Noscapine based oral colon SpecificNanoparticles by Kuldipsinh Thakor

SHREE NARANJIBHAI LALBHAI PATEL COLLEGE OF PHARMACY(268),
UMRAKH
Presented By:
Kuldipsinh N. Thakor
Enroll. No. : 152680808008
M.Pharm., Sem- III,
Pharmaceutics
Guided By:
Dr. Jitendra Singh Yadav
M.Pharm., Ph.D.,
Assistant Professor & HOD
Dept of Pharmaceutics
DESIGN,DEVELOPMENT AND EVALUATION OF
ORAL COLON SPECIFIC NANOPARTICLES OF
NOSCAPINE FOR TREATING CANCER

INTRODUCTION
DRUG PROFILE
NEED, OBJECTIVE AND PLAN OF RESEARCH WORK
LITERATURE REVIEW AND PATENT SEARCH
MATERIALS AND METHODOLOGY WITH RESULTS AND DISCUSSION
SUMMARY AND CONCLUSION
REFERENCES

• Drug delivery to the colon should be capable of protecting the drug en
route to the colon i.e. drug release and absorption should not occur in the
stomach as well as the small intestine and neither the bioactive agent
should be degraded in either of the dissolution sites but only released and
absorbed once the system reaches the colon.
• The colon specific drug delivery System (CDDS) is beneficial not only for
the oral delivery of proteins and peptide drugs (degraded by digestive
enzymes of stomach and small intestine) but also for the delivery of low
molecular weight compounds used to treat diseases associated with the
colon or large intestine such as ulcerative colitis, diarrhoea and colon
cancer.

The GI tract is divided into the stomach and the small and large intestine. The large
intestine extending from the ileocecal junction to the anus, is divided into three main
parts. These are the colon, the rectum and the anal canal. For the purpose of colonic drug
delivery, there are two important physiological factors to be considered these are pH and
GI transit time. The mid and left colon have pH values of approximately 6.6 and 7.0.
Interspecies variability in pH is a major concern when developing and testing colon-
specific delivery systems

• Colon is an ideal site for the delivery of agents to cure the
local diseases of the colon.
• Local treatment has the advantage of requiring smaller drug
quantities.
• Reduces dosage frequency. Hence, lower cost of expensive
drugs.
• Possibly leading to a reduced incidence of side effects and
drug interactions.
• Improve patient compliance.
• Targeted drug delivery system.

• Multiple manufacturing steps.
• The resident microflora could also affect colonic
performance via metabolic degradation of the drug.
• Incomplete release of drug Bioavailability of drug may
be low due to potentially binding of drug in a
nonspecific way to dietary residues, intestinal
secretions, mucus or faecal matter.
• An important limitation of the pH sensitive coating
technique is the uncertainty of the location and
environment in which the coating may start to dissolve.
Normal in patients with ulcerative colitis.

• Targeted drug delivery to the colon to ensure that direct treatment at
the disease site (local delivery),at lower dosing and fewer systemic
side effects.
• Site-specific or targeted drug delivery system would allow oral
administration of peptide and protein drugs, colon specific
formulation could also be used to prolong the drug delivery.
• Colon-specific drug delivery system is considered to be beneficial in
the treatment of colon disease.
• A number of others serious diseases of the colon, e.g. colorectal
cancer, might also be capable of being treated more effectively if
drugs were targeted to the colon.

• Colon Cancer
• Colorectal Polyps
• vomiting and Fever
• Traveler’s Diarrhoea
• Diarrhoea
• Hirschsprung’s disease
• Salmonellosis
• Colon Bleeding
• Diverticulitis
• Diverticulosis
• Amoebiasis
• Ulcerative Colitis
• bowel disease
• Crohn’s Disease

[A] Primary Approaches:-
I. pH sensitive polymer coated
drug delivery to colon
i) Enteric-coated time
release press coated tablets
I. Delayed (Time controlled
release system) release drug
delivery to colon
II. Microbial triggered drug
delivery system
i) Prodrug approach for
drug delivery to colon
ii) Polysaccharide based
delivery systems
[B] Newly Developed Approaches:-
I. Pressure controlled drug-delivery
systems
II. Pulsatile colon targeted drug
delivery
i) Pulsincap system
ii) Port system
III. Osmotically controlled colon
targeted drug delivery system
IV. CODES technology
V. Multi particulate system based
drug delivery

1) Physiological Factor:-
• Gastric emptying
• pH of colon
• Colonic microflora and
enzymes
2) Pharmaceutical Factor:-
• Drug candidates
• Drug carriers

• Polymer contain a large number of structural unit joined by same
type linkage, form into a chain like structure. These are nowadays
used in formulating various pharmaceutical products.
• Natural polymer
Guar gum, Insulin, Pectin, Cyclodextrin, Dextran, Amylase,
Chitosan, Chondrotin sulphate, Locust bean gum.
• Synthetic polymer
Shellac, Ethyl cellulose, Cellulose acetate phthalate, Hydroxy
propyl methyl cellulose, Eudragit, Poly vinyl acetate phthalate

1) In Vitro Evaluation:-
 In vitro dissolution test:-
Dissolution of controlled-release
formulations used for colon-specific
drug delivery are usually complex,
and the dissolution methods described
in the USP cannot wholly mimic in
vivo conditions such as those relating
to pH, bacterial environment and
mixing forces.
 In vitro enzymatic test:-
incubate carrier drug system in
fermenter containing suitable medium
for bacteria (Streptococcus faccium or
B.ovatus) amount of drug released at
different time intervals determined.
2) In Vivo Evaluation:-
A number of animals such as
dogs, guinea pigs, rats and pigs
are used to evaluate the delivery
of drug to colon because they
resemble the anatomic and
physiological conditions as well
as the microflora of human GIT

• Colon cancer is also known as colorectal cancer that involves the colon, the parts of
large intestine and rectum. Colorectal cancer is known as the third most common
cancer in the world and the second most common cause of deaths due to cancer.
Usually, the development of colorectal cancer takes place slowly over a period of
10-15 years from noncancerous polyps that develop on the lining of colon or
rectum.
• Various approaches have been used by researchers for targeting drugs successfully
to the colonic region by circumventing the upper part of gastrointestinal tract. These
approaches include pH-dependent, time-dependent and microflora-activated
systems. The most commonly used drug candidates for the treatment of colorectal
cancer comprises of 5-fluorouracil (5-FU), leucovorin, oxaliplatin and capecitabine.
Apart from these, other drug molecules that have been reported to play a significant
role in prevention of colorectal cancer are meloxicam, curcumin, valdecoxib,
resveratrol, indomethacin, celecoxib, methotrexate, gemcitabine, transcutol and
ginger extract.

• Classification of cancer by anatomic disease extent is one of the most important
factors for prognosis and therapeutic decision. Based on TNM (T: primary tumour
site; N: regional lymph node involvement; M: presence or otherwise of distant
metastatic) and UICC (Union for International Cancer Control) staging system,
CRC can be treated by surgery, chemotherapy, radiation, immunotherapy or
palliative care. Surgical resection offers high cure rates for CRC in early stages.

COLON
CANCER
STAGE I
(At this,
polyp has
progress)
STAGE II
(Spread to
Colon)
STAGE III
(Spread
outside of
colon)
STAGE IV
(Spread
other organ
of body)
STAGE 0
(Earlier
stage)

• A Nanoparticle is a solid submicron drug carrier system that may or may
not biodegradable. The drug or molecules of interest may dissolve into the
nanoparticles to entrap or adsorb or attach to them. Such targeting ability
improves patient’s comfort and standard of normal life. Targeting
nanoparticles to tumors cells have shown distinct advantages over other
delivery systems. Firstly, they can be directly attracted to the tumor cells by
using different strategies. Secondly, due to their nanosize, these colloidal
systems penetrate into rapid growing tumor mass and get concentrated
preferentially that can act as drug depot for controlled supply of
encapsulated therapeutic compound for extended period into tumor mass.
• The nanoscale size also allows for interaction with biomolecules on cell
surfaces and within cells in such a way that does not alter the behavior and
biochemical properties of these molecules.

• Among these systems, nanoparticle mediated delivery provides a number of
advantages including:
1) Small particle size,
2) Increased drug efficacy,
3) Lowered toxicity,
4) Enhanced drug solubility and stability, and
5) An ability to achieve steady-state therapeutic levels over an extended
time period.
• The passive tumor targeting involves modification or designing the
molecular weight of the system, size of the system, surface hydrophobicity,
and surface charge of the system in order to keep it long circulating in the
blood and target it to desired sites. In cancer tissues, the phenomenon of
“Enanced permeability and retention” (EPR) uniquely modifies the tumor
microenvironment promoting angiogenesis, hypervascularization, defective
vascular architecture, impaired lymphatic drainage recovery system and
increased production of permeability mediators. This passive mechanism
can be exploited for selective targeting of cancer tissue by nanoparticles.

• Stable in blood
• Non toxic
• Non thrombogenic
• Non immunogenic
• Non inflammatory
• No activation of neutrophils
• Biodegradable
• Avoidance of the reticulo-endothelial system
• Applicable to various molecules, such as small molecules,
proteins, peptides or nucleic acids (platform technology)
• Scalable and inexpensive manufacturing process

1 • Solvent Evaporation
2 • Nanoprecipitation
3 • Emulsification/solvent diffusion
4 • Salting out
5 • Dialysis
6 • Super critical fluid technology

ATTRIBUTE DETAILS
MOLECULAR STRUCTURE
MOLECULAR FORMULA C22H23NO7
MOLECULAR WIEGHT 413.42g/mol
IUPAC NAME (3S)-6,7-Dimethoxy-3-[(5R)-4-
methoxy-6-methyl-
5,6,7,8-tetrahydro[1,3]dioxolo[4,5-
g]isoquinolin-
5-yl]isobenzofuran-1(3H)-one

DESCRIPTION
Noscapine occurs as light yellowish crystals or
crystal-
line powder. It is odorless and tasteless.
SYNONYMS Narcotine, Nectodon, Nospen, Anarcotine Opiane
SOLUBILITY
very soluble in acetic acid, acetone and HCl
slightly soluble in ethanol and in diethyl ether,
practically insoluble in water.
MELTING POINT 174 -177 ˚C.
CATEGORY Centrally acting antitussive
CAS No. 128-62-1
LogP 5.0
pKa
7.8
242424

Mechanism of action :
•It is known that noscapine is centrally acting in the suppression of
cough.
•Noscapine inhibit bradykinin as the mode by which it functions.
•Noscapine’s effect in treating strokes has been attributed to its
bradykinin suppressive effect.
•Noscapine’s anti-cancer effect has been primarily attributed to its
microtubuleinterfering effect.
Pharmacokinetic:
Bioavailability -30%
Biological half-life 1.5 to 4h (mean 2.5)

USES:
• Cough suppressant.
• Usual doses 45 to 200 mg /day, usually in three
divided doses.
•1000-2250 mg/day in three divided doses for
cancer(from research paper)
• Off-label use of noscapine : Treating cancers of the
breast, lung, prostate ovaries and colon
• It gives pain-relief effects by combining it with lower
than usual doses of morphine(sometimes).

• Targeted drug delivery to the colon to ensure that direct treatment at the
disease site (local delivery),at lower dosing and fewer systemic side
effects. Site-specific or targeted drug delivery system would allow oral
administration of peptide and protein drugs. Colon-specific drug delivery
system is considered to be beneficial in the treatment of colon disease. A
number of others serious diseases of the colon, e.g. colorectal cancer,
might also be capable of being treated more effectively if drugs were
targeted to the colon.
• Noscapine is a very safe centrally acting cough suppressant (Antitussive), a
non-addictive, non-narcotic derivative of opium. Noscapine also used in
cancer treatment. It has long safety record, widespread availability and
ease of administration.

• Nanoparticles Fairly easy preparation. Due to their small size
Nanoparticles penetrate small capillary and are taken up by the cell which
allows for efficient drug accumulation at the target sites in the body.
Increased surface area results in a faster dissolution of active agents in an
aqueous environment. Faster dissolution generally equates with greater
bioavailability.
• Thus the present study focused on development of noscapine loaded
colon targeting Nanoparticles to improve bioavailability of drug and
reduced the dose by controlling the release of the drug to treat colorectal
cancer

• To perform preformulation study of drug for
identification and characterization
• To check compatibility study of drug with
excipient.
• To formulate and optimize Nanoparticle by
considering independent variables to characterize
Nanoparticles
• Targeting the drug to the colon to improve
efficiency and increasing bioavailability.
• To controlled the release to reduce the side effect

1. Identification and characterization of drug by different test
2. UV analysis of drug by:-
a. Selection of wavelength maxima
b. Calibration curve preparation in different media
3. Preformulation study
4. Saturated solubility of drug
5. Drug excipient compatibility study
6. Preliminary batches preparation for identification of independent
and dependent variables
7. Development of formulation using by apply 32 experimental designs
8. Preparation and Evaluation of Noscapine Based Nanoparticles
9. Accelerated stability study as per ICH
10. Cytotoxicity Study

Author’s
Name
Title Description Journal
Name
Chao Wang
et al. (2014)
Suppression of
colorectal
cancer
subcutaneous
xenograft and
experimental
lung metastasis
using
nanoparticle-
mediated drug
delivery to
tumor
neovasculature
The study was to investigate a new tumor
vessel targeting nanoparticulate drug
delivery system, F56 peptide conjugated
nanoparticles loading vincristine (F56-VCR-
NP), for the effective treatment of CRC
subcutaneous xenograft and experimental
lung metastasis model. About 18.3%
vincristine from F56-VCR-NP was released on
the first day, and then 65.5% was released in
the next 29 days. The initial fast release can
be ascribed to the drug near the particle
surface, while the slower and sustained
release may be from the diffusion of the drug
inside the nanoparticles or be facilitated by
the water-mediated erosion of the polymer
matrix.
Biomateria
ls
journal
homepage
:
www.elsev
ier.com

Regis
Coco et
al. (2013)
Drug delivery to
inflamed colon by
nanoparticles:
Comparison of
different
strategies
For inflammatory bowel disease (IBD)
treatment, local delivery of molecules
loaded in nanoparticles to the
inflamed colon could be a promising
strategy. The aim of this study was to
investigate how drugloaded polymeric
nanoparticles target the site of
inflammation and to analyse the
influence of different colon-specific
delivery strategies. Three different
polymeric nanoparticles were
formulated using ovalbumin (OVA) as
a model drug. pH-sensitive
nanoparticles were made with
Eudragit® S100.
International
Journal of
Pharmaceutics
Arvind
Gulbake
et al.
(2013)
COLON SPECIFIC
DELIVERY OF
MESALAZINE
USING
BIOCOMPATIBLE
POLYMERIC
NANOPARTICLES
Chitosan nanoparticles (CH-NPs)
bearing mesalazine (MSZ) were
developed by ionotropic gelation
method and encapsulated in Eudragit
S100 coated pellets for site specific
delivery to ulcerative colitis.

R. Seda Tigli
Aydin et al.
(2012)
5-Fluorouracil
Encapsulated
Chitosan
Nanoparticles
for pH-Stimulated
Drug Delivery:
Evaluation of
Controlled
Release Kinetics
5-fluorouracil (5-FU)
encapsulated chitosan
nanoparticles were
prepared in order to
investigate potentials of
localized drug delivery
for tumor environment
due to pH sensitivity of
chitosan nanoparticles.
Results suggested that 5-
FU encapsulated
chitosan nanoparticles
can be launched as pH-
responsive smart drug
delivery agents for
possible applications of
cancer treatments.
Journal of
Nanomaterials

Jingling Tang
et al. (2011)
Eudragit
nanoparticles
containing
genistein:
formulation,
development, and
bioavailability
assessment
To improve the oral bioavailability of genistein,
genistein nanoparticles were prepared by the
nanoprecipitation technique using EudragitR
E100 as carriers and an optimized formulation
of mass ratio (genistein:Eudragit E100, 1:10).
Encapsulation efficiency and drug loading of
the genistein nanoparticles were approximately
50.61%.These results suggested that a
nanoparticle system is a potentially promising
formulation for the efficient delivery of water-
soluble drugs by oral administration.
Internationa
l Journal of
Nanomedici
ne
Meltem Cetin
et al. (2010)
Formulation and
In vitro
Characterization
of Eudragit® L100
and Eudragit®
L100-PLGA
Nanoparticles
Containing
Diclofenac
Sodium
The aim of this study was to formulate and
characterize Eudragit® L100 and Eudragit®
L100-poly(lactic-co-glycolic acid) (PLGA)
nanoparticles containing diclofenac sodium.
Thus, nanoparticles were prepared to reduce
these drawbacks in the present study. These
nanoparticles were evaluated for surface
morphology, particle size and size distribution,
percentage drug entrapment, and in vitro drug
release in pH 6.8. The prepared nanoparticles
were almost spherical in shape, as determined
by atomic force microscopy
American
Association
of
Pharmaceuti
cal Scientists

Author’s Name Title Description Journal Name
Safaa Sebak at al.
(2010)
Human serum
albumin
nanoparticles as an
efficient
noscapine drug
delivery system for
potential use
in breast cancer:
preparation and in
vitro analysis
Drug delivery systems such as
nanoparticles can provide
enhanced efficacy for Anticancer
agents. Noscapine, a widely used
cough suppressant for decades
has recently been shown to
cause significant inhibition and
regression of tumor volumes
without any detectable toxicity
in cells or tissues. Nanoparticles
made of human serum albumin
(HSA) represent Promising
strategy for targeted drug
delivery to tumor cells by
enhancing the drug’s
bioavailability and distribution,
and reducing the body’s
response towards drug
resistance.
International
Journal of
Nanomedicine
2010

Ritu Aneja et al.
(2007)
p53 and p21
Determine the
Sensitivity of
Noscapine-
Induced
Apoptosis in
Colon Cancer
Cells
naturally occurring antitussive alkaloid
noscapine as a tubulin-binding agent that
attenuates microtubule dynamics and
arrests mammalian cells at mitosis via
activation of the c-Jun NH2-terminal kinase
pathway. It is well established that the p53
protein plays a crucial role in the control of
tumor cell response to chemotherapeutic
agents and DNA-damaging agents; however,
the relationship between p53-driven genes
and drug sensitivity remains controversial.
www.aa
crjourn
als.org
Ritu Aneja et al.
(2006)
Development of a
Novel Nitro-
Derivative of
Noscapine for the
Potential
Treatment of
Drug-Resistant
Ovarian Cancer
and
T-Cell Lymphoma
an antitussive plant alkaloid,noscapine,
binds tubulin, displays anticancer activity,
and has a safe pharmacological profile in
humans.
Thus, we conclude that 9-nitro-nos has great
potential to be a novel therapeutic agent for
ovarian and T-cell lymphoma cancers, even
those that have become drug-resistant to
currently available chemotherapeutic drugs.
The
Americ
an
Society
for
Pharma
cology
and
Experim
ental
Therap
eutics.

Jun Zhou et al.
(2002)
Paclitaxel-resistant
Human Ovarian
Cancer Cells
Undergo c-Jun
NH2-terminal
Kinase-mediated
Apoptosis in
Response to
Noscapine
the opium alkaloid noscapine as
a microtubule interacting agent
that binds to tubulin, alters the
dynamics of microtubule
assembly, and arrests
mammalian cells at mitosis
inhibition of JNK activity by
treatment with antisense
oligonucleotide or transfection
with dominant-negative JNK
blocks noscapine-induced
apoptosis. These findings thus
indicate a great potential for
noscapine in the treatment of
paclitaxel-resistant human
cancers. In addition, our results
suggest that the JNK pathway
plays an essential role in
microtubule inhibitor-induced
apoptosis.
THE JOURNAL
OF BIOLOGICAL
CHEMISTRY

Author Matrix Method Description
Indian
pharmacopoeia
Noscapine
Bulk
Potentiometric
titration
Solution:0.35 g + 40 m1 of anhydrous glacial acetic acid
Titrant :0.1 M perchloric acid 1 ml of 0.1 M perchloric acid is
equivalent to 0.04134 g of C22H23NO7
Indian
pharmacopoeia
Noscapine
linctus
UV
spectroscopi
method
ƛmax : 310 nm
Solvent : water
Specific absorbance :90.7
British
Pharmacopoeia
2013
Noscapine
Hydrochlori
de bulk
Potentiometric
titration
Solution: 0.400 g +5.0 ml of 0.01 M hydrochloric acid + 50 ml
of alcohol R.
Titrant : 0.1 M sodium hydroxide 1 ml of 0.1 M sodium
hydroxide is equivalent to 44.99 mg of C22H23NO7
European
pharmacopoeia
Noscapine
Bulk
Potentiometric
titration
Solution:0.35 g + 40 m1 of anhydrous glacial acetic acid
Titrant :0.1 M perchloric acid 1 ml of 0.1 M perchloric acid is
equivalent to 41.34 mg of C22H23NO7
Japanese
pharmacopoeia
Noscapine
Bulk
Potentiometric
titration
Solution:0.8 g + 30 mL of acetic acid (100)
Titrant :0.1 M perchloric acid VS 1 ml of 0.1 M perchloric acid
is equivalent to 41.34 mg of C22H23NO7
US
pharmacopoeia
Noscapine
Bulk
Potentiometric
titration
Solution:1.5g of Noscapine +25 mL of dioxin and Indicator : 5
drops of crystal violet.
Titrant :0.1 M perchloric acid VS 1 ml of 0.1 N perchloric acid
VS is equivalent to 41.34 mg of C22H23NO7

SR. NO PATENT APPLICATION
NUMBER
PATENT TITLE
1 10056913 Delivery systems and methods for noscapine
and noscapine derivatives, useful as anticancer
agents
2 13488621 Noscapine analogs and their use in treating
cancers
3 217344 colon-targeted delivery system
4 DE19906290 Orally administered medicament for treating
colon cancer comprises ursodesoxycholic acid
in gastric fluid resistant coating to provide
direct topical action at target
5 CN 201310340081 Biodegradable nanoparticle-entrapped oral
colon-targeted micro-capsule and preparation
method thereof

Sr.No Patent Application
No
Patent Title Description
1 10056913 Delivery systems
and methods for
noscapine and
noscapine
derivatives, useful
as anticancer
agents
Applicants have discovered that the
antitussive noscapine and its derivatives
are useful in the treatment of neoplastic
diseases. Noscapine is used as an
antitussive drug and has low toxicity in
humans. Noscapine arrests mammalian
cells at mitosis, causes apoptosis in cycling
cells, and has potent antitumor activity.
Noscapine is an alkaloid from opium, and
is readily available as a commercial by-
product in the commercial production of
prescription opiates. Applicants have
unexpectedly discovered that noscapine
promotes assembly of tubulin subunits.

No
2 13488621 Noscapine analogs
and their use in
treating cancers
The patent or application file contains at
least one drawing executed in color. Copies
of this patent or patent application
publication with color drawing(s) will be
provided by the Office upon request and
payment of the necessary fee.
Synthesis of 9-nitronoscapine from
noscapine, using silver nitrate in
acetonitrile and TFAA at 25° Cshows a
double reciprocal plot which gives a
dissociation constant (Kd) of 86±6 μM for
9-nitro-nos binding to tubulin
9-nitro-nos selectively kill cancer cells by
inducing apoptosis, as demonstrated by
devastating changes in the nuclear
morphology of these cells.

No
3 217344 colon-targeted
delivery system
The dual matrix tablet containing the core
and the erodible polymer layer may be
prepared by any conventionalmeans For
example, using a suitable tablet
compactionpress, half of the erodible
polymer layer matrix granulationmay be
placed in the die cavity, and the core,
previouslyprepared is centered in the die
cavity. The other half of theerodible
polymer layer matrix granulation is placed
in thedie cavity and the mass may then be
compressed at a suitablepressure, e.g.,
5,000 lbs. Alternatively dual matrix
tabletscould be prepared using
compression coating technique withaDri-
Coata Tablet Press

Sr.No Patent
Application No
Patent Title Discription
4 DE19906290 Orally
administered
medicament for
treating colon
cancer comprises
ursodesoxycholic
acid in gastric fluid
resistant coating to
provide direct
topical action at
target
The invention relates to pharmaceutical
compositions, particularly drugs with controlled
release profile, with ursodeoxycholic acid (UDCA),
pharmaceutically acceptable salts or derivatives
thereof as an active ingredient. Die Arzneimittel
dienen zur Behandlung bzw. Prävention von
Krebserkrankungen im Dickdarm. The medicaments
are used for treatment or prevention of cancers in the
colon. Im Unterschied zu bereits bekannten UDCA
enthaltenden Arzneimitteln wird bei den
erfindungsgemäßen Arzneimitteln der Wirkstoff erst
im terminalen Ileum und/oder Dickdarm freigesetzt
und zwar bevorzugt kontrolliert freigesetzt, so daß
der Wirkstoff nur wenig resorbiert wird. In contrast to
already known medicinal UDCA containing the active
ingredient is only released in the terminal ileum and /
or colon released and namely preferably controlled so
that the active agent is only slightly absorbed in the
pharmaceutical compositions of the invention. Damit
wirkt UDCA erfindungsgemäß nicht systemisch
sondern topisch direkt am Ort der Freisetzung. This
UDCA acts according to the invention not systemic
but topically directly at the site of release.

Sr.No Patent
Application No
5 CN
201310340081
Biodegradable nanoparticle
entrapped oral colon-targeted
micro-capsule and preparation
method thereof
The invention belongs to the
field of microcapsule
preparation technology, in
particular relates to an oral
colon targeting microcapsule
with a biodegradable
nanoparticles and a
preparation method thereof

Looking at above 05 patents, your Dissertation project is novel up to what extent?
Novelty grade: 50 -75%
Looking at the above 05 Patents, please explain about rational of selection of Your
dissertation project?
It is known that noscapine is centrally acting in the suppression of cough. Noscapine inhibit
bradykinin as the mode by which it functions.Noscapine’s effect in treating strokes has been
attributed to its bradykinin suppressive effect. Noscapine’s anti-cancer effect has been
primarily attributed to its microtubuleinterfering effect. Recently, noscapine has been found to
have antiangiogenic properties (preventing the formation of new blood vessels).
The Patent Search report comprise of preparation of nanoparticles along with the noscapine
nanoparticles to improve the bioavailability. nascapine is highly lipophilic compound
therefore it has a very low aqueous solubility. noscapine has shown to possess a wide range of
pharmacological activity including anticancer, antitussive, anticough, enhanced by
formulating it as microemulsion , solid lipid nanoparticle or microparticles. It is evidence that
noscapine is very used for anticancer activity. And colon targeted is best way to target the
drug to colon to use Nanoparticle.
Thus, the above patent search will be useful for preparing nanoparticles containing
noscapine for prevention against colon cancer.

SR.NO NAME OF MATERIAL NAME OF SUPPLIERS
1 Noscapine Bharat Parentals Pvt.Ltd
2 Eudragit S100 Chemdyes
3 Poly viny alcohol Sulab
4 Acetone Sulab
5 Methanol Sulab
6 Potassium dihydrogen phosphate Sulab
7 Sodium hydroxide Sulab
8 Hydrochloric acid Sulab

EQUIPMENT COMPANY/MODEL
Digital analytical balance Shimadzu Corporation
UV spectroscopy Shimadzu (UV-1700)
IR spectroscopy Shimadzu FTIR 8400S
Sonicator Trans-O-Sonic (D-compact)
Differential Scanning Calorimetery Shimadzu DSC EA-60
Rota evaporator Rocker 300
Zeta sizer nano ZS Microtrac
Liophilizer MAC Scientific Work
Centrifuge Lab Line
SEM HITACHI, S-3400N

PREFORMULATION STUDY
1.Organoleptic characteristics of Noscapine
The colour of noscapine was visualized light yellowish odourless or with a faint characteristic
odour having light yellowish crystalline powder appearance as shown in Table.
Parameters Observed Result
Colour Light yellowish powder
Odour Odourless
Appearance Light yellowish crystalline powder
Table : Organoleptic Characteristics of Noscapine

2. Determination of Melting Point of Noscapine:
Melting point was carried out to determine the purity of the sample using capillary method. The
drug sample has melting point of 174-175 °C which was near to standard value and indicate the
purity of sample as Noscapine
Table : Melting point of Noscapine
Drug Name
Standard Value
Observed Value
(Mean ± S.D.)
(n = 3)
Noscapine 174-178°C 174-175±1°C

3.Solubility Study of Noscapine:
Solubility is expressed in terms of parts per million of solvent in which 1 g of solid is soluble.
Solubility of the drug in different solvents are determined and expressed in mg/ml. The procedure
was follow by using different solvent like distilled water, acetone, ethanol, 0.1N HCL, acetic
acid, and diethyl ether.
Table : Solubility of Noscapine
Solvent Solubility of Noscapine
Parts of solvent
required
for part of solute
Measured
value
Distilled Water Insoluble
10,000 or more 1 mg in 1000 ml
Ethanol Slightly Soluble 100 to 1000 1 mg in 10 ml
Acetic acid Freely soluble
From 1 to 10 1 mg in 0.5 ml
Acetone Very Soluble
Less than 1 1 mg in 0.1 ml
Hydrochloric Acid Very Soluble
Less than 1 1 mg in 0.1 ml

4. Identification and Determination of Wavelength max (λmax) of Noscapine
Standard solution of Noscapine was scanned 200-400 nm against 0.1 N HCl as a blank reagent.
The spectrum of solution was obtained
Table : λmax Spectrum for Noscapine
Drug Name Observed λmax
Noscapine 213 nm

5.Preparation of Calibration Curve for Noscapine
Calibration curve of Noscapine in 0.1N HCl, Phosphate Buffer 6.8 pH, Phosphate Buffer
7.4 pH showed the linearity in different concentration range which followed Beer
Lambert’s law. The R2 value were found to be 0.996 for 0.1N HCL, 0.995 in P.B pH 6.8,
0.981 in P.B pH 7.4
Table : Calibration curve of Noscapine
Sr No. Concentration
(μg/mL)
Absorbance
(in 0.1N HCL)
(mean ± SD)
(n = 3)
Concentration
(μg/mL)
Absorbance
(in pH 6.8)
(mean ± SD)
(n = 3)
Concentration
(μg/mL)
Absorbance
(in pH 7.4)
(mean ± SD)
(n = 3)
1 1
0.177 ± 0.038
2 0.174 ± 0.027 2 0.170 ± 0.088
2 3
0.331 ± 0.026
4 0.361 ± 0.012 4 0.339 ± 0.012
3 5
0.524 ± 0.030
6 0.510 ± 0.008 6 0.495 ± 0.017
4 7
0.721 ± 0.026
8 0.660 ± 0.025 8 0.716 ± 0.017
5 9
0.935 ± 0.028
10 0.872 ± 0.020 10 0.917 ± 0.017

0
0.177
0.331
0.524
0.721
0.935
y = 0.099x
R² = 0.994
0
0.2
0.4
0.6
0.8
1
1.2
0 5 10
C.C in 0.1 N HCL
Series1
Linear (Series1)
Concentration((μg/ml)
a
b
s
o
r
b
a
n
c
e
0.174
0.361
0.51
0.66
0.872
y = 0.0857x
R² = 0.9957
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
0 5 10 15
C.C in P.B pH 6.8
absorbance
Linear (absorbance)
a
b
s
o
r
b
a
n
c
e
0.17
0.339
0.495
0.716
0.971
y = 0.0914x
R² = 0.9818
0
0.2
0.4
0.6
0.8
1
1.2
0 5 10 15
C.C in P.B pH 7.4
absorbance
Linear
(absorbance)
a
b
s
o
r
b
a
n
c
e
Figures : Calibration curve of Noscapine

Parameters 0.1N HCL
P.B pH 6.8 P.B pH 7.4
Wavelength (λmax) 213nm
213nm 213 nm
Beer’s limit (μg/ml) 1-9
2-10 2-10
Corrélation coefficient (R2) 0.994
0.995 0.981
Slope 0.061
0.065 0.063
Obeys Beer law in conc. range of
R2 value shows linearity
Table : Summary report of calibration curve for Noscapine

6.Identification of Drug-Noscapine by FTIR
Figure : Identification of Noscapine by IR Spectrum
Infrared spectrum of reference Noscapine (Japanese Pharmacopeia)
Sr. No. Functional group
stretching
Standard Wave
number (cm-1)
Wave number (cm-1)
1 C=O (ester) 1735-1750 1756
2 C-O 1000-1300 1037, 1005
3 C=C (aromatic) 1400-1600 1500, 1620
4 C-H(Alkane) 2850-3000 2946
5 C-N(amine) 1080-1360 1276
Table : Identification of IR peaks of Noscapine

7. Identification of Drug-Noscapine by DSC
Figure : DSC graph of Noscapine
The obtained FT-IR spectrum and DSC graph compiles with
standard data which further confirms Drug identity and purity.
Endothermic Peak
of Noscapine :
178.78 ̊C
50.00 100.00 150.00 200.00 250.00 300.00
Temp [C]
-20.00
-10.00
0.00
mW
DSC
178.78 x100
C
Thermal Analysis Result
Noscapine
The DSC thermo gram of noscapine analyses was conducted to explore the melting activities of drug. DSC
analysis showed a sharp endothermic peak at 178.78°C which is an indication melting point of noscapine.
Performed melting pointing capillary method get corresponded to the melting point of the pure drug (174-175°).
So, it was found same in authentic range of official standards.

Drug Excipients compatibility study by FTIR[39]
Noscapine + Ethyl Cellulose FTIR Spectrum
Table : IR peaks of Noscapine+ Eudragit S100
Noscapine + Eudragit S100 IR Spectrum:
Figure : FTIR Spectrum of Noscapine + Eudragit S100
stretching
Standard Wave
number (cm-1)
Wave number (cm-1)
1 C=O (ester) 1735-1750 1731
2 C-O 1000-1300 1037, 1005
3 C=C (aromatic) 1400-1600 1500, 1621
4 C-H(Alkane) 2850-3000 2949
5 C-N(amine) 1080-1360 1276

Figure : FTIR Spectrum of Noscapine + PVA
Table : IR peaks of Noscapine + PVA
Noscapine + PVA IR Spectrum:
Sr. No. Functional
group stretching
Standard Wave number
(cm-1)
Wave number (cm-1)
1 C=O (ester) 1735-1750 1756
2 C-O 1000-1300 1037, 1005
3 C=C (aromatic) 1400-1600 1500, 1621
4 C-H(Alkane) 2850-3000 2946
5 C-N(amine) 1080-1360 1276

62
Figure : FTIR Spectrum of Noscapine + Potassium dihydrogen Phosphate
Table : IR peaks of Noscapine + Potassium dihydrogen Phosphate
Noscapine + Potassium dihydrogen Phosphate IR Spectrum:
Sr. No. Functional
group stretching
Standard Wave number
(cm-1)
Wave number (cm-1)
1 C=O (ester) 1735-1750 1757
2 C-O 1000-1300 1037, 1005
3 C=C (aromatic) 1400-1600 1500, 1620
4 C-H(Alkane) 2850-3000 2946
5 C-N(amine) 1080-1360 1276

63
Figure : FTIR Spectrum of Noscapine + ALL Excipients
Table : IR peaks of Noscapine + ALL Excipients
Noscapine + ALL Excipients IR Spectrum:
stretching
Standard Wave
number (cm-1)
Wave number (cm-1)
1 C=O (ester) 1735-1750 1731
2 C-O 1000-1300 1037, 1005
3 C=C (aromatic) 1400-1600 1500, 1620
4 C-H(Alkane) 2850-3000 2948
5 C-N(amine) 1080-1360 1275

Drug Excipients compatibility study by DSC[39]
DSC graph of Noscapine + Eudragit S100:
50.00 100.00 150.00 200.00 250.00 300.00
Temp [C]
-10.00
-5.00
mW
DSC
178.29 x100
C
Therm al Analysis Res ult
DSC analysis showed a sharp endothermic peak at 178.29°C it was found near to authentic
range of official standards. So, conclude that there was no interaction drug is pure.

DSC graph of Noscapine + PVA:
50.00 100.00 150.00 200.00 250.00 300.00
Temp [C]
-8.00
-6.00
-4.00
-2.00
mW
DSC
179.02 x100
C
DSC analysis showed a sharp endothermic peak at 179.02°C it was found near to authentic range
of official standards. So, conclude that there was no interaction drug is pure.

DSC graph of Noscapine + Potassium dihydrogen Phosphate:
50.00 100.00 150.00 200.00 250.00 300.00
Temp [C]
-20.00
-15.00
-10.00
-5.00
0.00
mW
DSC
178.28 x100
C
236.84 x100
C
265.90 x100
C

DSC graph of Noscapine + All Excipients:
50.00 100.00 150.00 200.00 250.00 300.00
Temp [C]
-3.00
-2.00
-1.00
0.00
mW
DSC
178.05 x100
C
214.58 x100C
234.78 x100
C
As All The Graph Does Not Show Large Deviation As Compared With The DSC Graph of
Pure Drug, We Can Conclude That The Drug And Excipients Are Compatible With Each
Other

Dose of noscapine calculated using factor method. The dose by factor method applies an
exponent for bodysurface area (0.67), which account for difference in metabolicrate, to convert
doses between animals and humans. Thus, HED is determined by the equation:
• HED (mg / kg) = Animal NOAEL (mg / kg) × (Weightanimal[kg] /Weighthuman [kg])(1-0.67)
where, NOAEL- no observed adverse effect levels
Dose for Mice = 10 mg/kg
Weight of mice = 150 mg
Weight of human = 60 kg
HED = Human Equivalent Dose
HED = Animal dose (mg/kg) x (Wight Animal [kg] / WightHuman [kg])(1-.6)
= 10 x (0.15/60)(.33)
= 10 x (0.0025)(0.33)
= 10 x 0.1384
= 1.3845 mg/kg
= 83.075 mg
= 80 mg

Batch B1 B2 B3
Noscapine 80mg 80mg 80mg
Eudragit S100 200mg 600mg 1000mg
Acetone:Methanol(2:1) 40ml 40ml 40ml
PVA (%w/v) 0.1 0.2 0.3
Distilled water 80ml 80ml 80ml

To reduce particle size while maintaining a high amount of drug
inside the NPs, the concentration of PVA, and Eudragit S100 was
modified resulting in three additional formulations result are shown
in table. B1,B2,B3 which the amount of Eudragit S 100 and the PVA
concentration was increased, reduction in NP size relates to the
increase in PVA concentration and entrapment efficiency was
increases only up to an increasing amount of Eudragit S100. Trial
batch results reflect that higher amounts of PVA, significantly
reduced the Entrapment efficiency and by increasing the
concentration of the dissolved Eudragit S100 polymer resulting in
lower drug content and bigger emulsion droplets. So PVA and Drug:
Eudragit ratio were selected as critical parameter to control the
particle size, entrapment efficiency and release.

• Nanoparticles is prepare by a “water-in-oil-in water” method. Briefly, 10 ml of
solution of Noscapine is emulsify with 40 ml of a mix of acetone/methanol 2:1
containing polymer with an ultrasonic processor for 30 s at 70 W it formed primary
emulsion.
• The secondary emulsion was prepare by adding the first emulsion in 80 ml of
aqueous solution of polymer before ultrasonication for 1 min at 70 W.
• Finally organic phases were evaporated under reduced pressure in a rotary
evaporator at 800C. after evaporation of the solvent, Nanoparticle is recover by
centrifugation at 10,000 rpm for 40 min and wash with distilled water and
lyophilize overnight.
• All the batches is produce at least in triplicate.

Evaluation of Preliminary Batches
Batch
Code
Particle Size
(nm)
%E.E
(Mean ±S.D.)
(n=3)
Drug
Content(mg)
(Mean ±S.D.)
(n=3)
B1 115 nm 86.23±0.52 82.33±0.2
B2 123 nm 79.39±0.05 76.41±0.5
B3 140 nm 71.56±0.04 69.73±0.6
SEM Study of Preliminary Batch

Independent
Variable
Low(-1) Medium(0) High(+1)
Concentration
of Eudragit
S100 (X1)
1:1 1:2 1:3
Concentration
of
PVA (X2)
0.1 0.2 0.3
Dependent Variable
Y1=Particle Size
Y2= %CDR
Factorial Batches
Batch code Concentration of
(X1)
Concentration of
(X2)
N1 -1 -1
N2 -1 0
N3 -1 +1
N4 0 -1
N5 0 0
N6 0 +1
N7 +1 -1
N8 +1 0
N9 +1 +1
Compositions of Factorial Batches in Coded Form

Batch code Concentration of
(X1)
Concentration of
(X2)
N1 1:1 0.1
N2 1:1 0.2
N3 1:1 0.3
N4 1:2 0.1
N5 1:2 0.2
N6 1:2 0.3
N7 1:3 0.1
N8 1:3 0.2
N9 1:3 0.3
Formulation Design by 32 Factorial Designs
Batch B1 B2 B3 B4 B5 B6 B7 B8 B9
Noscapine(mg) 80 mg
Eudragit S100 (mg) 250 250 250 500 500 500 750 750 750
Acetone:Methanol(ml) 40 ml
PVA(%) 0.1 0.2 0.3 0.1 0.2 0.3 0.1 0.2 0.3
Distilled Water(ml) 80 ml
Formulation of Batches N1-N9

 Characterization of Batches N1-N9
Results And Discussion of 32 Full Factorial Design Formulations
Batch Code Particle Size(nm)
(Mean ±S.D.)
%E.E
(Mean ±S.D.)(n=3)
Drug Content(mg)
(Mean ±S.D.)
(n=3)
N1 116 83.21±0.6 80.39±0.9
N2 112 87.31±0.5 78.37±0.3
N3 107 89.32±0.3 84.83±0.5
N4 127 74.31±0.8 69.88±0.3
N5 125 76.54±0.7 70.32±0.5
N6 124 79.53±0.2 77.83±0.4
N7 143 65.31±0.3 64.42±0.5
N8 138 69.90±0.1 69.90±0.2
N9 131 73.73±0.2 71.39±0.7
Table.: Evaluation of Batch N1-N9
The parameter shows the result as given in table. The parameters including the
Particle Size, % E.E, and Drug Content of all the formulations are shown in
range of the limit.

 In-Vitro Drug Release Study
Table : Percent Cumulative Drug Release of batches N1-N9 with (Mean±S.D.) (n=3)
Time(Hr) N1 N2 N3 N4 N5 N6 N7 N8 N9
0 0 0 0 0 0 0 0 0 0
1 13.40±0.6 14.90±0.6 12.80±0.20 12.70±0.2 12.60±0.6 14.80±0.5 14.30±0.6 12.70±0.12 13.00±0.2
2 20.64±0.15 23.86±0.20 23.54±0.6 20.54±0.9 19.44±0.14 23.06±0.9 23.15±0.8 19.44±0.18 19.44±0.11
3 30.72±0.8 32.06±0.19 31.76±0.22 32.12±0.6 31.51±0.9 29.25±0.1 29.15±0.4 29.31±0.15 29.11±0.16
4 37.13±0.5 39.35±0.7 44.15±0.7 39.15±0.8 38.74±0.13 38.52±0.6 38.32±0.1 37.32±0.19 38.32±0.9
5 46.00±0.12 48.23±0.3 53.88±0.12 43.03±0.3 45.22±0.12 45.22±0.12 43.02±0.5 42.61±0.11 42.62±0.4
6 52.00±0.3 55.33±0.15 57.89±0.5 50.77±0.2 50.49±0.7 53.09±0.14 50.77±0.7 52.36±0.15 52.36±0.18
7 54.47±0.5 63.50±0.6 65.03±0.14 58.45±0.7 58.25±0.6 63.88±0.5 58.95±0.1 58.57±0.13 58.57±0.11
8 65.89±0.13 68.59±0.2 72.11±0.3 64.04±0.1 64.94±0.14 68.60±0.18 64.24±0.9 63.84±0.17 64.34±0.14
9 71.12±0.9 77.35±0.17 79.29±0.12 70.60±0.4 70.51±0.8 75.05±0.9 67.70±0.1 66.70±0.11 66.70±0.7
10 74.68±0.14 83.45±0.9 87.87±0.11 76.67±0.5 77.57±0.15 78.72±0.3 69.04±0.3 69.23±0.16 69.43±0.3
11 85.82±0.6 89.21±0.16 89.26±0.4 79.24±0.3 79.65±0.19 84.86±0.11 70.75±0.5 70.76±0.12 70.66±0.1
12 89.32±0.4 91.57±0.8 94.78±0.1 81.07±0.9 84.37±0.11 87.47±0.7 73.37±0.4 76.37±0.18 79.37±0.19

0
10
20
30
40
50
60
70
80
90
100
0 5 10 15
%CDR
Time (hrs)
Release profile of bathes N1-N3
N1
N2
N3
0
20
40
60
80
100
0 5 10 15
%CDR
Time (hrs)
Release profile of batches N4-N6
N4
N5
N6
0
20
40
60
80
100
0 5 10 15
%CDR
Time (hrs)
Release profile of batches N7-N9
N7
N8
N9
 There was %CDR found in batch N1-
89.32%, N2-91.57%, N3-94.78%,N4-81.07%,
N5-84.37, N6-87.47%, N7-73.37%, N8-76.37%
,N9 -79.37% in 12 hrs.
 Three was batch N3-94.78% found highest
%CDR in 12hrs.

 Statistical Analysis
Design expert software version 10.0.5 was used for Statistical analysis and produced first order polynomial
equations. From preliminary results, a 32 full factorial design was utilized in which two factors was evaluated,
separately at three levels and possible nine combinations was formulated. Three level factorial studies were
carried out using two different variables. In first factorial design, amount of Concentration of eudragit
S100(X1) and Concentration of PVA (X2) was taken as independent variables while Particle Size
(Y1),%CDR(Y2) was selected as dependent variables for both factorial designs.
 Effect on Particle Size (Y1) - Surface Response Study
Negative value of coefficient of A indicates decrease in Particle size. Positive value of coefficient B
indicates increase in Particle size. It indicates linearity of surface response and contour plot as shown
in figure and Full model was found significant for two independent variables and detailed ANOVA,
Response Surface Counter Plot and 3 D plots are as follows:
Particle Size = 124.78+12.83*A-4.00*B
Table.: ANOVA Table for Response Y1
Analysis of Variance Table [Partial of Squares – Type III]
Source
Sum of
Squares df
Mean
Square
F
value
p-value
Prob>F
Model 1084.17 2 542.08 139.6 <0.0001 Significant
A-Drug Polymer
Ratio
988.17 1 988017 253.50 <0.0025
B-Conc. of PVA 96.00 1 96.00 24.63
Residual 23.39 6 3.90
Cor Total 1107.56 8

Design-Expert® Software
Factor Coding: Actual
PARTICLE SIZE (nm)
Design Points
143
107
X1 = A: Drug polymer ratio
X2 = B: conc of PVA
250 350 450 550 650 750
0.1
0.15
0.2
0.25
0.3
PARTICLE SIZE (nm)
A: Drug polymer ratio (mg)
B:concofPVA(%)
110
120 130
140
PARTICLE SIZE (nm)
Design points above predicted value
Design points below predicted value
143
107
X2 = B: conc of PVA
0.1
0.15
0.2
0.25
0.3
250
350
450
550
650
750
100
110
120
130
140
150
PARTICLESIZE(nm)
A:Drug polymer ratio (mg)B: conc of PVA (%)
Figure .: 3D Surface Plot (A) Conc. Eudragit S100
of and (B) Conc. Of PVA on Particle Size (Y1)
Fig.: Response Surface Plot (A) Conc. of Eudragit
S100 and (B) Conc. Of PVA on Particle Size (Y1)

 Effect %CDR (Y2) - Surface Response Study
Negative value of coefficient of A indicates decrease in %CDR. Positive value of coefficient B indicates
increase in %CDR. It indicates linearity of surface response and contour plot as shown in figure and .
Full model was found significant for two independent variables and detailed ANOVA, Response Surface
Counter Plot and 3 D plots are as follows:
% CDR = 84.18-7.71*A+2.97*B
Table .: ANOVA Table for Response Y2
Analysis of Variance Table [Partial of Squares – Type III]
Source
Sum of
Squares df
Mean
Square
F
value
p-value
Prob>F
Model 409.80 2 204.90 2516.25 <0.0001 Significant
A-Drug Polymer
Ratio
356.82 1 356.82 4381.83 <0.0001
B-Conc. of PVA 52.98 1 52.98 650.67 <0.0001
Residual 0.49 6 0.081
Cor Total 410.29 8

CDR (%)
Design Points
94.7
73.45
X2 = B: conc of PVA
250 350 450 550 650 750
0.1
0.15
0.2
0.25
0.3
CDR (%)
B:concofPVA(%)
75
8085
90
CDR (%)
Design points above predicted value
Design points below predicted value
94.7
73.45
X2 = B: conc of PVA
0.1
0.15
0.2
0.25
0.3
250
350
450
550
650
750
70
75
80
85
90
95
CDR(%)
B: conc of PVA (%)
Fig.: Response Surface Plot (A) Conc. of Eudragit
S100 and (B) Conc. Of PVA on %CDR (Y2)
Figure .: 3D Surface Plot (A) Conc. Eudragit S100
of and (B) Conc. Of PVA on %CDR (Y2)

 Establishing Design Space and Control Strategy
The FDS curve indicates what % fraction
of design space has a given prediction
error or lower. A good design will have a
flatter and lower curve than a poor design
as shown in figure . Flatter means overall
prediction error will be constant. Lower
means overall prediction error will be
smaller. FDS should be at least 0.8 or 80%
for exploration, and 100% for robustness
testing. FDS was 0.96 or 96% which
indicating robust Standard error of
prediction relates to prediction interval
around a predicted response at a given
pair of factor levels.
Min Std Error Mean: 0.333
Avg Std Error Mean: 0.471
Max Std Error Mean: 0.667
Cuboidal
radius = 1
Points = 100000
t(0.05/2,6) = 2.44691
d = 1.46423, s = 1
FDS = 0.96
Std Error Mean = 0.598
0.00 0.20 0.40 0.60 0.80 1.00
0.000
0.200
0.400
0.600
0.800
1.000
FDS Graph
Fraction of Design Space
StdErrorMean

 Validation
From polynomial equations generated for
response, intensive grid and integrated examine
was performed over experimental field using
Design Expert Software (10.0.5). During
independent variable characterization study,
impact of parameters Concentration of Eudragit
S100(mg) and Concentration of PVA(%) were
assessed. Criteria considered of response
Particle Size (Y1), and %CDR (Y2) is between
100-150 nm and 70-95% respectively. Design
space shown in figure and also called as overlay
plot which is shaded region with yellow color
indicates that region of successful operating
ranges.
Overlay Plot
PARTICLE SIZE
TI Low
TI High
CDR
TI Low
TI High
Design Points
X2 = B: conc of PVA
250 350 450 550 650 750
0.1
0.15
0.2
0.25
0.3
Overlay Plot
B:concofPVA(%)
PARTICLE SIZE: 116.566
TI Low: 107.187
TI High: 125.945
CDR: 89.5751
TI Low: 88.1257
TI High: 91.0246
X1 399.403
X2 0.276163
Overlay Plot
PARTICLE SIZE
TI Low
TI High
CDR
TI Low
TI High
Design Points
X2 = B: conc of PVA
250 350 450 550 650 750
0.1
0.15
0.2
0.25
0.3
Overlay Plot
B:concofPVA(%)
PARTICLESIZE: 117.568
TI Low: 108.265
TI High: 126.87
CDR: 88.902
TI Low: 87.4644
TI High: 90.3396
X1 409.385
X2 0.26395
Figure : Overlay Plot

 Check point analysis of Validation Batches:
N10 & N11 formulation was made for check point analysis and predicted and experimental
values were compared.
Table.: Validation of Batches N10 & N11: Predicted Response
Batch no. Concentration of
Eudragit S100 (X1)
Concentration of PVA
(X2)
Particle Size (nm) %CDR
N10 399.4 0.27 116.90 89.57
N11 409.3 0.26 117.56 88.90
Batch no. Concentration of
Eudragit S100 (X1)
Concentration of PVA
(X2)
Particle Size (nm) %CDR
N10 399.4 0.27 116.02 89.47
N11 409.3 0.26 118.25 86.32
Table.: Validation Batches N10 & N11: Actual Response

 Optimized Batch Formulation From 32 Factorial Designs
Table.: Formulation of Optimized Batch N3
Sr.
No
.
Ingredients Optimized batch N3
1 Noscapine (mg) 80 mg
2 Eudragit S100 (mg) 250 mg
3 Acetone: Methanol (1:2)
(ml)
40 ml
4 PVA 0.3 %
5 Distilled Water 80 ml
 Characterization of Optimized Batch-N3 of
Noscapine Nanoparticles[43,44]
Table.: Evaluation of Optimized Batch N3
Parameters Results
Particle Size 107
% E.E 89.32±0.3
Drug Content 84.83±0.5
PDI Value 0.796
Zeta Potential 0.83 mv
 The parameter shows the result as given in table. The
parameters including the Particle Size, %E.E, Drug
Content, PDI Value and Zeta Potential and all the
formulations are shown in range of the standard
limit.

In - Vitro Drug release study
 There was found that %CDR of N3 batch in 12
hrs was 94.78%
Time (hr.) N3
0 0
1 12.80±0.7
2 23.54±0.2
3 31.76±0.16
4 44.15±0.2
5 53.88±0.15
6 57.89±0.6
7 65.03±0.9
8 72.11±0.14
9 79.29±0.6
10 87.87±0.8
11 89.26±0.3
12 94.78±0.5
0
12.8
23.54
31.76
44.15
53.88
57.89
65.03
72.11
79.29
87.8789.26
94.78
0
20
40
60
80
100
120
0 5 10 15
N3
N3
Linear (N3)
%
C
D
R
Time (Hr)

Figure.: Particle Size, PDI Value, Zeta Potential Report of
Optimized N3 Batch

PARAMETER Optimized Noscapine Based Nanoparticles (N10)
0 Days 15 Days 30 Days
Drug Content 72.15 67.45 70.29
%CDR (%) 84.52 87.21 82.08
 Accelerated Stability Analysis of Optimized Batch N3
Table.: Stability Analysis of optimized batch N3 for 1 Month
Results of stability studies shows that there is no significant changes occur in the
Drug Content and %CDR after storing the nanoparticles at 40ºC ± 2ºC, 75% ±
5% RH for one months. Thus it can be concluded that the formulation is stable.

 Cytotoxicity Study[45,46]
1.2
10.2
35.6
51.2
70.4
2.9
22.8
52.1
70.3
84.65
0
10
20
30
40
50
60
70
80
90
50 100 150 250 500
% PURE DRUG
% Nanoparticles
Concentration (µg/ml)
%
C
y
t
o
t
o
x
i
c
it
y
For noscapine loaded Eudragit S100/PVA nanoparticles, when the dosage concentration increases
from 50-500µg/ml, the %cytotoxicity increases dramatically from 2.9% to 84.65%, while the %
cytotoxicity for noscapine is increased from 1.2% to 70.4%. literature suggest that higher cell
viability was observed for Eudragit S100/PVA(75:25) nanoparticles due to higher lactide content of
PVA which was contributing to slower release. Therefore, the drug formulated in the Eudragit
S100/PVA nanoparticles exhibits a better performance on getting lower cell viability or equivalent
high cytotoxicity than that of pure therapeutic drug Noscapine.

• CHAPTER-1 Provides introduction to of Colon Targeted Drug Delivery, Nanoparticles-
methods of preparation, Disease and its Treatment, Drug profile.
• CHAPTER- 2 Provides the Needs, objective of Nanoparticles and Drug. and plan of work.
• CHAPTER-3 Discusses past work of colon targeted, colon cancer, nanoparticles from
Review of the literature.
• CHAPTER-4 Discusses in detail material and methodology of nanoparticles and drug.
Discussed in detail preliminary selection of formulation, Characterization of Noscapine
Based Nanoparticles. Further, this chapter describes preparation of Nanoparticles and
evaluation its. It also describes the results and discussion of the thesis. Which include
Preformulation study of drug (Noscapine). FTIR study of drug (Noscapine) and drug-
excipients shows the characteristic peaks and all other peaks observed with individual
compound have remained unaffected in formulations. DSC study of drug (Noscapine)
and drug-excipients shows that no interaction between drug and Excipients.
Formulation and Development of Noscapine Based Nanoparticles was carried out Using
32 Factorial Design approach, check point analysis of batches obtained from factorial
design and characterization of optimized batch. Which include evaluation parameter of
Nanoparticles like Particle Size, % E.E, Drug Content, % CDR, Zeta Potential, PDI, SEM
and Stability study of the Nanoparticles were also discussed.
• CHAPTER-5 Summary and Conclusion
• CHAPTER-6 Briefly explains the discussion of the whole thesis with brief summary and
conclusion.

In the in vitro drug release study for % CDR to be observed between Eudragit S100 and PVA to be
observed that drug release is give better in Eudragit S100 with lower concentration range for
optimization formulation. The % Entrapment Efficiency of the Nanoparticles ranged between 70 to
90, and increased with increase in the concentration of the polymer Eudragit S100 in nanoparticles.
The Particle Size of the nanoparticles ranged between 100 to 250 nm, is Decreased with increase in
the concentration of the polymer Eudragit S100 in the nanoparticles. The results of a 32 full factorial
design shown that the concentration of Eudragit S100 and PVA significantly affected on the
dependent variables like Particle Size (Y1), %CDR (Y2). The optimized formulation shown in-vitro
controlled drug release up to 12 hr. By considering all results of Check Point Analysis nanoparticles
formulations and Characterization of same. Stability study was carried out and no changes found
after stability analysis for a period of 1 month. Thus, colon specific nanoparticle system is increase
efficacy and patient compliance with lower dose of drug also promising approach for colon cancer
drug delivery of Noscapine.

ITD COMMENT SHEET
Typological error in Title has
been changed & Drug name
(Noscapine) has been added

1. Prasanth V.V, Jayaprakash. R, Sam T. Mathew “Colon Specific Drug Delivery
Systems: A Review On Various Pharmaceutical Approaches” Journal Of Applied
Pharmaceutical Science 02 (01); 2012: 163-169.
2. Singh Amritpal, Sharma Ankush, Pooja, Anju “Novel Approaches For Colon
Targeted Drug Delivery System” International Journal Of Research And
Development In Pharmacy And Life Sciences February - March, 2014, Vol. 3, No.2,
Pp 877 886 Issn: 2278-0238.
3. Ratnaparkhi Mukesh P., Somvanshi Fattesingh U., Pawar Sham A., Chaudhari Shilpa
P. , Gupta Jyoti P., Budhavant Kalyani A. “Colon Targeted Drug Delivery System”
International Journal Of Pharma Research & Review, August 2013; 2(8):33-42.
4. Altamash M. Qureshi*, Munira Momin, Sudha Rathod, Asish Dev, Chaitrali Kute
“Colon Targeted Drug Delivery System: A Review On Current Approaches” Indian
Journal Of Pharmaceutical And Biological Research 2013;1(4):130-147
5. Asha Patel1*, Nilam Bhatt1, Dr.K.R.Patel1, Dr.N.M.Patel1, Dr. M.R.Patel1, “Colon
Targeted Drug Delivery System: A Review System” Jpsbr: Volume 1, Issue 1: July-
August 2011 (37-49).
6. Jasvir Kaur1, Geeta Aggarwal2, Anu Mahajan*1 And Satvinder Kaur1 “Colon
Targeted Drug Delivery System To Treat Colorectal Cancer: A Review” World
Journal Of Pharmacy And Pharmaceutical Sciences Vol 5, Issue 01, 2016.

7. Congcong Lin 1, Huei Leng Helena Ng 1, Weisan Pan 2, Hubiao Chen 1, Ge Zhang 1,
Zhaoxiang Bian 1, Aiping Lu 1,3,* And Zhijun Yang 1,3,* “Exploring Different
Strategies For Efficient Delivery Of Colorectal Cancer Therapy” Int. J. Mol. Sci.
2015, 16, 26936–26952.
8. Fatima A. Haggar, M.P.H.,1,2 And Robin P. Boushey, M.D., Ph.D.1 “Colorectal
Cancer Epidemiology: Incidence, Mortality, Survival, And Risk Factors” Clinics In
Colon And Rectal Surgery/Volume 22, Number 4 2009.
9. Aarti P. Nikam*, Mukesh. P. Ratnaparkhiand, Shilpa P. Chaudhari “Nanoparticles –
An Overview” International Journal Of Research And Development In Pharmacy
And Life Sciences August - September, 2014, Vol. 3, No.5, Pp 1121-1127 Issn: 2278-
0238.
10. P. Venkatesan A, Nagaprasad Puvvada B, Rupesh Dash C, B.N. Prashanth Kumar A,
Devanand Sarkar C, Belal Azab C, Amita Pathak B, Subhas C. Kundu D, Paul B. Fisher
C, Mahitosh Mandal A,* “The Potential Of Celecoxib-Loaded Hydroxyapatite-
Chitosan Nanocomposite For The Treatment Of Colon Cancer” Biomaterials 32
(2011) 3794e3806.
11. Singh Davinder*, Harikumar S L, Nirmala “Nanoparticles: An Overview” Journal Of
Drug Delivery & Therapeutics; 2013, 3(2), 169-175.
12. Konwar Ranjit And Ahmed Abdul Baquee “Nanoparticle: An Overview Of
Preparation, Characterization And Application” International Research Journal Of
Pharmacy 2013 4 (4).

13. Abhishek Garg*, Sharad Visht, Pramod Kumar Sharma And Nitin Kumar
“Formulation, Characterization And Application On Nanoparticle
14. Noscapine Http://Www.Drugs.Com/International/Noscapine.Html, 20 september
2016
15. Indian Pharmacopoeia, Pharmacopoeia Commission-Ghaziabad, Govt Of India.
Ministry Of Health Family Welfare, 2010, 3, Pp 1798.
16. Chao Wang A, 1, Mei Zhao B, 1, Ya-Rong Liu A, Xin Luan A, Ying-Yun Guan A, Qin Lu
A, De-Hong Yu A, Fan Bai A, Hong-Zhuan Chen A, *, Chao Fang A, * “Suppression Of
Colorectal Cancer Subcutaneous Xenograft And Experimental Lung Metastasis
Using Nanoparticle-Mediated Drug Delivery To Tumor Neovasculature”
Biomaterials, Biomaterials 35 (2014) 1215e1226.
17. R. Seda Tigli Aydın1, 2 And Mehlika Pulat2 “5-Fluorouracil Encapsulated Chitosan
Nanoparticles For Ph-Stimulated Drug Delivery: Evaluation Of Controlled Release
Kinetics” Journal Of Nanomaterials Volume 2012.
18. Régis Cocoa, Laurence Plapieda, Vincent Pourcelleb, Christine Jérômec, David J.
Braydend, Yves-Jacques Schneidere, Véronique Préata, “Drug Delivery To Inflamed
Colon By Nanoparticles: Comparison Of Different Strategies” International Journal
Of Pharmaceutics 440 (2013) 3– 12.
19. Arvind Gulbake1 & Sanjay Kumar Jain2 “Colon Specific Delivery Of Mesalazine
Using Biocompatible Polymeric Nanoparticles” International Journal Of
Pharmacology And Pharmaceutical Technology, Issn-2277-3436, Vol-2, Iss-1, 2013.

20. Jingling Tang2 Na Xu1,2 Hongyu Ji1 Hongmei Liu1 Zhiyong Wang1 Linhua Wu1,2
“Eudragit Nanoparticles Containing Genistein: Formulation, Development, And
Bioavailability Assessment” International Journal Of Nanomedicine 2011:6 2429–
2435.
21. Meltem Cetin,1,3 Alptug Atila,2 And Yucel Kadioglu2 “Formulation And In Vitro
Characterization Of Eudragit® L100 And Eudragit® L100-Plga Nanoparticles
Containing Diclofenac Sodium” American Association Of Pharmaceutical Scientists
Pharmscitech, Vol. 11, No. 3, September 2010.
22. Safaa Sebak Maryam Mirzaei Meenakshi Malhotra Arun Kulamarva Satya Prakash
“Human Serum Albumin Nanoparticles As An Efficient Noscapine Drug Delivery
System For Potential Use In Breast Cancer: Preparation And In Vitro Analysis”
International Journal Of Nanomedicine 2010:5 525–532.
23. Ritu Aneja,1 Amr M. Ghaleb,2 Jun Zhou,3 Vincent W. Yang,2 And Harish C. Joshi1
“P53 And P21 Determine The Sensitivity Of Noscapine-Induced Apoptosis In Colon
Cancer Cells” Cancer Res 2007; 67: (8). April 15, 2007.
24. Ritu Aneja, Surya N. Vangapandu, Manu Lopus, Ramesh Chandra, Dulal Panda, And
Harish C. Joshi “Development Of A Novel Nitro-Derivative Of Noscapine For The
Potential Treatment Of Drug-Resistant Ovarian Cancer And T-Cell Lymphoma” The
American Society For Pharmacology And Experimental Therapeutics Mol
Pharmacol 69:1801–1809, 2006 Vol. 69, No. 6.

25. Jun Zhou‡§, Kamlesh Gupta¶, Joyce Yao‡§, Keqiang Ye_, Dulal Panda¶,
Paraskevi Giannakakou**, And Harish C. Joshi§‡‡ “Paclitaxel-Resistant
Human Ovarian Cancer Cells Undergo C-Jun Nh2-Terminal Kinase-
Mediated Apoptosis In Response To Noscapine” The Journal Of Biological
Chemistry Vol. 277, No. 42, Issue Of October 18, Pp. 39777–39785, 2002.
26. Indian Pharmacopoeia, Pharmacopoeia Commission-Ghaziabad, Govt Of
India. Ministry Of Health & Family Welfare, 2010, 3, Pp 1800.
27. British Pharmacopoeia 2011. British Pharmacopoeia Commission, Vol I, Pp
1573.
28. British Pharmacopoeia 2011. British Pharmacopoeia Commission, Vol I, Pp
1574.
29.European Pharmacopoeia Vii. European Directorate For Quality Of
Medicines And Health Care, 2011, Vol Ii, Pp 2529.
30. European Pharmacopoeia Vii. European Directorate For Quality Of
Medicines And Healthcare, Vol Ii, Pp 2530.
31. Japanese Pharmacopoeia. Ministry Of Health, Labour And Welfare, 2009,
Pp 936.
32. Japanese Pharmacopoeia. Ministry Of Health, Labour And Welfare, 2009,
Pp 937, 1475.
33.United States Pharmacopoeia 35-National Formulary-30, The United States
Pharmacopeial Convention Rockville, 2012, Pp 3120.

34. Joshi Harish C. Et Al. “Delivery Systems And Methods For Noscapine And
Noscapine Derivatives, Useful As Anticancer Agents”10056913.
35. Joshi Harish C. Aneja Ritu Vangapandu Surya N. “Noscapine Analogs And
Their Use In Treating Cancers” 13488621.
36. Navnit H. Shah, Clifton; Wantanee ,Phuapradit, Kearny; Aruna Railcar
,Clifton, “Colon-Targeted Delivery System” 217344.
37. Otterbecknorbert “Orally Administered Medicament For Treating Colon
Cancer Comprises Ursodesoxycholic Acid In Gastric Fluid Resistant Coating
To Provide Direct Topical Action At Target” 19906290.
38. Liu Jie, Jiang Qing, Luo New Roots, Shuya Jun, Gaudi “Biodegradable
Nanoparticle-Entrapped Oral Colon-Targeted Micro-Capsule And
Preparation Method Thereof” CN-201310340081.
39. Singh Davinder*, Harikumar S L, Nirmala “Nanoparticles: An Overview”
Journal Of Drug Delivery & Therapeutics; 2013, 3(2), 169-175.
40. Anroop B. Nair, Shery Jacob “A simple practice guide for dose conversion
between animals and human” Journal of Basic and Clinical Pharmacy, Vol.
7 Issue 2 March-May 2016.

41. Meng Liu, Xiao-Juan Luo, Fei Liao, Xiao-Fei Lei, Wei-Guo Dong “Noscapine
induces mitochondria-mediated apoptosis in gastric cancer cells in vitro and
in vivo” Cancer Chemother Pharmacol (2011) 67:605–612
42. Régis Cocoa, Laurence Plapieda, Vincent Pourcelleb, Christine Jérômec,
David J. Braydend, Yves-Jacques Schneidere, Véronique Préata, “Drug
Delivery To Inflamed Colon By Nanoparticles: Comparison Of Different
Strategies” International Journal Of Pharmaceutics 440 (2013) 3– 12.
43. Konwar Ranjit And Ahmed Abdul Baquee “Nanoparticle: An Overview Of
Preparation, Characterization And Application” International Research
Journal Of Pharmacy 2013 4 (4).
44.Singh Davinder*, Harikumar S L, Nirmala “Nanoparticles: An Overview”
Journal Of Drug Delivery & Therapeutics; 2013, 3(2), 169-175
45. Zhang ZP, Feng SS. The drug encapsulation efficiency, in vitro drug
release,cellular uptake and cytotoxicity of paclitaxel-loadedpoly (lactide)-
tocopherylpolyethylene glycol succinate nanoparticles. Biomaterials 2006;
27: 4025–33.
46. Evans D.F, Pye G, Bramley R, Clark A.G, Dyson T.J, Hardcastle J.D.
1988.Measurement of gastrointestinal pH profiles in normal ambulant
humansubjects. Gut 1988; 29: 1035–1041.

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