P15-04

Formulation and Characterization of Curcumin Nanoparticles Sricharan Gumudavelli * Research Intern Ascent Pharmaceuticals Inc., www.ascentpharm.com, 550 S Research Place, Central Islip, NY 11722 *Corresponding Author: srinija2004@yahoo.com

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OBJECTIVE The objective of this work is to formulate, develop and characterize Curcumin Nanoparticles using wet milling technology.

INTRODUCTION Curcumin is chemically [(E,E)-1,7-bis(4-hydroxy-3methoxy-phenyl)-1,6- heptadiene-3,5-ione] (Figure1) phenolic pigment of turmeric, the powdered rhizome of Curcuma longa.

Description Molecular Formula Molecular Weight Melting Range Solubility

Figure 1: Molecular Structure of Curcumin

Dark orange colored crystalline powder C21H20O6 368.379 170-175 Insoluble in water, Soluble in Ethanol (10 mg/mL), DMSO (70 mg/mL), chloroform, acetone

2 References: http://www.chemspider.com/Chemical-Structure.839564.html:

Therapeutic Uses of Curcumin

3 Reference: http://www.google.com/imgres?imgurl=http://www.curcuminresearch.org/Images/inhibits.jpg&imgrefurl=http://www.curcuminresearch.org/

Nanoparticle Development & Manufacturing Particle size reduction is one of the most commercially successful Drug Delivery System for poorly water soluble compounds like BCS Class II

compounds.

Particle Size Reduction Methods: * High Pressure Homogenization * Dry Milling * Ultra Sonication * Wet Milling/grinding/media milling

Top-Down methods of nanoparticle production are extremely effective in increasing the bioavailability of the active ingredient thereby increasing

absorption rate.

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Wet Milling

Dry Milling

• Size reduction possible up to nanosize • Increased stabilization of the particle due to protective liquid coating

• Size reduction possible up to microns • Process wide range of materials

Ultra Sonication • Size reduction possible up to microns • Sound used to agitate particles

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Wet Milling Wet milling of substances offers a multitude of advantages

• Friction is bonded directly into suspension • Substances to be ground remain in system • Reduces cost for extensive filtration

• Results in finer particle sizes Time and cost efficient as it allows wet milling to occur simultaneously with other processes.

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MEDIA MILLING TECHNOLOGY

Dyno Mill

Grinding Chamber

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Figure 2. Principle of Media Milling

Composition of Curcumin Nanoparticles

Ingredient

% w/w

Functionality

Curcumin Powder

10.00 Active Pharmaceutical Ingredient

Hydroxypropylmethyl Cellulose (5 cP)

2.50

Electrostatic Stabilizer

Docusate Sodium, USP

0.18

Steric Stabilizer

Sodium Benzoate, NF

0.2

Preservative

Purified Water, USP

87.12

Solvent/Vehicle

Total

100.0 8

• CRITICAL PROCESS PARAMETERS & CRITICAL QUALITY ATTRIBUTES Critical Formulation Variables Solid Content

Critical Process Variables Size of Bead

Type of Stabilizer

Amount of Bead (Load) Homogeneity

Concentration of Stabilizer

Temperature

Solvent (Aqueous/Nonaqueous) Morphology of Active Molecule Order of Addition of Stabilizer

Feed Rate

Critical Quality Attributes Size Stability

Rate of Mixing Mill Speed

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MANUFACTURING PROCEDURE

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PHYSICAL CHARACTERIZATION A. Measurement of Viscosity Viscosity of Curcumin Nanosuspension was determined using a

Brookfield Type DVII+ programmable viscometer with temperature controlling bath and small sample adaptor 3R or equivalent with “spindle SC4-21” at a constant speed of 80 rpm. The temperature

was kept constant at 25.0°C ± 0.1°C throughout the testing process.

B. Measurement of pH Suspension pH was measured using an Oakton pH/MV/°C meter.

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C. Sedimentation Volume The sedimentation volume of the Curcumin nanosuspension product was measured by placing nanosuspension in a 100 mL measuring cylinder. The sedimentation volume was recorded every 48 hours for 14 days at room temperature. The sedimentation volume was calculated using the following formula.

Sedimentation Volume (F)

=

Final height of sediment (Hu) Original height of suspension (Hi)

D. Freeze-Thaw study Freeze-thaw or temperature cycling was performed by placing samples in a freezer

(-20째C) for a 24-hour period, then held at ambient conditions for the next 24 hours.

This was defined as 1 cycle. Three cycles were performed on Curcumin nanosupension . At the end of each cycle, samples were evaluated for particle size

distribution.

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E. Centrifugation Test The physical stability of the product was evaluated by subjecting the suspension to centrifugation at room temperature at 15,000 rpm for 15 minutes using Fisher Scientific,

Model 228 instrument. After centrifugation, samples were evaluated for sedimentation and phase separation.

F. Density The density of the Curcumin nanosuspension was determined by transferring 50 mL

of suspension into a pre-weighed volumetric flask and recording the weight. The suspension was deaerated to remove any air from the suspension and more suspension was added, if needed, to ensure a volume of 50 mL. The flask was re-

weighed. Density was calculated using the following formula: Weight of the suspension Density = (weight of flask with suspension - Weight of the empty flask) (d) Volume of suspension (50 mL) 13

G. Particle Size Distribution Particle size distribution of Curcumin nanosuspension was determined using Nicomp

380 ZLS particle size analyzer equipped with Autocorrelator software CW 388. Light Scattering was monitored at 25째C at a 90째 angle, after external standardization with Spherical 90 nm diameter polystyrene beads.

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Results Testing Parameter(s) Physical Appearance

Results Milky white, lemon-lime suspension

Viscosity (cP)

4.95

pH

4.01

Density (g/ml)

1.055

Sedimentation Volume

flavored

1.0

25%

<171.8

Particle Size

50%

<205.0

Distribution**

75%

<244.7

(nm)

80%

<255.6

90%

<286.8 15

Results

We can see from the graph of diameter vs. % volume that following each subsequent cycle of milling, the product showed an increasingly homogenous mixture with the highest percentage of particles .1 to 1 micron.

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CONCLUSIONS

• Curcumin Nanoparticles were successfully manufactured using Media Milling Technology.

• Curcumin Nanoparticles were produced in range of 100120 nm.

• The developed nanoparticulate formulations did not show an increase in particle size at RT and 5°C thereby indicating acceptable stability.

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Future Plan of Action Improve bioavailability/reduce dose administration

â&#x20AC;˘ Functionalization of particles with hyaluronic acid to improve uptake

â&#x20AC;˘ The effect of functionalization will be observed in rheumatoid arthritis through cell culture studies

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