Methods of preparing lyophilized spherical-shaped pellets of biological materials

Patent metadata

Field	Value
Publication number	US-10183968-B2
Application number	US-201214355405-A
Country	US
Kind code	B2
Filing date	Oct 26, 2012
Priority date	Oct 31, 2011
Publication date	Jan 22, 2019
Grant date	Jan 22, 2019

Abstract

Methods for preparing lyophilized pellets of biological materials are described. The pellets have a substantially spherical shape and are prepared by freezing droplets of a liquid composition of a desired biological material on a flat, solid surface, in particular, a surface that does not have any cavities, followed by lyophilizing the frozen droplets. These methods are useful for preparing lyophilized pellets having a high concentration of a desired biological material, in particular a therapeutic protein or vaccine, and which have a faster reconstitution time than lyophilized powder cakes prepared in vials.

First claim

What is claimed:

1. A method of preparing a lyophilized pellet of a biological material, comprising:
providing a vessel which contains a liquid composition comprising the biological material;
providing a metal plate which comprises a top surface that is solid and flat and does not contain a hydrophobic film and a bottom surface that is in physical contact with a heat sink adapted to maintain the top surface of the metal plate at a temperature of −90° C. or below;
positioning a dispensing tip above the top surface of the metal plate, the dispensing tip having an open end configured to dispense liquid droplets and another end connected to the vessel, wherein there is a gap of at least 0.1 cm between the top surface of the metal plate and the open end of the dispensing tip;
dispensing an aliquot of the liquid composition through the open end of the dispensing tip as a single droplet onto the top surface of the metal plate in a manner that maintains the droplet as a single droplet having a substantially spherical shape as it contacts and freezes on the top surface; and lyophilizing the frozen droplet to produce a dried pellet of substantially spherical shape.

2. The method of claim 1 , wherein the dispensing is performed at a speed and at a gap distance that prevents freezing of any portion of the aliquot in the dispensing tip and maintains the droplet in simultaneous contact with the top surface of the metal plate and the open end of the dispensing tip until the droplet surface touching the plate is frozen.

3. The method of claim 2 , wherein the dispensing speed is selected from the group consisting of: 3 ml/min to 75 ml/min; 5 ml/min to 75 ml/min; 3 ml/min to 60 ml/min, 20 ml/min to 75 ml/min; and 20 ml/min to 60 ml/min.

4. The method of claim 2 , wherein the aliquot is 250 μl and the dispensing speed is between 5 ml/min to 75 ml/min, or wherein the aliquot is 100 μl and the dispensing speed is between 3 ml/min to 60 ml/min.

5. The method of claim 1 , wherein the top surface temperature of the metal plate is below −150° C. and a gap distance between the open end of the dispensing tip and the top surface of the metal plate is between 0.1 cm and 0.5 cm or between 0.1 cm and 1 cm or between 0.1 cm and 0.75 cm.

6. The method of claim 5 , wherein the surface temperature of the metal plate is between −180° C. and −196° C. or between −180° C. and −273° C.

7. The method of claim 1 , wherein the heat sink comprises a plurality of metal fins having first and send ends and arranged perpendicularly to the metal plate, with the first end of each fin touching the bottom surface of the metal plate and the second end of each fin immersed in liquid nitrogen.

8. The method of claim 1 , wherein the biological material is selected from the group consisting of a purified antibody at a concentration in the liquid composition of at least 50 mg/ml or 100 mg/ml; a vaccine, a fusion protein, a polypeptide, and a peptide.

9. The method of claim 1 , wherein the liquid composition comprises a total solute concentration of at least 25 on a weight by weight basis.

10. The method of claim 1 , further comprising measuring the reconstitution time of the lyophilized pellet.

11. A container containing at least one lyophilized pellet, wherein the lyophilized pellet is prepared by:
providing a vessel which contains a liquid composition comprising a biological material;
providing a metal plate which comprises a top surface that is solid and flat and does not contain a hydrophobic film and a bottom surface that is in physical contact with a heat sink adapted to maintain the top surface of the metal plate at a temperature of −90° C. or below;
positioning a dispensing tip above the top surface of the metal plate, the dispensing tip having an open end configured to dispense liquid droplets and another end connected to the vessel, wherein there is a gap of at least 0.1 cm between the top surface of the metal plate and the open end of the dispensing tip;
dispensing an aliquot of the liquid composition through the open end of the dispensing tip as a single droplet onto the top surface of the metal plate in a manner that maintains the droplet as a single droplet having a substantially spherical shape as it contacts and freezes on the top surface; and lyophilizing the frozen droplet to produce a dried pellet of substantially spherical shape.

12. The container of claim 11 , wherein the lyophilized pellet has a reconstitution time of less than 3 minutes or less than 2 minutes or less than 1 minute.

13. The container of claim 11 , wherein the container is a glass vial.

14. The container of claim 11 , wherein the container comprises first and second compartments, with the at least one lyophilized pellet present in the first compartment and a reconstitution liquid present in the second compartment.

15. The container of claim 11 , wherein the lyophilized pellet comprises a TNFRII-Fc fusion protein, and is prepared from a liquid composition comprising 25 mg/ml of a TNFRII-Fc fusion protein, 40 mg/ml mannitol, 10 mg/ml sucrose, and 1.2 mg/ml tromethamine in sterile water, pH 7.

16. A container comprising at least two dried pellets that are substantially spherical in shape, wherein one of the two dried pellets comprises a first biological material and the other dried pellet comprises a second biological material that is different than the first biological material, wherein each of the two dried pellets in the container is prepared by:
providing a vessel which contains a liquid composition comprising the first and second biological materials;
providing a metal plate which comprises a top surface that is solid and flat and does not contain a hydrophobic film and a bottom surface that is in physical contact with a heat sink adapted to maintain the top surface of the metal plate at a temperature of −90° C. or below;
positioning a dispensing tip above the top surface of the metal plate, the dispensing tip having an open end configured to dispense liquid droplets and another end connected to the vessel, wherein there is a gap of at least 0.1 cm between the top surface of the metal plate and the open end of the dispensing tip;
dispensing an aliquot of the liquid composition through the open end of the dispensing tip as a single droplet onto the top surface of the metal plate in a manner that maintains the droplet as a single droplet having a substantially spherical shape as it contacts and freezes on the top surface; and lyophilizing the frozen droplet to produce a dried pellet of substantially spherical shape.

17. The container of claim 16 , wherein each the first and second biological materials are components of a multi-component vaccine.

18. The method of claim 1 , wherein the metal plate is selected from the group consisting of gold, silver, stainless steel, aluminum or copper.

19. The container of claim 11 , wherein the metal plate is selected from the group consisting of gold, silver, stainless steel, aluminum or copper.

20. The container of claim 16 , wherein the metal plate is selected from the group consisting of gold, silver, stainless steel, aluminum or copper.

Assignees

• Merck Sharp & Dohme

Inventors

• Barr Colleen
• Bhambhani Akhilesh
• Evans Robert
• Isopi Lynne
• Krah David
• Kriss Jennifer
• Sinacola Jessica

Classifications

• F26B5/06

  the process involving freezing · CPC title

• C07K14/7151

  for tumor necrosis factor [TNF], for lymphotoxin [LT] · CPC title

• A61K39/39591 Primary

  Stabilisation, fragmentation · CPC title

• C07K16/00

  Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies · CPC title

• C07K2319/30

  Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto · CPC title