Formulation and Evaluation of Dexketoprofen Trometamol Loaded Extended Release Cubosomal Topical Gel

Abstract

Objective-The research work comprises formulation and evaluation of cubosome as a potential drug loading carrier and extended release formulation. Dexketoprofen Trometamol is a NSAID drug used in some cases of acute pain and its formulation in cubosome hypothesizes that its entrapment efficiency is enhanced and there is also enhancement in its spread ability and to show extended release so that it may be easily used in topical formulations. Method- The cubsosome was prepared by dispersion method. The method used was the method based on dispersion based on GMO, in which melted lipids are added drop-wise in aqueous medium at high temperature of about 80oC and at very high speed stirring of 6000-8000rpm followed by addition of ethyl alcohol. The mixture was stirred continuously and then polymer polyvinyl alcohol (PVA) was added in drop-wise manner, Thus, A total of 5 formulations were prepared. Result- The formulated cubosome showed regular particle size ranging from 137-225nm (F2>F1>F3>F4>F5) and had a ph range of 6.3 to 6.7 which showed its slightly acidic nature. The different formulation showed drug entrapment ranging from 74.34%-80.50% (F2<F1<F3<F4<F5) and showed drug content from 62.14%-67.30% (F2<F3<F1<F4<F5). The formulations showed a fantastic spreadability capability of 8.12-9.37(F2<F3<F4<F1<F5). It was found that the percentage cumulative drug release of Dexketoprofe trometamol ranges from 79.24%-92.24% at 24-hour duration, in which formulation F5 showed maximum drug release of 92.24% while formulation F2 showed least release of 79.24%. Conclusion– After the formulation and evaluation of cubosomal gel prepared by dispersion-based technique we found that the formulation had sufficient entrapment capacity of drug into it. The drug content of prepared formulations found to be greater than conventional dosage form of the drug and also higher entrapment than other nanoparticles. In 5 formulations prepared the formulation F5 was better among all with grater drug content, greater drug entrapment and with lower particle size and a better spreadability. Thus, it was concluded that formulation F5 was optimized formulation. As conventional dosage form (tablet) of Dexketoprofen trometamol had shorter half-life (1.3hr) and having gastric irritability and higher first-pass metabolism the cubosome topical gel formulation of Dexketoprofen trometamol passes these drawbacks and showed extended drug release over 24 hour of time interval.

Keywords

Introduction

Cubosome are liquid crystalline nanoparticle, the original observation of cubic liquid crystal comes in appearance during the study on polar lipids i.e. Monoolein, which are used as food stabilizer ^{1, 2, 3}. Cubosome are discrete, nanostructured particles of bicontinuous cubic crystalline phase^{4, 5}. In this continuous phase a continuous lipid bilayer separates two water channels^{6, 7}. Cubosome are liquid but have solid crystal-like symmetry (cubic crystal-like symmetry). Cubosome are highly Important in nanotechnology-based drug delivery⁵.Cubosome are derived from words ‘cube’ which means cubic in structure and prefixes with ‘some’ means phase⁹. Liquid crystals are intermediate states that have both solid and liquid properties¹⁰. They have molecular arrangement like solid and have flow property like liquids ¹¹. Liquid crystals are mainly of two types –

1. Thermotropic
2. Lyotropic (Cubosome)¹²

Cubosome comprise of curved lipid bilayer arranged in three-dimensional honeycomb-like structure that separates two internal aqueous channels with large interfacial area ¹³. The term ‘Cubosome’ was coined by Larsan^6,14,15The term Cubosome is USPTO (United states patent and trademark office) trademark for GS development AB corporation, Sweden¹⁶. Cubosome comes under category of ISA some (Internally self-assembled somes – phases) they envelope inverse non-lamellar liquid crystalline phases¹⁷.Cubosome are biocompatible and biodegradable carrier due to their lipid phase. These are available as non-parenteral medication in UK while it is also included in USFDA list of guidelines for inactive ingredient ¹⁸. Cubosome can accommodate three times the concentration of hydrophobic substances in it than traditional liposome. Appropriate 60% of outer surface of cubosome is in contact with water so it is highly suitable vehicle for delivering hydrophilic compounds ¹⁸. Due to amphiphilic natured it can encapsulate various active molecules of different nature like hydrophilic,hydrophobic,Amphiphilicand Lipophilic ¹⁹. Generally, cubosome size ranges from 100-300nm ²⁰. Cubosome are usually composed of either unsaturated monoglyceride (GMO) or phytantrial because both forms bicontinuous cubic phase in water²¹.

ADVANTAGES:

Figure 01: -Flow chart diagram representing Advantages of Cubosome ³²

DISADVANTAGES

Figure 02: -Diagrammatic view of disadvantages of cubosome ³²

Application of Cubosome

Figure 03: - Chart showing preparation method of GMO based Dexketoprofen Trometamol Cubosome ³²

Evaluation Parameters of Cubosome

1. Particle size
2. Surface topography
3. Ph
4. Drug content
5. Entrapment efficiency
6. Thermal stability
7. Drug release rate through skin permeation
8. Drug release time

1. Particle size and particle shape- The mean particle size of Dexketoprofen Trometamol loaded cubosome was determined using an optical microscope. The microscope was fitted with a stage micrometer to calibrate the eyepiece micrometer. The particle size of formulated cubosome was determined using microscopy. Simple microscope under 100x resolutions was used to determine size of the cubosome. The average particle size was determined using the following formula:

D?mean=∑nd∑n

2. Surface topography- Surface topography and surface morphology was examined using high resolution scanning electron microscopy (HR-SEM) under resolution 100x to 5000x.
3. Ph- The ph of the formulation was determined using normal ph paper.
4. Drug content-1 g of prepared formulation containing drug equivalent to 10mg was extracted with 30ml of ethanol. The volume was made up to 100ml with phosphate buffer (pH 7.4). The solution was filtered. The absorbance of the resulting solution was measured at 259nm using UV spectrophotometer after suitable dilutions. The drug content was determined by using phosphate buffer (pH 7.4) with the help of UV spectrophotometer by dissolving the formulation in phosphate buffer for 24hrs and then sample was taken and analyzed in UV-spectrophotometer. The drug content of the formulation was determined using the following equation:

%Drug content=Actual concentration of drug in the formulationTheoractiucal concentration of drug×100

5.  Drug Entrapment Efficiency-Accurately weighed quantity of prepared Dexketoprofen Trometamol loaded cubosome were taken and crushed in a mortar and pestle. 5ml of ethanol was added and transferred contents to a 100ml standard flask and made up to the volume with acetate buffer pH 5.5. Kept aside for 1h with frequent shaking for extracting the drug from the cubosome. Then it was filtered and the absorbance of filtrate was measured at 259nm after suitable dilutions. The drug content was calculated from the calibration curve and expressed as actual drug content in cubosome. The drug entrapment efficiency (%) of the cubosome was calculated according to following equation:

DEE%=Experiment drug loadingTheoratical drug loading×100

6. Thermal stability-The thermal stability of formulation was determined using Differential Scanning Calorimetry (DSC). DSC measures heat flow associated with phase transitions and thermal events. The thermal efficiency of microsponge are analyzed by differential scanning calorimetry (DSC). The DSC analysis was performed from temperature range of 0^oC to 600^oC and the effect of temperature was noted and graph was prepared to show chart of effect of temperature on enthalpy change of the formulations.

Procedure of differential scanning calorimetry:

• Sample Preparation: Place a small amount (1–10 mg) of the sample in an aluminum or standard pan; seal it properly. Leave an empty pan as a reference.
• Instrument Setup: Load both pans into the DSC instrument. Set the desired temperature range and heating/cooling rate.
• Analysis: The instrument heats or cools the pans, measuring the difference in heat flow between the sample and reference.
• Data Interpretation

          Detector Type: DSC-60

          Atmosphere: Nitrogen

          Gas Flow: 100 ml/min

          Pan Name: Aluminum

          Sample Weight: 5.000mg

7. Spreadability- spread ability is the measurement of time in seconds taken by two glass slides to slip off from gel which was placed in between the glass slides by applying certain load. Less the time taken for the separation of two glass slides, better is the spread ability. 2g of the formulation was placed on a ground glass slide fixed on a wooden block. The gel formulation was sandwiched between this slide and the second slide having same dimensions. Second slide was provided with a hook. Measured quantity of weight (30g) was placed in a pan attached to the pulley with the help of hook. Time (in seconds) required by the top slide to separate from ground slide was noted. Shorter the interval, better the spreading coefficient.
8. Drug release from egg membrane- In- vitro release study of Dexketoprofen trometamol loaded Glycerol monooleate based cubosomal topical gel was carried out by using Franz diffusion cell. The formulation was taken in the donor compartment and Phosphate buffer pH 7.4 was taken in the receptor compartment. The egg membrane, previously soaked overnight in the diffusion medium (Phosphate buffer pH 7.4) was placed between the donor and receptor compartment. 1g of the Dexketoprofen trometamol loaded Glycerol monooleate based cubosomal topical gel was spread uniformly on the egg membrane, which is in contact with the receptor medium. The whole assembly was placed on the thermostatically controlled (constant temperature) magnetic stirrer with continuous stirring and the temperature of the medium was maintained at room temperature. At specific intervals, 2ml of sample was withdrawn from the receptor compartment and replaced with an equal volume of Phosphate buffer pH 7.4. Absorbance of the sample was determined after suitable dilutions at 259nm using UV-visible spectrophotometer.

RESULT AND DISCUSSION

Drug- Excipient interaction compatibility study of Dexketoprofen trometamol and GMO: -

 The compatibility study of drug and excipient shows no interaction. The graph complies with the graph of standard graph of Dexketoprofen Trometamol.

Figure 04: - FTIR spectrum of Dexketorofen trometamol, Glycerol monooleate, Ethanol and Polyvinyl alcohol

Evaluation of Formulated Cubosome

Figure 05: - Optical microscopic image of GMO cubosome containing Dexketoprofen trometamol

Figure 06: - HR-SEM image of GMO based Dexketoprofen trometamol cubosome under 100000 x Resolution

Figure 07: - Bar graph showing Ph distribution of various formulations of Dexketoprofen trometamol loaded cubosomes

Figure 08: - Bar graph showing percentage drug content of 5 formulation of dexketoprofen trometamol loaded cubosome

Figure 09: - Bar graph showing percentage drug entrapment efficiency of Various formulations of dexketoprofen trometamol loaded cubosome

Figure 10: -Bar graph showing spreadability of formulations of dexketoprofen trometamol loaded cubosomal Gels

Figure 11: - Bar graph showing pH, %Drug content, %DEE, Spreadability of Dexketoprofen trometamol loaded cubosome Gel

Figure 12: - Graph showing Differential scanning calorimetry for cubosome

Figure 13: - Graph showing percentage cumulative drug release of 5 formulations of Dexketoprofen trometamol loaded cubosomal topical gel

CONCLUSION:

The research work aims to demonstrate the potential of this cubosomal nanoparticle as an effective treatment for acute pain with improved drug delivery. The research work comprises formulation and evaluation of cubosome as a potential drug loading carrier.  There are many dosage forms like tablets and capsules available in the market for the treatment of inflammation, but still there is a need for new dosage forms which acts effectively. NSAIDs were mainly used for the treatment of Inflammation. Dexketoprofen Trometamol is a NSAID drug used in some cases of acute pain and its formulation in cubosome hypothesizes that its entrapment efficiency is enhanced and there is also enhancement in its spread ability and also there is increment in drug release time so that it may be easily used in topical formulations. Cubosomes can be prepared by simple combination of biologically compatible lipids (GMO) and water and are thus more suited for pharmaceutical and body tissue. The formulated cubosome showed regular particle size ranging from 137-225nm (F2>F1>F3>F4>F5) and had a ph range of 6.3 to 6.7 which showed its slightly acidic nature. The different formulation showed drug entrapment ranging from 74.34%-80.50% (F2<F1<F3<F4<F5) and showed drug content from 82.14%-87.30% (F2<F3<F1<F4<F5). The formulations showed a fantastic spreadability capability of 8.12-9.37(F2<F3<F4<F1<F5). We found that the formulation had sufficient entrapment capacity of drug into it. The drug content of prepared formulations found to be greater than conventional dosage form of the drug and also higher entrapment than other nanoparticles. In 5 formulations prepared the formulation F5 was better among all with grater drug content, greater drug entrapment and with lower particle size and a better spreadability. As conventional dosage form (tablet) of Dexketoprofen trometamol had shorter half life (1.3hr) and having gastric irritability and higher first-pass metabolism The cubosome topical gel formulation of Dexketoprofen trometamol passes these drawbacks and showed extended drug release over 24 hour of time interval with a magnificent drug release of upto 92.24%.

ACKNOWLEDGEMENT

The formulation was done in Institute of pharmacy, Vikram University, Ujjain and DSC, FTIR procedures were done by central instrument facility, Indian institute of technology, Banaras Hindu University (IIT BHU), Varanasi. The research will never be completed without spiritual support of Baba Vishwanath and Baba mahakaal. The research was also supported by my family, my friend and colleague.

CONFLICT OF INTEREST

Nil

REFERENCES

1. Prashar Deepak and Sharma Dharmesh, review article Cubosome: A Sustained drug delivery carrier, Asian J.Res.Pharm.Sci. volume 1 issue 3 July September 2011;page 59-62.
2. Linderstorm M, Ljusberg- Warden H, Larsson K and Borgstorm B. Aqueous Lipid phases of Relevance to intestinal Fat Digestion and Absorption. Lipids 16: 749-754.
3. Anderson S Jacob M, Ladin S and Larsson K, Structure of the cubosome – a Closed Lipid Bilayer aggregate, Zeitschrift fur Kristallographie 210: 1995: 315-318.
4. Hyde S Andersson A, Larsson K, Blunn Z, Landh T, Lidin S and Ninham BW. The Language of shape. Elsevier, New York 1997.
5. Naveentaj S and Muzib Indita Y, A review on crystalline nanoparticles (Cubosome): Emerging Nanoparticulate Drug Carrier, International Journal of Current Pharmaceutical Research vol 12 Issue 1, 2010 page 5-9.
6. Rao S, Sravya B and Padmalatha K: A review on cubosome: The novel drug delivery system. GSC Bio and Pharm sci 2018 :05 (01), 0076-081.
7. Salah S, Mahmoud A and Kamel A:Etodolac transdermal cubosome for the treatment of rheumatoid arthritis:ex-vivo permeation and in-vivo pharmacokinetic studies. Drug Delivery 2017: 24 (1) ,846-856.
8. Tekade A.R. and Avhad G.D., A Review on Cubosome: A Novel Approach for Drug Delivery, IJPSR 2022 Volume 13 Issue 2, 579-588.
9. Chong JYT, Drummond XMBBCJ: Steric stabilizer for cubic phase lyotropic liquid crystal Nano dispersions (Cubosome) Advances in Planar Lipidic Bilayer and Liposomes 2015 ;21 :131-187
10. Mo J, Mileret G and Nagaraj M;Liquid nanoparticles for commercial drug delivery, Journal of liquid Crystals Reviews 2017; 5(2) 69-81.
11. Andrieko D: Introduction to liquid crystals; Journal of molecular Liquids 2018
12. Hiltrop K: Liquid crystals, Lyotropic liquid crystals Springer book Archive, Chapter 4 Page 143-162.
13. Bhosale RR, Osmani R, Harkare BR and Ghodake PP: Cubosome the immutable nanoparticle drug carrier, Scholars academic Journal of pharmacy (SAJP) 2013 vol 2 issue 6, page 481-486.
14. Xin P, Ke H, Xinsheng P, Zhiwen Y, Lingzhen Q and Chune Z: Nanostructured cubosome as advanced drug delivery system, Curr Pharm Design 2013; vol 19 issue 35; page 6290-6297.
15.  Larsson K: Cubic lipid – water phases: Structure and bio membrane aspects. Journal of Chemistry 1989 vol 93 issue 21: page 3304-7314.
16. Spicer PT, Small WBII, Lynch ML and Burns JL: Dry powder precursor of cubic liquid crystalline nanoparticle (Cubosome). Journal of Nanoparticle Research 2002 4: page 297-311.
17. Mu Huiling, Yanghmur Anan, Recent advances in drug delivery application of cubosome, hexosome and solid lipid nanoparticle – ScienceDirect vol 11 issue 4 April 2021 page 871-885.
18. Gowda Jaswanth B.H, Ahmed gulzar mohammad, Alshehri AliSaad, Wahab shadma, Vora K Lalitkumar, Thakur Singh Raghu raaj, Kesharwani Prashant: The cubosome – based nanoplatforms in cancer therapy: seeking new paradigms for cancer theranostics.-ScienceDirect volume 237 part 1, 15 Nov 2023.
19. Zakaria Fazila, Ashari Efliza siti, Azmi mat Diana Intan, Rahman Abdul Basyaruddin Mohd: Recent advances in encapsulation of drug delivery (active substances) in cubosome for skin disease -ScienceDirect; volume 68 February 2022.
20. Victorelli Damiani Francesca, Manni Salvati Livia, Biffi Stefania, Bartot Barbara, Buzza harb Hilde, Lutz-Bueno Viviane, Handschin Stephan, Calixto Giovana, Murgia Sergio, Chorilli Marlus, Mezzenga Raffaele; Potential of curcumin – loaded cubosome for topical treatment of cervical cancer; ScienceDirect volume 620, 15 Aug 2022, page 419-430.
21. Bryant J Saffron, Bathke K. Elly, Elder J. Karen: Bottom -up cubosome synthesis without organic solvents – ScienceDirect volume 601, Nov 2021 pages 98-105.
22. Bei D,Meng J and Youan BC: Engineering Nanomedicine for Improved Melanoma therapy: Progress and Premises, Nanomedicine (London, England) 2010; volume 5 issue 9, page 1385-1399.
23. Spicer PT: Cubosome Processing Industrial Nanoparticle Technology Development, Chemical Engineering Research and Design 2005 83(A11) page 1283-1286.
24. Tilekar KB, Khade PH, Shitole MH, Jograna MB and Patil RY: Cancer oriented cubosome – a review, International Journal for Pharmaceutical Research and Scholar (IJPRS) 2014, 3, page 198-210.
25. Yingchoncharoen P. Kalinowski DS and Richardson DR Lipid-based drug delivery systems in cancer therapy: what is available and what is yet to come Pharmacol Rev 2016, 68 701-787.
26. Sastri KT, Radha GV, Pidikiti S and P Vajghala. Solid Lipid nanoparticles preparation techniques, their characterization, and an update on recent studies J Appl Pharmaceut Sci 2020, 10: 126-141.
27. Rizwan SB, Dong YD, Boyd BJ, Rades T and Hook S Characterization of bicontinuous cubic liquid crystalline systems of phytantrial and water using cryo field emission scanning electron microscopy Micron 2007; volume 38 page 478-485.
28. Tilekar KB, Khade PH, Kakade S, Kotwal S and Patil R: Cubosomes a drug delivery system. International Journal of Chemical and Biochemical Science, 2014, 4 page 812-824.
29. Nanjwade BK, Hundekar YR, Kamble MS and Srichana T. Development of cuboidal nanomedicine by nanotechnology. Austin J Nanomed Nanotechnol 2014; 2:  1023.
30. Karami Z and Hamidi M Cubosomes: Remarkable drug delivery potential Drug Discovery Today 2016, 21: 789-801.
31. Barriga HMG, Ces O, Law RV, Seddon JM and Brooks NJ: Engineering Swollen Cubosomes Using Cholesterol and Anionic Lipids. Langmuir 2019, 35: 16521-16527.
32. Singh Aashish, Dr. Dashora Kamlesh, Dr. Bhargava Tanu, Dr. Sisodiya Dharmesh, Dr. Khirwadkar Praveen, Tomar Anju, “A Comprehensive Review on Cubosome: A Novel Versatile Nanocarrier”, International Journal of All Research Education and Scientific Methods (IJARESM), ISSN: 2455-6211, Volume 12, Issue 7, July-2024, Page no 94-105.