This research proposal summarizes work on developing lonidamine-loaded liposomes for breast cancer treatment. Liposomes will be prepared using soya lecithin and cholesterol via thin film hydration. Lonidamine inhibits glycolysis in tumor cells. Formulations will be optimized and characterized for size, drug content, release. In vivo studies will evaluate toxicity reduction and bioavailability enhancement. The proposal describes preparation methods, characterization tests, and expected outcomes of enhanced solubility, stability and reduced toxicity of lonidamine for breast cancer treatment.

1. A
Research Proposal
on
“Mitocondrial Targeted Lonidamine Loaded Liposomes
for the Treatment of Breast Cancer”
1
Submitted by:
Hemendra Kumar
RET Reg.No: 21000490
Email id: hemendrak96@gmail.com

2. CONTENT:-
1.INTRODUCTION
2.DRUG &EXCIPIENTS PROFILE
3.PREPARATION OF LIPOSOMES
5.PLAN OF WORK
7.REFERENCES
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3. INTRODUCTION:-
• Breast cancer is a systemic disease. It has become the most frequently diagnosed cancer
and the leading cause of cancer-related death among females worldwide, with an
stimated 2.1 million cases and 627,000 deaths in 2018.
Signs and symptoms of breast cancer may include-
• A breast lump or thickening that feels different from the surrounding tissue.
• Change in the size, shape or appearance of a breast.
• Changes to the skin over the breast, such as dimpling.
• Peeling, scaling, crusting or flaking of the pigmented area of skin surrounding the nipple
or breast skin.
• Redness or pitting of the skin over your breast, like the skin of an orange
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4.  A liposome is an artificial vesicle composed of one or more concentric phospholipid
bilayers and used especially to deliver microscopic substances (such as drugs or DNA)
to body cells.
The liposome is designed for the following optimized properties:
• Drug loading and control of drug release rate
• Overcoming the rapid clearance of drug
• Intracellular delivery of drugs
• Receptor-mediated endocytosis of ligand-targeted liposomes
• Reduce the toxicity
• Delivery of nucleic acids and DNA
4
Contd..

5. • Lonidamine is a derivative of indazole-3- carboxylic acid and is known to
inhibit aerobic glycolysis and energy metabolism selectively in tumor cells.
• Lonidamine (LND), first introduced in 1979 as an antispermatogenic agent,
has limited antineoplastic activity. Current evidence indicates that LND
inhibits glycolysis by the inactivation of hexokinase. Hexokinase is an
enzyme that catalyzes glucose, the first step in glycolysis.
• Lonidamibe also inhibits lactate export by the proton-linked
monocarboxylate transporter(s) (MCT) and pyruvate uptake into
mitochondria via the mitochondrial pyruvate carrier (MPC), whereas
inhibition of respiration involves both diminished mitochondrial uptake of
pyruvate.
5
Contd..

6. DRUG PROFILE
• Chemical Name- Lonidamine
• Synonyms- AF 1890, DICA, Diclondazolic Acid
• Chemical Formula- - C15H10Cl2N2O2
• Molecular Weight- 321.2 g/mol
• Chemical Structure-
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7. • Color- white
• Taste- Sour
• Solubility- Freely soluble in water, almost chloroform & methanol, insoluble in oil.
• Storage temperature – cold
• Stability- ≥2 Years
• Mechanisms of Action- LND inhibits glycolysis by the inactivation of hexokinase.
Hexokinase is an enzyme that catalyzes glucose, the first step in glycolysis.
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8. EXCIPIENT PROFILE
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Cholesterol
Synonym – Cholesterin, Cholesteryl alcohol, Cholest-5-en-3beta-ol.
Molecular weight – 386.7 g/mol
Structural formula –

9. • Description- In its pure state it is a white, crystalline substance that is odourless and
tasteless
• Solubility: insoluble in water, slightly soluble in alcohol, freely soluble in acetone,
chloroform, benzene
• Applications-
• A major component of the cell membrane
• Precursor in hormone biosynthesis.
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10. Soya lecithin
• Lecithin is a group of yellow-brownish fatty substance occurring in animal and
plant tissues, and in egg yolk, composed of phosphoric acid, choline, fatty acids,
glycerol, glycolipids, triglycerides, and phospholipids .
• Structural formula –
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11. Description:
• Colour: Yellowish brown
• Molecular Formula: C36H72NO8P
• Molecular Weight: 677.93gm
• Iodine value: 85-95
• Solubility: Lecithin is soluble in both aqueous and organic phase. Hence it can be
used as emulsifier in food industry and it is also capable of forming vesicles
thereby it is used in pharmaceutical industry. It gives clear or slightly opalescent
solutions with both phases.
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12. PLAN OF WORK
1.Preformulation studies-
Identification of drug-
 Physical appearance
 Melting point
 Solubility Analysis
 HPLC assay
 Fourier Transform infrared radiation Measurement (FT-IR)
2. Screening & Optimization of Suitable Method to Form Liposomes
 Solvent Evaporation Method
 Thin Film Hydration Method
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13. 3. Formulation of Pharmaceutical Liposmes by Best Selected Method.
4. Characterization-
• Percentage yield (% yield)
• Drug content estimation
• Drug entrapment Study
• Determination of size distribution of Liposmes
• In-vitro drug release
• Scanning electron microscopy
• In-vivo study
5. Stability Studies (as per ICH Guidelines)
6. Computation & Compilation of Work
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14. Method for preparation of Liposomes-
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Accurately weighed Cholestrol and Soya Lecithin was transferred
in solvent
Solvent was evaporated by using Rotatory Evaporator and a thin
film formed
Liposomal dispersion is formed
Liposome of reduced size is obtained
Stirring till dissolve
Phosphate Buffer + DRUG added for the
hydration of film
Sonication for size reduction
Vortex untill the formation of milky solution

15. Formulation and Development :-
S.No. Formulation code Soya Lecithin (mg) Cholesterol (mg) Sonication Time (Min)
1 F1 200 10 15
2 F2 200 10 10
3 F3 200 20 15
4 F4 100 10 10
5 F5 200 20 10
6 F6 100 10 15
7 F7 100 20 15
8 F8 100 20 10
9 F9 150 15 12.5
 By using DesignExpert-12
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DoE of the formulation

16. Expected Outcome:
• Enhance the Bioavailability of lonidamide.
• Enhance Solubility
• Increase Stability
• Reduce the toxicity
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17. REFERENCES
• Rangarajan Bharath, Shet tanuja, Wadasadawala tabassum, Nair S. Nita “Breast Cancer: An
overview of published indian data” South Asian Journal of Cancer; sept 2016; vol 5: issue 3.
• Sharma N. Ganesh, Dave Rahul, Sanadya Jyotsana, Sharma Piyush “Various types and
Management of Breast Cancer: An overview” Journal of Advanced Pharmaceutical Technology &
research; 2010 Apr-Jun; 1(2): 109-126.
• Sharma Amarnath, Sharma S. Uma, ”Liposome in drug delivery: progress and limitation”,
International journal of Pharmaceutics 154 (1997) 123-140
• Dua J.S.,Prof. Rana A.C.,Dr.Bhandari A.K. “LIPOSOME:Methods of prepration and Applications”
International journal of Pharmaceutical studies and research Vol. III/Issue II/April-June,2012/14-
20
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18. • Eugenia, Cruz M, Manuela, Gaaspar M, Lusia Corovo M, Manuela Carvalheiro.
Applications of lposomes in drug delivery. Novas Formas de Agents Bioactivos 1996; 22:
1649-1652.
• Nath Kavindra, Guo Lili, Nancolas Bethany et al. ”Mechanism of antineoplastic activity of
lonidamine”, Elsevier’s Biochimica et Biophysica Acta 2016; 152-156
• Cheng Gang, Zhang Qi, Pan Jing, Lee Yongik, Ouari Olivier et al, ”Targetting lonidaminen
to mitochondria mitigates lung tumorigenesis and brain metastasis” Nature
Communications (2019) 10:2205.
• www.Wikipedia. Org/wiki/lecithin
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