SVKM’S NMIMS

Shobhaben Pratapbhai Patel School of Pharmacy & Technology Management,

Mumbai- 400056

SYNOPSIS

Project Title: Design and development of salts / cocrystals /

inclusion complexes of niclosamide, to improve its
solubility

Name of the Student : Deepank Gupta

Program and Specialization : M. Pharm. (Industrial Pharmacy) + MBA
Name of the Guide : Dr. Sonali S. Bharate, Associate Professor

PROJECT- M.Pharm. + MBA

Academic Year- (2020-2023)

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1. Introduction:
Niclosamide (1) is a US FDA-approved anthelmintic drug. Currently, it is being investigated
in 16 clinical trials for treating mild to moderate COVID-19. It is administered orally at a
dose of ~2000 mg/day, making it challenging in terms of patient compliance. It is available as
tablets and in India, is sold by Western remedies, India (Brand name: Niclesone containing 1
g niclosamide; b.i.d.) and GlaxoSmithKline Pharmaceuticals Ltd. (Brand name: Niclosan
containing 500 mg niclosamide; 4-times a day). Niclosamide is insoluble in water and
categorized as a BCS class 2 drug (low solubility and high permeability) (1). Salification and
crystallization have a successful track record in modulating the biopharmaceutical properties
of drugs. In the last 80-years, ~40% of FDA-approved drugs are available as pharmaceutical
salts (2, 3). Niclosamide is an ionizable compound, and its ethanolamine salt is reported.
However, salt screening of niclosamide has never been performed to explore other salt forms.

The concept of cocrystals emerged in the late 1980s and early 1990s. The
pharmaceutical cocrystals are defined as ‘co-crystals that are formed between a molecular or
ionic active pharmaceutical ingredient (API) and a co-crystal former that is solid under
ambient conditions’. Cocrystallization is an alternative technique to the salt formation, which
is particular used for the neutral compound. Cocrystals are a crystalline mixture of an active
pharmaceutical agent bonded non-covalently with an inactive pharmaceutical agent called a
‘conformer’. Inclusion complexes are considered as a subgroup of co-crystalline material.
Hence complexation with cyclodextrins will also be helpful to modulate the
biopharmaceutical properties of drugs (4, 5). The cocrystals commendably satisfy
patentability criteria, namely, novelty, non-obviousness, and utility (6-8). For instance,
carbamazepine: saccharin co-crystals are reported and evaluated at a multi-gram scale,
physical stability, in vitro dissolution, and oral bioavailability. The developed cocrystals were
also compared with commercially available formulations and were advantageous over the
immediate release product (9).

The ionization constant (pKa) plays a critical role in forming stable salt or co-crystal
of a drug substance. The minimum difference of pKa value of three units between counterion
and the ionizable drugs provides a stable salt. On the other hand, if this difference (ΔpKa) is
less than two units, it will form cocrystals of the drug.

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 Inhibit virus specific targets
Cl NO2
O
Cl
N
H
OH
Niclosamide (1)
Swater = 8.0 g/mL
CLogP: 4.34
pKa = 5.6
Oral bioavailability = 10%
Mild inhibitor of 3CL protease

Herein, we propose to develop salts/ cocrystals/ inclusion complexes of niclosamide

with counterions or coformers or complexing agents to enhance its solubility, dissolution, and
oral pharmacokinetics.

2. Objectives:

The present proposal aims to improve the solubility of niclosamide through the formation of
salts or cocrystals, or inclusion complexes to enhance its oral bioavailability.
The present proposal envisages,
 Salt screening of niclosamide using suitable counterions
 Development of cocrystals of niclosamide with coformers
 Preparation of inclusion complexes of niclosamide
 Evaluation and characterization of optimized formulations using 1H NMR, 13C NMR,
FT-IR, DSC, SEM analysis, powder X-ray diffraction studies, in vitro dissolution
studies, in vitro PAMPA assay, etc.
 Oral pharmacokinetic profile of optimized formulation/s in rats

3. Plan of work:

3.1. Analytical method development

A UV-visible method will be developed to determine niclosamide in formulations and in
vitro dissolution studies.

3.2. Primary screening for selection of suitable counterion/coformer/complexing agent:

Based on the pKa of niclosamide and counterions, salt/cocrystal screening experiments will

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be conducted to identify suitable counterion demonstrating solubility improvement of the
drug. Counterions such as maleic acid, succinic acid, fumaric acid, hippuric acid, 4-
aminobenzoic acid, malonic acid, oxalic acid, citric acid, NaOH, Ca(OH)2, Mg(OH)2, KOH,
meglumine, tromethamine, t-butylamine, etc. will be screened. Their effect on the solubility
improvement of niclosamide will be determined using our previously published protocols
(10).

3.3. Secondary screening to identify optimum counterion ratio and reaction solvent/s: From
the primary screening results, the different molar ratios viz. 1:1, 1:2, and 1:4 of niclosamide
and counterions will be conducted to identify the optimum ratio of counterion/complexing
agent.

3.4. Scale-up of optimum salt/cocrystal/inclusion complex: The identified

salt/cocrystal/inclusion complex will be scaled up, initially at 1 g level and further at 10 g
scale.

3.5. 1H NMR and 13C NMR Analyses: The chemical interaction, if any, among niclosamide,
counterions/complexing agent, and their respective salt/cocrystal/inclusion complex will be
studied by 1H NMR and 13C NMR.

3.6. Characterization of salt/cocrystals/inclusion complexes: The optimum solid form/s will

be evaluated /characterized using FT-IR, DSC, SEM, Powder XRD studies, etc. Additionally,
in vitro PAMPA assay will also be done to identify any change in the permeability across the
membrane.

3.7. Dissolution studies: Dissolution studies will be conducted at 37 ± 0.5 °C at 100 rpm
using a USP dissolution apparatus (Lab-India Dissolution Tester, Model: DS
8000; Type 2 – Paddle) in water, HCl buffer (pH 1.2), and phosphate buffer (pH 6.8). The
data will be compared with the parent molecule.

3.7. Oral pharmacokinetic study: The performance of identified solid form of niclosamide
against the plain API will be determined using pharmacokinetic studies. The objective behind
this study is to evaluate increased exposure (AUC) of formulation against API. A
pharmacokinetic study of optimized formulations will be performed perorally in SD rats.
Mean plasma concentration will be calculated, and data will be further analyzed to determine
PK parameters using WinNonlin 5.3 software package.

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4. Time Line of the Project:

Sr. No. Nature of work Duration in weeks

1. Literature, patent search, and review article 4
2. Listing and procuring chemicals 2
Authentication and Characterization of chemicals: API,
3. excipients 2
4. Preformulation studies 2
5. Formulation and optimization 12
6. Evaluation and spectral characterization of formulations 6
7. Oral pharmacokinetic studies 6
th
8. Research and review publication and thesis preparation 25 week onwards
Total 40 weeks

5. Budget:
Sr. No. Requirements Approximate cost in Rs.
Chemicals/materials/reagents/media/culture
Methnaol HPLC grade: 2.5 L
Acetonitrile HPLC grade: 2.5 L
Salts for preparation of buffers
1. Various counterions
2. Glassware
Animals (SD rats: the protocol will be
3. submitted in December 2021)
Funds for characterization data: pXRD, SEM
4. studies
Total

6. Application of the Work:

The oral route is mostly preferred for this drug administration. However, solubility and
dissolution are limiting factors while developing the formulation. This liability of the drug
candidate can be overcome by salification/cocrystallization/inclusion complexation. Through
the proposed research, the following are expected outcomes:
• New salt form/ cocrystals/ inclusion complex of niclosamide with improved oral
bioavailability
• Proof of concept for future research/ may lead to new lead molecule for further
development

7. References (Including Patents)

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1. Al-Hadiya BM. Niclosamide: comprehensive profile. Profiles Drug Subst Excip Relat Methodol.
2005;32:67-96. DOI: 10.1016/S0099-5428(05)32002-8.
2. Bharate SS. Recent developments in pharmaceutical salts: FDA approvals from 2015 to 2019. Drug Discov
Today. 2021;26(2):384-98. DOI: 10.1016/j.drudis.2020.11.016.
3. Bharate SS. Carboxylic Acid Counterions in FDA-Approved Pharmaceutical Salts. Pharm Res. 2021. DOI:
10.1007/s11095-021-03080-2.
4. Szejtli J, Szente L. Elimination of bitter, disgusting tastes of drugs and foods by cyclodextrins. Eur J Pharm
Biopharm. 2005;61(3):115-25. DOI: 10.1016/j.ejpb.2005.05.006.
5. Sohi H, Sultana Y, Khar RK. Taste masking technologies in oral pharmaceuticals: recent developments and
approaches. Drug Dev Ind Pharm. 2004;30(5):429-48. DOI: 10.1081/ddc-120037477.
6. Desiraju GR. Crystal engineering: structure, property, and beyond. IUCrJ. 2017;4(Pt 6):710-1. DOI:
10.1107/S2052252517014853.
7. Maity DK, Paul RK, Desiraju GR. Drug-Drug Binary Solids of Nitrofurantoin and Trimethoprim: Crystal
Engineering and Pharmaceutical Properties. Mol Pharm. 2020;17(12):4435-42. doi:
10.1021/acs.molpharmaceut.0c00090.
8. Vishweshwar P, McMahon JA, Bis JA, Zaworotko MJ. Pharmaceutical co-crystals. J Pharm Sci.
2006;95(3):499-516. DOI: 10.1002/jps.20578.
9. Hickey MB, Peterson ML, Scoppettuolo LA, Morrisette SL, Vetter A, Guzman H, et al. Performance
comparison of a co-crystal of carbamazepine with marketed product. Eur J Pharm Biopharm.
2007;67(1):112-9. DOI: 10.1016/j.ejpb.2006.12.016.
10. Kumar V, Bharate SB, Vishwakarma RA, Bharate SS. Selection of a Water-Soluble Salt Form of a
Preclinical Candidate, IIIM-290: Multiwell-Plate Salt Screening and Characterization. ACS Omega.
2018;3(7):8365-77. DOI: 10.1021/acsomega.8b00801.

Signature of Student Signature of Guide

Signature of HOD

Approved by

Dr. Bala Prabhakar

Dean, SPPSPTM