.
  • Drying chamber design and air flow rate provide a droplet residence time in the chamber, so that the desired droplet moisture removal is completed and product removed from the dryer before product temperatures can rise to the outlet drying air temperature. Hence, there is little likelihood of heat damage to the product.
  • Two systems are used to separate the product from the drying medium. First, primary separation of the drying product takes place at the base of the drying chamber, and second, total recovery of the dried product in the separation equipment.
  • a cyclone is used to collect the material. Based on inertial forces, the particles are separated to the cyclone wall as a down-going strain and removed.
  • Other systems such as electrostatic precipitators , textile (bag) filters or wet collectors like scrubbers, may also be used to collect the dried product.
  • spray drying offers advantages over other drying methods such as lyophilization (freeze drying).
  • Use of spray drying produces a product that is more consistent, less clumpy, and better dispersed than freeze drying methods.
  • the highly dispersed particles produced by spray drying also allow for a rapid rehydration rate, which is likely a result of a larger available surface area.
  • the clumped nature of a freeze dried product results in substantially longer rehydration times for the blood products that are dried in the method of the invention. Since many transfusions and other uses of blood products can be highly time- sensitive, this higher rate of rehydration can be a significant advantage in battlefield or emergency treatment situations.
  • spray dried fixed blood platelets of the invention can be rehydrated to form a rehydrated fixed blood platelet composition, and the composition has a turbidity (A 500 ) value less than that of a comparable rehydrated lyophilized composition of fixed blood platelets.
  • the spray-dried products of the method of the invention may be used as topical treatments in treating wounds.
  • the products may be used directly on a wound to assist clotting, or may be applied to a bandage or surgical aid or covering to assist in wound healing.
  • the rehydrated forms of the spray- dried products of the method of the invention may be administered via intravenous injection as therapeutic treatments to patients afflicted with blood-related disorders such as thrombocytopenia (including washout thrombocytopenia), hemorrhagic platelet dysfunction, and trauma victims experiencing severe bleeding.
  • blood-related disorders such as thrombocytopenia (including washout thrombocytopenia), hemorrhagic platelet dysfunction, and trauma victims experiencing severe bleeding.
  • Porcine and human plasma can be spray-dried in a Buchi B- 270 research spray-dryer at a flow rate of 415 liters N 2 per hour at 140 0 C, 130 0 C, 120 0 C, 110 0 C, and lower if dehydration can be obtained. Runs are preferably performed three times at each temperature and with each type (i.e., porcine and human) plasma. The final product can be analyzed for moisture content and microparticles imaged with scanning electron microscopy. Portions of pig and human plasma may also be lyophilized at -20 0 C for three days from a 4 mm layer to obtain a "lyophilization control" cake. As shown in the accompanying Figures, spray-dried material is observed to be a fine powder, and appear as microspheres under the microscope, while lyophilized material forms a cake.
  • Turbidity and rate of solubilization - Optical measurement of the light absorption at 700 nm can assess turbidity as a function of time after initiation of the rehydration reaction.
  • Viscosity can be estimated with a falling ball viscometer.
  • Coagulation factor levels (including FII, FV, FVII, FVIII, FIX, FX, FXII, FXII, FXIII, protein S, protein C, von Willebrand factor) are measured with ELISA analysis.
  • Coagulation pathway turnover - Prothrombin times and activated partial thromboplastin times are measured with concentrated plasmas after dilution of the hyper- concentrated solutions to Ix. Final clots are examined with scanning electron microscopy to assess fiber thickness and density.
  • a concentrated solution preferably will have the appropriate rheology for standard transfusion practice in which coagulation factor levels and activities are within normal intra- and inter-individual ranges of variation. This solution can be utilized for the "most concentrated" infusions in porcine studies described below.
  • thermo dilution catheter is inserted via the external jugular vein into a pulmonary artery; micromanometer- tipped catheters are positioned via the left femoral vessels into the right atrium and thoracic aorta; a .22 gauge catheter is inserted into the left femoral artery and connected to a withdrawal pump. Patterns of blood flow are measured by placing Doppler flow probes on the cephalic and mesenteric arteries; this procedure can be supported by carotid artery cut down and laparotomy.
  • Hemorrhagic shock can be induced by withdrawing 40% of total blood volume over a one-hour period. After withdrawal of blood and verification of hemorrhagic shock (mean arterial blood pressure ⁇ 40 mm Hg, shift in cephalic, splanchnic blood flow pattern), the animals are infused with multiple doses of Ix spray-dried plasma or hyper-concentrated spray-dried plasma at an intermediate and high level of concentration (to be determined as described above). Each infusion is preferably a volume equivalent to 1/lOth of the animal's blood volume, and is preferably performed over a three minute period with a Harvard syringe pump.
  • Hemodynamic and other physiological parameters can be measured, and infusions can be stopped when two successive boluses result in worsening hemodynamic stability. Animals are then be sacrificed for autopsy and histological analysis. The number of animals and the infused products used in this Example are shown in Table 1.
  • Microvasculopathologies and hemolytic disorders After sacrifice, selected renal, hepatic, pulmonary, splenic, lung and other tissue are prepared for light microscopic analysis. The histological analysis focuses on identifying signs of macroscopic or disseminated intravascular coagulation or premature induction of selected organ failure. Data analysis. Comparisons between plasma groups are made with the Wilcoxon Signed Rank Test, and directionality assessed using the Sign test.
  • Example 1 Spray- Drying Of Plasma and Preservation of Coagulation Protein Activities
  • Solvent-detergent pooled plasma was subjected to standard spray-drying (415 liters N 2 per hour at 120 0 C in Butchi, Inc. B-270) to obtain the product depicted in Figure 1.
  • the spherical-dimpled geometry of the resulting microparticles is similar to the shapes obtained when other proteins are spray-dried, indicating that a protein surface shell forms as a result of the initial kinetics of water removal and concentration (e.g., see Vehring 16 ).
  • blood products may be dried and rehydrated in accordance with the description above.
  • Virtually any treated or untreated blood product may be used in the method of the invention.
  • blood products include whole blood, blood plasma, blood platelets, red blood cells, blood serum, as well as combinations of these.
  • the blood products may be used in the method of the invention in their naturally occurring state, or may be modified in any way. Examples of modifications of these blood products include fixation with a fixing agent such as formaldehyde or paraformaldehyde as described in U.S. Patent Nos.
  • STASIX derivatized dried blood platelets
  • Entegrion, Inc. Research Triangle Park, NC
  • the goal of this example is to rehydrate spray-dried derivatized blood platelets
  • a "reference A 2 so value" for the bulking medium used for the pre- spray-dried suspension is obtained. This is an A 2 so nm value for the pre- spray-dry after the platelets are spun out, reflecting the supernatant protein concentration, which is largely human serum albumin bulking agent.
  • a trial rehydration with the post- spray-dried powder is performed at 10% (w/v), then the optical density at 280 nm (A 280 ) of the bulking agent (human serum albumin) is measured.
  • the pre- spray-dried supernatant A 280 and 10% supernatant A 280 values are compared (ratioed) to determine how far off the 10% rehydration approximation was.
  • This ratio is then used to calculate the exact weight percentage of dried powder that is needed to match the bulking agent protein concentration of the pre-spray dried suspension.
  • the platelet count of the post-rehydration particles are then measured two ways. First with a Hiska cell counter and second by measuring the optical turbidity. These values, and related rehydration volumes, form the starting point for all the particle characterization assays.
  • weight percentage can be in units of mg/ml, e.g., 8.9% (w/v) is equivalent to 89 mg/ml.
  • Electron micrographs of rehydrated spray-dried derivatized blood platelets are shown in Figure 6 and Figure 7.
  • Group 1 - vehicle (buffer) control Five groups of monkeys were used - Group 1 - vehicle (buffer) control; Group 2 - IX therapeutic STASIX dose; Group 3 - 5X therapeutic STASIX dose; Group 4 - 1OX therapeutic STASIX dose; and Group 5 - human serum albumin (500 mg/kg). Dosages were respectively 0.0, 2.1 x 10 9 , 1.05 x 10 10 , 2.1 x 10 10 , and 0.0 platelets/kg in group 1, 2, 3, 4, and 5.
  • a IX dose is the estimated therapeutic STASIX dose in a human patient, i.e., an additional 30,000 platelet particles per microliter of blood.
  • Example 4 Spray-Drying of Aldehyde Stabilized Platelets
  • the utility of spray-drying as an alternative to lyophilization for the dehydration of aldehyde-stabilized platelets is examined in this example.
  • Human apheresis platelets were stabilized using the procedure of Read et al. described in U. S,. Patent No. 5,651,966, which is herein incorporated by reference in its entirety.
  • Figure 8 depicts spray-dried platelets after rehydration, exchange into normal human plasma (as a von Willebrand factor source) and addition of ristocetin to 1 mg/ml (Panel B) or a corresponding volume of control buffer (Panel A). Large aggregates were noted with ristocetin addition, indicating that spray-drying preserved glycoprotein IB
  • Cynomolgus monkeys (1 or 2/sex/group) received a single 5 -minute intravenous infusion of the spray-dried platelets at doses of 2.IxIO 9 , 1.05xl0 10 , or 2.IxIO 10 platelets/kg.
  • Control animals (2/sex) received vehicle (5.375 mM sodium citrate and 2 mM cysteine in physiological saline) and an additional group received 500 mg/kg human serum albumin (HSA). The dose volume was 2 niL/kg/min for all groups. Animals were observed for 1 or 7 days post-dose. One day after dose administration, 1 animal/sex/group was euthanized and necropsied.
  • Plasma separated from fresh porcine blood was either stored as fresh frozen plasma (FFP) or preserved as freeze dried plasma (FDP) or spray-dried plasma (SDP, prepared as detailed in previous examples).
  • FFP fresh frozen plasma
  • FDP freeze dried plasma
  • SDP spray-dried plasma
  • IxSDP distilled water which was either equal (IxSDP) or one-third (3xSDP) the original volume of FFP.
  • Coagulation profiles (PT, PTT, thromboelastography) were measured at baseline (BL), post-shock (PS), post crystalloid (PC), treatment (MO), and during 4 hours of monitoring (M 1-4).

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ES10762281.3T ES2684130T3 (es) 2009-04-09 2010-04-06 Productos de la sangre secados por aspersión y métodos para elaborar los mismos
AU2010234607A AU2010234607B2 (en) 2009-04-09 2010-04-06 Spray-dried blood products and methods of making same
JP2012504757A JP2012523414A (ja) 2009-04-09 2010-04-06 噴霧乾燥した血液製剤とその製造方法
CN2010800227674A CN102448475A (zh) 2009-04-09 2010-04-06 喷雾干燥的血液产品和制造其的方法
BRPI1006722A BRPI1006722A2 (pt) 2009-04-09 2010-04-06 "método de preparação de hemoderivados desidratado, hemoderivados desidratados, bandagem ou auxílio cirúrgico, método para a preparação de plaquetas sanguíneas fixas desidratadas, plaquetas sanguíneas fixas desidratadas, método para o tratamento de um paciente que sofre de um distúrbio sanguíneo e plaquetas de sangue fixas secas por atomização tendo geometria com cavidades esféricas"
MX2011010633A MX2011010633A (es) 2009-04-09 2010-04-06 Productos sanguineos deshidratados por aspersion y metodos para elaborar los mismos.
EP10762281.3A EP2416790B1 (en) 2009-04-09 2010-04-06 Spray-dried blood products and methods of making same
US13/262,931 US9867782B2 (en) 2009-04-09 2010-04-06 Spray-dried blood products and methods of making same
CA2757961A CA2757961C (en) 2009-04-09 2010-04-06 Spray-dried blood products and methods of making same
US15/872,727 US11213488B2 (en) 2009-04-09 2018-01-16 Spray-dried blood products and methods of making same
US17/566,226 US12414920B2 (en) 2009-04-09 2021-12-30 Spray-dried blood products and methods of making same
US19/204,890 US20250332106A1 (en) 2009-04-09 2025-05-12 Spray-dried blood products and methods of making same

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