Method for producing sterile polynucleotide based medicaments

• US 8,435,557 B2
• Filed: 04/25/2008
• Issued: 05/07/2013
• Est. Priority Date: 12/23/2002
• Status: Expired due to Fees

• Current Assignee: Vical, Inc. (Fresh Tracks Therapeutics, Inc.)
• Original Assignee: Vical, Inc. (Fresh Tracks Therapeutics, Inc.)
• Inventors: Enas, Joel, Geall, Andrew
• Primary Examiner(s): SCHNIZER, RICHARD A
• Application Number:
  US12/109,944
• Publication Number:
  US 20080287387A1
• Time in Patent Office: 1,838 Days
• Field of Search: None
• US Class Current: 424/450
• CPC Class Codes: A61K 9/1075 Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers A61K9/0026 takes precedence
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07C 211/63 having quaternised nitrogen atoms bound to acyclic carbon atoms
• CPC Class Codes: C07C 217/28 having one amino group and at least two singly-bound oxygen atoms, with at least one being part of an etherified hydroxy group, bound to the carbon skeleton, e.g. ethers of polyhydroxy amines
• CPC Class Codes: C07C 219/06 having the hydroxy groups esterified by carboxylic acids having the esterifying carboxyl groups bound to hydrogen atoms or to acyclic carbon atoms of an acyclic saturated carbon skeleton
• CPC Class Codes: C07C 219/14 having at least one of the hydroxy groups esterified by a carboxylic acid having the esterifying carboxyl group bound to a carbon atom of a six-membered aromatic ring
• CPC Class Codes: Y10S 977/797 Lipid particle

Abstract

The present invention relates to a novel method for producing formulations comprising a polynucleotide, block copolymer and cationic surfactant. The formulations produced by the current method are suitable for use in polynucleotide based medicaments. A suitable method of production disclosed herein additionally comprises cold filtering a mixture of a polynucleotide, block copolymer and cationic surfactant, thereby sterilizing the formulation. The method of the present invention also eliminates the need for thermal cycling of the formulation, thereby reducing the time and expense required to produce large quantities of a formulation during commercial manufacturing. The present invention also relates to novel cationic lipids.

Claims

1. A method of producing a sterile formulation consisting of:

(a) mixing

(i) benzalkonium chloride (BAK), wherein the final concentration of BAK is from about 0.01 mM to about 0.3 mM;

(ii) a polyoxyethylene (POE) and polyoxypropylene (POP) block copolymer having a cloud point of about −

2°

C. to about 8°

C.; and

(iii) a polynucleotide;

at a temperature below the cloud point of said block copolymer to form a mixture; and

(b) cold filtering the mixture to produce a sterile formulation;

wherein said method does not include raising the temperature of the mixture above the cloud point of said block copolymer.

2. The method of claim 1, wherein said block copolymer is of the general formula:

HO(C₂H₄O)_x(C₃H₆O)_y(C₂H₄O)_xH;

wherein (y) represents a number such that the molecular weight of the hydrophobic POP portion (C₃H₆O) is up to approximately 20,000 Daltons and wherein (x) represents a number such that the percentage of the hydrophilic POE portion (C₂H₄O) is between approximately 1% and 50% by weight.

3. The method of claim 2, wherein said block copolymer is the poloxamer CRL-1005.

4. The method of claim 1, wherein said block copolymer is of the general formula:

HO(C₃H₆O)_y(C₂H₄O)_x(C₃H₆O)_yH wherein (y) represents a number such that the molecular weight of the hydrophobic POP portion (C₃H₆O) is up to approximately 20,000 Daltons and wherein (x) represents a number such that the percentage of hydrophilic POE portion (C₂H₄O) is between approximately 1% and 50% by weight.

5. The method of claim 1, wherein said cold filtering is performed using a filter with a pore size of about 0.01 microns to about 2 microns.

6. The method of claim 1, wherein the final concentration of said BAK present in said formulation is about 0.3 mM.

7. The method of claim 1, wherein the final concentration of said block copolymer present in said formulation is from about 1 mg/mL to about 50 mg/mL.

8. The method of claim 1, wherein the final concentration of said polynucleotide present in said formulation is from about 1 ng/mL to about 10 mg/mL.

9. A method of producing a sterile formulation consisting of:

(a) mixing

(i) benzalkonium chloride (BAK), wherein the final concentration of BAK is from about 0.01 mM to about 0.3 mM;

(ii) a polyoxyethylene (POE) and polyoxypropylene (POP) block copolymer having a cloud point of about −

2°

C. to about 8°

C. and of the general formula;

HO(C₃H₆O)_y(C₂H₄O)_x(C₃H₆O)_yH wherein (y) represents a number such that the molecular weight of the hydrophobic POP portion (C₃H₆O) is up to approximately 20,000 Daltons and wherein (x) represents a number such that the percentage of hydrophilic POE portion (C₂H₄O) is between approximately 1% and 50% by weight; and

(iii) a polynucleotide;

at a temperature below the cloud point of said block copolymer to form a mixture; and

(b) cold filtering the mixture to produce a sterile formulation;

wherein said method does not include raising the temperature of the mixture above the cloud point of said block copolymer.