Biomaterials 24 (2003) 33333343

Review

Pectin-based systems for colon-specic drug delivery via oral route

LinShu Liua,*, Marshall L. Fishmana, Joseph Kostb, Kevin B. Hicksa
a

US Department of Agriculture, ARS, Eastern Regional Research Center, 600 East Mermaid, Lane, Wyndmoor, PA 19038, USA b Department of Chemical Engineering, Ben-Gurion University, Beer-Sheva 84105, Israel Received 25 November 2002; accepted 24 March 2003

Abstract Pectin-derived matrices are now being examined and tested for controlled drug delivery. Pectin is intact in the upper gastrointestinal tract and degraded by colonic microora. The composition of this microora remains relatively consistent across a diverse human population. Thus, pectin-derived drug carriers provide promising potential for colon-specic drug delivery. This paper reviews recent developments in pectin-derived formulations. Subjects reviewed include gelation of pectin, calcium cross-linked pectinate, composites of pectin and other polymers, technologies to fabricate pectin into useful drug delivery vehicles, and methods to evaluate release kinetics of incorporated drugs. This article discusses advantages, limitations, and possible future developments in pectin-based formulations with particular emphasis on the eld of colon-specic drug delivery. Published by Elsevier Science Ltd.
Keywords: Pectin; Controlled release; Oral administration; Colon-specic drug delivery

Contents 1. 2. 3. 4. 5. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3333 Physiology of human GI tract and strategies to deliver drugs to the colon site . . . . . . . . . . . . . . 3334 Gelation of pectin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3335 Calcium pectinate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3336 Combination of pectin and other polymers . . . . . . 5.1. Pectin and ethylcellulose . . . . . . . . . . . . . 5.2. Combination of pectin and hydroxypropylmethyl 5.3. Polyelectrolyte of pectin with polycations . . . . . . . . . . . . . . cellulose . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3338 3339 3339 3339

6.

Current status and challenges . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3341

References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3341

1. Introduction For most of civilized history, there was no clear difference in the way in which humans consumed food and medicine. To date, oral delivery is still the preferred route of drug administration, especially for chronic therapies where repeated administration is required. Oral administration offers patients less pain, greater convenience, higher likelihood of compliance, and

*Corresponding author. Tel.: +1-215-233-6486; fax: +1-215-2336406. E-mail address: lsliu@arserrc.gov (L. Liu). 0142-9612/03/$ - see front matter Published by Elsevier Science Ltd. doi:10.1016/S0142-9612(03)00213-8

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reduced risk of cross-infection and needlestick injuries [1,2]. Thus, formulations of oral drug delivery continue to dominate more than half of the drug delivery market share [3]. Despite these advantages, the oral route is not amenable to the administration of most protein and polypeptide drugs available today, due to their high susceptibility to digestive enzymes in the gastrointestinal (GI) tract, poor absorption, and their limited ability to transport across the intestinal epithelial barrier. As a result, new strategies of drug delivery have been developed to overcome obstacles encountered by oral delivery. Among these strategies, colon-specic delivery has been extensively studied for the last two decades. Colon, an area where protein drugs are free from the attack of numerous proteases, is thought to be an ideal location to direct drugs into the bloodstream and the immune system [46]. New formulations of different delivery vehicles from synthetic and natural polymers, which are either hydrophilic or hydrophobic, have been tested for these purposes. The challenge in the design of oral drug delivery vehicles which effectively carry drugs to the colon site is to meet a certain criteria. Firstly, they need to remain intact when traveling through the upper GI tract in order to protect the incorporated drugs from chemical and enzymatic degradation. Secondly, they should be able to release the incorporated drugs immediately upon reaching the colon segment of the lower GI tract. Furthermore, the released drugs need to be absorbed at an efcient rate in the GI tract in order to be therapeutically effective. Several carbohydrate polymers are able to satisfy these requirements to some extent, having demonstrated their potential as starting materials for the construction of oral drug delivery vehicles. Pectin, a structural plant polysaccharide, remains as aggregates of macromolecules in acid environments. At neutral solution pH pectin aggregates tend to dissociate and expand. Also, pectin is resistant to proteases and amylase which are active in the upper GI tract, whereas they are digested by a large number of microora of the colon [4,5,7]. Due to these properties it is highly possible that pectin could function as a delivery vehicle to escort protein and polypeptide drugs from the mouth to the colon. Although research on pectin-based oral drug deliver systems is proceeding at a rapid pace, most of the studies focus on the construction of vehicles by using pectin as a shield material. In this review article we have summarized advances in research on pectin-based, oral drug delivery systems and discussed potential future research strategies in this eld. We have emphasized matrix preparation and drug formulation. 2. Physiology of human GI tract and strategies to deliver drugs to the colon site The GI tract, also called the alimentary canal, is a muscular digestive tube that winds through the body

Fig. 1. Schematic summary of GI tract (modied from Ref. [8]).

(Fig. 1). The GI is a selective barrier between the environment and the systemic circulation, which functions to digest dietary food, to absorb nutrients, electrolytes and uid, and to prevent the absorption of potentially harmful substances [8]. To perform these complex functions the GI tract is differentiated into organs which possess unique characteristics along its length. The small intestine is the longest part of the GI tact. There most enzymatic digestion and virtually all absorption occurs. Most digestive enzymes that operate within the small intestine are secreted by the pancrease. Peristalsis propels chyme through the small intestine in about 36 h. The large intestine is the last major subdivision of the GI tract. The digested materials that reach the large intestine contain few nutrients, but the residues remain here for 1224 h. The pH in the stomach ranges from 1.5 to 3.5, and increases to 5.56.8 in the small intestine. The pH is 6.4 in the ascending colon, rises in the transverse colon, and approaches neutrality in the descending colon. The variety of pHs within the colon seems to be an important factor in determining the nature of the ora in the GI tract. The upper GI tract is sparsely populated with acid-resistant species, such as aerobic lactobacilli and streptococci. They are mainly Gram-positive, facultative microorganisms. The colon is the site of the most abundant microora in the GI tract. It consists predominantly anaerobic bacteria, such as Bacteroides, Bidobacteria, Eubacteria, and Enterobacteria, etc. [9]. These microora fulll their energy needs by fermenting various substances that are left undigested in the small intestine, such as di- trioligo- saccharides and polysaccharides. Based on transit time from the mouth to the colon, individual variations in pH and the types of bacteria and

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enzymes along the GI tract, two strategies to deliver drugs to the colon site via oral route have often been investigated. One strategy is to fabricate a delivery system that releases the preloaded drugs by a predetermined time schedule. Examples of this approach are Pulsincaps [10] and OROS-CT [11]. The limitation of this strategy is that the release of drugs is preprogrammed, based on the statistics of a normal situation. Variations in individuals are ignored. The second strategy is to develop a system that releases the preloaded drugs in response to the stimuli of local environments, such as pH and enzymes [12]. Coating the drug core with pH-sensitive polymers, such as methylacrylate-co-methylmethacrylate, has been suggested for this purpose. These polymers are insoluble in acidic environments and are dissolved in the media with a pH equal to or higher than the value of 6. Such a system is expected to preserve the drugs in the core of the formulation in the upper GI tract and release them when it enters the colon. However, similar to timedependent delivery systems, the pH of the GI tract is also subject to both internal and external individual variations, such as health, physical and emotional situations, diet, and whether with fed or fasted state. These methodologies are based on a few simple principles: (1) Construct a physical environment using the formulation itself to isolate the incorporated drugs from outside conditions when traveling the upper GI tract. (2) Turn to an active system to deliver the drugs at the colon site. The drug delivery efciency is estimated

by the difference between the drug released at the colon site and the initial dosage of the drug. The smaller the difference, the more effective the delivery system. For such applications, pectin-derived drug delivery systems of various types have been developed. These systems take advantage of the specic activity of colon inhibiting microora. The microora that ferment pectin only exist in the colon and are qualitatively consistent in various populations. Together with other properties of pectin, such as its pH-sensitivity and lm forming ability, it allows pectin-based drug delivery systems to be more reliable and reproducible for colon-specic drug delivery than other drug delivery systems.

3. Gelation of pectin Pectin is a cell wall structural carbohydrate present in all higher plants. Commercially available pectin is obtained from edible plants. Like many naturally occurring polysaccharides, pectin is heterogeneous with respect to chemical structure and molecular weight [13,14]. Primarily, pectin contains large amounts of poly(d -galacturonic acid) bonded via a 1; 4-glycosidic linkage (Fig. 2). Pectin also contains neutral sugars such as l-rhamnose, which are either inserted in or attached to the main chains. In pectin from sugar beet, feruloylester substituents attach on the side chains. In pectin from all sources, the carboxyl groups are partially in the methyl ester form. The degree of esterication

Fig. 2. Structure of pectin.

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(DE) varies depending on the source of the pectin and enzymatic activity in the process of ripening and maturation, and the conditions under which the isolation is conducted. Some of the carboxyl groups may be converted to carboxamide groups, when ammonia is used in the process of de-esterication, producing amidated pectin. The DE and the degree of amidation (DA) determine the content of carboxylic acid in pectin chains. The most attractive property of pectin for industrial applications is its gelling activity. Studies on the congelative property of pectin were initiated by Vauquelin in 1790 [15,16]. These led to applications of pectin in the food industry as gelling or thickening agent in the beginning, and then, as an excipient for pharmaceutical purposes. Factors which determine whether gellation can occur and which inuence gel characteristics are the types and concentrations of pectin, DE, DA, the modications of hydroxyl group, solution pH, temperature and the presence of cations. All these parameters are interdependent. In general, under similar conditions, degree of gelation, gel strength and gelling temperature increase proportionately. Also, each of these properties is proportional the molecular weight and inversely proportional to the DE of pectin [7,14]. Pectins of high DE, used alone, have been developed into hydrogels for drug delivery [5,17]. For high DE pectins, the formation of hydrophobic areas parallel to the helix axes can expand to such an extent as to dramatically reduce the solubility of pectin. High DE pectins also gel in the presence of large concentrations of sugar. Aqueous solutions of univalent salts of pectins exhibit low viscosity at physiological pH [18]. Univalent salts of low DE pectins are highly water soluble and gel only at extremely low solution pH or in the presence of divalent cations. The introduction of amide groups in low DE pectin reduces the hydrophilic property with an increasing tendency to form gels. Those pectins of low DE, while having high DA, have received signicant attention in the development of colon-specic drug delivery systems [4,5,1921]. Pectins can gel in various ways depending on the type and structure of the pectin molecule. Gelling can be induced by acid, by cross-linking with calcium ion, through the oxidization of the feruloylester substituents on sugar beet pectin [22,23], or by synergistic reaction with alginate [24,25]. When solution pH is lowered, i.e. titrated with acid, the ionization of carboxylate groups on pectins is repressed. Pectin molecules no longer repel each other over their entire chains, and thus can associate over a portion of their chains to form acidpectin gels. For acid-induced pectin gels, the hydration of pectin is reduced and there is less water incorporated into interchain entanglements. When solution pH is raised, the polycarboxylate groups are ionized, and able to react with calcium ions to form calcium-pectinate

Fig. 3. Cross-linking reaction for sugar-beet pectins (modied from Ref. [23]).

gels. The interaction of calcium ions and the carboxylate groups in pectin involves intermolecular chelate binding of the cation leading to the formation of macromolecular aggregates [26]. Pectins extracted from sugar beet contain feruloylester substituents on the side chains. Pectin of sugar beet can be cross-linked via the oxidization of those groups (Fig. 3). The reaction can be initiated by using peroxide or peroxidase or with persulfate; gels thus obtained have a remarkable water absorbing capacity ranging from 50 to 160 times the weight of the dry gel. When pectin is mixed with alginate in the presence of d -glucono-delta-lactone, gelling occurs by a synergistic reaction, which is referred to as the strong interchain contact between the protonated GG-blocks in alginate and the methoxy groups in pectin [24,25]. Of these four systems, acid-inducing and calcium cross-linking pectin gels have been used most often for the development of pectin-based drug delivery systems.

4. Calcium pectinate Major efforts have been focused on looking for pectin derivatives which are more water resistant, while still enzymatically degradable. Calcium salts of pectins have been studied for this purpose. The basic poly-d galacturonate structure of pectin shows close structural analogy to the poly-l -guluronate chain sequences of alginate, being almost the mirror image [27]. A similar behavior has been reported for the interaction of polygalacturonate and polyguluronate with calcium ions and it was attributed to the site-binding of calcium ions to dimers of chains of 21 helical symmetry [28]. It was suggested that the binding of calcium ions to polyguluronate or polygalacturonate molecules was through egg-box complexes with the polysaccharide chains in analogous 21 conformations. Calcium binding to pectin reduces the solution solubility and induces non-covalent associations of the carbohydrate chains.

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The study of the interaction of polygalacturonate gels with calcium ions as principal or sole counterions suggested changes in chain conformation and in the manner of chains packing, as determined by circular dichroism and stoichiometric characterization [28,29]. The calcium-induced molecular associations of carbohydrate chains, as expected, are stable in low solution pH and able to resist extensive hydration in vivo in the GI tract. In addition, the incorporation of calcium salts to pectin matrices should enhance the susceptibility of the polysaccharides to enzymes of the bacterial ora existing in the human large bowel, since many pectinases have been shown to be stimulated by or have an absolute requirement for calcium ions for their activity [30]. Although the presence of calcium ions also enhances the activity of Gram-positive facultative bacteria, those bacteria do not digest pectins [9,31]. The potential of calcium pectinate as a drug carrier for colon-specic delivery has been evaluated in vitro and in vivo by the use of drug markers, both water soluble and water insoluble, either small organic compounds or active protein drugs [17,3242]. In these studies, calcium pectinate was formulated into lms, gels, droplets, microsphere, and most often, in the form of compressed tablets. These studies demonstrated the tolerance of calcium pectinate in the upper GI tract and its enzymatic susceptibility in the lower GI tract. In one of the earliest studies [36], a calcium pectinatebased delivery system was prepared from highly deesteried pectin (DE 16%) and a drug marker of low solubility, indomethacin. The design of the experiment minimizes the drug dissolution and diffusion during their controlled delivery, and it enables one to verify that the kinetics of drug delivery occurred due to the degradation of the vehicle and not because of diffusion through the vehicle. Fine powders of calcium pectinate and indomethacin were well-mixed and compressed into tablets. The matrices were evaluated for degradation in vitro by incubating them in buffer solutions containing Pectinex 3XL, a typical pectinolytic enzyme mixture, and in the presence of the human colonic bacterium Bacteroides ovatus. It was found that the degradation of matrices was proportional to the enzyme concentration in solution. Increasing the concentration of the enzyme resulted in a higher release rate, hence, faster completion of the release process (Fig. 4). The amount of calcium ions added into the pectin gels also has an impact on the strength of the gels and the release of incorporated drugs [21,32,43,44]. The incorporation of calcium into pectin gel increased the magnitude of hysteresis loops in apparent viscosity [21] and shear stress of the gels [32]. When the amount of calcium exceeded a certain value, poor quality gels with complex ow patterns were observed (Fig. 5). This phenomenon occurs frequently with most polysacchar-

Fig. 4. Percentage cumulative amounts of indomethacin released from pectin tablets, with the pectinolytic enzymes mixture Pectinex 3XL: 120 FDU/ml (open circle), 12 FDU/ml (solid circle), and without enzymes (open triangle). Data shown are the mean of three sets of experiments 7SD (adopted from Ref. [36]).

Fig. 5. The effect of calcium concentration on the yield stress of pectin (adopted from Ref. [32]).

ide gels which are stabilized by a large number of secondary bonds, and has been referred to as shear degradation [43,44]. Fell and others [21,32] conrmed that there is an optimum level of calcium content in calcium pectinate gels for minimal drug release [21], which corresponds to the maximum of gel strength. The authors concluded that a carefully controlled amount of calcium is necessary to provide optimal protection for drugs in calcium pectinate matrices. The values of DE and DA are other factors having considerable effect on the formation of calcium induced pectin gels. Morris and co-workers have demonstrated that the extent of calcium ions binding decreased dramatically with increasing content of esteried residues randomly distributed along the polymer chain [28]. The formation of stable interchain junction zones requires participation of a minimum critical

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uninterrupted sequences length. For high DE or high DA pectin, the uninterrupted sequence of free carboxyl groups may be too short to extend the egg-box nests in which each calcium ion is xed by two carboxyl groups. A minimum DE of 50% has been suggested [45]. The presence of calcium ions with the concentrations up to the optimal amount decreases the release of drug from matrices with low DE pectin in the absence of enzymes, but increases the drug release from matrices with high DE pectin [21,45]. The sites for egg-box formation will be relatively unaffected by the smaller amide groups in comparison with methoxyl groups, but it can be further limited as the amide groups tend to be distributed in blocks with a random distribution of carboxyl and methylated groups [46]. The cross-linking of pectin with calcium ions inhibits the release of incorporated drug from the pectin tablets by suppressing the dissolution and swelling of pectin macromolecules. Nevertheless, it does not inhibit the diffusion of incorporated drugs from the surfaces of compressed tablets to the surrounding medium upon the drugs hydration. The release of drugs is affected by the mechanisms of drug diffusion and solvent activation. For tablets incorporated with water soluble drugs, the dissolution of drugs from the surfaces of the tablets is fast. As a result, water migrates into the matrix to replace dissolved drugs and create pores and channels. Fluid ingress promotes the extent and the rate of matrix swelling and creates a large surface area, which in turn, enhances the release of incorporated drugs. Therefore, as we can see in the following examples, the use of calcium pectinate matrices for colon-specic drug delivery seems highly restricted to low water soluble drugs. In one experiment, tablets consisting of calcium pectinate and insulin, a water soluble protein drug, showed no ability to delay the release of incorporated insulin [37]. The tablets started to release their insulin content right after the administration and prior to reaching the colon. For water soluble drugs such as insulin, an additional protective coating, which functions as a physical barrier to isolate the drug from the surrounding water, was suggested. Another development has been to apply calcium pectinate under high pressure as a layer to coat the surface of a core material. The process is known as the compression-coating method [17,37]. By this mode, additives and drugs are compressed into tablets and serve as core materials. The core tablet is located in the center of a layer of calcium pectinate powders, which is followed by an additional compression-coating procedure to form a thick and tough outer layer. The water penetration is retarded by the coating layer and the physical contact between drugs and uid can be inhibited till the drug carriers reach the colon and the outer layer is degraded by colon enzymes. The tablets were evaluated by testing the release of incorporated

insulin in vivo on dogs [37]. When the weight ratio of calcium pectinate between the core and the coating layer was adjusted to 50/50, a delayed release of insulin was reached. A sharp elevation in blood insulin concentration appeared 58 h after the oral administration. This period of time is presumed to be sufcient for tablets to travel from the stomach to the large bowel of dogs. In another study, drug release from compression-coated tablets of pectins was evaluated in vivo by gamma scintigraphic method on human volunteers and in vitro under the conditions mimicking closely the mouth to colon transit [17]. In determination of the minimum coat thickness required for protection, the coated tablets, varying in the thickness of the coating layer and the sources of pectins, were prepared [17]. It was found that the coat integrity decreased with decreasing thickness of coating layer; at the lower end, the coat dissolves to such an extent that it can no longer protect the enclosed core materials. The coat thickness of 0.92.0 mm for pectin USP (a commercially available pectin with the DE of 70%) is essential to satisfactorily prevent the incorporated drug from early release. The threshold is raised further for pectins with lower DE. The drawback of compression-coating technology comes from the technical difculty to center the core tablet within solid powders and to compress it to a formulation with even thickness and density in every direction. Regions with a thin or slack coat swell more easily and lead to the collapse of the coat. As gastric emptying is the most variable part of mouth-to-colon transit, compression-coating tablets may suffer from the alteration of the GI transit time, presenting another limitation in clinical applications. In addition, these coats are necessarily bulky and not suitable for all dosage forms.

5. Combination of pectin and other polymers Pectin-based colon-specic drug delivery vehicles have been developed from pectin, either plain or chemically modied, in combination with other polymers, either naturally occurring or synthetic. The composites take the advantages of their parent polymers and/or create useful new properties. For example, the composite of pectin and ethylcellulose combines the enzymatic susceptibility of pectin and the protective properties of ethylcellulose. Composites of pectin and chitosan or pectin and the polymethylmethacrylate derivatives carrying various amounts of NH+ 4 groups (Eudragits RL, RS) are enzymatically degradable and more water resistant, although the three polymers in origin are water soluble. These composite matrices are often used for tablet coating in the forms of aqueous lm-coating dispersion or blended powder, or for drug loading in the form of coacervate. In the later case, a

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drug can be loaded into the composite matrix either by the diffusion method or mixed with one of the components prior to coacervation. And, sometimes the composites are used as excipients in tablet formation. 5.1. Pectin and ethylcellulose An aqueous dispersion of ethylcellulose and pectin is prepared by blending the two polymers in double distilled water. It has been tested as a lm-coating composite by dispersing it on the surface of tablets to test its stability under conditions which mimick the GI tract and for its ability to deliver drugs in the presence of pectinolytic enzymes [4750]. Ethylcellulose functions in masking the inherent solubility of pectin in the composite. The combinational lm-coating composite is relatively impermeable to drugs, providing protection for drugs in the core tablets from the early release in the upper GI tract, while it begins to release drugs in the presence of bacterial ora of the colon. Increasing the proportion of ethylcellulose or increasing the coat thickness reduced the drug release to media as tested at pH 1 and pH 7.4. The addition of pectinolytic enzymes to media at pH 6 increased the release of drugs [47]. To get suitable mechanical properties, the optimal pectin content of less than 20% in the composite has been recommended [5154]. Coating techniques also affect the release kinetics and the stability of the dosage form [5557]. Careful control of coating temperature and the viscosity of the dispersion solution is essential. When the proportion of pectin is high in the ethylcellulose/pectin composite, the coating lm only slightly retards the release of incorporated drugs [47,58]. It was found that the molecular coalescence of pectin and ethylcellulose was observed in aqueous phase, but not in dry lms. Instead, dry lms prepared from pectin and ethylcellulose were characterized by the presence of pores. The porosity in dry lm-coatings was governed mainly by the composition of the aqueous composite dispersion, from which the lm was formed. For the lm-coating composite of Aquacoats ECD 30 (30% by weight aqueous dispersion of ethylcellulose polymer) containing high methoxyl pectin, up to 80% of pectin leached from the lms after incubation with 0.05 m acetatephosphate buffer (pH 4.5, 37 C) for 4 h in the absence of pectinolytic enzyme. These results suggested the need to further improve their water resistance in order to be used for colon-specic drug delivery. When calcium pectinate was included in the lms, a suppressed release was achieved, because the alcium pectinate is less hydrophilic and bigger in molecular size than uncrosslinked pectin [58]. A similar result was obtained by the study on the system composed of pectin and Eudragits NE 30D, an insoluble and exible polymer of crosslinked methyl methacrylate. The release of pectin from the composite in the absence of pectinolytic enzymes

was reduced by the addition of calcium. The release delay was found dependent on the weight ratio of calcium/pectin and appeared to be optimal at a value of 30 mg Ca2+/g of pectin. The release of pectin from the composite in the presence of enzymes was enhanced and dependent on pectin content. Interestingly, a followup study [59,60] found that the release of theophylline, a hydrophilic drug, from the composite of pectin, calcium pectinate, and Aquacoats was suppressed in the presence of pectinolytic enzymes, in comparison to that measured in the absence of the enzymes. This was explained by the hypothesis that pectin channels were formed and destroyed. In the absence of pectinolytic enzymes, pectin is retained in the coating layers, rapidly absorbs surrounding water, and swells to form water channels. These facilitate the dissolution of incorporated theophylline. In the presence of pectinolytic enzymes, presumably pectin is degraded and leached quickly, creating pores in the coating layer. However, re-structure in the lm coating follows and causes the pores to plug, which then slows down drug release. Possibly, this phenomenon is governed by the elasticity and softness of the insoluble polymer in the composite lm. 5.2. Combination of pectin and hydroxypropylmethyl cellulose Another cellulose derivative used in combination with pectin for tablet coating is hydroxypropylmethyl cellulose (HPMC) [6164]. Tablets from the mixtures of methoxyl pectin and HPMC have been prepared and tested [61,62]. The study suggests that a thick layer of high methoxyl pectin coating may considerably delay the release of tracers from tablets to dissolution media. Increasing the rate of composite layer breakdown by the presence of pectinolytic enzymes makes the pectin/ HPMC composite a candidate for colon-specic drug delivery. In another experiment, pectin-HPMC compression coated core tablets were tested for the controlled delivery of 5-aminosalicylic acid under the conditions mimicking the GI tract. It was found that the optimum HPMC concentration was 20% and such system would protect the cores up to 6 h that corresponded to 2535% erosion and after that under the inuence of pectinase the system would degrade faster and deliver the drugs to the colon [63]. 5.3. Polyelectrolyte of pectin with polycations Pectin is an anionic biopolymer with the ultimate propensity to bind with oppositely charged surfaces and to associate to form complexes with oppositely charged polymers. It forms coacervates with polycations, such as chitosan, poly(lysine), and polyacrylate derivatives carrying quaternary ammonium groups, Eudragits RL

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and Eudragits RS. The coacervation can be monitored by measuring the change in solution viscosity, while titrating one polymer solution into the other. When Eudragits RL 30D was titrated with pectin solution, the viscosity of the mixture solution increased initially, and decreased after exceeding a specic ratio of the two polymers. This ratio was about 5% by weight for high methoxyl pectin. Five percent is sufcient pectin to neutralize the positive charges in Eudragits RL 30D and to form a pectinEudragits RL complex. The capability of pectinEudragits RL complex to prevent the release of incorporated drugs has been demonstrated. Nevertheless, a lm-coating composite made of 5% high methoxyl pectin and Eudragits RL is impermeable to pectinolytic enzymes, resulting in an extremely slow leaching of pectin and subsequent slower drug release. Studies showed that the optimal weight ratio of high methoxyl pectin in the composite is 10 15%. This weight percent of pectin provides the best solute release kinetics with respect to colon-specic drug delivery. The attack of pectinolytic enzymes initiates pectin degradation and its release. Consequently, the pectinEudragits RL complex dissociates. The restored Eudragits RL recovers its original physical and physiological properties, and it is eliminated from the GI tract. The excessive pectin in the pectinEudragits RL composite will lead to a faster release of pectin in the absence of enzyme and a slower release of pectin in the presence of enzyme, similar to the case of pectin Aquacoats ECD 30 aqueous dispersion system, described above, where pores generated by pectin release were relled [59,60]. Chitosan is a polycationic derivative of the polysaccharide, chitin. It has demonstrated favorable biological properties, such as non-toxicity, biodegradability, tissue-adhesive activity, and anti-inammatory response. Chitosan and alginate form coacervates, which have been used in drug delivery, tissue repair, and other biomedical applications [65]. The combination of pectin with chitosan has also been prepared and tested in vitro as a potential drug carrier [66,67]. The mixture of pectin and chitosan (10:1, w/w), in the form of blended powder, was compressed-coated to pellets. The in vitro evaluation of the release of incorporated chloropromazine HCl showed promise for the use in colon-specic drug delivery [66]. In another experiment, compressed tablets containing indomethacin or paracetamol were coated with pectin or a pectin/chitosan blend at the weight ratio of 5:1 by the compression-coating method [67]. As expected, the release of the water-soluble paracetamol from tablets coated with pectin alone was faster than the release from those coated with pectin/ chitosan mixtures, and was much faster than the release of poorly water soluble indomethacin. Furthermore, a signicant release of paracetamol was detected in vitro at pH 6, even in the absence of enzymes. There is,

therefore, considerable work to do, especially in the regulation of the porosity in the coating layer of pectin/ chitosan mixtures, to develop these matrices from experimental to practical delivery systems for colonspecic drug delivery. The inclusion of hydroxypropylmethyl cellulose into pectin, chitosan mixtures as a compression-coating substance has been studied for the potential application in colon-specic drug delivery [68,69]. The compressed lm is water insoluble, but degradable in the presence of pectinolytic enzymes. The degree of lm swelling varied with the concentrations of pectin and chitosan. The in vitro experiments suggested that swelling and hydration of the composite lm is necessary before enzymatic breakdown occurs; and also suggested a system with a release prole which is tailorable by altering the ratio of the compositions to meet the particular requirements of individual drugs. Gamma scintigraphy for GI transit found that radiolabelled (99 m Tc) tablets, which were coated with a pectin:chitosan:hydroxypropylmethyl cellulose composite lm (P/C/H, 3:1:1), were able to pass through the stomach and small intestine. Disintegration of the tablets commenced upon entering the colon, where a signicant decrease in the percentage activity remaining in the tablets was detected, due to the degradation of the coat by colonic bacteria. On the other hand, it was reported that the inclusion of chitosan into the pectin/HPMC lm rendered it more stable at all physisological pH values [64]. HPMC initially increased the permeability of the lms and subsequently reduced it at higher concentrations. The minimum permeability was obtained at pH 3 and at an HPMC level of 5% where the potential for polyelectrolyte complex formation between pectin and chitosan was supposed to exist. The permeability of the lm increased when they were exposed to pectinolytic enzymes (Table 1). It is often observed that the delivery of single-unit drug formulations exhibits delays at the ileocecal junction, which leads to drug loss prior to reaching the lower GI tract. In an attempt to overcome this drawback, a multiparticulate system of amidated pectin
Table 1 Effect of pectinolytic enzymes (2 ml/l) on the permeation of paracetamol through pectin/chitosan/HPMC lms into pH 6.4 Sorensens phosphate buffer (mean7SD, n 3) % HPMC Permeability coefcient 106 (mg/min/cm) No enzyme 0 5 10 15 20 5.1370.38 6.7270.14 8.1170.21 6.1370.40 6.1970.22 With enzyme 9.4170.21 9.7270.19 10.7270.64 10.2470.20 9.3870.47

Modied from Ref. [64].

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beads was prepared by injecting amidated pectin solution into a calcium solution containing chitosan. Calcium ions are allowed to react with amidated pectin to form hydrogel beads. The polycationic chitosan is entrapped in these beads and coupled to the pectin chains through electrostatic interaction to form a polyelectrolyte complex lm on the wall of pores inside the gel beads and around the beads [19,20]. In comparison with the dry compression-coating method used to prepare tablets [68,69], this method is simpler, easier to operate, and is able to provide a complete and even coating layer. Since the interaction between the amidated pectin and chitosan is favored at low pH, coated beads function better as a protective vehicle in a lower pH environment, such as in the upper GI tract. Experimental results showed that the pectinolytic enzymes can still attack the beads despite cross-linking with chitosan. Drug release kinetics depends on media pH and drug loading. However, no in vivo studies were reported.

6. Current status and challenges Considerable efforts have been devoted to the development of pectin-based drug delivery systems, which can effectively and specically deliver therapeutics at the colon-site via the oral route. However, existing evidence for the efcacy of these drug carriers is lack of consistency, partly due to the large diversity of pectin sources and partly coming from the variety of methodologies adopted for different experiments. A systematic comparison study among selective pectin structures using a reliable in vitro and in vivo model is required to overcome the complexities associated with the choice of polymer, dosage formulation, and intestinal absorption barrier. It will be crucial for the optimization of pectin-based drug delivery systems. Many approaches have been evaluated in the aim to create an effective pectin-based drug delivery system. These include the attempt to reduce matrix swelling by calcium cross-linking, retard water penetration by thickening the coating layer, and inhibit the activity of protease or isolate them from the incorporated protein drugs by using a physical barrier. Among these approaches, the combination of pectin with waterinsoluble polymers as lm-coating materials appears especially promising. Apparently, the techniques applied for the preparation of dosage formulation play a role in the control of the drug release prole. The method of spread coating by the use of aqueous dispersion of pectinpolymer composite appears to be more effective than that of compression-coating using a blend of pectinpolymer composite. However, the most successful examples by far are related to water insoluble drugs, such as anti-inammatory drugs for lower bowel

diseases. Relatively little progress on the delivery of polypeptide or protein drugs has been reported. The search for pectin derivatives, which are more water resistant and still enzymatically labile, clearly is a continuing effort. Nevertheless, many potentially useful pectin derivatives can be synthesized via traditional chemical methods or enzymatic methods. Further studies on the chemical and enzymatic modication of commercially available pectins, aimed at tailoring their physical and chemical properties, functionality and bioavailability for different therapeutics are essential in developing more delicate and effective delivery systems for the release of protein and polypeptide drugs to the colon site. Besides pectin, there are a number of other polysaccharides commercially available that have been used or tested in pharmaceutical applications, such as dextran, guar gum, and chitosan. They are stable, nontoxic, and enzymatically degradable in the colon. A sideby-side comparison of pectin with other polysaccharides for various types of drugs remains to be conducted. This will be critical for exploring a unique market for pectinbased drug delivery systems. Finally, drug release is not the end point of oral delivery. The bioavailability of protein drugs delivered at the colon site should be addressed [7072]. The inclusion of drug absorption enhancers into the drug delivery systems is likely necessary to achieve therapeutic efciency. Studies on drug absorption by the intestinal system have focused on drug transporters that mediate drug inux and efux [7176] and ingredients which can enhance drug absorption [7782]. The colon segment is designed mainly to expel metabolism products rather than to absorb nutrients. Therefore, more research focused on the specicity of drug uptake at the colon site is necessary. For example, one such study is the inuence of P-glycoprotein mediated secretion and metabolism products on the absorption of protein drugs. These kinds of studies will be signicant in both academic study and practical applications.

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