Fusogenic properties of saposin C and related proteins and peptides for application to transmembrane drug delivery systems

• US 9,271,932 B2
• Filed: 04/27/2007
• Issued: 03/01/2016
• Est. Priority Date: 04/28/2006
• Status: Active Grant

• Current Assignee: Children's Hospital & Medical Center
• Original Assignee: Children's Hospital & Medical Center
• Inventors: Qi, Xiaoyang
• Primary Examiner(s): Sang, Hong
• Application Number:
  US11/741,323
• Publication Number:
  US 20090142267A1
• Time in Patent Office: 3,231 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/18 Growth factors; Growth regulators
• CPC Class Codes: A61K 9/0014 Skin, i.e. galenical aspects of topical compositions non-active ingredients are additionally classified in A61K47/00; A61K9/0009, A61K9/0021, A61K9/7015, A61K9/7023 take precedence; cosmetic preparations A61K8/00, A61Q; preparations for wound dressings or bandages A61L26/00
• CPC Class Codes: A61K 9/0043 Nose
• CPC Class Codes: A61K 9/006 Oral mucosa, e.g. mucoadhesive forms, sublingual droplets; Buccal patches or films; Buccal sprays
• CPC Class Codes: A61K 9/007 Pulmonary tract; Aromatherapy
• CPC Class Codes: A61K 9/127 Liposomes
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4705 stimulating, promoting or activating activity

Abstract

The present invention comprises a method for delivering pharmaceutical and/or imaging agents within and/or through the dermal, mucosal and other cellular membranes, and across the blood-brain barrier, utilizing a fusogenic protein. The fusogenic protein is associated with a phospholipid membrane, such as a liposome. The liposome may include dioleoylphosphatidylserine, a negatively charged long-chain lipid. Alternatively, the liposome is comprised of a mixture of negatively charged long-chain lipids, neutral long-chain lipids, and neutral short-chain lipids. Preferred fusogenic proteins include saposin C and other proteins, polypeptides and peptide analogs derived from saposin C. The active agent contained within the liposome may comprise biomolecules and/or organic molecules. This technology can be used for both cosmetic and medicinal applications in which the objective is delivery of the active agent within and/or beneath biological membranes or across the blood-brain barrier and neuronal membranes.

Claims

1. A method of delivering an anti-cancer agent or an imaging agent to a lesion of the nervous system, said method comprising the step of systemically administering to a patient a composition comprising:

a) one or more phospholipids selected from the group consisting of anionic long-chain lipids, neutral long chain lipids, neutral short chain lipids, anionic short-chain lipids, and mixtures thereof;

b) a safe and effective amount of the agent; and

c) a saposin c polypeptide;

wherein the composition forms nanoparticles having a mean diameter between 200 and 350 nanometers,

wherein the overall charge of the nanoparticle is negative, and

wherein the lesion is a brain cancer.

2. The method of claim 1 wherein the phospholipids comprise a mixture of anionic long-chain lipids and neutral long-chain lipids.

3. The method of claim 1 wherein the phospholipids comprise a mixture of anionic long-chain lipids, neutral long-chain lipids and neutral short-chain lipids.

4. The method of claim 3 wherein the nanoparticle contains cationic phospholipids.

5. The method of claim 3 wherein the amount of the anionic long-chain lipids, the neutral long-chain lipids and the neutral short-chain lipids is governed by a formula of ([neutral long chain lipids]+[anionic long chain lipids])/(neutral short chain lipids), and wherein the result of the formula equals about 4.

6. The method of claim 1 wherein the nanoparticle comprises dioleoylphosphatidylserine, dipalmitoyl phosphatidylcholine and hexanoyl phosphatidylcholine, wherein the amount of the anionic long-chain lipids, the neutral long-chain lipids and the neutral short-chain lipids is governed by a formula of ([neutral long chain lipids]+[anionic long chain lipids])/(neutral short chain lipids), and wherein the result of the formula equals about 4.

7. The method of claim 1, wherein the pH of the composition is between about 5.5 and about 2.

8. The method of claim 1 wherein the concentration of the phospholipids is in at least about 1 to about 10-fold excess, by molar ratio, to that of the polypeptide.

9. The method of claim 6 wherein the molar ratio of dioleoylphosphatidylserine to dipalmitoyl phosphatidylcholine is from about 10 to about 1.

10. The method of claim 1, wherein the composition further comprises a pharmaceutically acceptable carrier.

11. The method of claim 1, wherein the brain cancer is neuroblastoma.

12. The method of claim 1, wherein the agent is an imaging agent.

13. The method of claim 12 wherein the imaging agent is selected from the group consisting of magnetic resonance detectable label, fluorescence, and CT/PET detectable label.

14. The method of claim 12 wherein the imaging agent has two or more imaging properties.

15. The method of claim 14 wherein the imaging agent is a PTIR dye containing both a fluorophore and a Gd(Ill) moiety that can be detected via magnetic resonance imaging (MRI) or confocal fluorescence microscopy.