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Developing a Dissociative Nanocontainer for Peptide Drug Delivery.

Kelly P, Anand P, Uvaydov A, Chakravartula S, Sherpa C, Pires E, O'Neil A, Douglas T, Holford M - Int J Environ Res Public Health (2015)

Bottom Line: The potency, selectivity, and decreased side effects of bioactive peptides have propelled these agents to the forefront of pharmacological research.Recent efforts have demonstrated that P22 nanocontainers can effectively encapsulate analgesic peptides and translocate them across blood-brain-barrier (BBB) models.Our results demonstrate initial attempts to characterize the ROMP-triggered release of cargo peptides from P22 nanocontainers.

View Article: PubMed Central - PubMed

Affiliation: Hunter College and The Graduate Center, City University of New York, Belfer Research Building, 413 E. 69th Street, New York, NY 10021, USA. mkelly3@gc.cuny.edu.

ABSTRACT
The potency, selectivity, and decreased side effects of bioactive peptides have propelled these agents to the forefront of pharmacological research. Peptides are especially promising for the treatment of neurological disorders and pain. However, delivery of peptide therapeutics often requires invasive techniques, which is a major obstacle to their widespread application. We have developed a tailored peptide drug delivery system in which the viral capsid of P22 bacteriophage is modified to serve as a tunable nanocontainer for the packaging and controlled release of bioactive peptides. Recent efforts have demonstrated that P22 nanocontainers can effectively encapsulate analgesic peptides and translocate them across blood-brain-barrier (BBB) models. However, release of encapsulated peptides at their target site remains a challenge. Here a Ring Opening Metathesis Polymerization (ROMP) reaction is applied to trigger P22 nanocontainer disassembly under physiological conditions. Specifically, the ROMP substrate norbornene (5-Norbornene-2-carboxylic acid) is conjugated to the exterior of a loaded P22 nanocontainer and Grubbs II Catalyst is used to trigger the polymerization reaction leading to nanocontainer disassembly. Our results demonstrate initial attempts to characterize the ROMP-triggered release of cargo peptides from P22 nanocontainers. This work provides proof-of-concept for the construction of a triggerable peptide drug delivery system using viral nanocontainers.

No MeSH data available.


Related in: MedlinePlus

Conjugation of Norbornene-COOH to the Capsid Surface. (A) Norbornene-COOH is activated with EDC and sulfo-NHS, leading to the formation of an amide bond with the ε-NH2 of surface-exposed lysine residues. (B) Mass spectrum of unmodified capsid coat protein (m/z = 47,192). (C) Mass spectrum of coat protein subunit conjugated with five norbornene subunits (m/z = 47,792).
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ijerph-12-12543-f003: Conjugation of Norbornene-COOH to the Capsid Surface. (A) Norbornene-COOH is activated with EDC and sulfo-NHS, leading to the formation of an amide bond with the ε-NH2 of surface-exposed lysine residues. (B) Mass spectrum of unmodified capsid coat protein (m/z = 47,192). (C) Mass spectrum of coat protein subunit conjugated with five norbornene subunits (m/z = 47,792).

Mentions: The P22-GFP-norbornene conjugation reaction was monitored by MS analysis. Compounds with masses corresponding to that of the P22 coat protein (expected mass 46,596 Da, observed mass 47,192 Da) plus an integral number (between 2 and 6) of 120 Da adducts was detected (Table 1). The 120 Da mass corresponds to a 5-norbornene-2-carboxylic acid moiety conjugated to a free amine to form an amide bond (Figure 3). A weighted average of the number of norbornene adducts per coat protein monomer (with the weight determined by the relative abundance by volume of each compound cited in Table 1) suggests there are slightly more than four norbornene moieties per coat protein monomer, or approximately (4 norbornene adducts per monomer) × (420 coat protein monomers per nanocontainer) equates to 1700 norbornenes per nanocontainer. Although the P22 coat protein contains 19 lysine residues that could potentially couple with an activated 5-Norbornene-2-carboxylic ester, not all of these are surface-exposed, which would explain why not all 19 lysines are conjugated with norbornene.


Developing a Dissociative Nanocontainer for Peptide Drug Delivery.

Kelly P, Anand P, Uvaydov A, Chakravartula S, Sherpa C, Pires E, O'Neil A, Douglas T, Holford M - Int J Environ Res Public Health (2015)

Conjugation of Norbornene-COOH to the Capsid Surface. (A) Norbornene-COOH is activated with EDC and sulfo-NHS, leading to the formation of an amide bond with the ε-NH2 of surface-exposed lysine residues. (B) Mass spectrum of unmodified capsid coat protein (m/z = 47,192). (C) Mass spectrum of coat protein subunit conjugated with five norbornene subunits (m/z = 47,792).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4626985&req=5

ijerph-12-12543-f003: Conjugation of Norbornene-COOH to the Capsid Surface. (A) Norbornene-COOH is activated with EDC and sulfo-NHS, leading to the formation of an amide bond with the ε-NH2 of surface-exposed lysine residues. (B) Mass spectrum of unmodified capsid coat protein (m/z = 47,192). (C) Mass spectrum of coat protein subunit conjugated with five norbornene subunits (m/z = 47,792).
Mentions: The P22-GFP-norbornene conjugation reaction was monitored by MS analysis. Compounds with masses corresponding to that of the P22 coat protein (expected mass 46,596 Da, observed mass 47,192 Da) plus an integral number (between 2 and 6) of 120 Da adducts was detected (Table 1). The 120 Da mass corresponds to a 5-norbornene-2-carboxylic acid moiety conjugated to a free amine to form an amide bond (Figure 3). A weighted average of the number of norbornene adducts per coat protein monomer (with the weight determined by the relative abundance by volume of each compound cited in Table 1) suggests there are slightly more than four norbornene moieties per coat protein monomer, or approximately (4 norbornene adducts per monomer) × (420 coat protein monomers per nanocontainer) equates to 1700 norbornenes per nanocontainer. Although the P22 coat protein contains 19 lysine residues that could potentially couple with an activated 5-Norbornene-2-carboxylic ester, not all of these are surface-exposed, which would explain why not all 19 lysines are conjugated with norbornene.

Bottom Line: The potency, selectivity, and decreased side effects of bioactive peptides have propelled these agents to the forefront of pharmacological research.Recent efforts have demonstrated that P22 nanocontainers can effectively encapsulate analgesic peptides and translocate them across blood-brain-barrier (BBB) models.Our results demonstrate initial attempts to characterize the ROMP-triggered release of cargo peptides from P22 nanocontainers.

View Article: PubMed Central - PubMed

Affiliation: Hunter College and The Graduate Center, City University of New York, Belfer Research Building, 413 E. 69th Street, New York, NY 10021, USA. mkelly3@gc.cuny.edu.

ABSTRACT
The potency, selectivity, and decreased side effects of bioactive peptides have propelled these agents to the forefront of pharmacological research. Peptides are especially promising for the treatment of neurological disorders and pain. However, delivery of peptide therapeutics often requires invasive techniques, which is a major obstacle to their widespread application. We have developed a tailored peptide drug delivery system in which the viral capsid of P22 bacteriophage is modified to serve as a tunable nanocontainer for the packaging and controlled release of bioactive peptides. Recent efforts have demonstrated that P22 nanocontainers can effectively encapsulate analgesic peptides and translocate them across blood-brain-barrier (BBB) models. However, release of encapsulated peptides at their target site remains a challenge. Here a Ring Opening Metathesis Polymerization (ROMP) reaction is applied to trigger P22 nanocontainer disassembly under physiological conditions. Specifically, the ROMP substrate norbornene (5-Norbornene-2-carboxylic acid) is conjugated to the exterior of a loaded P22 nanocontainer and Grubbs II Catalyst is used to trigger the polymerization reaction leading to nanocontainer disassembly. Our results demonstrate initial attempts to characterize the ROMP-triggered release of cargo peptides from P22 nanocontainers. This work provides proof-of-concept for the construction of a triggerable peptide drug delivery system using viral nanocontainers.

No MeSH data available.


Related in: MedlinePlus