Granted Patent B2

US 12,702,682 · App. 18/052,709 · Granted Aug 11, 2026

Inventors: Joshua Donald Montgomery (Silver Spring, MD); Braden Carl Ishler (Montgomery Village, MD); Stephen Edward Amos (Buckeystown, MD); Keith Andrew Moskowitz (Westfield, IN); Amber Nicole Lee (Montgomery Village, MD); Rafael Jorda (Merignac, FR); Glen Michael Fitzpatrick (North Potomac, MD); Michael Alexander Mathews (Arlington, VA)

Assignee: Cellphire, Inc.

A61K35/19A61K9/19A61K47/26

Abstract

Claims (28)

1 . A method for restoring hemostasis in a subject,

wherein said method comprising:

rehydrating a platelet derivative composition in the form of a powder to form a rehydrated platelet derivative composition; and

administering intravenously an effective amount of platelet derivatives in the rehydrated platelet derivative composition to the subject,

wherein the platelet derivatives in the rehydrated platelet derivative composition:

have a CD41 percent positivity of at least 70% when measured using flow cytometry,

have a phosphatidylserine percent positivity of at least 70%, when measured using Annexin V binding in flow cytometry,

have a concentration of at least 6,000×10 3 platelet derivatives/μl,

are capable of generating thrombin in an in vitro thrombin formation assay, and

are capable of producing an occlusion by attaining a pressure of 80 kPa in less than 14 minutes when used at a concentration of about 70×10 3 platelet derivatives/μL in platelet-reduced citrated whole blood in an in vitro total thrombus-formation analysis system (T-TAS), and

wherein less than 3.5% of the platelet derivatives are microparticles.

2 . The method of claim 1 , wherein the platelet derivatives

have a CD41 percent positivity of at least 80%, when measured using flow cytometry, and

have a phosphatidylserine percent positivity of at least 80%, when measured using Annexin V binding in flow cytometry.

3 . The method of claim 1 , wherein the microparticles are present in an amount in the range of 0.1% to 3% when measured using scattering intensity in flow cytometry.

4 . The method of claim 1 , wherein the rehydrated platelet derivative composition comprises trehalose, polysucrose, one or more salts, and a buffer, and wherein the trehalose is present in an amount in the range of 50 mM to 200 mM, and the polysucrose is present in the range of 3% w/v to 10% w/v.

5 . The method of claim 3 , wherein the platelet derivatives when analyzed using flow cytometry for the binding of anti-thrombospondin (TSP) antibody in the absence of an agonist, exhibit at least 10-fold higher mean fluorescent intensity (MFI) as compared to the MFI of binding of the anti-TSP antibody to resting platelets in the absence of the agonist.

6 . The method of claim 5 , wherein the platelet derivatives in the rehydrated platelet derivative composition have less than 2% crosslinking of platelet membranes via proteins and/or lipids present on the platelet membranes, and wherein the platelet derivatives in the rehydrated platelet derivative composition are capable of generating thrombin in the in vitro thrombin formation assay such that the platelet derivatives when present at a concentration of at least 4.8×10 3 particles/μL generate a thrombin peak height (TPH) of at least 25 nM in the presence of a reagent containing tissue factor and phospholipids.

7 . The method of claim 4 , wherein at least 90% of the platelet derivatives are in the diameter range of 0.5 μm to 2.5 μm.

8 . The method of claim 7 , wherein the microparticles are present in an amount in the range of 0.1% to 3% when measured using scattering intensity in flow cytometry, wherein the platelet derivatives have a phosphatidylserine percent positivity of at least 90% when measured using Annexin V binding in flow cytometry, wherein the platelet derivatives have a CD41 percent positivity of at least 95%, when measured using flow cytometry.

9 . The method of claim 8 , wherein the platelet derivatives when analyzed using flow cytometry for the binding of anti-thrombospondin (TSP) antibody in the absence of an agonist, exhibit at least 10-fold higher mean fluorescent intensity (MFI) as compared to the MFI of binding of the anti-TSP antibody to resting platelets in the absence of the agonist.

10 . The method of claim 9 , wherein the platelet derivatives when analyzed using flow cytometry for the binding of anti-von Willebrand factor (vWF) antibody in the absence of an agonist, exhibit at least 2-fold higher mean fluorescent intensity (MFI) as compared to the MFI of binding of the anti-vWF antibody to the resting platelets in the absence of the agonist.

11 . The method of claim 10 , wherein the platelet derivatives in the rehydrated platelet derivative composition are present in a concentration in the range of 6,000×10 3 to 11,000×10 3 platelet derivatives/μl of the rehydrated platelet derivative composition.

12 . The method of claim 11 , wherein the platelet derivative composition is in a vial, and the volume of the rehydrated platelet derivative composition in the vial after the rehydrating is 10-20 ml.

13 . The method of claim 10 , wherein the platelet derivatives in the rehydrated platelet derivative composition are in a vial and are present in a concentration in the range of 10,000×10 3 to 20,000×10 3 platelet derivatives/μl of the rehydrated platelet derivative composition.

14 . The method of claim 13 , wherein the platelet derivative composition is in a vial, and the volume of the rehydrated platelet derivative composition in the vial after the rehydrating is 10-20 ml.

15 . The method of claim 10 , wherein the platelet derivative composition in the form of the powder is in a vial, wherein the method further comprises selecting the vial from a plurality of vials comprising the platelet derivative composition from at least 2 different lots in separate vials, wherein each lot comprises platelet derivatives from different subjects, wherein the rehydrated platelet derivative composition comprises plasma protein in a concentration in a range of 5%-50%, and wherein the percent of plasma protein in the powder from any two vials chosen from different lots, differs by less than an absolute 20% plasma protein in the powder.

16 . The method of claim 15 , wherein the percent of microparticles in the powder from any two vials chosen from different lots, differs by less than an absolute 2% microparticles in the powder.

Assignments (1)

Continuity (8)

References Cited (400)