Network


Latest external collaboration on country level. Dive into details by clicking on the dots.

Hotspot


Dive into the research topics where Charles Pak is active.

Publication


Featured researches published by Charles Pak.


Biochimica et Biophysica Acta | 1998

Triggerable liposomal fusion by enzyme cleavage of a novel peptide–lipid conjugate

Charles Pak; Shaukat Ali; Andrew S. Janoff; Paul Meers

A novel peptide-lipid sensitive to enzyme cleavage was designed to generate liposomes that could be triggered to fuse by enzymatic activation. Covalent linkage of dioleoyl phosphatidylethanolamine (DOPE) to an elastase substrate, N-acetyl-ala-ala-, resulted in a cleavable peptide-lipid (N-Ac-AA-DOPE) with no intrinsic fusogenic activity. Cleavage of N-Ac-AA-DOPE and concomitant conversion to the fusogenic lipid DOPE could be detected after treatment with human leukocyte elastase or proteinase K, two proteases with similar substrate specificities. A strategy to utilize this cleavage to trigger fusogenicity was tested by modeling the fusion of liposomes containing the expected product of complete cleavage. Based on these results liposomes were designed to contain N-Ac-AA-DOPE, DOTAP, and PE in the ratio of 15/15/70. These liposomes exhibited lipid mixing with acceptor liposomes after elastase or proteinase K protease treatment. Activation of fusion, as monitored by a lipid mixing assay, appeared to be dependent on protease activity, as (1) heat inactivated enzyme did not activate liposomal fusion, and (2) the time and concentration dependence of proteinase K mediated cleavage of N-Ac-AA-DOPE correlated with membrane mixing. Liposomes could also be formulated that exhibited lipid mixing and transfer of aqueous fluorescent probe with erythrocyte ghosts. These observations demonstrate fusogenic lipids conjugated to enzyme substrates serve as triggerable fusion systems that may be useful for gene and drug delivery.


Biochimica et Biophysica Acta | 1998

Cation-dependent fusogenicity of an N-acyl phosphatidylethanolamine

Tong Shangguan; Charles Pak; Shaukat Ali; Andrew S. Janoff; Paul Meers

N-acyl phosphatidylethanolamines (NAPEs) are natural lipid components of many organisms. N-acylation of unsaturated phosphatidylethanolamines with a saturated fatty acid converts them from non-lamellar organizing lipids into lamellar organizing, acidic lipids which can interact with cations and potentially return to non-lamellar structures. These special properties make NAPEs candidates for fusogens. We tested the fusogenicity of one of the NAPEs, N-dodecanoyl-di-oleoylphosphatidylethanolamine (N-C12-DOPE) mixed with dioleoylphosphatidylcholine (DOPC) in liposomes. Binding and fusion to erythrocyte ghosts in the presence of 3 mM Ca2+ required at least 60 mol% of N-C12-DOPE. Fusion was not observed when phosphatidylglycerol or phosphatidylserine was substituted for N-C12-DOPE, indicating specificity for properties of this lipid. Binding of N-C12-DOPE/DOPC (70:30) liposomes required 1 mM Ca2+ while 1.25 mM Ca2+ and Mg2+ were sufficient for lipid mixing and delivery of encapsulated dextrans to erythrocyte ghosts. These liposomes also bound and possibly mixed lipid with nucleated U-937 cells in a Ca2+ -and endocytosis-dependent manner. Low pH-dependent fusion with ghosts was observed in the absence of any divalent cation, indicating that fusion with U-937 cells could result after endocytosis into the acidic endosomes. The possible mechanisms for N-C12-DOPE mediated binding and fusion and the potential application of these liposomes as delivery vehicles for therapeutic agents are discussed.


Biochimica et Biophysica Acta | 1999

Elastase activated liposomal delivery to nucleated cells

Charles Pak; Ravi K. Erukulla; Patrick L. Ahl; Andrew S. Janoff; Paul Meers

The specific activation of liposomes for delivery has been explored by enzyme mediated cleavage of a peptide substrate covalently conjugated to a fusogenic lipid. We have previously shown an elastase sensitive peptide conjugated to 1, 2-dioleoyl-sn-glycero-3-phosphoethanolamine [corrected] (DOPE) could be activated by enzymatic cleavage, triggering liposome-liposome lipid mixing and fusion with erythrocyte ghosts (Pak et al., Biochim. Biophys. Acta, 1372 (1998) 13-27). Further optimization of this system has been aimed at obtaining substrate cleavage at or below physiological elastase levels and to demonstrate triggered delivery to living cells. Therefore a new peptide-lipid, MeO-suc-AAPV-DOPE (N-methoxy-succinyl-Ala-Ala-Pro-Val-DOPE), has been developed that exhibits greater sensitivity and selectivity for elastase cleavage and subsequent conversion to DOPE. This peptide-lipid was used with DODAP (dioleoyl dimethylammonium propane, a pH dependent cationic lipid) in a 1:1 mol ratio with the expectation that endocytosis would lead to a liposome with an overall positive charge if enzymatic cleavage had occurred. Elastase treated liposomes displayed pH dependent enhancement of binding, lipid mixing, and delivery of 10000 MW dextrans, relative to untreated liposomes, when incubated with HL60 human leukemic cells. Heat denatured elastase did not activate DODAP/MeO-suc-AAPV-DOPE liposomes, indicating enzymatic activity of elastase is necessary. Liposomes bound to ECV304 endothelial cells at physiological pH could be activated by elastase to deliver an encapsulated fluorescent probe, calcein, into the cell cytoplasm. These results suggest enzyme substrate peptides linked to a fusogenic lipid may be used to elicit specific delivery from liposomes to cells.


Archive | 1997

Peptide-lipid conjugates, liposomes and lipsomal drug delivery

Paul Meers; Charles Pak; Shaukat Ali; Andrew S. Janoff; J. Craig Franklin; Ravi K. Erukulla; Donna Cabral-Lilly; Patrick L. Ahl


Archive | 1997

Peptide-lipid conjugates

Paul Meers; Charles Pak; Shaukat Ali; Andrew S. Janoff; J. Craig Franklin; Ravi K. Erukulla; Donna Cabral-Lilly


Archive | 1997

N-acyl phosphatidylethanolamine-mediated liposomal drug delivery

Paul Meers; Tong Shangguan; Shaukat Ali; Andrew S. Janoff; Charles Pak


Archive | 1998

Liposomal peptide-lipid conjugates and delivery using same

Paul Meers; Charles Pak; Shaukat Ali; Andrew S. Janoff; J. Craig Franklin; Ravi K. Erukulla; Donna Cabral-Lilly


Archive | 2000

Peptid-lipid-konjugate, liposomen und liposomale medikamentenverabreichung

Paul Meers; Charles Pak; Shaukat Ali; Andrew S. Janoff; J. Craig Franklin; Ravi K. Erukulla; Donna Cabral-Lilly; Patrick L. Ahl


Archive | 2000

Conjugues peptides-lipides, liposomes et apport de medicaments liposomiques

Paul Meers; Charles Pak; Shaukat Ali; Andrew S. Janoff; J. Craig Franklin; Ravi K. Erukulla; Donna Cabral-Lilly; Patrick L. Ahl


Biochimica et Biophysica Acta | 2000

Corrigendum to: 'Elastase-activated liposomal delivery to nucleated cells'.

Charles Pak; Ravi K. Erukulla; Patrick L. Ahl; Andrew S. Janoff; Paul Meers

Collaboration


Dive into the Charles Pak's collaboration.

Top Co-Authors

Avatar
Top Co-Authors

Avatar
Top Co-Authors

Avatar
Top Co-Authors

Avatar
Top Co-Authors

Avatar
Top Co-Authors

Avatar
Top Co-Authors

Avatar
Top Co-Authors

Avatar
Researchain Logo
Decentralizing Knowledge