Proteins and peptides: The need to improve them as promising therapeutics for ulcerative colitis

Figures & data

Figure 1. Schematic diagram showing the process of inflammation and colitis.

Table I. Types of ulcerative colitis (Conrad et al. 2014, Travis et al. 2008, Bokemeyer et al. 2012, Bitton 2001, Limbergen et al. 2008).

		Prevalence onset
Types of UC	Parts affected	Adult	Childhood
Proctitis	Rectum	40–50%	1.4%
Left-side colitis [Most prevalent subtype]	Descending colon or Splenic flexure	30–40%	16%
Extensive colitis/Pancolitis	Hepatic flexure or entire colon	25–30%	82%

Figure 2. Factors involved in pathogenesis of ulcerative colitis.

Figure 3. Integrated process of absorption mechanism and hindrance factor of peptide and protein in intestinal epithelial cell. [A, B, C and D stand for receptor-mediated, paracellular, transcellular and carrier-mediated transport of protein drug respectively, whereas E, F, G and H stand for luminal enzyme, mucin barrier, brush border enzyme and intracellular enzyme respectively].

Table II. Various strategies for biotherapeutics of protein and peptide.

Strategy	Method	Outcomes	References
I. Chemical modification
Modification of peptide at N- and C-terminal	Gly-Gly in methionyl enkephalin is replaced by D-Ala. Replacement of L-amino acid by D-form at C-terminal inhibits the degradation by blocking carboxypeptidases	Half-life of modified peptide is increased significantly compared to that of the parent which is due to low cleavage power of aminopeptidases on Tyr-D-Ala amide bond	(Pauletti et al. 1996)
Cyclic analog of linear one	In this method, an amide bond is formed in between various ways such in N- and C- terminal, in side chain or N- or C-termini or in two side chains	Increased half-life is noticed such as in case of α-melanotropin (α-MSH), it shows 100-fold greater half-life than its linear parent	(Al-Obeidi et al. 1989)
II. Polymeric carrier
Ionically (Fe^3+) crosslinked alginate carboxymethyl cellulose (AC) beads	AC solution is dropped into ferric chloride which contains protein, so that beads containing therapeutic protein are formed	Bioactivity and fragility of protein is preserved in such type of control release beads	(Kim et al. 2012)
New graft copolymers containing NPs using salmon calcitonin as peptide	It has hydrophilic branches and hydrophobic backbone prepared by the dispersion copolymerization method of hydrophilic polyvinyl macromonomers using polar solvent with styrene	Absorption of peptide is enhanced because of bioadhesion of nanoparticle in GI mucosa	(Sakuma et al. 1997)
N-trimethyl chitosan chloride (TMC) for peroral delivery of peptides like buserelin and octreotide	Chitosan undergoes reductive methylation to form TMC which is a partially quaternized derivative of chitosan. TMC, being an excellent absorption enhancer shows its action in neutral and basic environments	In vitro studies show that the transepithelial electrical resistance (TEER) is reduced, which opens the tight junction between the epithelial cells. This results in enhancement of paracellular absorption	(van der Merwe et al. 2004)
III. Vesicular Carrier
Archeosomes	Sulfolobus acidocaldarius extracted polar lipid fraction E (PLFE) is used for preparation which is suitable for oral insulin delivery. Fluorescent labelling of peptide is done for the absorption study	Blood glucose level is decreased significantly as compared to conventional liposome, which proves that it could be a potential carrier	(Li et al. 2010)
Nanosphere containing calcitonin-loaded PA (polyacrylamide)	Prepared according to Birrenbach and Speiser method in which inverse w/o microemulsion undergoes polymerization. Particles are obtained by washing with ethanol several times and then finally freeze dried	Nanosphere of size below 50 nm is formed with good peptide release profile, having good antiprotease activity	(Allemann et al. 1998)
Coated calcium alginate gel bead-entrapped liposome for bee-venom peptide	Solvent evaporation method is used for liposome preparation. Liposome so formed is suspended in sodium alginate solution, and then added dropwise in calcium chloride solution to form beads	In vivo study is done by γ-scintigraphy which reveals that liposomes so formed have protection of peptides from the physiological environment in the GIT	(Xing et al. 2003)
IV. Other approaches
Mini tablet and granule of Desmopressin (1-(3-mercaptopropionic acid)-8-D-arginine) vasopressin monoacetate (DDAVP)	Tablets and granules are prepared with the aid of N-trimethyl chitosan chloride (TMC) as absorption enhancer. Granules are prepared by wet granulation method	Paracellular transport of peptide increases by the action of TMC on epithelial tight junction, which is opened	(Paulsen et al. 1993)

Table III. Proteins and peptides potentially used in colitis.

Proteins/Peptides	Dosage forms	Dose	Route of administration	Comments	References
Vasoactive intestinal peptide (VIP)	Solution	20 μM	Ex vivo (Male ddY mice colon)	Dysfunction of VIP receptors in muscles and neuronal pathways are involved in colitis	(Kato et al. 2012)
Protamex hydrolysate from C. gigas	Solution	50 mg/kg	Oral (mice) and RAW264.7 cell cultures are also used	It has anti-inflammatory effect and reduces production of potent pro-inflammatory mucosal cytokines	(Hwang et al. 2012)
Trefoil factor family 1 (TFF1) peptide	Solution	100 μg/kg	Subcutaneous (Male, Sprague Dawley rats)	Systemic administration of TFF1 can reduce DSS-induced colitis and shows synergistic effect with co-administered epidermal growth factor (EGF)	(FitzGerald et al. 2004)
Polyethylene glycol (PEG)-porcine glucagon-like peptide-2 (pGLP-2)	Solution	30 μg	Intraperitoneally (male BALB/c mice)	Resistant to degradation, thus increased biological potency and reduced severity of colonic injury	(Qi et al. 2014)
Tumor necrosis factor alpha (TNF-α) and TNF receptor-binding cyclic peptide (TRBCP)	Solution	2.5 mg/kg	Intragastrically (male Wistar rats)	Combination treatment effectively reduces the inflammation of colitis by neutralization of TNF-α and blockage of TNFR1	(Yin et al. 2011)
P-317 peptide [Dmt-c(D-Lys-Phe D-Pro-Asp)NH2], a cyclic analog of morphiceptin	Solution	0.1 mg/kg, 1 mg/kg	Intraperitoneally, oral (Male BALB/c mice)	Shows anti-inflammatory effect in an opioid receptor-dependent manner, which is due to decreased expression of mRNA for proinflammatory cytokines	(Sobczak et al. 2014)
Egg white peptides (EWP)	Solution	150 mg/kg	Intra-gastric infusion through catheter (Yorkshire piglets)	Significantly decreases the expression of pro-inflammatory cytokines TNF-α, IL-6, IL-1β, IFN-γ, IL-8 and IL-17	(Lee et al. 2009)
Nuclear factor κB (NF-κB) essential modulator or NEMO Binding Domain Peptide1	Solution	10 mg/kg	Intraperitoneally (IL-10-deficient (IL-10–/–) mouse)	Histological improvement of chronic colitis with, decreased levels of inflammatory cytokines	(Dave et al. 2007)
L-cysteine	Suspension	0.144 g/kg	Intragastrically, through catheter (Yorkshire piglets)	Improvement of colon histology by reducing local chemokine expression and neutrophil influx. TNF-α, IL-6, IL-12p40, IL-1β expression is also reduced	(Kim et al. 2009)
PepT1-transportable soy tripeptide VPY	Solution	10 and 100 mg/kg	Oral (female BALB/c mice) Caco-2 cell and HT-29 cells are also used	Reduction in MPO activity and gene expression of the pro-inflammatory cytokines TNF-α, IL-6, IL-1β, IFN-γ and IL-17 in the colon	(Nolan et al. 2012)
L-tryptophan	Solution	110 mg/kg	Oral (Yorkshire piglets)	Weight gain and reduction in gut permeability. Decreased expression of IL-6, TNF-α, IFN-γ, IL-12p40, IL-1β and IL-17 and increase in expression of apoptosis initiators caspase-8 and Bax	(Kim et al. 2010)

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