Int J Curr Pharm Res, Vol 16, Issue 4, 59-64Original Article

IN SILICO DOCKING STUDY OF HERBAL COMPOSITION AND DEVELOPMENT OF MOUTH DISSOLVING TABLETS FOR COMMON RESPIRATORY DISEASES THEREOF

ANKITA CHANDWANIa, KARUNA MODIa, KAUSHIK SHAHb, MAMTA SHAHa*

aDepartment of Pharmacognosy, L. M. College of Pharmacy, Navrangpura, Ahmedabad-380009, Gujarat, India. bSushen Medicamentos Pvt. LTD., Ahmedabad-382213, Gujarat, India
*Corresponding author: Mamta Shah; *Email: mbshah2007@rediffmail.com

Received: 14 Apr 2024, Revised and Accepted: 05 Jun 2024

ABSTRACT

Objective: The present study aims to develop a novel mouth dissolving tablets containing a combination of herbal extracts and a bioactive constituent and evaluating it for activity against common respiratory diseases (in silico studies).

Methods: Docking study was done to provide a scientific foundation, keeping the traditional knowledge as a base. Four trial batches were developed. The final batch was then formulated and various pre and post-compression and assays were performed to evaluate the formation of good quality of product. The final batch was prepared by the method of direct compression and taken for accelerated stability studies.

Results: The final batch containing 10 % active ingredients, 7.5 % super-disintegrant and 47 % diluent was found to be stable, easily producible and economic.

Conclusion: This research work grasps possibilities for researchers in the development and evaluation of mouth-dissolving tablets with significant bioactive potential against common respiratory diseases.

Keywords: Accelerated stability study, Bioactive, Direct compression, Docking, In silico studies, Super-disintegrant

© 2024 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/)
DOI: https://dx.doi.org/10.22159/ijcpr.2024v16i4.5010 Journal homepage: https://innovareacademics.in/journals/index.php/ijcpr

INTRODUCTION

Respiratory diseases are wide spread and one of the major reasons of morbidity and mortality, predominantly in the developing world. These diseases are categorised as acute (pneumonia and influenza); chronic (chronic obstructive pulmonary disease (COPD) and asthma); occupational lung (byssinosis, asbestosis, and coal worker's pneumoconiosis); and other parenchymal lung diseases (immune-related lung diseases) [1]. Through decades, almost 4 million people die every year due to acute respiratory tract infections globally. Approximately 4.6 percentage people having moderate to severe COPD die every year. Children are the most affected population due to asthma, a common chronic disease affecting 14 % of children worldwide. Respiratory disorders contributes towards over all 10 % of disability-adjusted-life-years (DALY’s), a matrix estimating the amount of dynamic and productive life loss [2-8].

Mouth dissolving tablets are a type of novel buccal cavity tablets that disintegrate and dissolve rapidly in the buccal cavity (saliva) and does not require chewing. They generally dissolve in 15 sec to 3 min in the buccal cavity and so mostly includes super-disintegrants and several taste masking agents [9-11]. These dosage forms are favourable for paediatric, geriatric or disabled persons facing problem in gulping tablets and capsules.

Fig. 1: Herbal components of formulation (A) A. galanga (B) M. oleifera (C) C. limon (D) Apocynin

Alpinia galanga (greater galangal or kulinjan) is traditionally used to treat asthma, bronchitis, heart diseases and rheumatism. It consists hydroquinone and hydroxymethylfurfural, galangoisoflavonoids, 1’-acetoxychavicol, galangin and galanganol B [12–16] (fig. 1A).1’-acetoxychavicol has a potential effect against asthma [15]. Moringa oleifera (drumstick tree or senjana) is widely used for anaemia, bronchitis, catarrh, chest congestion, asthma, cough, fever, respiratory disorders, TB and diabetes in the traditional system of medicine. It is a rich source of various vitamins, minerals, flavonoids such as niazirin and quercetin, and benzyl isothiocyanates [17-23] (fig. 1B). It treats the four basic symptoms of asthma i. e. dyspnoea, wheezing, chest tightness and cough [24]. Citrus limon (lemon or nimbu) is rich in vitamin C and flavonoids such as bergapten, eriocitrin, hesperidine and oxypeucedanin. Traditionally it is used to treat cold, cough, scurvy, sore throat and fever [25-29] (fig. 1C). Apocynin (4’-hydroxy-3’-methoxyacetophenone or acetovanillone) a naturally originating methoxy-substitute of catechol, is promisingly used for its bronchodilatory activity. It reduces influenza A virus induced lung inflammation and viral titres [30]. It is also reported to possess anti-arthritic, anti-asthmatic, anti-atherosclerotic and ant-oxidant activities [31-35] (fig. 1D).

Even in the present age of science and technology, the popularity of complementary and integrative therapies is on rise and people all over the world still opt for traditional systems of healthcare majorly due to less side effects, as compared to the modern allopathic medicines [36]. In the present work mouth dissolving tablets of dried extracts of Alpinia galanga, Moringa oleifera, Citrus limon and a bioactive apocynin were prepared and evaluated.

MATERIALS AND METHODS

Procurement of herbal extracts and bioactive

The dried plant extracts and the bioactive were procured form Bhagwati Herbal Pvt. ltd, Vapi, Gujarat and Sigma Aldrich, India, respectively. Percentage of phenolics and flavonoids are estimated in procured extracts.

Docking studies

Docking study was done using four different proteins against some of the common respiratory diseases i. e. influenza, asthma and COPD. The software used were Autodock Vina version 1.1.2 [37], MGL tools, Pymol and Open babel. The PDB IDs of the 4 different proteins used are 5EFA (influenza), 4FQH (influenza), 3WZE (asthma) and 3Q76 (COPD). The proteins were taken from RSCB PDB online and the ligands were obtained from PubChem. Protein and ligand were then prepared for docking using Autodock Vina. Grid box was formed around the appropriate receptor site and the docking was performed. The compatibility of ligand-protein interaction was visualized through the visualization software that is Pymol. The details of H-bond and Π-bond were then visualized using Autodock Vina.

Preliminary dosage form designing

Designing of the four trial batches was done at Sushen Medicamentos Pvt. ltd., Ahmedabad. Different batches were prepared with different concentration of extracts and excipients (table 1).

Table 1: Formulation of preliminary trial batches

S. No. Ingredient Quantity (g) Batch 1 Quantity (g) Batch 2 Quantity (g) Batch 3 Quantity (g) Batch 4
1 A. galanga extract 12 12 24 7.2
2 M. oleifera extract 12 12 24 7.2
3 C. limon extract 12 12 24 7.2
4 Avicel 102 108 185.1 108 108
5 β-cyclodextrin - 87.5 - -
6 Crospovidone 27 27 27 27
7 Sodium saccharin 3.6 7.2 3.6 3.6
8 Talc 7.2 3.6 7.2 7.2
9 Mannitol SD 200 164.6 - 128.55 179
10 Magnesium stearate 3.6 3.6 3.6 3.6
Total 350 350 350 350

Formulation of the final batch

Tablets were prepared by direct compression method with a weight of 350 mg each. Formula of the same is given table 2. All the required ingredients were weighed as per the suggested quantities. Except that of talc and magnesium stearate which were sieved 60 # size separately, rest of the other ingredients were passed through 30 # sieve. All the ingredients except magnesium stearate were blended in the V-blender at 15 RPM for 10 min. Following this magnesium stearate was added in the same and mixed at 15 RPM for 5 min. This blend is used for the blend analysis and the tablets were compressed using Elisa press punching machine with 11 mm round punches at 10 RPM speed by means of B tooling. The tablets were packed and utilized for further evaluations.

Table 2: Formulation of the final batch

S. No. Ingredient % Taken Quantity taken for 1 tablet (mg) Quantity taken for 1000 tablets (gm)
1 Apocynin 1.43 5 5
2 Alpinia galanga 2.94 10.3 10.3
3 Moringa oleifera 2.94 10.3 10.3
4 Citrus limon 2.97 10.4 10.4
5 Avicel 102 30.86 108 108
6 Crospovidone 7.71 27 27
7 Sodium saccharine 1.03 3.6 3.6
8 Talc 2.06 7.2 7.2
9 Mannitol SD 200 46 161 161
10 Magnesium stearate 1.03 3.6 3.6
11 Orange flavour 1.03 3.6 3.6
Total 100 350 350

Evaluation of the tablets

The evaluation studies were done in 2 phases i. e., pre-compression evaluation and post-compression evaluation.

Pre-compression evaluation

This includes loss on drying and testing of flow properties of the powder mixture such as bulk density, tapped density, Carr’s index, Hausner ratio, angle of repose, etc [38, 39].

Post-compression evaluation

It includes testing of appearance, weight variation, uniformity of weight, hardness test, friability test, disintegration test, wetting test, content assay, etc [38-41].

Stability studies

The aim of performing the stability parameters was to achieve a product which is stable and complies with safety and efficacy aspects as per regulations. The optimized formulation of mouth dissolving tablets was kept at room temperature condition 40̊ °C±2̊ °C and humidity 75 % RH±5 % RH for a period of three month [42]. The study was carried out at the end of 0, 1 mo, 3 mo and tested for disintegration time and hardness and assays were performed. The TLC fingerprinting was developed for mixture of extracts and bioactive as well as for the tablets of final batch using mobile phase, toluene: ethyl acetate: formic acid: acetone (5: 4: 1: 0.1, V/V).

RESULTS AND DISCUSSION

Analysis of the procured extracts

The phyto-chemical analysis of the extracts revealed presence of flavonoids, phenolic and saponins. Percentages of phenolic and flavonoid content in procured extracts are given in table 3.

Table 3: Assay of the procured extracts

S. No. Extract Flavonoid content (% w/w) Phenolic content (% w/w)
1 Alpinia galanga 8.12±0.63 % 2.4±0.31 %
2 Moringa oleifera 10.53±0.22 % 2.23±0.12 %
3 Citrus limon 13.52±0.15 % 2.89±0.47 %

Docking study

Docking was done using four proteins targeting influenza, asthma and chronic obstructive pulmonary disorder (COPD) against various phytoconstituents of the above-mentioned plants and the bioactive. The docking score was found to be ranging from-10.5 kcal/mol (hesperidin of C. limon with 3WZE) to-3.9 kcal/mol (2-deoxy-d-ribose of A. galanga with 5EFA). Apocynin possess highest docking score of-9.0 kcal/mol with 4FQH and 3WZE. 1’-acetochavicol acetate from A. galanga possess maximum binding affinity of-6.2 with 3WZE. Binding energy of value of quercetin from M. oleifera is-8.4 with 4FQH and that of hesperidin from C. limon is-10.5 with 3WZE (table 4, fig. 2).

Fig. 2: Interaction diagram of protein with ligand, 5EFA with 2-deoxy-d-ribose (B) 5EFA with Kaempferol (C) 4FQH with Hydroquinone (D) 3WZE with Hesperidin (E) 3Q76 with eriocitrin (F) 3Q76 with quercetin

Table 4: Docking score of various phytoconstituents and bioactive

Plant/Bioactive Phytoconstituent Docking score
5EFA 4FQH 3WZE 3Q76
Apocynin
-6.9 -9.0 -9.0 -7.4
Alpinia galanga Cinnamic acid -4.9 -5.7 -6.1 -5.4
Hydroquinone -4.2 -4.5 -5.1 -5.0
Hydroxymethylfurfural -4.3 -5.3 -5.4 -4.8
2-deoxy-d-ribose -3.9 -4.3 -4.1 -4.5
1’-acetoxychavicol acetate -4.7 -5.7 -6.2 -5.5
Moringa oleifera Kaempferol -6.1 -7.7 -8.1 -6.4
Myrecetin -6.3 -8.3 -8.0 -7.0
Niazirin -5.3 -6.8 -6.8 -6.4
Phthalic acid -4.9 -5.6 -6.5 -5.2
Quercetin -6.4 -8.4 -8.1 -7.5
Citrus lemon Ascorbic acid -4.5 -5.6 -5.7 -5.5
Bergapten -5.2 -6.4 -6.5 -6.5
Eriocitrin -7.6 -9.4 -10.4 -8.7
Hesperidin -7.2 -9.8 -10.5 -8.8
Oxypeucedanin -5.9 -7.1 -7.4 -6.7

Preliminary batches

The formulated four trial batches were evaluated for pre and post-compression parameters and various assays were performed. The disintegration times of batch 1-4 are 35, 28, 75 and 30 sec, respectively (table 5-7). Batch 1 showed good flow properties, hardness and disintegration time over other batches, so selected for final batch preparation.

Table 5: Pre-compression evaluation of trial batches

S. No. Parameter Batch 1 Batch 2 Batch 3 Batch 4 Final batch
1 Bulk density (gm ml-1) 0.516 0.463 0.476 0.44 0.476
2 Tapped density (gm ml-1) 0.622 0.561 0.595 0.561 0.588
3 Carr’s index 16.98 17.43 20.00 21.64 19.04
4 Hausner ratio 1.204 1.211 1.250 1.276 1.235
5 Angle of repose 25̊ 30̊ 35̊ 30̊ 20̊
6 Loss on Drying 2.51 6.83 2.98 2.22 2.19

Table 6: Post-compression evaluation of trial batches

S. No. Parameter Batch 1 Batch 2 Batch 3 Batch 4 Final batch
1 Average weight (g) 3.512 3.482 3.505 3.532 3.499
2 Weight variation (mg) 351.32 349.56 350.83 353.63 349.87
3 Thickness (mm) 4.27 4.657 4.321 4.309 4.19
4 Diameter (mm) 10.92 10.89 10.93 10.92 10.95
5 Hardness (N) 78.6 93.5 80.6 96.5 89
6 Friability (%) 0.209 Nil Nil Nil 0.26
7 Wetting test (sec) 20 12 55 15 20
8 Dispersion test (sec) 12 10 42 11 20
9 Disintegration test (sec) 35 28 75 30 35

Table 7: Assay of trial batches

S. No. Assay Batch 1 Batch 2 Batch 3 Batch 4 Mixture of 3 extracts and bioactive Final batch
1 Flavanoid content (% w/w) 16.43±0.28 16.02±0.32 22.32±0.31 7.61±0.41 20.42±0.24 19.94±0.28
2 Phenolic content (% w/w) 3.01±0.25 2.95±0.32 4.92±0.28 2.25±0.29 3.25±0.23 3.11±0.25

Table 8: Data of stability study

S. No. Parameter Limit 0 day 1 month 3 months
1 Appearance Light brown colour No change No change No change
2 Average weight 3.5 g 3.499 g 3.495 g 3.498 g
3 Shape Round No change No change No change
4 Hardness 80-90 N 89 N 88 N 89 N
5 Disintegration time 15 sec – 3 min 30-40 sec 30-35 sec 30-35 sec
6 Flavonoid content (% w/w) - 19.94±0.38 18.93±0.26 18.43±0.42
7 Phenolic content (% w/w) - 3.11±0.25 3.23±0.56 3.23±0.72

Formulation and evaluation of final batch

The formulated tablets were tested for pre and post-compression parameters and they passed all the tests significantly. The disintegration time of final batch was found 35 sec (table 5-7).

Stability studies

The tablets were found stable under the accelerated conditions 40̊ °C±2̊ °C and humidity 75 % RH±5 % RH for a period of three month as shown in table 8. TLC fingerprint of the mixture of extracts and bioactive and that of tablets of final batch showed same resolution after three month also (fig. 3).

Fig. 3: TLC study A. Mixture of extracts and bioactive B. Final batch

CONCLUSION

The developed formulation was evaluated and the results indicated good flow properties, strength of hardness, disintegration, dispersion and wetting time. The final batch passed all the tests sufficiently. Thus, the proposed formulation is easily preparable, stable, economical and possesses a bronchodilatory effect.

The current research work will be supportive in developing efficacious and potent polyherbal formulations for the treatment of common respiratory diseases. This investigation can also be significantly used as an important tool for uncovering of possible mechanism for the development of herbal drugs, optimization and their evaluation for future researchers.

FUNDING

Nil

AUTHORS CONTRIBUTIONS

All the authors have contributed equally

CONFLICTS OF INTERESTS

Declared none

REFERENCES

  1. Respiratory diseases of adults. In: Speizer FE, Horton S, Batt J, Slutsky AS, Jamison DT, Breman JG, Measham AR, Alleyne G, Claeson M, Evans DB, Jha P, Mills A, Musgrove P. editors. Disease control priorities in developing countries. 2nd ed. Washington, (DC): International Bank for Reconstruction and Development/the World Bank; 2006. p. 1128-59.

  2. European Respiratory Society. The global impact of respiratory disease. 2nd ed. Forum of International Respiratory Societies; 2017. p. 7-24.

  3. GBD 2015 Mortality and Causes of Death Collaborators. Global, regional, and national life expectancy, all-cause mortality, and cause-specific mortality for 249 causes of death, 1980-2015: a systematic analysis for the global burden of disease study 2015. Lancet. 2016;388(10053):1459-544. doi: 10.1016/S0140-6736(16)31012-1, PMID 27733281.

  4. Bousquet J, Kaltaev N. Global surveillance, prevention and control of chronic respiratory diseases: a comprehensive approach. Geneva: World Health Organization; 2007. p. 146.

  5. Burney PG, Patel J, Newson R, Minelli C, Naghavi M. Global and regional trends in COPD mortality, 1990-2010. Eur Respir J. 2015;45(5):1239-47. doi: 10.1183/09031936.00142414, PMID 25837037.

  6. Global Asthma Network. The Global Asthma Report 2018. Auckland, New Zealand; 2018.

  7. Pearce N, Ait Khaled N, Beasley R, Mallol J, Keil U, Mitchell E. Worldwide trends in the prevalence of asthma symptoms: phase III of the international study of asthma and allergies in childhood (ISAAC). Thorax. 2007;62(9):758-66. doi: 10.1136/thx.2006.070169, PMID 17504817.

  8. World Health Organization, The United Nations Children’s Fund. Pneumonia: the forgotten killer of children. Geneva: United Nations Children’s Fund (United Nations Children’s Fund)/World Health Organization (WHO); 2006.

  9. Kaur T, Gill B, Kumar S, Gupta GD. Mouth dissolving tablets: a novel approach to drug delivery. Int J Curr Pharm Res. 2011;3(1):1-7.

  10. Chauhan K, Solanki R, Sharma S. A review on fast dissolving tablet. Int J App Pharm. 2018;10(6):1-7. doi: 10.22159/ijap.2018v10i6.28134.

  11. Rathod S, Phansekar M, Bhagwan A, Surve G. A review on mouth dissolving tablets. Indian Drugs. 2013;50(11):5-14. doi: 10.53879/id.50.11.p0005.

  12. Subramanian P, Nishan M. Biological activities of greater galangal, Alpinia galanga-a review. Res Rev J Bot Sci. 2015;5(2):28-41.

  13. Zhou YQ, Liu H, He MX, Wang R, Zeng QQ, Wang Y. A review of the botany, phytochemical, and pharmacological properties of galangal. In: Grumezescu AM, Holban AM. editors. Natural and artifiial flavoring agents and food dyes, Handbook of food bioengineering. Vol. 7. Elsevier Inc.; 2018. p. 351-96.

  14. Kaushik D, Yadav J, Kaushik P, Sacher D, Rani R. Current pharmacological and phytochemical studies of the plant Alpinia galanga. Zhong Xi Yi Jie He Xue Bao. 2011;9(10):1061-5. doi: 10.3736/jcim20111004, PMID 22015185.

  15. Seo JW, Cho SC, Park SJ, Lee EJ, lee JH, Park DH, Kim BH. 1′-Acetoxychavicol acetate isolated from Alpinia galanga ameliorates ovalbumin-induced asthma in mice. Plos One. 2013;8(2):4-11.

  16. Chouni A, Paul S. A review on phytochemical and pharmacological potential of Alpinia galanga. Pharmacogn J. 2017;10(1):9-15. doi: 10.5530/pj.2018.1.2.

  17. Gopalakrishnan L, Doriya K, Kumar DS. Moringa oleifera: a review on nutritive importance and its medicinal application. Food Sci Hum Wellness. 2016;5(2):49-56. doi: 10.1016/j.fshw.2016.04.001.

  18. Rani NZ, Husain K, Kumolosasi E. Moringa genus: a review of phytochemistry and pharmacology. Front Pharmacol. 2018;9(108):1-26.

  19. Meireles D, Gomes J, Lopes L, Hinzmann M, Machado J. A review of properties, nutritional and pharmaceutical applications of Moringa oleifera: integrative approach on conventional and traditional Asian medicine. Adv Tradit Med. 2020;20(4):495-515. doi: 10.1007/s13596-020-00468-0.

  20. Paikra BK, Dhongade HK, Gidwani B. Phytochemistry and pharmacology of Moringa oleifera lam. J Pharmacopuncture. 2017;20(3):194-200. doi: 10.3831/KPI.2017.20.022, PMID 30087795.

  21. Kasolo JN, Bimenya GS, Ojok l, Ochieng J, Ogwal Okeng JW. Phytochemicals and uses of Moringa oleifera leaves in Ugandan rural communities. J Med Plants Res. 2010;4(9):753-7.

  22. Rachmawati I, Rifa’i M. In vitro immunomodulatory activity of aqueous extract of moringa oleifera lam. Leaf to the CD4 +, CD8+ and B220+ Cells in Mus musculus. J Exp Life Sci. 2014;4(1):15-20. doi: 10.21776/ub.jels.2014.004.01.03.

  23. Anudeep S, Prasanna VK, Adya SM, Radha C. Characterization of soluble dietary fiber from Moringa oleifera seeds and its immunomodulatory effects. Int J Biol Macromol. 2016;91:656-62. doi: 10.1016/j.ijbiomac.2016.06.013, PMID 27283233.

  24. Agrawal B, Mehta A. Antiasthmatic activity of Moringa oleifera lam: a clinical study. Indian J Pharmacol. 2008;40(1):28-31. doi: 10.4103/0253-7613.40486, PMID 21264158.

  25. Okwu DE. Citrus fruits: a rich source of phytochemicals and their roles in human health. Int J Chem Sci. 2008;6(2):451-71.

  26. John S, Monica SJ, Priyadarshini S, Arumugam P, Gupta P. Investigation on phytochemical profile of Citrus limonum peel extract. Int J Food Sci Nutr. 2017;2(1):65-7.

  27. Chauhan N, Saxena J. Phytochemical screening of yellow and green Citrus limon peel extracts in different solvents. Int Res J Pharm. 2019;10(4):121-5. doi: 10.7897/2230-8407.1004136.

  28. Ali J, Das B, Saikia T. Antimicrobial activity of lemon peel (Citrus limon) extract. Int J Curr Pharm Sci. 2017;9(4):79-82. doi: 10.22159/ijcpr.2017v9i4.20962.

  29. Oikeh EI, Omoregie ES, Oviasogie FE, Oriakhi K. Phytochemical, antimicrobial, and antioxidant activities of different citrus juice concentrates. Food Sci Nutr. 2016;4(1):103-9. doi: 10.1002/fsn3.268, PMID 26788316.

  30. Oostwoud LC, Gunasinghe P, Seow HJ, Ye JM, Selemidis S, Bozinovski S. Apocynin and ebselen reduce influenza a virus-induced lung inflammation in cigarette smoke-exposed mice. Sci Rep. 2016;6:20983. doi: 10.1038/srep20983, PMID 26877172.

  31. Stefanska J, Pawliczak R. Apocynin: molecular aptitudes. Mediators Inflamm. 2008;2008:106507. doi: 10.1155/2008/106507, PMID 19096513.

  32. Hart BA, Copray S, Philippens I. Apocynin, a low molecular oral treatment for neurodegenerative disease. BioMed Res Int. 2014;2014:298020. doi: 10.1155/2014/298020, PMID 25140304.

  33. Soares MP, Silva DP, Uehara IA, Ramos ES Jr, Alabarse PV, Fukada SY. The use of apocynin inhibits osteoclastogenesis. Cell Biol Int. 2019;43(5):466-75. doi: 10.1002/cbin.11110, PMID 30761659.

  34. Heumuller S, Wind S, Barbosa Sicard E, Schmidt HH, Busse R, Schroder K. Apocynin is not an inhibitor of vascular NADPH oxidases but an antioxidant. Hypertension. 2008;51(2):211-7. doi: 10.1161/Hypertensionaha.107.100214, PMID 18086956.

  35. Stefanska J, Sarniak A, Wlodarczyk A, Sokolowska M, Pniewska E, Doniec Z. Apocynin reduces reactive oxygen species concentrations in exhaled breath condensate in asthmatics. Exp Lung Res. 2012;38(2):90-9. doi: 10.3109/01902148.2011.649823, PMID 22296407.

  36. Semenya SS, Maroyi A. Data on medicinal plants used to treat respiratory infections and related symptoms in South Africa. Data Brief. 2018;21:419-23. doi: 10.1016/j.dib.2018.10.012, PMID 30364644.

  37. Trott O, Olson AJ. AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J Comput Chem. 2010;31(2):455-61. doi: 10.1002/jcc.21334, PMID 19499576.

  38. Bagul U, Bagul N, Kulkarni M. Manufacturing technologies for mouth dissolving tablets. Pharm Rev. 2006.

  39. Sahu V, Bakade B. Formulation and evaluation of mouth dissolving tablets. Int J Pharm Sci Res. 2012;3(12):4831-7.

  40. Baba SA, Malik SA. Determination of total phenolic and flavonoid content, antimicrobial and antioxidant activity of a root extract of Arisaema jacquemontii blume. J Taibah Univ Sci. 2015;9(4):449-54. doi: 10.1016/j.jtusci.2014.11.001.

  41. Chandra S, Khan S, Avula B, lata H, Yang MH, ElSohly MA. Assessment of total phenolic and flavonoid content, antioxidant properties, and yield of aeroponically and conventionally grown leafy vegetables and fruit crops: a comparative study. Evid Based Complement Alternat Med. 2014;2014:253875. doi: 10.1155/2014/253875, PMID 24782905.

  42. International Conference on Harmonization. ICH. Q1A(R2) harmonised tripartite guideline. Stab Test New Drug Subst Prod; 2003. p. 1-18.