CHAPTER1: INTRODUCTION 2022

Chapter 1
INTRODUCTION

1.1. Inflammatory bowel disease (IBD)

Inflammatory bowel disease refers to two diseases that are related but not the
same: ulcerative colitis and Crohn's disease. These diseases cause chronic intestine
inflammation, resulting in a different type of symptoms. Inflammation can also cause
organs other than the intestines to become involved. IBD is a lifetime disorder that
varies between durations of active disease and periods of disease control. IBD is
often confused with irritable bowel syndrome, but they are not the same thing. In
the United States of America, there are over 1 million people with IBD, with new
cases being diagnosed at a rate of 10 per 100,000 people. These diseases account for
700,000 physician visits and 100,000 hospitalizations in the United States each year.
Surgery can cure ulcerative colitis, but it cannot cure Crohn's disease [Baumgart DC
et al., 2007].

In developing countries, IBD has become more common reversing the

previously held belief that these were Western diseases. Inflammatory bowel
disease (IBD) is on the rise in India, which now ranks first among Southeast Asian
countries. In 2010, India's estimated IBD population was 1.4 million, making it the
world's second-largest after the United States (with 1.64 million). As a result, while
disease prevalence in India is lower than in the West, India has one of the world's
largest IBD populations, with more than 120 million people. The general public
needs to know what the disease is all about and when to seek help in order to avoid
the serious medical problems that this disease can cause.
[https://www.timesnownews.com/health/article/incidence-and-prevalence-
of-inflammatory-bowel-disease-in-india].

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1.1.1. Crohn’s disease

Crohn's infection can distress whole gastrointestinal tract, from the mouth to
anus. It usually affects the ileum, which links the small intestine's end to the colon's
beginning. Crohn's disease can manifest itself in "patches," affecting some areas of
the gastrointestinal tract while leaving others completely unaffected. The
inflammation in Crohn's disease can spread throughout the entire thickness of the
affected bowel wall [Ardizone S et al., 2003].

1.1.2. Ulcerative colitis

The colon and the rectum are the only parts of the body affected by ulcerative
colitis. Only the innermost layer of the intestine's lining is affected by inflammation.
It usually starts in the rectum and lower colon, but it can strike at any point in the
colon [Ardizone S et al., 2003].

1.1.3. Causes of IBD

IBD's actual cause is unknown, but it is thought to be related to protective

immune cells found in the intestine lining. This immune system normally activates
and deactivates in order to combat harmful substances such as bacteria and viruses
that pass through the intestines. In IBD, it appears that the immune system is
activated by an initial trigger, such as an infection or something ingested from the
diet or the surrounding environment. The difference in those who develop IBD is
that the immune system does not shut down once the initial trigger has been
removed. This causes uncontrolled inflammation and the destruction of normal
intestinal cells. The precise contributions of such factors are unknown and difficult
to define [Klotz U et al., 2005].

1.1.4. Symptoms of IBD [Collnot E et al., 2006]

• Anaemia
• Fatigue
• Weight loss
• Loss of appetite
• Rectal bleeding

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Table 1.1. Key features of ulcerative colitis and crohn’s disease [Rebecca T et
al., 2007]
Parameters Ulcerative colitis Crohn’s disease

Area affected Colon only Mouth to anus

Distribution Continuous Skip lesions
Histological Mucosa/sub mucosa, crypt Transmural granulomas,
features abscesses; superficial aphthoid ulcers
ulcers
Rectal bleeding Diarrhoea Abdominal pain, diarrhoea,
Typical symptoms often bloody, abdominal fever, weight -loss, fistulas
pain, weight loss

1.1.5. Diagnosis of ulcerative colitis and colorectal cancer

The key to controlling and preventing ulcerative colitis and Crohn's disease is early
detection. As represent in table 1.2, diagnostic tools such as patient history and
visual observation aid in detecting disease conditions. Clinical examination of body
fluid and blood components is advised in mild to moderate inflammatory conditions.
A microbiological test of the stool is sometimes required to determine the strain of
bacteria responsible for the inflammation. Microscopic imaging is also used to
ascertain the degree of colitis.

1.1.6. Changes in the GI tract's physiology during active IBD

When considering oral delivery strategies for IBD, the change in the
physiological condition of the GI tract caused by chronic inflammation is an often
overlooked confounder. In Mucosal inflammation in IBD leads to pathophysiological
changes like (i) a disrupted intestinal barrier caused by alterations in mucosal
surface, crypt distortions, and ulcers, (ii) enhanced mucus production, and (iii)
immune cell infiltration. Neutrophils, macrophages, lymphocytes, and dendritic cells
are examples of these cells [Li AC et al., 2003 & Antoni L et al., 2014]. Patients
with severe mucosal inflammation during IBD relapse may experience altered GI
motility and diarrhea, affecting intestinal volume, pH, and mucosal integrity. The

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inflammatory effect on the mucosa, combined with severe diarrhoea, will disrupt the
resident microbiome, altering metabolism of microbes in the GI tract. As a result,
active inflammation causes physiological changes in the GI tract (Figure 1.1), which
can impair the efficacy of traditional approaches to colon targeted drug delivery.

Table 1.2. Modes of clinical diagnosis [Taghipour et al., 2016]

S.No Method Example

Age of onset, Type of stool, lifestyle,

1 History previous bowel infection and medication,
smoking.
Pulse, BP, temperature, weight, height, BMI,
2 Visual observation abdominal pain, rectal inspection,
examination of eyes, skin, joints and muscles
Electrolytes, urea, creatinine, complete
blood count, ESR, liver enzymes, bilirubin,
3 Clinical investigation
alkaline phosphatase, transferrin, ferritin,
vitamin B12, B folic acid, CRP, urinalysis

4 Faecal investigation Stool testing for bacterial organisms

5 Microscopic Endoscopy and colonoscopy

observation
6 Imaging Ultrasound and MRI

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Tight junction

Mucus layer

Tight junction
disassembly

Epithelial
leakage

Damage
mucus layer

Figure 1.1: Representative diagram of healthy and inflamed colon. Tight

junctions and the mucus layer are fully intact in healthy tissue, whereas in the
inflamed colon this arrangement of tight junctions is disrupted giving rise to
epithelial leakage and loss of the mucus layer.

1.1.6.1. Transit time and microbial considerations

Changes in GI physiology in disease states are frequently dynamic and inter-
related, making it difficult to investigate in isolation. In both CD and UC patients, the
time it takes for food to reach the cecum has been shown to be delayed when
compared to healthy controls [Rana SV et al., 2013]. However, IBD patients with
dysbiotic conditions – small intestinal bacterial overgrowth – have significantly
faster OCTT. Following dietary manipulation of the gut microflora, these results
were confirmed experimentally in humanised mice [Kashyap PC et al., 2013].
Changes in the composition of the microbiome (dysbiosis) are common in GI
disease, and can be caused by changes in physiology, an inflammatory state, or
treatment regimens [Sartor RB et al., 2008]. While it is widely recognized that the
bacterial load in IBD is relatively stable, the diversity of the microbiota is decreasing
as the number of main species like Bacteroides, Eubacteria, and Peptostreptococcus

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CHAPTER1: INTRODUCTION 2022

increases, to the detriment of other populations [Linskens RK et al., 2001]. It is

unknown what causes the initial dysbiosis or whether dysbiosis is a symptom of
disease. However, there is some evidence that physiological factors such as
dysmotility and increased luminal fluid (diarrhoea) may be involved. Extensive
secretion of water into the bowel, resulting in shorter colonic transit times, has been
shown in animal models to change the colonic microbiota [Keely S et al., 2012 &
Musch MW et al., 2013]. As a result, not only can the microbiome affect intestinal
transit times, but transit times themselves may change the microbiome.
In contrast to OCTT, colonic transit in IBD patients is significantly faster, most
likely due to the disease's diarrhea [Hebden JM et al., 2002]. Transit times in UC
patients can be twice as fast as in healthy people, making it difficult to target specific
of action with traditional formulations. In studies using traditional delayed release
formulations, asymmetric biodistribution of drug in the colon was observed, with
maximum drug retention in the proximal colon and substantially lower drug
concentrations in the distal colon [Podolsky DK et al., 2000]. As a result, In IBD,
transit time may not be a successful way of site-specific delivery of drug.

1.1.6.2. Changes in colonic pH

Although there is little indication to suggest that IBD patients have significant
changes in small intestinal pH [Barkas F et al., 2013 & Lucas ML et al., 1978] in
both UC and CD patients, the pH of the colon is significantly lower. Intestinal pH is
influenced by microbial fermentation processes, bile acid metabolism of fatty acids,
bicarbonate and lactate secretions, and intestinal volume and transit times [Nugent
SG et al., 2001]. Variation in colonic luminal pH is not unexpected when all of these
factors are disrupted during active IBD. While the normal pH is 6.8 in the proximal
colon and rises to 7.2 in the distal colon, in active UC patients, it can range from 5.5
to as low as 2.3 [Fallingborg J et al., 1993]. Similarly, regardless of disease activity,
observed colon pH for CD patients is around 5.3 [Sasaki Y et al., 1997]. These pH
shifts are likely to alter the colonic microbiota composition and, As a result, colonic
transit times potentially affecting drug release from formulations requiring bacterial

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CHAPTER1: INTRODUCTION 2022

fermentation or enzymatic activity. Similarly, changes in pH can affect compound

release from pH-dependent release coatings.

1.1.6.3. Intestinal volume

In disease, the constituents of intestinal biomass changes as a result of
microbial metabolism, gastrointestinal transit time, and luminal pH. Increased fluid
secretion and decreased reabsorption, in particular, can dilute the digestive
enzymes that regulate intestinal transit, allowing nutrients to be absorbed [Van
Citters et al., 2006]. This, in turn, may have an impact on the intestinal microbiome,
altering carbohydrate and polysaccharide digestion and contributing to changes in
intestinal transit times [Yang L et al., 2008]. These variations in the volumes of
intestinal fluid may have an effect on how conventional dosage forms are processed
in the gastrointestinal tract and successive local delivery of drug to the colon.

1.1.6.4. Mucosal integrity

The epithelial barrier selectively regulates transport from the lumen to the
underlying tissue compartments [Laukoetter MG et al., 2008], limiting small
molecule transport while virtually eliminating macromolecule transit. Tight
junctions, which are apical transmembrane protein complexes, determine this
selectivity. These multi-protein complexes have direct interactions with actomycin
rings in the underlying epithelium, influencing physiological and pathophysiological
stimuli such as ion transport, luminal glucose transport, water secretion, and
cytokine and leukocyte transport [Cunningham KE et al., 2012]. While these
characteristics make TJ an appealing pharmacological target for improving drug
absorption [Wang X et al., 2010], abnormal TJ complex formation is linked to a loss
of epithelial integrity in inflammatory bowel diseases like IBD [Laukoetter MG et
al., 2008]. Active inflammation not only affects intestinal mucosal integrity, but also
mucosal metabolism as the tissue tries to halt further damage and repair [Goggins
BJ et al., 2013]. For example, to compensate for the loss of intestinal epithelial
integrity caused by inflammation and tissue ischemia, several endogenous defensive
pathways are triggered subtly modifying the composition of the mucosa. The

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oxygen-sensing transcription factor hypoxia-inducible factor (HIF) mediates

increased expression of intestinal mucins and trefoil factors to augment intestinal
barrier function and possibly compensate for the loss of mucous-gel integrity with
fluid secretion [Goggins BJ et al., 2013]. The permeability of lipophilic drugs and
mucoadhesive formulations is likely to be affected by the viscosity of the mucous-gel
layer. HIF also regulates the transcription of MDR1, which codes for the xenobiotic
drug efflux pump P-glycoprotein (P-gp), which is involved in actively transporting
substrate compounds back into the lumen [Comerford KM et al., 2002].
Glucocorticoids, for example, are P-gp substrates and have been shown to stimulate
MDR1 expression, potentially contributing to steroid resistance in IBD [Creed TJ et
al., 2007]. Nano-delivery methods have been shown to target both pharmacological
and biological pathways to overcome multidrug resistance by inhibiting P-gp and
depleting ATP [Dong X et al., 2009].

1.1.7. Treatment of IBD

The majority of inflammatory bowel disease (IBD) treatments are either
medical or surgical in nature. Both Crohn's disease and ulcerative colitis are treated
with a variety of medications. Surgery is more specific to the type of IBD when it is
used to treat it. Surgery is usually reserved for those who have exhausted all other
medical options. Sulfasalazine (Azulfadine), mesalazine (Asacol, Pentasa),
azathioprine (Imuran), 6-MP (Purinethol), cyclosporine, methotrexate, infliximab
(Remicade), and corticosteroids are some of the more popular medications to treat
IBD.

Sulfasalazine (Azulfidine):- Sulfasalazine is a 5-aminosalicylate acid (5-ASA)

substance that decreases inflammation in the colon by combining two drugs
(sulfapyridine and an aspirin-like compound [5-ASA]. Nausea, headaches, diarrhoea,
and abdominal pain are all common side effects.

Mesalazine (Asacol, Pentasa) and Olsazine (Dipentum):- Because they do not

contain the sulfa component, these newer 5-ASA medications are equally effective as

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CHAPTER1: INTRODUCTION 2022

sulfasalazine but have fewer side effects. Mesalazine is most effective in patients
with mild to severe Crohn's disease, and it may reduce the chances of a relapse
following surgery.

Azathioprine (Imuran), 6-mercaptopurine (Purinethol, 6-MP) and

cyclosporine A (Sandimmune, Neoral):- Immunosuppressants, such as
azathioprine, 6-MP, and cyclosporine, are drugs by which the immune system was
compromised. IBD is an autoimmune disease, and suppressing the immune system
may help to alleviate symptoms, but it also makes the body more susceptible to
infection. Azathioprine and 6-MP are slow-acting drugs that are frequently
combined with a faster-acting medication like a corticosteroid. Cyclosporine has a
faster action, but it comes with serious side effects. These drugs are most commonly
used to treat Crohn's disease, with ulcerative colitis being used less frequently.

Budesonide (Entocort EC):- Budesonide is a medication that is used to treat

Crohn's disease that affects the ileum and/or ascending colon. Budesonide is a non-
systemic corticosteroid that has fewer side effects than other corticosteroids
because the majority of the drug is released in the intestine rather than the blood
stream. Headache, respiratory infection, and nausea are some of the side effects.
Both types of IBD are treated with budesonide.

1.1.8. Side effects of treatments prescribed in ulcerative colitis and crohn’s

disease
The side effects of long-term exposure and drug interactions of commonly
prescribed ulcerative colitis drugs are summarised in Table 1.3 [Carter M et al.,
2007]. These treatments necessitate multiple long-term exposures, which disrupts
daily life. Patients with ulcerative colitis do not stick to these treatments because of
the negative side effects, which increase the risk of disease remission, colon cancer,
and colon surgery [Kane SV et al., 2006].

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Table 1.3: Different ulcerative colitis and crohn’s disease treatments with side
effects

Treatment Side effects Reference

5-aminosalicylic Headache, epigastric pain, (Loftus et al.,
acid
diarrhoea, idiosyncratic reactions, 2004., Van Staa et
Renal impairment al., 2004)
Corticosteroids Mood disturbance, dyspepsia, glucose (Fraser, 2003),
intolerance, posterior sub-capsular
cataracts, osteoporosis, osteonecrosis
of the femoral head, myopathy, and
susceptibility to infection, adrenal
insufficiency, myalgia, malaise, and
arthralgia, raised intracranial

Methotrexate Nausea, vomiting, diarrhoea, (Te et al., 2000)

stomatitis, hepatotoxicity, and
pneumonitis
Cyclosporine Tremor, paraesthesia, malaise, (D'Haens et al.,
headache, abnormal liver function, 2001., Van Assche
gingival hyperplasia, and hirsutism, et al., 2003)
renal impairment, infections, and
neurotoxicity
Infliximab Joint pain and stiffness, fever, myalgia, (Ljung et al., 2004.,
and malaise Colombel et al.,
2004)

1.2. Anatomy and physiology of colon

Irrespective of therapy preferred for local (colonic) or systemic delivery of
drug, the development and aim of the drug delivery to colon remains same [Roop K
et al., 2006] that is

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• Other parts of the GI must not absorb the medication.

• In the lumen of intestine, it should only undergo minor degradation.
• The drug should be released in the colon at a quantitatively controlled rate, and
the drug should be absorbed without significant degradation from the lumen of
the large intestine.
In order to meet these properties, a thorough knowledge of the anatomy and
physiology of GIT is requisite. In GIT, large intestine starts from the ileocecal
junction to the anus with a length of about 1.5 meters (adults) and is alienated into
three parts; they are colon, rectum and anal canal. The colon is made up of the upper
five feet of the large intestine and is mostly found in the belly. The upper five feet of
the large intestine make up the colon, which is usually situated in the belly. The
colon is a cylindrical mucosa-lined tube. The cecum, colon ascends, colon
transversale, colon descendens, and recto sigmoid colon make comprise the colon.
Serosa, muscularis externa, sub mucosa, and mucosa are the four layers that make
up the colon. Although the colon lacks villi, the intestinal surface of the colon is
increased to approximately 1300 cm2 due to the existence of plicaesemilunares
(cresentic folds) [Roop K et al., 2006 & Vincent H et al., 2002].

1.2.1. Colonic absorption of drugs

The colon has a much smaller surface area than the small intestine, which is
compensated by the lack of endogenous digestive enzymes and the colon's long
residence time (10-24 hours). Vincent H et al., 2002, reported on various factors
that affect colonic absorption.
 Passes through colonocytes (Trans-cellular transport)
 Passes between adjacent colonocytes (Para-cellular transport)

1.2.2. Drugs candidates for colon specific drug delivery systems

Based on literature review, the following different categories of drugs are
suitable for colon drug delivery.

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CHAPTER1: INTRODUCTION 2022

• Drugs used to treat irritable bowel disease (IBD), such as sulfasalazine, olsalazine,
mesalazine, and steroids like fludrocortisone, budesonide, prednisolone, and
dexamethasone, require local delivery to the colon.
• Drugs like 5-fluorouracil, doxorubicin, and methotrexate, which are used to
treat colon cancer, must be delivered locally.
• Protein and peptide drugs - preventing drug degradation, for example, growth
hormones, calcitonin, insulin, interleukin, interferon, and erythropoietin.
• In order to cure infectious diseases (amoebiasis and helminthiasis),
metronidazole, mebendazole, and albendazole must be delivered to specific sites.
• Due to cicardian cycles, rheumatoid arthritis (NSAID S ), nocturnal asthma, and
angina necessitate a delay in absorption.
• Due to the small extent of paracellular transport, drugs such as glibenclamide and
diclofenac show more selective absorption in the colon than the small intestine.

1.2.3. Therapeutic benefits of colon-specific drug delivery system (Girish et al.,

& Vyas S et al., 2006)
• Reducing the adverse effects in the treatment of colonic diseases (ulcerative
colitis, colorectal cancer, crohn’s disease etc.)
• When contrasted to the upper gastrointestinal system, it creates a 'friendlier'
environment for peptides and proteins.
• Reducing widespread first pass metabolism of steroids.
• Avoiding the gastric irritation caused by administration of oral NSAIDS.
• Drugs used to treat angina, asthma, and rheumatoid arthritis are released in
delayed manner.
To achieve effective colon targeting it should overcome the below limitations
(Jack A et al., 2006)
• The colon is difficult to reach due to its position at the distal end of the alimentary
canal.
• For delivery to be successful, the medicine must be in solution before reaching
the colon, but the colon's fluid level is lower and more viscous than the upper GIT,

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CHAPTER1: INTRODUCTION 2022

which is the constraint for pharmaceuticals that are poorly soluble.

• Lower surface area and relative tightness of tight junctions in the colon can
impede drug transport across the mucosa into the systemic circulation.

1.2.4. The feature of the colon that make it suitable for targeting various drug
including protein & peptide (Jack A et al., 2006).
• Its lower metabolic activities.
• Longer residence time (20-30 hrs).
• Responsiveness to absorption enhancers.
• Targeting opportunities offered by colonic bacterial enzyme.
• Transmucosal membrane potential difference that is significant in the
absorption of the ionized & unionized drugs.
• Possibility that bulk water absorption in this region may provide scope for
solvent drag.
Table1.4 Drug metabolising enzyme in the human colon that catalyze
reductive reaction (Binder HJ et al., 1987)

Enzyme Microorganism Metabolic reaction catalysed

Reduce aromatic and heterocyclic
Nitroreductase E-coli, Becteroides
compound.
Clostridia spp, Reductive cleavage of aromatic
Azoreductase
Lactobacilli, E-coli compound.
N-oxide
E-coli Reduce N-oxide.
reductase
Clostridia spp, Reduce carbonyle group and
Hydrogenase
Lactobacilli aliphatic double bond.
Estrase and E-coli, P.vulgaris, Cleavage of ester and amide group
Amidase B.subtilis of carboxilic acid.
Cleavage of β-glycosidase of
Gulucosidase Clostridia, Eubacteria
alcohol and phenol.
Clostridia, Eubacteria, Cleavage of O-sulfate and
Sulfatase
Streptococci sulfamate.

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1.2.5. Approaches in colon specific drug delivery system

The most commonly used targeting mechanism is:
 Drug release based on variation of pH.
 Drug release based on the presence of micro flora.
 Time controlled drug delivery system.
 Pressure controlled drug delivery system.

1.2.5.1. Drug release based on variation of pH (Evans DF et al., 1988 Bussemer

T et al., 2001 & Rodríguez M et al., 1998)
The different pH of the human GIT is exploited in pH controlled release
systems by coating the dosage form with pH dependent polymers that remain intact
in the upper GIT and degrade in the large intestine where the pH is high, 7-8. This
method can be applied to any dosage form, including tablets, capsules, and pellets.
The active drug is protected from gastric fluid and a delayed release is obtained by
coating the dosage forms with pH sensitive polymers. Delivery systems are designed
to target drugs to desired locations by gathering the most information about
polymers and their solubility at various pH levels. The most commonly used
polymers for colonic drug delivery are methacrylic acid and methyl methacrylate.
Following an in vitro comparison of Eudragit S and Eudragit FS, it was discovered
that the latter is more suitable for ileocolonic drug delivery. The dissolution rate of
Eudragit is affected by a variety of factors, including polymer combinations, coating
level, and media pH. Some medications, such as mesalazine (5 ASA) (Asacol and
Salafalk) and budesonide (Budenofalk and Entrocort) for the ulcerative colitis and
crohn's disease treatment, are commercially available in pH-controlled systems as
shown in Table 1.5.

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Table 1.5 Threshold pH of commonly used enteric polymer

Enteric polymers Threshold pH

Polyvinyl acetate phthalate(PVAP) 4.5-5.0

Cellulose acetate phthalate (CAP) 5.0

Eudragit L 100 6.0

Eudragit S 100 7.0

Eudragit L 100-55 5.5

Eudragit L 30-D 5.6

Hydroxypropyl methylcellulose phthalate ≥ 5.5

Cellulose acetate trimelliate 5.5

Hydroxypropyl methylcellulose acetate ≥6

Succinate
Eudragit FS 30 D 6.8

1.2.5.2. Drug release based on the presence of microflora (Sinha V R et al.,

2003, Schacht E et al., 1996 & Takaya T et al., 1995)

The principle at work in this system is that the microbiome of the colon
degrades the polymers coated on the dosage form, releasing the drug load. The colon
has a diverse microflora that meets its energy demands by digesting the substrate
found in the intestine, such as polysaccharides. This microflora produces a wide
range of enzymes that can metabolise substrates such as carbohydrates and
proteins that are not degraded in the upper GI tract. Polymers are commonly used in
pharmaceutical formulations and are commonly considered as safe excipients. When
used alone to control the release of drug from the dosage form, polymer pectin was
required in large quantities. However, when pectin was combined with chitosan and

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hydroxyl propyl methyl cellulose in sufficient amounts, it proved to be very effective

in preventing drug release in the stomach and colon. The first bacteria-sensitive
system to deliver drug to the colon was sulphasalazine, a prodrug of mesalazine.
Chitosan, Pectins, Guar Gum, Dextrans, Inulin, Lactulose, Amylose, Cyclodextrins,
Alginates, Locust bean gum, Chondroitin sulphate, and Boswellia gum are among the
microbially degradable polymers. The three microbially triggered approaches listed
below are examples of microbially triggered approaches:

a. Prodrug approach: A prodrug is a biologically inactive compound of a parent

drug that requires in vivo spontaneous or enzymatic transformation to release the
therapeutics. A covalent bond exists between the drug and its carrier in this
approach, which remains intact in the upper GIT before breaking down in the colon,
releasing the drug. A number of drug linkages containing hydrophobic moieties such
as amino acids, glucoronic acid, glucose, galactose, cellulose, and others that are
susceptible to hydrolysis in the colon have been developed. The major drawback of
the prodrug approach is that the functional group presents on the drug moiety plays
a critical role in chemical linkage in its design and development. 5-ASA, for example,
was conjugated with glycine via an amide linkage that was found to be stable in the
upper GIT and hydrolyzed by cecal contents to release 5-ASA.

b. Azo - polymeric prodrugs: The use of different polymers as drug carriers for
colonic delivery is a newer technique. Sub synthetic polymers are used to make
polymeric prodrugs with an azo bond between the polymer and the drug moiety.
When polymers cross linked with an azo aromatic group were coated on a drug, they
protected it from degradation in the upper GIT and allowed it to be released in the
colon, where the azo bonds were reduced, allowing it to be released. An example of
an azo polymer-based drug delivery system is segmented polyurethane, which was
coated over budesonide pellets and evaluated in vivo and in vitro to result in drug
delivery to the colon (Mooter GV et al., 1995).

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c. Polysaccharide based approach: Naturally occurring polysaccharides are widely

in use for drug targeting because of their abundance, easy availability, and also they
are inexpensive. They're extremely stable, nontoxic, hydrophilic, gel-forming, and
biodegradable. Chitosan, Pectin, Chondroitin sulphate, Alginates, and other natural
polysaccharides derived from plants, animals, algae, or microbes are being studied.
These polysaccharides can be broken down into simpler ones by colonic microflora.
Chitosan is primarily used in colonic delivery as a capsule forming material [Fukui E
et al., 2001].

1.2.5.3. Time controlled drug delivery system [Arora S et al., 2006, & Jung YJ et
al., 1998]
The drug is released after a predetermined lag time at the desired site of action and
time of release in time controlled systems. For colon targeting, a five-hour lag time is
thought to be sufficient. The thickness of the coated polymer or mixture of polymers
affects the time it takes for the dosage form to release the drug in the colon. The
colon arrival time of drug formulation cannot be determined reliably because the
gastric emptying timing of dosage forms differs from person to person. These
systems, on the other hand, can be used to treat diseases that are affected by
circadian rhythms. Capsules and bilayer tablets are used to create a time-controlled
system. The drug's release time from the dosage form is determined by the balance
of the thickness of the water-insoluble membrane and the quantity of swellable
excipient. L-HPC, sodium starch glycolate, and other swellable excipients could be
used. As a result, combining pH and time release systems in a formulation may
improve targeted delivery of therapeutics to the colon as well as dosage form transit
time in the small intestine, which is approximately 3±1 hours. Hydroxyl Propyl
Methyl Cellulose, Hydroxy Ethyl Cellulose, Ethyl Cellulose, Microcrystaline Cellulose,
Hydroxy Propyl Methyl Cellulose Acetate Succinate, Lactose/Behinic acid, and other
polymers are used to develop time-dependent systems.

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1.2.5.4. Pressure controlled drug delivery system (Spraycar M et al., 1995 &
Fukui E et al., 2001).
Contractile and peristaltic movements occur within the GIT to propel intestinal
contents forward. Forcible peristaltic movements, known as mass peristalsis, occur
in the large intestine, moving intestinal contents from one location to another. These
peristaltic waves last only a few seconds and occur only 3 to 4 times per day. The
drug is released after the water insoluble polymer capsule is disintegrated by the
luminal pressure of the colon. The disintegration of the dosage form is influenced by
the thickness of the membrane. Because of peristaltic motion, which is caused by the
viscosity of luminal contents, the luminal pressure in the colon is higher. Takaya et
al., 1995 used ethyl cellulose, which is water insoluble, to create pressure-controlled
colon specific capsules. The size of the capsule is also a factor in this system. When
the pressure controlled capsules were given to humans, there was a 3 to 5 hour
delay in drug absorption. The capsule shells are made of ethyl cellulose, and the
collapse time can be controlled by varying the thickness of the shell. A capsule wall
thickness of 35-60 micrometres is considered satisfactory.

1.3 Prebiotics in crohn’s disease & ulcerative colitis

Non digestible oligo-saccharides are prebiotics, which are defined as
“Ingredients that have been selectively fermented to allow specific changes in the
characteristics and/or activity of the gastrointestinal microflora, resulting in
benefits to the host's well-being and health" [M. Robertfroid et al., 2007]. When
substances meet the following criteria, they are classified as prebiotics: 1) In the
upper gastrointestinal tract, it cannot be hydrolyzed or absorbed; 2) be selectively
fermented in the intestine by one or a small number of potentially beneficial
bacteria; and 3) be able to change the colonic microflora toward a healthier
composition[G. R. Gibson et al., 1995].

Beta-fructans oligosaccharides are commonly used prebiotics Inulin and

oligofructose are natural food components or dietary fibres found in plants that act
as storage carbohydrates. Wheat, chicory, bananas, onions, leek, artichoke,

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CHAPTER1: INTRODUCTION 2022

asparagus, and garlic all contain prebiotics. The majority of commonly available
prebiotics are made from sucrose or extracted from chicory roots. They are used in
candies, confectioneries, bakery products, fermented products, fruit juices,
desserts, spreads, fat replacers, etc [Swennen K et al., 2006].

Inulins consist of multiple fructose units connected by a glucose molecule at

the end; glycosidic links connect the fructose units. Inulin is made up of 2–140 beta-
fructans units in nature. Oligofructose is a decomposition product of inulin,
consisting of fructose polymers with a degree of polymerization (DP) of less than
10. Inulin with a greater DP is less soluble and can be used as a fat substitute.
Lower-DP oligofructose is more soluble and is used to substitute sugar.

Prebiotics are not digested in the upper gastrointestinal tract and instead
reach the colon intact, where they are fermented by the local microbiota into short
chain fatty acids (SCFAs) and lactate. Prebiotics, like probiotics, need to be
consumed on a regular basis to maintain their positive effects [Bouhnik Y et al.,
2004 & Kleessen Bet al., 2001].

According to several studies prebiotic effects are influenced by a variety of

factors, including luminal pH, prebiotic dosage and concentration, duration of
intake, fermentation site, and the initial configuration of the intestinal microflora
[Tuohy KM et al., 2001].

Several compounds have been identified as prebiotics, but according to

Roberfroid and Cherbut [Roberfroid M et al., 2007 & Cherbut C et al., 2001]. All
three criteria listed above have been met by inulin. Other prebiotic candidates are
promising, but there is only preliminary data on their health-promoting effects, and
they don't meet all of the prebiotic criteria listed above, so they can't be designated
as prebiotics.

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CHAPTER1: INTRODUCTION 2022

1.3.1. Protective mechanisms of prebiotics

Prebiotics gastrointestinal defense mechanisms are still relatively unknown.
Variations in the intestinal microflora, improved intestinal barrier, regulation of the
mucosal and systemic immune response, and increased production of intestinal
SCFAs are among the prevailing hypotheses on prebiotics defense mechanisms as
shown in Figure 1.2.

changes in
the intestinal
microflora

increasing
the improving
production of Prebiotics intestinal
intestinal barrier
SCFAs.

regulating
the mucosal
and systemic
immune
response

Figure1.2: Protective mechanisms of prebiotics

1.3.2 Prebiotics and the intestinal microflora

Intestinal bacteria are thought to play a significant role in IBD pathogenesis,
particularly in CD. Prebiotics change the intestinal microbial composition by
promoting the growth of commensal protective bacteria and increasing resistance

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CHAPTER1: INTRODUCTION 2022

to disease-causing bacteria, resulting in a reduction in colitis [Gibson GR et al.,

1999 & Sartor RB et al., 2004].

Prebiotics transform the intestinal microflora toward a favourable one in

both animal models and human studies, increasing the number of defensive
bacteria like bifidobacteria and lactobacilli to the detriment of disease-causing
bacteria, according to several research findings. Prebiotic milk-oligosaccharides are
found in breast milk. Several researches has found that bifidobacteria and lactic
acid bacteria dominate the intestinal microbiota of breast-fed infants, whereas the
microflora of formula-fed infants includes fewer of these protective bacteria and
much more bacteroides, clostridia, and enterobacteriaceae [Bruzzese E et al.,
2006 & Forchielli ML et al., 2005].

The recent literature offers some explanations for how prebiotics

selectively stimulate the growth of protective bacteria. Specific enzymes in some
protective organisms can hydrolyze prebiotic oligosaccharides, resulting in the
emergence of these protective bacteria. Bifidobacterium infantis, for example, has
been found to have this mechanism [Perrin S et al., 2001].

Furthermore, Bacteroides spp. cannot grow at pH below 5.5, so

fermentation of non - digestible carbohydrates in the proximal colon results in a
lower luminal pH, which prevents their growth [Walker AW et al., 2005] and the
mechanism of prebiotic action, was represented in Figure 1.3.

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CHAPTER1: INTRODUCTION 2022

Intestinal Epithelium

GUT SCFA’s
PREBIOTICS MICROFLORA
Fermentation
Acetic Acid COLONIC AND
(PROBIOTICS) end product Propionic Acid
Inulins SYSTAMIC
Fructans Butyric Acid
Bifidobacteria SCFA’s HEALTH
Lactobacilii EFFECTS

pH
Drop

Figure 1.3: Mechanism of prebiotic action.

Prebiotics can also prevent pathogenic bacteria from adhering to the gut epithelium
and colonising it. Prebiotic oligosaccharides' terminal sugars can interfere with
disease-causing bacteria's receptors, restricting them from adhering to the
intestinal epithelium. Hopkins and colleagues discovered that
Fructooligosaccharide (FOS), Galactooligosaccharide (GOS), and inulin inhibit
Clostridium difficile colonisation in the intestine in vitro [Hopkins MJ et al., 2003]

1.4. Inulin in IBD

Inulin is a fructose-containing polymer that belongs to the fructans family.
Many members of the Composite family, including Cichoriumintybus (chicory),
Inulahelenium (elecampane), Taraxacumofficinalis (dandelion), and Helianthus
tuberosus, use fructans as storage polymers (Jerusalem artichoke). Chicory inulin is
a naturally occurring polydisperse carbohydrate [Goudberg A et al., 1913]. It is a
fructan composed primarily of linear chains of 1, 2-[3-1inked d-fructofuranose
units linked to a terminal glucose moiety by a (od-132) type linkage (as in sucrose).
This is the type of fructan that can be found in dicotyledons and some

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CHAPTER1: INTRODUCTION 2022

monocotyledons. Fructans are primarily made up of linear fructose units linked by

a [3-(2---6) glycosidic bond.

Because of its bioactivity, inulin is now a promising molecule with

research and applications in medicine, pharmacology, and the food industry
(Mensink MA). In medicine, inulin serves a variety of purposes, including
maintaining colon microflora, improving metabolism, and maintaining
gastrointestinal health (Carlson JL). In addition, Smecuol E [Smecuol E et al.,
2013] discovered that inulin-type fructans make a significant contribution to the
emergence of Bifidobacterium infantis probiotic strains that have a beneficial
impact on subjects with active coeliac disease, a chronic irritation of the intestinal
mucosa caused by a severe immune response to dietary gluten in genetically
vulnerable individuals. Colonic disorders are harder to treat because oral
medications are absorbed in the stomach and intestine but do not reach the colon,
whereas intravenous therapeutics are eliminated from the body before reaching
the colon. Inulin is not broken down or absorbed in the stomach or small intestine,
but it is broken down by bacteria in the colon [Jain AK et al., 2014 & López-
Molina D et al., 2015]. Previous research has shown that digestion of the fibre
inulin by bacteria in the large bowel (colon) stimulates the production of
metabolites known as short chain fatty acids (SCFAs). SCFAs have been shown to
have anti-inflammatory properties, and we recognize that patients with IBD have
lower levels of these beneficial SCFAs. As a result, we hypothesise that prebiotic
inulin supplementation will support IBD patients by increasing the abundance of
healthy SCFA-producing bacteria and their metabolites, resulting in less gut
inflammation.

Inulin is a multifunctional polysaccharide which could be used as a drug

delivery vehicle, a diagnostic tool, or a dietary fiber with numerous health benefits.
There has been a lot of research done on inulin and its use in drug delivery. Inulin is
most commonly used for targeted drug delivery in the colon. The main reason is its
ability to survive in an acidic environment in the stomach. This unique stability and

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CHAPTER1: INTRODUCTION 2022

strength is being used in our current research to safely deliver drugs to the colon
where they can be easily absorbed or produce a local effect for the effective
management of gastrointestinal disorders. In short, we used inulin as a vehicle for
drug delivery to the desired site of action as well as a potential prebiotic for
promoting the growth of beneficial intestinal microflora.

1.5. Chitosan in IBD

Chitosan is a polysaccharide made up of D-glucosamine and N-acetyl-D-
glucosamine crosslinked together by - (1,4) linkages. It is produced by
deacetylation of chitin. Chitosan is a biodegradable, biocompatible, nontoxic, and
slightly allergenic cationic polymer. It has antitumor, antimicrobial, and antioxidant
properties, which make it a popular research topic for pharmaceutical and
biomedical applications. Chitosan can protect drugs from chemical and enzymatic
degradation during oral administration when used as a matrix and/or coat
material. Chitosan high binding to mucus and improves mucosal permeation,
allowing drugs to pass through intestinal epithelial cells more easily. For the
prevention and treatment of colon cancer, ulcerative colitis, Crohn's disease,
diverticulitis, irritable bowel syndrome, Hirschsprung's disease, antibiotic-
associated colitis, and other colon diseases, oral colon-specific delivery systems
have been investigated. This polymer is distinguished from other polysaccharides
by the presence of nitrogen in specific properties such as its molecular structure,
cationicity, and, most importantly, its ability to form polyelectrolyte complexes. The
cationic nature of the polymer allows it to become water-soluble after the
formation of carboxylate salts such as formate, particularly acetate, lactate, and
malate, and to a lesser extent, ascitrate, glyoxylate, pyruvate, glycolate, and
ascorbate. Chitosan is an excellent excipient because it is non-toxic, stable,
biodegradable, and sterilizable. Chitosan is a very versatile material with a wide
range of applications in biomedical and biotechnological fields [Dash M et al.,
2011]. The polymer's appealing characteristics also make it an excellent candidate
for controlled release preparations.

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CHAPTER1: INTRODUCTION 2022

Chitosan was chosen as a therapeutic agent for colon-selective drug

delivery because of its precise biodegradability by the specified enzyme, lysozyme,
which is thought to be highly concentrated in the mucosa, and with the help of
enzymes produced naturally by colonic bacteria, as well as its mucoadhesive
properties. Chitosan improves absorption in the colon by extending the time that
dosage forms spend at mucosal sites. Furthermore, Chitosan has the benefit of
being widely accepted as a food ingredient, implying that it could be used as a new
excipient for oral administration [Şenel S et al., 2010].

1.5.1. Mucoadhesion from chitosan

Two key characteristic features in orally administered dosage forms are
the ability to efficiently traverse the epithelium and delayed GI transit in the
duodenum. Chitosan-based NP has been shown to have these properties in this
regard [Shaikh R et al., 2011]. Mucoadhesion is the adhesion of two materials, one
of which is mucosal. It can be used to improve bioavailability by extending the GI
transit of dosage forms in the duodenum. Within mucin, interactions of positively
charged chitosan moieties with negatively charged moieties in sialic acid cause
delayed transit [Garg UI et al., 2019]. Chitosan can also physically penetrate the
mucous membranes. Increased net drug flux across the GIT membrane is the result
of prolonged GI residence. Drug flux is the result of a combination of passive
diffusion and whole-NP uptake by Peyer's patches [Boddupalli BM et al., 2010].
Furthermore, chitosan allows for controlled drug release from the matrix via
diffusion. Yin et al. prepared thiolated trimethyl chitosan NPs for the oral delivery
of insulin and found that increasing mucoadhesion increased insulin transport
through the rat intestine and uptake by Peyer's patches in comparison to controls.
The formation of a disulfide bond between the NPs and the mucin was attributed to
these findings [Yin L et al., 2009]. Overall, the method of preparation of chitosan
NP is critical for achieving the desired properties of interest, such as particle size,
particle size distribution, and application area.

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CHAPTER1: INTRODUCTION 2022

1.6. Multiparticulate systems

Oral dosage forms made up of a large number of small discrete units, each
with its own set of desired properties, are known as multi-particulate drug delivery
systems. The drug substance dosage is divided into a number of subunits in these
systems, which are typically thousands of spherical particles with diameters ranging
from 0.05 to 2.00 mm. Multiparticulate dosage forms are pharmaceutical
formulations in which the active ingredient is present as a number of small,
individual subunits. These subunits are loaded into a sachet and encapsulated or
compressed into a tablet to deliver the recommended total dose. A multiple unit
system is made up of discrete particles called multiparticulates. They have
numerous advantages over single-unit systems due to their small size.
Multiparticulates are less reliant on gastric emptying, resulting in less
gastrointestinal transit time variability between and within subjects. They are also
more evenly spread and less likely to irritate localised areas. Because of their
promising benefits such as increased bioavailability, reduced risk of systemic
toxicity, decreased risk of local irritation, and predictable gastric emptying,
multiparticulate dosage forms have recently received a lot of attention in
comparison to single unit dosage form [Tang E et al., 2005, Shaji J et al., 2007 &
Laila F et al., 2006].

1.6.1. Advantages of multi -particulate system

• To facilitate drug delivery to the desired location.
• The pharmacokinetics behaviour of this system is more consistent than that of
conventional systems. (monolithical).
• Individual subunit particles pass quickly through the GI tract after
disintegration.
• Even if the pylorus is closed, these subunits pass through the stomach
continuously if their diameters are less than 2 mm.
• These result in lower inter and intra individual variability in plasma levels.

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CHAPTER1: INTRODUCTION 2022

1.6.2. Drug safety in multi -particulates system

Drug safety is also improved by using these dosage forms, particularly for
modified release systems. If the film coat of a single-unit (monolithic) enteric
coated tablet is damaged, the entire dose will be released into the site, potentially
causing pain, ulceration, or reduced efficacy, depending on why the enteric coating
was chosen. Destruction to the film coating of a tablet with a sustained release
formulation can cause "dose dumping," which can be dangerous. In a
multipariculate formulation, however, the release characteristics are incorporated
into each individual subunit, and any damage only affects the release behaviour of
the involved subunit, which accounts for a small portion of the total dose, lowering
the risk of safety concerns [Dey NS et al., 2008].

1.6.3. Mechanism of drug release from multi -particulates system [Dey NS et

al., 2008].
The release of drugs from multiparticulates can take place in a variety of ways:
 Diffusion: Water diffuses into the interior of a particle when it comes into contact
with aqueous fluids in the gastrointestinal tract (GIT). The drug solutions dissolve
and diffuse to the release coat's outer surface.
 Erosion: Some coatings can be developed to erode over time, allowing the drug
contained within the particle to be released.
 Osmosis: Water enters the particle, causing osmotic pressure to build up inside
the particle. Through the coating, the drug is forced out of the particle and into
the environment.

1.6.4. Nano particles in IBD

Nanotechnology has had a huge impact on medicine in the last decade, thanks
to advances in drug delivery [Juliano R et al., 2013]. Drug delivery employing
nanotechnology aims to deliver the drug payload to the desired site, at the right
time, and at the right (optimal) dose [Bourzac K et al., 2012] Submicron-sized
particles are dispersed, adsorbed, or covalently bound with one or more therapeutic
agents in encapsulated vesicles, capsules, or polymer matrices in nanodrug delivery

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CHAPTER1: INTRODUCTION 2022

systems. [Cho K et al., 2008]. Nanodrug delivery technologies improve each drug's
bioavailability, decreasing the negative side effects induced by related toxicities [De
Jong et al., 2008]. The majority of NPs are biodegradable and nontoxic to humans,
thus there are no negative effects. Within the core, NPs can be loaded with the
desired medications, which will then be released locally into a target cell, enabling
for lower drug doses to be administered. In this way, NPs therapy maintains the
same efficacy as the traditional formulation while reducing the negative side effects
of free medication delivery. Natural nanoparticles (phospholipids, lipids, lactic acid
and chitosan) and chemical nanoparticles (polymers carbon, silica, and metals) are
examples of natural nanoparticles [The Royal Society 2004]. NPs that have been
properly created should remain in the bloodstream and interact less with the
reticuloendothelial system (RES). Indeed, RES sequestration can interfere with NPs
blood circulation, limiting their ability to reach target tissues. Furthermore, NPs
must avoid macrophage phagocytosis and filtering organ sequestration (ie, liver,
spleen, and kidney). In order to produce the most effective NPs, researchers have
proposed several changes to the canonical formulation of NPs, such as changing the
size, zeta potential, composition, shape, or surface. The main challenge for NPs in the
treatment of IBD is to reach and penetrate the gastrointestinal mucosa in order to
target the underlying inflammatory cells. Due to the biochemical and physiological
features of the gastric mucosa, adding a hydrophilic polymer like poly-ethylene-
glycol (PEG) to the NP surface is one of the most effective modifications. PEG inhibits
serum protein binding, restricts macrophage uptake, and allows NPs to penetrate
the mucus layer of the gastrointestinal mucosa [Cu Y et al., 2009]. In the
development of NPs, other polymers are frequently used. These can be natural or
synthetic; in either case, they must be biodegradable, and the degradation products
must not be toxic to humans. Natural polymers include proteins (gelatin, albumin)
and polysaccharides (alginate, chitosan, pectin), with the latter having the benefit of
being degraded by colonic bacteria [Laroui H et al., 2011]. Poly(D,L-lactide) (PLA),
poly(glycolide) (PGA), and poly(lactide-co-glycolide) (PLGA), which deteriorate into
lactic acid and glycolic acid in the body, or poly—caprolactone (PCL), a synthetic

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CHAPTER1: INTRODUCTION 2022

polymer degraded by lipase, are examples of synthetic polymers that are easily
modifiable in size and weight. Surface decoration can also be done with
methylcellulose, which increases NPs contact with the mucosa due to its muco-
adhesive properties. Other factors that influence the success of NP-based therapies
include hydrophobicity, porosity, and surface charge. Surface charge, for example,
reduces NPs aggregation and increases absorption, with different affinity in
inflamed and healthy colonic mucosa [Jani P et al., 1989].

1.6.4.1 Nanopharmacology in the treatment of IBD

Sulfasalazine, mesalazines, olsalazine, and balsalazide are 5-ASA drugs that
act as free radical scavengers, inhibitors of leukotriene production and leukocyte
recruitment, and inhibitors of nuclear factor-B (NF-B), which stimulates receptors
that manage inflammation and cell proliferation [Greenfield SM et al., 1993]. These
medications are primarily used to treat mild to moderate disease and maintain
remission in UC patients. In contrast, 5-ASAs are quickly absorbed in the small
intestine, with only a minor fraction reaching the colon. To address this problem,
nanotechnologists have developed various formulations of NPs loaded with 5-ASAs
that have a controlled and sustained release. In an in vitro cellular assay, Varshosaz
et al demonstrated that 5-ASA chitosan NPs could deliver 80 percent of the drug to
the colon [Varshosaz J et al., 2006]. In vivo studies on mice given 2,4,6-
trinitrobenzenesulfonic acid (TNBS) shown that 5-ASA-loaded NPs were present
primarily in the colon, with minimal systemic bioavailability [Mladenovska K et al.,
2007]. Furthermore, these patients receive free 5-ASA treatment, providing an
alternative for IBD drug dosing to avoid medication side effects [Pertuit D et al.,
2007].

Because of its safe pharmacological profile, silica is an excellent candidate

for drug delivery. 5-ASA can be loaded into silica nanoparticles (SiNPs) to keep it
encapsulated until it is selectively delivered to inflamed colonic tissue. According to
Moulari et al., 5-ASA/SiNPs at 25 or 50 mg/kg caused a greater decrease in MPO
activity than free 5-ASA at 100 mg/kg. Overall, 5-ASA/SiNPs were more effective
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CHAPTER1: INTRODUCTION 2022

than traditional 5-ASA formulations for reducing inflammation at lower doses

[Moulari B et al., 2008]. Rectal mesalazine was recently infused into the coating
agent hydroxypropyl methylcellulose (HPMC K4M) in order to achieve a controlled
release of the anti-inflammatory drug in the management of distal UC. The improved
preparation was examined on mice treated with acetic acid, which had considerably
lower Disease Activity Index scores after treatment than mice treated with a
polymer-free formula. The mucosal lining of the HPMC K4M mesalazine group was
intact, with only minor cellular inflammatory infiltrates, whereas the polymer-free
group had an eroded mucosal lining and a mild inflammatory infiltrate, as evidenced
by histopathology. As a film-former, hydroxypropyl methylcellulose provided muco-
adhesive properties to the formula, increasing the contact time between mesalazine
and the damaged mucosa, increasing its therapeutic efficacy [Ali HS et al., 2016].

1.6.4.2 Preparation of nanoparticles

The appropriate method for preparing nanoparticles is determined by the drug to be
loaded and the polymer's physicochemical properties. The primary preparation
methods of nanoparticles include:
• Solvent evaporation method:
• Spontaneous emulsification or solvent diffusion method:
• Solvent Displacement/Precipitation method
• Polymerization method
• Ionic gelation method
We will limit our discussions to these aspects because our current research work is
only concerned with the ionotropic gelation method.

1.7. Ionotropic gelation method

Ionotropic gelation is a technology that, under certain conditions, allows the
production of nanoparticles by electrostatic interactions between two ionic species.
A polymer must be present in at least one of the species. When a drug or bioactive
molecule is added to the reaction, it can become trapped between the polymeric
chains, resulting in the drug or bioactive molecule being trapped inside the
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CHAPTER1: INTRODUCTION 2022

nanoparticle structure [Koukaras EN et al., 2012]. This type of formulation allows

for controlled drug release as well as other benefits such as molecule co-
encapsulation, particle site-specific functionalization, and longer drug bioactivity
times.
Because the process is very simple and mild, the use of complexation
between oppositely charged macromolecules to formulate nanoparticles has gotten
a lot of attention. To avoid reagent toxicity and other undesirable effects, reversible
physical cross-linking by electrostatic interaction was used instead of chemical
cross-linking. The polyanion tripolyphosphate (TPP) interacts with cationic chitosan
via electrostatic forces. Since Bodmeier et al., reported the preparation of TPP–
chitosan complex by adding chitosan droplets into a TPP solution, many researchers
have looked into the potential pharmaceutical applications of TPP–chitosan
complex. To obtain the chitosan cation, the ionotropic gelation technique involves
dissolving chitosan in an aqueous acidic solution. This solution is then added to the
polyanionic TPP solution and stirred constantly. To form cross-linked chitosan
nanoparticles, the abundant NH3+ group in chitosan molecules reacts with the
negatively charged phosphoric ions in TPP. During the cross-linking and hardening
process, water was extruded from the particles, which may aid in the drug's long-
term release.

Beyond the significant improvements of encapsulation for drug formulations

in terms of bioavailability, the ionotropic gelation method has other advantages.
High encapsulation efficiency, close to 100%, is achieved when interactions among
the drug and polymer are optimal [Hoo CM et al., 2008]. The use of natural and
biodegradable biocompatible polymers provides the formulation with muco-
adhesion and other relevant biological properties. Ionotropic gelation does not
necessitate the use of expensive equipment or reagents and it is a relatively
inexpensive and quick method.

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