All posts by jppres

About jppres

The Journal of Pharmacy & Pharmacognosy Research (JPPRes) is an international, specialized and peer-reviewed open access journal, which publishes studies in the pharmaceutical and herbal fields concerned with the physical, botanical, chemical, biological, toxicological properties and clinical applications of molecular entities, active pharmaceutical ingredients, devices and delivery systems for drugs, vaccines and biologicals, including their design, manufacture, evaluation and marketing. This journal publishes research papers, reviews, commentaries and letters to the editor as well as special issues and review of pre-and post-graduate thesis from pharmacists or professionals involved in Pharmaceutical Sciences or Pharmacognosy.

Syzygium polyanthum bioactive compounds in polycystic ovary syndrome

J. Pharm. Pharmacogn. Res., vol. 10, no. 4, pp. 725-736, July-August 2022.

Original Article

Anti-inflammatory and antioxidant potential of Syzygium polyanthum (Wight) Walp. bioactive compounds in polycystic ovary syndrome: An in silico study

[Potencial anti-inflamatorio y antioxidante de compuestos bioactivos de Syzygium polyanthum (Wight) Walp. en el síndrome de ovario poliquístico: Un estudio in silico]

Renny Aditya1,4, Budi Santoso2*, Widjiati Widjiati3

1Doctoral Program of Medical Science, Faculty of Medicine, University of Airlangga, Surabaya, Indonesia.

2Department of Obstetrics and Gynecology, Faculty of Medicine, University of Airlangga, Surabaya, Indonesia.

3Department of Veterinary Anatomy, Faculty of Veterinary Medicine, University of Airlangga, Surabaya, Indonesia.

4Department of Obstetrics and Gynecology, Faculty of Medicine, Universitas Lambung Mangkurat, Banjarmasin, Indonesia.



Context: Polycystic ovary syndrome (PCOS) is significantly associated with inflammation and oxidative stress. Syzygium polyanthum is a plant rich in pharmacological properties. Aims: To evaluate the anti-inflammation and antioxidant potential of S. polyanthum bioactive compounds using in silico approach.

Methods: The S. polyanthum was extracted using the ultrasound-assisted extraction (UAE) method, and the bioactive compounds were screened using Liquid Chromatography–High Resolution Mass Spectrometry (LC-HRMS) analysis. This study predicted the biological activity of S. polyanthum compounds using PASS Online server. Before docking, we analyzed the protein-protein interactions (PPIs) network of TNFα, NF-kB, SOD, and KEAP1. The molecular docking was done using Autodock Vina in PyRx software and visualized using Discovery Studio. Probability to be active (Pa) was determined.

Results: The bioactive compounds found in S. polyanthum and used in this study were deoxyphomalone, NCGC00169066-01, and phloretin with retention times [min] of 0.886, 0.907, and 8.323, respectively. The predicted biological activity of compounds and controls were anti-inflammatory, immunosuppressant, TNF expression inhibitor, immunomodulatory and HIF1α expression inhibitor (Pa>0.5 for all S. polyanthum compounds and Pa<0.5 for SPD304, MG-132, and MDF). Based on PPIs network analysis, TNFα, NF-kB, SOD, and KEAP1 are associated. The molecular docking analysis showed that deoxyphomalone, NCGC00169066-01, and phloretin had inhibition potential against TNFα and NF-kB, and activation potential against SOD, due to several residues involved in the interaction of compounds-protein was the same as the interaction of inhibitor (SPD-304 and MG-132) and activator (gallic acid) control against the protein. The residues may have the same inhibition or activation mechanism as the control. However, S. polyanthum bioactive compounds may still have inhibition potential against KEAP1 through Ala548 residue that is also involved in the interaction of DMF-KEAP1.

Conclusions: The bioactive compounds of S. polyanthum showed anti-inflammation and antioxidant potential, which may have a good effect in the treatment of PCOS, yet still need to be confirmed in vitro or in vivo research.

Keywords: antioxidant; inflammation; molecular docking; polycystic ovary syndrome; Syzygium polyanthum.


Contexto: El síndrome de ovario poliquístico (SOP) está significativamente asociado con la inflamación y el estrés oxidativo. Syzygium polyanthum es una planta rica en propiedades farmacológicas. Objetivos: Evaluar el potencial anti-inflamatorio y antioxidante de los compuestos bioactivos de S. polyanthum utilizando un enfoque in silico.

Métodos: S. polyanthum se extrajo mediante el método de extracción asistida por ultrasonido (UAE), y los compuestos bioactivos se seleccionaron mediante análisis de cromatografía líquida-espectrometría de masas de alta resolución (LC-HRMS). Este estudio predijo la actividad biológica de los compuestos de S. polyanthum utilizando el servidor PASS Online. Antes del acoplamiento, analizamos la red de interacciones proteína-proteína (PPI) de TNFα, NF-kB, SOD y KEAP1. El acoplamiento molecular se realizó con Autodock Vina en el software PyRx y se visualizó con Discovery Studio. Se determinó la probabilidad de estar activo (Pa).

Resultados: Los compuestos bioactivos encontrados en S. polyanthum y utilizados en este estudio fueron desoxifomalona, ​​NCGC00169066-01 y floretina con tiempos de retención [min] de 0,886; 0,907 y 8,323, respectivamente. La actividad biológica predicha de los compuestos y controles fue anti-inflamatoria, inmunosupresora, inhibidora de la expresión de TNF, inmunomoduladora e inhibidora de la expresión de HIF1α (Pa>0,5 para todos los compuestos de S. polyanthum y Pa<0,5 para SPD304, MG-132 y MDF). Según el análisis de red de PPI, se asocian TNFα, NF-kB, SOD y KEAP1. El análisis de acoplamiento molecular mostró que la desoxifomalona, ​​NCGC00169066-01 y la floretina tenían potencial de inhibición contra TNFα y NF-kB, y potencial de activación contra SOD, debido a que varios residuos involucrados en la interacción de compuestos-proteína eran los mismos que la interacción del inhibidor (SPD-304 y MG-132) y activador (ácido gálico) controlan contra la proteína. Los residuos pueden tener el mismo mecanismo de inhibición o activación que el control. Sin embargo, los compuestos bioactivos de S. polyanthum aún pueden tener un potencial de inhibición contra KEAP1 a través del residuo Ala548 que también está involucrado en la interacción de DMF-KEAP1.

Conclusiones: Los compuestos bioactivos de S. polyanthum mostraron potencial anti-inflamatorio y antioxidante, lo que puede tener un buen efecto en el tratamiento del SOP, pero aún debe confirmarse en investigaciones in vitro o in vivo.

Palabras Clave: acoplamiento molecular; antioxidante; inflamación; síndrome de ovario poliquistico; Syzygium polyanthum.

Citation Format: Aditya R; Santoso B; Widjiati W (2022) Anti-inflammatory and antioxidant potential of Syzygium polyanthum (Wight) Walp. bioactive compounds in polycystic ovary syndrome: An in silico study. J Pharm Pharmacogn Res 10(4): 725–736.

Amini L, Tehranian N, Movahedin M, Tehrani FR, Ziaee S (2015) Antioxidants and management of polycystic ovary syndrome in Iran: A systematic review of clinical trials.  Iran J Reprod Med 13(1): 1-8.

Arulselvan P, Fard MT, Tan WS, Gothai S, Fakurazi S, Norhaizan ME, Kumar SS (2016) Role of antioxidants and natural products in inflammation. Oxid Med Cell Longev 2016: 5276130.

Gao L, Gu Y, Yin X (2016) High serum tumor necrosis factor-alpha levels in women with polycystic ovary syndrome: a meta-analysis. PloS One 11(10): e0164021

González F (2012) Inflammation in polycystic ovary syndrome: underpinning of insulin resistance and ovarian dysfunction. Steroids 77(4): 300-305.

Hartanti L, Yonas SMK, Mustamu JJ, Wijaya S, Setiawan HK, Soegianto L (2019) Influence of extraction methods of bay leaves (Syzygium polyanthum) on antioxidant and HMG-CoA reductase inhibitory activity. Heliyon 5(4): e01485.

He Z, Wang Y, Zhuan L, Li Y, Tang ZO, Wu Z, Ma Y (2021) MIF-mediated NF-κB signaling pathway regulates the pathogenesis of polycystic ovary syndrome in rats. Cytokine 146: 155632.

Ibáñez L, Oberfield SE, Witchel S, Auchus RJ, Chang RJ, Codner E, Dabadghao P, Darendeliler F, Elbarbary NS, Gambineri A, Garcia Rudaz C, Hoeger KM, López-Bermejo A, Ong K, Peña AS, Reinehr T, Santoro N, Tena-Sempere M, Tao R, Yildiz BO, Alkhayyat H, Deeb A, Joel D, Horikawa R, de Zegher F, Lee PA (2017) An international consortium update: pathophysiology, diagnosis, and treatment of polycystic ovarian syndrome in adolescence. Horm Res Paediatr 88: 371-395.

Ismail A, Ahmad WANW (2019) Syzygium polyanthum (Wight) Walp: A potential phytomedicine. Pharmacogn J 11(2): 429-438.

Kalliolias GD, Ivashkiv LB (2016) TNF biology, pathogenic mechanisms and emerging therapeutic strategies. Nat Rev Rheumatol 12(1): 49-62.

Lagunin AA, Dubovskaja VI, Rudik AV, Pogodin PV, Druzhilovskiy DS, Gloriozova TA, Filimonov DA, Sastry NG, Poroikov VV (2018) CLC-Pred: A freely available web-service for in silico prediction of human cell line cytotoxicity for drug-like compounds. PLoS One 13(1): e0191838.

Li M, Huang W, Jie F, Wang M, Zhong Y, Chen Q, Lu B (2019) Discovery of Keap1− Nrf2 small− molecule inhibitors from phytochemicals based on molecular docking. Food Chem Toxicol 133: 110758.

Liu T, Zhang L, Joo D, Sun SC (2017) NF-κB signaling in inflammation. Signal Transduct Target Ther 2: 17023.

Ma B, Lucas B, Capacci A, Lin EYS, Jones JH, Dechantsreiter M, Richter K (2020) Design, synthesis and identification of novel, orally bioavailable non-covalent Nrf2 activators. Bioorg Med Chem Lett 30(4): 126852.

Mascret A, Mouhsine H, Attia G, Cabrera D, Benchekroun M, Gizzi P, Zerrouki C, Fourati N, Zagury JF, Veitía MS, Port M (2021) New contributions to the drug profile of TNFα inhibitor SPD304: Affinity, selectivity and ADMET considerations. Eur J Pharmacol 907: 174285.

Mohammadi M (2019) Oxidative stress and polycystic ovary syndrome: a brief review. Int J Prev Med 10: 86.

Nafisah W, Pinanti HN, Christina YI, Rifa’i M, Djati MS (2021) Computational biological activity and pharmacological properties analysis for anti-cancer Cyperus rotundus bioactive compounds. AIP Conf Proc 2353: 030118.

Palazon A, Goldrath AW, Nizet V, Johnson RS (2014) HIF transcription factors, inflammation, and immunity. Immunity 41(4): 518-528.

Pandey PK, Ahmed B, Khan HA, Bala M, Prasad J (2019) In silico molecular docking and comparative in-vitro analysis of ethyl 3, 4, 5-trihydroxybenzoate and its derivative isolated from Hippophae rhamnoides leaves as free radical scavenger and anti-inflammatory compound. Pharmacogn Mag 15(64): 313.

Prabhu YD, Borthakur A, Subeka AG, Vellingiri B, Gopalakrishnan AV (2021) Increased pro-inflammatory cytokines in ovary and effect of γ-linolenic acid on adipose tissue inflammation in a polycystic ovary syndrome model. J of Reprod Immunol 146: 103345.

Rani R, Hajam YA, Kumar R, Bhat RA, Rai S, Rather MA (2022) A landscape analysis of the potential role of polyphenols for the treatment of polycystic ovarian syndrome  (PCOS). Phytomedicine Plus 2(1): 100161.

Regidor PA, Mueller A, Sailer M, Gonzalez Santos F, Rizo JM, Moreno Egea F (2020) Chronic inflammation in PCOS: The potential benefits of specialized pro-resolving lipid mediators (SPMs) in the improvement of the resolutive response. Int J Mol Sci 22(1): 384.

Rosenfield RL, Ehrmann DA (2016) The pathogenesis of polycystic ovary syndrome (PCOS): The hypothesis of PCOS as functional ovarian hyperandrogenism revisited. Endocr Rev 37(5): 467-520.

Rudnicka E, Suchta K, Grymowicz M, Calik-Ksepka A, Smolarczyk K, Duszewska AM, Meczekalski B (2021) Chronic low grade inflammation in pathogenesis of pcos. Int J Mol Sci 22(7): 3789.

Sever MJ, Janež A, Dolžan V (2019) Interplay between oxidative stress and chronic inflammation in PCOS: The role of genetic variability in PCOS risk and treatment responses. In (Ed.), Polycystic Ovarian Syndrome. IntechOpen. intechopen.88698.

Shao Y, Cheng Z, Li X, Chernaya V, Wang H, Yang XF (2014) Immunosuppressive/anti-inflammatory cytokines directly and indirectly inhibit endothelial dysfunction-a novel mechanism for maintaining vascular function. J Hematol Oncol 7(1): 80.

Sulaiman MA, Al-Farsi YM, Al-Khaduri MM, Saleh J, Waly MI (2018) Polycystic ovarian syndrome is linked to increased oxidative stress in Omani women. Int J Womens Health 10: 763-771.

Suzuki K, Tominaga T, Ruhee RT, Ma S (2020) Characterization and modulation of systemic inflammatory response to exhaustive exercise in relation to oxidative stress. Antioxidants 9(5): 401.

Tosatti JAG, Sóter MO, Ferreira CN, Silva IFO, Cândido AL, Sousa MO, Reis FM, Gomes KB (2020) The hallmark of pro-and anti-inflammatory cytokine ratios in women with polycystic ovary syndrome. Cytokine 134: 155187.

Uçkan K, Demir H, Turan K, Sarıkaya E, Demir C (2022) Role of oxidative stress in obese and nonobese PCOS patients. Int J Clin Pract 2022: 4579831.

Victor VM, Rovira-Llopis S, Bañuls C, Diaz-Morales N, Martinez de Marañon A, Rios-Navarro C, Alvarez A, Gomez M, Rocha M, Hernández-Mijares A (2016) Insulin resistance in PCOS patients enhances oxidative stress and leukocyte adhesion: Role of myeloperoxidase. PLoS One 11(3): e0151960.

Wang Y, Chen Y, Zhang X, Lu Y, Chen H (2020) New insights in intestinal oxidative stress damage and the health intervention effects of nutrients: A review. J Funct Food 75: 104248

Witchel SF, Oberfield SE, Peña AS (2019) Polycystic ovary syndrome: pathophysiology, presentation, and treatment with emphasis on adolescent girls. J Endocr Soc 3(8): 1545-1573.

Zhang W, Xu W, Chen W, Zhou Q (2018) Interplay of autophagy inducer rapamycin and proteasome inhibitor MG132 in reduction of foam cell formation and inflammatory cytokine expression. Cell Transplant 27(8): 1235-1248.

Zuo T, Zhu M, Xu W (2016) Roles of oxidative stress in polycystic ovary syndrome and cancers. Oxid Med Cell Longev 2016: 8589318.

© 2022 Journal of Pharmacy & Pharmacognosy Research (JPPRes)

Resistant hypertension in end-stage kidney disease

J. Pharm. Pharmacogn. Res., vol. 10, no. 4, pp. 717-724, July-August 2022.

Short Communication

Usefulness of American College of Cardiology/American Heart Association treatment-resistant hypertension criteria among end-stage kidney disease patients – A case series

[Utilidad de los criterios de hipertensión resistente al tratamiento del Colegio Americano de Cardiología/Asociación Americana del Corazón entre pacientes con enfermedad renal en etapa terminal: Una serie de casos]

Shorouq Khader Abujadallah1, Seeba Zachariah1*, Rajaram Jagdale2,3

1College of Pharmacy, Gulf Medical University, Ajman, United Arab Emirates.

2Department of Nephrology, Thumbay University Hospital, Ajman, United Arab Emirates.

3College of Medicine, Gulf Medical University, Ajman, United Arab Emirates.



Context: American College of Cardiology/American Heart Association (ACC/AHA) antihypertensive treatment-resistance criteria have been not used in the hemodialysis (HD) population.

Aims: To assess interim Blood Pressure (BP) measures in End-Stage Kidney Disease (ESKD) patients on maintenance hemodialysis after classifying them using the ACC/AHA criteria.

Methods: The study was a case series assessing blood pressure readings of ESKD patients undergoing maintenance HD at a tertiary-level care hospital in the United Arab Emirates. Blood pressure readings before, during, and after HD sessions were collected and analyzed. SPSS version 26 was used for data analysis.

Results: Out of 55 patients with HD, 10 were classified as having treatment-resistant hypertension, and 6 had controlled hypertension in borderline based on mean post-dialysis Systolic BP (SBP) data of a month. Altogether, the patients underwent 183 dialysis sessions in a month with over 1000 BP readings. The treatment-resistant population showed a p-value less than 0.05 with elevated Pre-SBP, Post-SBP, Mean Intra-SBP, Mean Intra-diastolic BP (DPB), Pre-DBP, Post-DBP, Post-pulse pressure (PP), and Post-mean arterial pressure (MAP).

Conclusions: Use of ACC/AHA criteria for treatment-resistant hypertension might guide setting better treatment goals among ESKD patients on maintenance HD.

Keywords: antihypertensive; blood pressure control; hemodialysis; end-stage kidney disease; resistant hypertension.



Contexto: Los criterios de resistencia al tratamiento antihipertensivo del American College of Cardiology/American Heart Association (ACC/AHA) no se han utilizado en la población de hemodiálisis (HD).

Objetivos: Evaluar las medidas provisionales de presión arterial (PA) en pacientes con enfermedad renal en etapa terminal (ESKD) en hemodiálisis de mantenimiento después de clasificarlos utilizando los criterios ACC/AHA.

Métodos: El estudio fue una serie de casos que evaluó las lecturas de presión arterial de pacientes con ESKD que se sometieron a HD de mantenimiento en un hospital de atención terciaria en los Emiratos Árabes Unidos. Se recogieron y analizaron las lecturas de presión arterial antes, durante y después de las sesiones de HD. Para el análisis de datos se utilizó SPSS versión 26.

Resultados: De los 55 pacientes con HD, 10 se clasificaron con hipertensión resistente al tratamiento y 6 con hipertensión controlada en el límite en función de los datos de PA sistólica (PAS) media posdiálisis de un mes. En total, los pacientes se sometieron a 183 sesiones de diálisis en un mes con más de 1000 lecturas de PA. La población resistente al tratamiento mostró un valor de p inferior a 0,05 con pre-PAS, post-PAS, media intra-PAS, media intradiastólica (PAD), pre-PAD, post-PAD, presión post-pulso elevadas (PP), y post-presión arterial media (MAP).

Conclusiones: El uso de los criterios ACC/AHA para hipertensión resistente al tratamiento podría guiar el establecimiento de mejores objetivos de tratamiento entre los pacientes con ESKD en HD de mantenimiento.

Palabras Clave: antihipertensivos; control de la presión arterial; enfermedad renal terminal; hemodiálisis; hipertensión resistente.


Citation Format: Abujadallah SK, Zachariah S, Jagdale R (2022) Usefulness of American College of Cardiology/American Heart Association treatment-resistant hypertension criteria among end-stage kidney disease patients – A case series. J Pharm Pharmacogn Res 10(4): 717–724.

Aftab RA, Sellappans R, Ming CK, Shaik I (2020) Taking a step further in identifying ideal blood pressure range among hemodialysis patients: A systematic review and a meta-analysis. Front Pharmacol 11: 729.

Agarwal R, Sinha AD, Pappas MK, Abraham TN, Tegegne GG (2014) Hypertension in hemodialysis patients treated with atenolol or lisinopril: a randomized controlled trial. Nephrol Dial Transplant 29(3): 672-681.

Agarwal R, Nissenson AR, Batlle D, Coyne DW, Trout JR, Warnock DG (2003) Prevalence, treatment, and control of hypertension in chronic hemodialysis patients in the United States. Am J Med 115(4): 291-297.

Bakris GL, Burkart JM, Weinhandl ED, McCullough PA, Kraus MA (2016) Intensive hemodialysis, blood pressure, and antihypertensive medication use. Am J Kidney Dis 68(5, Supplement 1): S15-S23.

Davenport A, Cox C, Thuraisingham R (2008) Achieving blood pressure targets during dialysis improves control but increases intradialytic hypotension. Kidney Int 73(6): 759-764.

de Jager DJ, Grootendorst DC, Jager KJ, van Dijk PC, Tomas LM, Ansell D, Collart F, Finne P, Heaf JG, De Meester J, Wetzels JF (2009) Cardiovascular and noncardiovascular mortality among patients starting dialysis. JAMA 302(16): 1782-1789.

Flythe JE, Chang TI, Gallagher MP, Lindley E, Madero M, Sarafidis PA, Unruh ML, Wang AY, Weiner DE, Cheung M, Jadoul M (2020) Blood pressure and volume management in dialysis: conclusions from a Kidney Disease: Improving Global Outcomes (KDIGO) Controversies Conference. Kidney Int 97(5): 861-876.

Georgianos PI, Agarwal R (2021) Antihypertensive therapy in patients receiving maintenance hemodialysis: A narrative review of the available clinical-trial evidence. Curr Vasc Pharmacol 19(1): 12-20.

Karavetian M, Salhab N, Rizk R, Poulia KA (2019) Malnutrition-inflammation score vs phase angle in the Era of GLIM criteria: A cross-sectional study among hemodialysis patients in UAE. Nutrients 11(11): 2771.

Kramer HJ, Townsend RR, Griffin K, Flynn JT, Weiner DE, Rocco MV, Choi MJ, Weir MR, Chang TI, Agarwal R, Beddhu S (2019) KDOQI US Commentary on the 2017 ACC/AHA Hypertension Guideline. Am J Kidney Dis 73(4): 437-458.

K/DOQI (2005) Clinical practice guidelines for cardiovascular disease in dialysis patients. Am J Kidney Dis 45: 16-153.

Mallamaci F, Torino C, Sarafidis P, Loutradis C, Karpetas A, Raptis V, Papagianni A, Ekart R, Siamopoulos K, Del Giudice A, Aucella F (2019) Apparent treatment-resistant hypertension in the hemodialysis population: An Ambulatory BP Monitoring (ABPM) based study. Hypertension 37: e15.

Maruyama T, Takashima H, Abe M (2020) Blood pressure targets and pharmacotherapy for hypertensive patients on hemodialysis. Expert Opin Pharmacother 21(10): 1219-1240.

McCallum W, Sarnak MJ (2019) Blood pressure target for the dialysis patient. Semin Dial 32(1): 35-40.

Morais JG, Pecoits‐Filho R, Canziani ME, Poli‐de‐Figueiredo CE, Cuvello Neto AL, Barra AB, Calice‐Silva V, Raimann JG, Nerbass FB (2020) Fluid overload is associated with the use of a higher number of antihypertensive drugs in hemodialysis patients. Hemodial Int 24(3): 397-405.

Santos SFF, Mendes RB, Santos CA, Dorigo D, Peixoto AJ (2003) Profile of interdialytic blood pressure in hemodialysis patients. Am J Nephrol 23(2): 96-105.

Sars B, van der Sande FM, Kooman JP (2020) Intradialytic hypotension: Mechanisms and outcome. Blood Purif 49(1-2):158-167.

Stidley CA, Hunt WC, Tentori F, Schmidt D, Rohrscheib M, Paine S, Bedrick EJ, Meyer KB, Johnson HK, Zager PG (2006) Changing relationship of blood pressure with mortality over time among hemodialysis patients. J Am Soc Nephrol 17(2): 513-520.

Reeves PB, Mc Causland FR (2018) Mechanisms, Clinical implications, and treatment of intradialytic hypotension. Clin J Am Soc Nephrol 13(8): 1297-1303.

Rohrscheib MR, Myers OB, Servilla KS, Adams CD, Miskulin D, Bedrick EJ, Hunt WC, Lindsey DE, Gabaldon D, Zager PG (2008) Age-related blood pressure patterns and blood pressure variability among hemodialysis patients. Clin J Am Soc Nephrol 3(5): 1407-1414.

Tanaka S, Ninomiya T, Hiyamuta H, Taniguchi M, Tokumoto M, Masutani K, Ooboshi H, Nakano T, Tsuruya K, Kitazono T (2019) Apparent treatment-resistant hypertension and cardiovascular risk in hemodialysis patients: Ten-year outcomes of the Q-Cohort study. Sci Rep 9(1): 1043.

Taniyama Y (2016) Management of hypertension for patients undergoing dialysis therapy. Ren Replace. Ther 2(1): 21.

Tsuruya K, Kanda E, Nomura T, Iseki K, Hirakata H (2020) Postdialysis blood pressure is a better predictor of mortality than pre-dialysis blood pressure in Japanese hemodialysis patients: the Japan Dialysis Outcomes and Practice Patterns Study. Hypertens Res 43(8): 791-797.

Van Buren PN, Inrig JK (2017) Special situations: Intradialytic Hypertension/Chronic Hypertension and Intradialytic Hypotension. Semin Dial 30(6): 545-552.

Wang Z, Yu D, Cai Y, Zhao B, Zhang X, Zhao Z (2019) Optimal cut-off threshold in pulse pressure predicting cardiovascular death among newly diagnosed end-stage renal disease patients. Medicine (Baltimore) 98(27): e16340.

© 2022 Journal of Pharmacy & Pharmacognosy Research (JPPRes)

Oryza sativa hydroxypropylated crosslinked starch as disintegrant in tablet

J. Pharm. Pharmacogn. Res., vol. 10, no. 4, pp. 701-716, July-August 2022.

Original Article

Application of hydroxypropylated crosslinked starch from the grains of Oryza sativa L. as potential disintegrant in compressed solid dosage form

[Aplicación de almidón reticulado hidroxipropilado de los granos de Oryza sativa L. como desintegrante potencial en forma de dosificación sólida comprimida]

Elizabeth A. Pagtalunan1,2,3*, Judson Lowie T. Ramos4, Gerard Q. De Guzman5

1Graduate School, Adamson University, Manila, Philippines.

2Belo Medical Group, San Juan, Manila, Philippines.

3School of Pharmacy, Saint Jude College, Manila, Philippines.

4School of Chemical, Biological and Materials Engineering, and Sciences, Mapua University, Manila, Philippines.

5Department of Pharmacology and Toxicology, College of Medicine, University of the Philippines, Manila, Philippines.



Context: The modification of starch from the grains of NSICRc222 rice variety may provide a new material with enhanced disintegrating functionality in compressed solid dosage forms.

Aims: To synthesize hydroxypropylated crosslinked rice starch that may possess characteristics of a good disintegrant.

Methods: Starch was isolated by the wet milling method. The crosslinking was performed using trisodium trimetaphosphate as crosslinking agent, and successively hydroxypropylated by using phosphorous oxychloride. The evidences of chemical modifications were determined by FTIR, TGA, and DSC. The application of modified rice starch as a disintegrant was evaluated by determining the dissolution profile, and was compared to the performance of commercially available disintegrants. The compatibility of modified rice starch was employed using FTIR, and the stability of the formulation was subjected to an accelerated stability study.

Results: The changes in infrared spectra and thermogram confirmed successful chemical modification of rice starch. Allopurinol and modified starch mixture staged from 0 to 3 months showed no significant changes in the infrared spectra, which suggest compatibility of HCR with allopurinol. The dissolution rate of allopurinol tablets at pH 1.2, pH 4.5, and pH 6.8 with various superdisintegrants were comparable to the dissolution rate that utilized hydroxypropylated crosslinked rice starch as disintegrant. The accelerated stability study showed no significant changes from 0th to 3rd month, and no degradation products were detected in the HPLC analysis.

Conclusions: The chemical modification of rice starch through crosslinking and hydroxypropylation yielded a novel material comparable to the commercially available superdisintegrants.

Keywords: crosslinking; disintegrant; hydroxypropylation; immediate release; NSIC Rc222-Tubigan 18; rice starch.



Contexto: La modificación del almidón de los granos de la variedad de arroz NSIC RC 222 puede proporcionar un nuevo material con una funcionalidad de desintegración mejorada en formas de dosificación sólidas comprimidas.

Objetivos: Sintetizar almidón de arroz reticulado hidroxipropilado que pueda poseer características de un buen desintegrante.

Métodos: El almidón se aisló por el método de molienda húmeda. El entrecruzamiento se realizó con trimetafosfato trisódico como agente de entrecruzamiento, y sucesivamente se hidroxipropiló utilizando oxicloruro de fósforo. Las evidencias de modificaciones químicas fueron determinadas por FTIR, TGA y DSC. La aplicación de almidón de arroz modificado como desintegrante se evaluó determinando el perfil de disolución y se comparó con el desempeño de los desintegrantes disponibles comercialmente. La compatibilidad del almidón de arroz modificado se empleó mediante FTIR y la estabilidad de la formulación se sometió a un estudio de estabilidad acelerado.

Resultados: La caracterización por FTIR reveló una modificación exitosa del almidón de arroz según los cambios de espectros. Los cambios en los termogramas confirmaron la modificación exitosa del almidón de arroz. La mezcla de alopurinol y almidón modificado en etapas de 0 a 3 meses no mostró cambios significativos en los espectros infrarrojos, lo que sugiere compatibilidad de HCR con alopurinol. La velocidad de disolución de las tabletas de alopurinol a pH 1,2, pH 4,5 y pH 6,8 con varios superdesintegrantes fue comparable a la velocidad de disolución de HCR. El estudio de estabilidad acelerada no mostró cambios significativos del mes O al 3, y no se detectaron productos de degradación en el análisis HPLC.

Conclusiones: La modificación química del almidón de arroz a través de la reticulación y el hidroxipropilado produjo un material novedoso comparable a los superdesintegrantes disponibles comercialmente.

Palabras Clave: almidón de arroz; desintegrante; hidroxipropilado; liberación inmediata; NSIC Rc222-Tubigan 18; reticulado.


Citation Format: Elizabeth AP, Judson Lowie TR, Gerard QDG (2022) Application of hydroxypropylated crosslinked starch from the grains of Oryza sativa L. as potential disintegrant in compressed solid dosage form. J Pharm Pharmacogn Res 10(4): 701–716.

ASEAN Guideline on Stability of Drug Product (2005) In Update (Issue February): 6-9.

Bindar Y, Efan A, Rahmi (2013) Sodium hydroxide (NaOH) concentration and steeping time duration effects on starch production from dry-milled low quality rice IR 64 grade 3 flour using alkaline-protease enzyme digestion method. Int Food Res J 20(3): 1353–1358.

Casper Pharma LLC (2018) Product information. Zyloprim® (allopurinol). Reference ID: 4357411. 1-9.

Changdeo JS, Vinod M, Shankar KB, Rajaram CA (2011) Physicochemical characterization and solubility enhancement studies of allopurinol solid dispersions. Braz J Pharm Sci 47(3): 513–523.

Chen CR, Lin YH, Cho SL, Yen SY, Wu HLS (1997) Investigation of the dissolution difference between acidic and neutral media of acetaminophen tablets containing a superdisintegrant and a soluble excipient. Chem Pharm Bull 45(3): 509-512.

Colussi R, Pinto VZ, Halal SLME, Vanier NL,Villanova FA, Silva RM, Zavareze EDR, Dias ARG (2014) Structural, morphological, and physicochemical properties of acetylated high-, medium-, and low-amylose rice starches. Carbohydr Polym 103: 405-413.

Dai XL, Yao J, Wu C, Deng JH, Mo YH, Lu TB, Chen, J M (2020) Solubility and permeability improvement of allopurinol by cocrystallization. Cryst Growth Des 20(8): 5160–5168.

Desai PM, Liew CV, Heng PWS (2016) Review of disintegrants and the disintegration phenomena. J Pharm Sci 105(9): 2545–2555.

Feng K, Wen G (2017) Absorbed Pb2+ and Cd2+ ions in water by crosslinked starch xanthate. Int J Polymer Sci 2017: 6470306.

Furrer P (2013) The central role of excipients in drug formulation. Eur Pharm Rev: 1–13.

Granza AG, Travalini AP, Farias FO, Colman TAD, Schnitzler E, Demiate IM (2015) Effects of acetylation and acetylation–hydroxypropylation (dual-modification) on the properties of starch from Carioca bean (Phaseolus vulgaris L.). J Therm Anal Calorim 119: 769–777.

Hausler O, Rowe R, Shesky P, Quinn ME (2009) Starch. Handbook of Pharmaceutical Excipients. 6th Edition. Pharmaceutical Press, pp. 685-691.

Hu X, Xie Y, Jin Z, Xu X, Chen H (2014) Effect of single- , dual- , and triple-retrogradation treatments on in vitro digestibility and structural characteristics of waxy wheat starch. Food Chem 157: 373–379.

Iftikhar SA, Dutta H (2019) Status of polymorphism, physicochemical properties and in vitro digestibility of dual retrogradation-annealing modified rice starches. Int J Biol Macromol 132: 330–339.

Kar M, Chourasiya Y, Maheshwari R, Tekade RK (2019) Chapter 2 – Current Developments in Excipient Science: Implication of Quantitative Selection of Each Excipient in Product Development, pp. 29-83.

Karthik V (2016) Excipients used in the formulation of tablets. Research and reviews. J Chem 5(2): 143–154.

Kunle O (2019) Starch source and its impact on pharmaceutical applications. In: Volume Chemical Properties of Starch, Emeje M, ed. DOI: 10.5772/intechopen.89811.

Lawal OS (2004) Composition, physicochemical properties and retrogradation characteristics of native, oxidised, acetylated and acid-thinned new cocoyam (Xanthosoma sagittifolium) starch. Food Chem 87(2): 205–218.

Lenaerts V, Beck RHF, Bogaert EV, Chouinard F, Höpcke R, Désévaux C (2003) Crosslinked high amylase starch for use in controlled-release pharmaceutical formulations and processes for its manufacture. United States Patent US 6,607,748 B1: 13-15.

Li HH, Gao YH, Zhang LH, Li N, Liu ZF (2020) Synthesis and adsorption properties of crosslinked carboxymethyl starch (CMS). J Phys Conf Ser 1605: 012130.

Liu J, Wang B, Jin L, Zhang J, Liu W, Xie J (2013) Functional, physicochemical properties and structure of crosslinked oxidized maize starch. Food Hydrocoll 36: 45-52.

Liu J, Wang B, Lin L, Zhang J, Liu W, Xie J, Ding Y (2014) Functional, physicochemical properties and structure of cross-linked oxidized maize starch. Food Hydrocoll 36: 49–51.

Liu X, Lan C, Al A, Yu L, Zhou S (2016) Preparation of cross-linked high amylose corn-starch and its effects on self-reinforced starch films. Int J Food Eng 12(7): 673–680.

NDA – New Drug Application (2017) Multidisciplinary Review and Evaluation Lesinurad/Allopurinol (Duzallo). Application Number: 209203Orig1s000. U.S. FDA Center for Drug Evaluation and Research: 40.

Neha D (2017) Immediate release tablets. A review. World J Pharm Pharm Sci 10(8): 595–611.

Pahwa R, Gupta N (2011) Superdisintegrants in the development of orally disintegrating tablets: A review. Int J Pharm Sci Res 2(11): 2767–2780.

Prathyusha CH, Murthy TEGK (2013) Compatibility studies of donepezil with different excipients by using HPLC and FTIR. J Adv Pharm Educ Res 3(3): 273–278.

Radhakrishnan A, Palanisamy V, Sanphui P (2020) Organic molecular salts of allopurinol with improved solubility. Mater Today Proc 40(suppl. 1): S210–S215.

Rahaju MR, Fardiaz D, Kusnandar F, Candra Sunarti T (2014) Characterization of chemical and physical properties of hydroxypropylated and crosslinked arrowroot (Marantha arundinacea) starch. J Eng Technol Sci 45(3): 207–221.

Ravenelle F, Rahmouni M (2006) Contramid: High-amylose starch for controlled drug delivery. ACS Symposium Series 934: 79-104.

Runa F, Sarkar MR, Sultana R, Jahan K, Labu ZK (2013) Study on dissolution improvement of allopurinol by co-grinding and fusion method using solid dispersion technique. J Biomed Pharm Res 2(3): 1–7.

Shangraw R, Mitrevej A, Shah M (1980) A new era of tablet disintegrants. Pharm Tech 4(10): 49–57.

Shen Y, Yao Y, Wang Z, Wu H (2021) Hydroxypropylation reduces the gelatinization temperature of corn starch for textile sizing. Cellulose 28: 5123–5134.

Singh AV, Nath LK (2011) Synthesis and evaluation of physicochemical properties of crosslinked Phaseolus aconitifolius starch. Starch 63: 655–660.

Tiwari A (2015) Practical Biochemistry: A Student Companion. LAP Lambert Academic Publishing, p. 6.

Troy D, Beringer P (2006) Tablet Characteristics. Remington: The Science and Practice of Pharmacy. 21st Edition. Lippincott Williams & Wilkins, pp. 916-918.

US FDA – United States Food and Drugs Administration (2019) Guidance for Industry: Dissolition Testing of Immediate Release Solid Oral Dosage Forms. [Updated October 2019].

USP-NF – United States Pharmacopeia and National Formulary (2021). Allopurinol Tablets. [Updated October 2019].

Usman M, Ishfaq MT, Malik SR, Iqbal, M, Ishfaq B (2014) Alkaline extraction of starch from broken rice of Pakistan. Int J Innov Appl Res 7(1): 146–152.

Xiao HX, Lin QL, Liu GQ, Yu FX (2012) A comparative study of the characteristics of crosslinked, oxidized and dual-modified rice starches. Molecules 17(9): 10946–10957.

Xie Y, Zhang B, Li MN, Chen HQ (2019) Effects of cross-linking with sodium trimetaphosphate on structural and adsorptive properties of porous wheat starches. Food Chem 289: 187–194.

Zaidul ISM, Absar N, Kim S, Suzuki T (2008) DSC study of mixtures of wheat flour and potato, sweet potato, cassava, and yam starches. J Food Eng 86: 68–73.

Zarmpi P, Flanagan T, Meehan E., Mann J, Fotaki N (2020) Biopharmaceutical understanding of excipient variability on drug apparent solubility based on drug physicochemical properties. Case study: Superdisintegrants. AAPS J 22: 46.

Zhao X, Li Z, Wang L, Lai X (2008) Synthesis, characterization, and adsorption capacity of crosslinked starch microspheres with N,N1-Methylene bisacrylamide. J Appl Polym Sci 109: 2571–2575.

© 2022 Journal of Pharmacy & Pharmacognosy Research (JPPRes)

Orthodontic force: Immunohistochemistry versus hematoxylin-eosin

J. Pharm. Pharmacogn. Res., vol. 10, no. 4, pp. 695-700, July-August 2022.

Short Communication

Histological analyses of orthodontic force in Cavia porcellus: Comparison between immunohistochemistry and hematoxylin-eosin

[Análisis histológicos de la fuerza ortodóncica en Cavia porcellus: Comparación entre inmunohistoquímica y hematoxilina-eosina]

Erliera Sufarnap1,3*, Syafruddin Ilyas2, Nazruddin Nazruddin3, Deddi P. Putra4, Aditya Rachmawati3

1Doctorate Program, Faculty of Dentistry, Universitas Sumatera Utara, Medan, Indonesia.

2Department of Biology, Faculty of Mathematics and Natural Science, Universitas Sumatera Utara, Medan, Indonesia.

3Department of Orthodontic, Faculty of Dentistry, Universitas Sumatera Utara, Medan, Indonesia.

4Faculty of Pharmacy, Universitas Andalas, Padang, Indonesia.



Context: Histological quantification of osteoclasts and osteoblasts can evaluate biological responses to orthodontic tooth movement. Histological analysis of bone samples can be technically challenging.

Aims: To evaluate the differences between hematoxylin and eosin (HE) staining and immunohistochemistry (IHC) in quantifying osteoblast and osteoclast cells following the application of static orthodontic force.

Methods: Orthodontic force was applied using a rubber separator around the maxilla incisor of Cavia porcellus. Tooth samples were taken at 0, 4, 8, 14, 21, and 28 days after applying orthodontic force. HE and IHC staining quantify osteoblast and osteoclast cells in the alveolar bone. IHC staining, i.e., Tartrate-resistant acid phosphatase (TRAP) staining, was used to identify osteoclasts, and osteocalcin (OCN) staining was used to identify osteoblasts.

Results: Significantly higher numbers of osteoclasts and osteoblasts were observed with IHC compared to HE staining (p<0.05). Significant positive linear correlations in the numbers of osteoclasts (r = 0.757) and osteoblasts (r = 0.622) identified were observed between IHC and HE staining.

Conclusions: The results of this study indicate HE staining may represent an acceptable alternative method of quantifying osteoclasts and osteoblasts in the preliminary research of orthodontic tooth movement (OTM).

Keywords: hematoxylin; immunohistochemistry; orthodontic; osteocalcin; tartrate-resistant acid phosphatase.



Contexto: La cuantificación histológica de osteoclastos y osteoblastos puede evaluar las respuestas biológicas al movimiento dental ortodóncico. El análisis histológico de muestras de hueso puede ser técnicamente desafiante.

Objetivos: Evaluar las diferencias entre la tinción con hematoxilina y eosina (HE) y la inmunohistoquímica (IHC) en la cuantificación de células de osteoblastos y osteoclastos después de la aplicación de fuerza ortodóncica estática.

Métodos: Se aplicó fuerza de ortodoncia utilizando un separador de goma alrededor del incisivo maxilar de Cavia porcellus. Se tomaron muestras de dientes a los 0, 4, 8, 14, 21 y 28 días después de aplicar la fuerza de ortodoncia. La tinción con HE e IHC cuantifica las células de osteoblastos y osteoclastos en el hueso alveolar. Se usó tinción IHC, es decir, tinción con fosfatasa ácida resistente a tartrato (TRAP), para identificar osteoclastos, y tinción con osteocalcina (OCN) para identificar osteoblastos.

Resultados: Se observaron números significativamente más altos de osteoclastos y osteoblastos con IHC en comparación con la tinción con HE (valor de p<0,05). Se observaron correlaciones lineales positivas significativas en el número de osteoclastos (r = 0,757) y osteoblastos (r = 0,622) identificados entre la tinción IHC y HE.

Conclusiones: Los resultados de este estudio indican que la tinción HE puede representar un método alternativo aceptable para cuantificar osteoclastos y osteoblastos en la investigación preliminar del movimiento dental ortodóncico (OTM).

Palabras Clave: fosfatasa ácida tartrato resistente; hematoxilina; inmunohistoquímica; ortodoncia; osteocalcina.


Citation Format: Sufarnap E, Ilyas S, Nazruddin N, Putra DP, Rachmawati A (2022) Histological analyses of orthodontic force in Cavia porcellus: Comparison between immunohistochemistry and hematoxylin-eosin. J Pharm Pharmacogn Res 10(4): 695–700.

Ariffin S, Yamamoto Z, Abidin I, Wahab R, Ariffin Z (2011) Cellular and molecular changes in orthodontic tooth movement. Sci World J 11: 1788–1803.

Bennett JH, Moffatt S, Horton M (2001) Cell adhesion molecules in human osteoblasts: Structure and function. Histol Histopathol 16: 603–611.

Blumer MJF, Hausott B, Schwarzer C, Hayman AR, Stempel J, Fritsch H (2012) Role of tartrate-resistant acid phosphatase (TRAP) in long bone development. Mech Dev 129: 162–176.

Dang V, Bao S, Ault A, Murray C, McFarlane-Mills J, Chiedi C, Dillon M, Todd JP, DeTolla L, Rao S (2008) Efficacy and safety of five injectable anesthetic regimens for chronic blood collection from the anterior vena cava of guinea pigs. J Am Assoc Lab Anim Sci 47: 56–60.

de Araujo RMS, Oba Y, Moriyama K (2007) Identification of genes related to mechanical stress in human periodontal ligament cells using microarray analysis. J Periodontal Res 42: 15–22.

Downey PA, Siegel MI (2006) Bone biology and the clinical implications for osteoporosis. Phys Ther 86: 77–91.

Fawcett A (2012) Guideline 22: Guidelines for the Housing of Mice in Scientific Institutions. Orange NSW: Animal Welfare Branch, pp. 1–143.

Fedchenko N, Reifenrath J (2014) Different approaches for interpretation and reporting of immunohistochemistry analysis results in the bone tissue – a review. Diagn Pathol 9: 221.

Fischer AH, Jacobson KA, Rose J, Rolf Z (2017) Hematoxylin and eosin staining of tissue and cell sections. CSH Protoc 2008: pdb.prot4986.

Florencio-Silva R, Sasso GR, Sasso-Cerri E, Simões MJ, Cerri PS (2015) Biology of bone tissue: structure, function, and factors that influence bone cells. Biomed Res Int 2015: 421746.

Grosset AA, Loayza-Vega K, Adam-Granger É, Birlea M, Gilks B, Nguyen B, Soucy G, Tran-Thanh D, Albadine R, Trudel D (2019) Hematoxylin and eosin counterstaining protocol for immunohistochemistry interpretation and diagnosis. Appl Immunohistochem Mol Morphol 27: 558–563.

Hamilton PW, van Diest PJ, Williams R, Gallagher AG (2019) Do we see what we think we see? The complexities of morphological assessment. J Pathol 218(3): 285-291.

Holland R, Bain C, Utreja A (2019) Osteoblast differentiation during orthodontic tooth movement. Orthod Craniofac Res 22: 177–182.

Jonsdottir SH, Giesen EBW, Maltha JC (2012) The biomechanical behaviour of the hyalinized periodontal ligament in dogs during experimental orthodontic tooth movement. Eur J Orthod 34: 542–546.

Kirschneck C, Bauer M, Gubernator J, Proff P, Schröder A (2020) Comparative assessment of mouse models for experimental orthodontic tooth movement. Sci Rep 10: 12154.

Matos LL, Trufelli DC, de Matos MG, da Silva Pinhal MA (2010) Immunohistochemistry as an important tool in biomarkers detection and clinical practice. Biomark Insights 5: 9-20.

Meikle MC (2006) The tissue, cellular, and molecular regulation of orthodontic tooth movement: 100 years after Carl Sandstedt. Eur J Orthod 28: 221-240.

Mukaka (2012) Statistics corner: A guide to appropriate use of correlation coefficient in medical research. Malawi Med J 24: 69–71.

Phan TCAA, Xu J, Zheng MH (2004) Interaction between osteoblast and osteoclast: impact in bone disease. Histol Histopathol 19: 1325–1344.

Ruifrok AC, Johnston DA (2001) Quantification of histochemical staining by color deconvolution. Anal Quant Cytol Histol 23: 291–299.

Sufarnap E, Siregar D, Lindawati Y (2020) Effect of vitamin E supplementation on orthodontic tooth movement in Wistar rats: A prelimary study. F1000Res 9: 1093.

Taddei SR, Moura AP, Andrade I Jr, Garlet GP, Garlet TP, Teixeira MM, da Silva TA (2012) Experimental model of tooth movement in mice: A standardized protocol for studying bone remodeling under compression and tensile strains. J Biomech 45: 2729–2735.

Taylor CR, Levenson RM (2006) Quantification of immunohistochemistry? Issues concerning methods, utility and semiquantitative assessment II. Histopathology 49: 411–424.

Vandevska-Radunovic V, Murison R (2010) Emotional stress and orthodontic tooth movement: Effects on apical root resorption, tooth movement, and dental tissue expression of interleukin-1 alpha and calcitonin gene-related peptide immunoreactive nerve fibres in rats. Eur J Orthod 32: 329–335.

Wahab RMA, Dasor MM, Senafi S, Abdullah AAA, Jemain AA, Kasim NA, Yamamoto Z, Ariffin SHZ (2011) Crevicular tartrate resistant acid phosphatase activity and rate of tooth movement under different continuous force applications. African J Pharm Pharmacol 5: 2213–2219.

Watts NB (1999) Clinical utility of biochemical markers of bone remodeling. Clin Chem 45: 1359–1368.

© 2022 Journal of Pharmacy & Pharmacognosy Research (JPPRes)

Metal ion and cytokines trigger dental metal allergy in silico

J. Pharm. Pharmacogn. Res., vol. 10, no. 4, pp. 687-694, July-August 2022.

Original Article

Computational study of Cu2+, Fe2+, Mn2+, Mn3+, Fe3+, CrO42-, Si4+, and Hg+ binding sites identification on cytokines to predict dental metal allergy: An in silico study

[Estudio computacional de Cu2+, Fe2+, Mn2+, Mn3+, Fe3+, CrO42-, Si4+ y Hg+ e identificación de sitios de unión a citocinas para predecir la alergia dental a metales: Un estudio in silico]

Titiek Berniyanti1, Alexander Patera Nugraha2,3*, Novi Nurul Hidayati2, Viol Dhea Kharisma4, Albertus Putera Nugraha5, Tengku Natasha Eleena Binti Tengku Ahmad Noor6,7

1Dental Public Health Department, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia.

2Graduate Student of Dental Health Science, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia.

3Department of Orthodontics, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia.

4Graduate Student of Biology, Department of Biology, Faculty of Mathematics and Natural Science, Universitas Brawijaya, Malang, Indonesia.

5Faculty of Medicine, Universitas Airlangga, Surabaya, Indonesia.

6Membership of Faculty of Dental Surgery, Royal College of Surgeon, Edinburgh University, United Kingdom.

7Malaysian Armed Forces Dental Officer, 609 Armed Forces Dental Clinic, Kem Semenggo, Kuching, Sarawak, Malaysia.



Context: Metal allergy is a general term to describe allergic diseases due to the release of metal ion reactions in the body which are mediated by T cells and involve inflammatory cytokines that can cause morbidity and mortality. Molecular docking is an analysis that can be used to assess the interaction of ligand bonds with target proteins that are used to predict metal allergies caused by metal ions that stimulate cytokines.

Aims: To analyze the binding sites of Cu2+, Fe2+, Mn2+, Mn3+, Fe3+, CrO42-, Si4+, and Hg+  ions on cytokines to predict dental metal allergy through a bioinformatics approach, in silico.

Methods: Metal ion particles consisting of Cu2+, Fe2+, Mn2+, Mn3+, Fe3+, CrO42-, Si4+, and Hg+  were predicted to bind tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin (IL) IL-1b, IL-2, IL-4, IL-10, IL-13, IL-17, IL-23, and IL-33 act as target proteins were examined.

Results: The blind docking simulation succeeded in identifying the comparison of the binding activity of metal ion particles on cytokines target proteins. The docking simulation results show that the metal ion with the most negative binding affinity value binds to the IL-17 protein. Conclusions: Metal ion particles consisting of Cu2+, Fe2+, Mn2+, Mn3+, Fe3+, CrO42-, Si4+, and Hg+ have the most negative binding affinity values for binding to IL-17 protein, which can cause allergic reactions predicted by molecular docking, in silico.

Keywords: allergy; dentistry; good health and well-being; medicine; orthodontics.



Contexto: La alergia a los metales es un término general para describir las enfermedades alérgicas debidas a la liberación de reacciones de iones metálicos en el cuerpo que están mediadas por células T e involucran citocinas inflamatorias que pueden causar morbilidad y mortalidad. El acoplamiento molecular es un análisis que se puede utilizar para evaluar la interacción de los enlaces de ligandos con proteínas diana que se utilizan para predecir alergias a metales causadas por iones metálicos que estimulan las citocinas.

Objetivos: Analizar los sitios de unión de los iones Cu2+, Fe2+, Mn2+, Mn3+, Fe3+, CrO42-, Si4+ y Hg+ en citocinas para predecir la alergia dental a metales mediante un enfoque bioinformático, in silico.

Métodos: Partículas de iones metálicos que consisten en Cu2+, Fe2+, Mn2+, Mn3+, Fe3+, CrO42-, Si4+ y Hg+ fueron predichas para unirse al factor de necrosis tumoral-α (TNF-α), interferón-γ (IFN-γ), interleucina (IL) Se examinaron IL-1b, IL-2, IL-4, IL-10, IL-13, IL-17, IL-23 e IL-33 que actúan como proteínas diana.

Resultados: La simulación de acoplamiento ciego logró identificar la comparación de la actividad de unión de las partículas de iones metálicos en las proteínas diana de las citocinas. Los resultados de la simulación de acoplamiento muestran que el ion metálico con el valor de afinidad de unión más negativo se une a la proteína IL-17.Conclusiones: Las partículas de iones metálicos que consisten en Cu2+, Fe2+, Mn2+, Mn3+, Fe3+, CrO42-, Si4+ y Hg+ tienen los valores de afinidad de unión más negativos para unirse a la proteína IL-17, lo que puede causar reacciones alérgicas predichas por acoplamiento molecular, in silico.

Palabras Clave: alergia; buena salud y bienestar; medicamento; odontología; ortodoncia.


Citation Format: Berniyanti T, Nugraha AP, Hidayati NN, Kharisma VD, Nugraha AP, Tengku NEBTAN (2022) Computational study of Cu2+, Fe2+, Mn2+, Mn3+, Fe3+, CrO42-, Si4+, and Hg+ binding sites identification on cytokines to predict dental metal allergy: An in silico study. J Pharm Pharmacogn Res 10(4): 687–694.

Agnihotri R, Gaur S, Albin S (2020) Nanometals in dentistry: Applications and toxicological implications-a systematic review. Biol Trace Elem Res 197(1): 70–88.

Ardani IGAW, Nugraha AP, Suryani NM, Pamungkas RH, Vitamamy DG, Susanto RA, Sarno R, Fajar A, Kharisma VD, Nugraha AP, Noor TNEBTA (2022) Molecular docking of polyether ether ketone and nano-hydroxyapatite as biomaterial candidates for orthodontic mini-implant fabrication. J Pharm Pharmacogn Res 10(4): 676–686.

Ashraf T, Taneez M, Kalsoom S, Irfan T, Shafique MA (2021) Experimental calculations of metals content in skin-whitening creams and theoretical investigation for their biological effect against tyrosinase enzyme. Biol Trace Elem Res 199(9): 3562–3569.

Bonaventura P, Lamboux A, Albarède F, Miossec P (2017) Regulatory effects of zinc on cadmium-induced cytotoxicity in chronic inflammation. PLoS One 12(7): e0180879.

Chandu GS, Hema BS, Mahajan H, Mishra S (2014) Dental metal allergy: An update. J Res Adv Dent 3(3): 156–163

Christensen TJ, Samant SA, Shin AY (2017) Making sense of metal allergy and hypersensitivity to metallic implants in relation to hand surgery. J Hand Surg Am 42(9): 737–746.

Dundu MAJ, Aditya G (2017) The effect of 50% betel leaf extract (Piper betle L.) on the release of metal ions (Ni, Cr and Fe) in orthodontic brackets. Odonto Dent J 4(1): 32–37.

FDA – Food and Drug Administration (2019) Biological Responses to Metal Implants: Clinical Response to Metal Implants.  

Haddad SF, Helm MM, Meath B, Adams C, Packianathan N, Uhl R (2019) Exploring the incidence, implications, and relevance of metal allergy to orthopaedic surgeons. J Am Acad Orthop Surg Glob Res Rev 3(4): e023.

Her M, Kavanaugh A (2016) Alterations in immune function with biologic therapies for autoimmune disease. J Allergy Clin Immunol 137(1): 19–27.

Hosoki M, Nishigawa K, Tajima T,  Ueda M, Matsuka Y (2018) Cross-sectional observational study exploring clinical risk of titanium allergy caused by dental implants. J Prosthodont Res  62(4): 426–431.

Itoh E, Furumura M, Furue M (2020) Rate of actual metal allergy prior to dental treatment in subjects complaining of possible metal allergy. Asian Pac J Allergy Immunol 38(3): 186–189.

Jukka (2018) Dental Materials (Priciples And Applications): Dental Amalgam 3rd Ed. Paramount Books (Pvt.) Ltd.

Kharisma VD, Ansori ANM,  Nugraha AP (2020) Computational study of ginger (Zingiber officinale) as E6 inhibitor in human papillomavirus type 16 (HPV-16) infection. Biochem Cell Arch 20(1): 3155–3159.

Kharisma VD, Widyananda MH, Ansori ANM, Nege AS, Naw SW, Nugraha AP (2021) Tea catechin as antiviral agent via apoptosis agonist and triple inhibitor mechanism against HIV-1 infection: A bioinformatics approach. J Pharm Pharmacogn Res 9(4): 435–445.

Kitagawa M, Murakami S, Akashi Y, Oka H, Shintani T, Ogawa I,  Inoue T, Kurihara H (2019) Current status of dental metal allergy in Japan. J Prosthodont Res 63(3): 309–312.

Liu J, Wang Y, Zhao H, Mu M, Guo M, Nie X, Sun Y, Xing M (2020) Arsenic (III) or/and copper (II) exposure induce immunotoxicity through trigger oxidative stress, inflammation and immune imbalance in the bursa of chicken. Ecotoxicol Environ Saf  190: 110127.

Maeno M, Tamagawa-Mineoka R, Arakawa Y, Masuda K, Adachi T, Katoh N (2021) Metal patch testing in patients with oral symptoms. J Dermatol 48(1): 85–87.

Nakasone Y, Kumagai K, Matsubara R, Shigematsu H, Kitaura K, Suzuki S, Satoh M, Hamada, Suzuki R RS (2018) Characterization of T cell receptors in a novel murine model of nickel-induced intraoral metal contact allergy. Plos One 13(12): e0209248.

Nugraha AP, Rahmadhani D,  Puspitaningrum MS, Rizqianti Y,  Kharisma VD, Ernawati DS (2021) Molecular docking of anthocyanins and ternatin in Clitoria ternatea as coronavirus disease oral manifestation therapy. J Adv Pharm Technol Res 12(4): 362.

Pazzini C, Pereira L, Peconick A,  Marques LS, Paiva SM (2016) Nickel allergy: Blood and periodontal evaluation after orthodontic treatment. Acta Odontol Latinoam 29(1): 42–48.

Prahasanti C, Nugraha AP,  Kharisma VD, Ansori ANM, Ridwan RD, Putri TPS, Ramadhani NF, Narmada IB, Ardani IGAW, Ramadhani NF, Noor TNEBA (2021) A bioinformatic approach of hydroxyapatite and polymethylmethacrylate composite exploration as dental implant biomaterial. J Pharm Pharmacogn Res 9(5): 746–754.

Ramadhani NF, Nugraha AP, Rahmadhani D, Puspitaningrum MS, Rizqianti Y, Kharisma VD, Noor TNEBTA, Ridwan RD, Ernawati DS, Nugraha AP (2022) Anthocyanin, tartaric acid, ascorbic acid of roselle flower (Hibiscus sabdariffa L.) for immunomodulatory adjuvant therapy in oral manifestation coronavirus disease-19: An immunoinformatic approach. J Pharm Pharmacogn Res 10(3): 418–428.

Rantam FA, Kharisma VD, Sumartono C, Nugraha J, Wijaya AY, Susilowati H, Kuncorojakti S, Nugraha AP (2021) Molecular docking and dynamic simulation of conserved B cell epitope of SARS-CoV-2 glycoprotein Indonesian isolates: An immunoinformatic approach. F1000Res 10: 813.

Rose PW, Prlić A, Altunkaya A, Bi C,  Bradley AR, Christie CH, Costanzo LD, Duarte JM, Dutta S, Feng Z, Green RK, Goodsell DS, Hudson B, Kalro T, Lowe R, Peisach E, Randle C, Rose AS, Shao C, Tao YP, Valasatava Y, Voigt M, Westbrook JD, Woo J, Yang H, Young JY, Zardecki C, Berman HM, Burley SK (2017) The RCSB protein data bank: integrative view of protein, gene and 3D structural information. Nucleic Acids Res 45(D1): D271–D281.

Saito M, Arakaki R, Yamada A, Tsunematsu T, Kudo Y, Ishimaru N (2016) Molecular mechanisms of nickel allergy. Int J Mol Sci 17(2): 202.

Sakaguchi R, Ferracane JPJ (2019) Craig’s Restorative Dental Materials. 14th Ed. Elsevier.

Sitalaksmi RM, Ito K, Ogasawara K, Suto Y, Itabashi M, Ueda K, Hirasawa N, Narushima T, Hendrijantini N, Kresnoadi U, Sasaki K (2019) COX-2 induces T cell accumulation and IFN-γ production during the development of chromium allergy. Autoimmunity 52(5–6): 228–234.

Susanto H, Kharisma V, Listyorini D, Taufiq A, Sunaryono, Aulanni’am (2018) Effectivity of black tea polyphenol in adipogenesis related IGF-1 and its receptor pathway through in silico based study. J Phys Conf Ser 1093(1): 012037.

Takaoka Y, Akiba Y, Nagasawa M,  Ito A, Masui Y, Akiba N, Eguchi K, Miyazawa H, Tabeta K, Uoshima K (2021) The relationship between dental metal allergy, periodontitis, and palmoplantar pustulosis: An observational study. J Prosthodont Res. doi: 10.2186/jpr.JPR_D_20_00307

Teo Wendy ZW, Schalock PC (2016) Hypersensitivity reactions to implanted metal devices: Facts and fictions.  J Investig Allergol Clin Immunol 26(5): 279–294.

Younas, Khan A, Shehzad O, Seo EK, Onder A, Khan S (2021) Anti-allergic activities of umbelliferone against histamine- and picryl chloride-induced ear edema by targeting Nrf2/iNOS signaling in mice. BMC Complement Med Ther 21(1): 215.

© 2022 Journal of Pharmacy & Pharmacognosy Research (JPPRes)

Molecular docking of polyether ether ketone and nano-hydroxyapatite in orthodontics

J. Pharm. Pharmacogn. Res., vol. 10, no. 4, pp. 676-686, July-August 2022.

Original Article

Molecular docking of polyether ether ketone and nano-hydroxyapatite as biomaterial candidates for orthodontic mini-implant fabrication

[Acoplamiento molecular de poliéter éter cetona y nano-hidroxiapatita como biomateriales candidatos para la fabricación de mini-implantes de ortodoncia]

I Gusti Aju Wahju Ardani1,2, Alexander Patera Nugraha1,2,3*, Monika Nilam Suryani1, Ryan Hafidz Putra Pamungkas1, Devani Githa Vitamamy1, Rizky Alif Susanto1, Riyanarto Sarno4, Aziz Fajar4, Viol Dhea Kharisma5, Albertus Putera Nugraha6, Tengku Natasha Eleena binti Tengku Ahmad Noor7,8

1Orthodontics Department, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia.

2Dental Implant Research Group, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia.

3Graduate Student of Dental Health Science, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia.

4Department of Informatics, Institut Teknologi Sepuluh Nopember, Surabaya, Indonesia.

5Department of Biology, Faculty of Mathematics and Natural Science, Universitas Brawijaya, Malang, Indonesia.

6Faculty of Medicine, Universitas Airlangga, Surabaya, Indonesia.

7Membership of Faculty of Dental Surgery, Royal College of Surgeons, Edinburgh University, United Kingdom.

8Malaysian Armed Forces Dental Officer, 609 Armed Forces Dental Clinic, Kem Semenggo, Kuching, Sarawak, Malaysia.



Context: Modified polyether ether ketone (PEEK) by adding nano-hydroxyapatite (HA) material on its fixture for mini-implant fabrication may increase resistance force through osseointegration.

Aims: To analyze the binding molecular docking of PEEK incorporated with HA as a biomaterial candidate for orthodontic mini-implant fabrication through a bioinformatic approach, an in silico study.

Methods: 3D ligand structure consisting of HA, PEEK and target proteins consisting of osteopontin, osteocalcin, osteonectin, bone morphogenetic protein 4 (BMP4), bone morphogenetic protein 2 (BMP2), bone morphogenetic protein 7 (BMP7), alkaline phosphatase (ALP),  runt-related transcription factor 2 (RUNX2), Insulin growth factor-1 (IGF-1), osterix, tartrate-resistant acid phosphatase (TRAP), collagen alpha-1 (COL1A1) obtained from RCSB-PDB. It was analyzed the binding affinity of a single HA, PEEK, and HA + PEEK complex to twelve target proteins related to osseointegration. The types of chemical interactions produced by the ligands in the target protein domain consisted of Van der Waals, hydrogen, hydrophobic, pi, and alkyl.

Results: The blind docking simulation succeeded in identifying the most negative binding affinity; it was found in the HA + PEEK molecular complex compared to HA and PEEK in the single condition. The type of chemical interaction formed consisted of hydrogen, van der Waals, pi, and alkyl. HA+PEEK showed the most negative binding affinity with ALP and IGF-1, as much as -8.7 binding affinity.

Conclusions: The molecular docking of PEEK with HA exhibited a prominent binding affinity with osteogenic markers like ALP and IGF-1 in silico, allowing it to have a higher potential than nano-HA or PEEK as a single biomaterial for osseointegration as the fabrication of mini-implants that may support orthodontic treatment.

Keywords: dentistry; good health and well-being; in silico; medicine; temporary anchorage device.

This image has an empty alt attribute; its file name is jppres_pdf_free.png


Contexto: La poliéter éter cetona modificada (PEEK) puede aumentar la fuerza de resistencia a través de la osteointegración mediante la adición de material de nanohidroxiapatita (HA) para la fabricación de mini-implantes.

Objetivos: Analizar el acoplamiento molecular de PEEK incorporado con HA como candidato a biomaterial para la fabricación de miniimplantes de ortodoncia a través de un enfoque bioinformático, un estudio in silico.

Métodos: Estructura de ligando 3D que consiste en HA, PEEK y proteínas diana como osteopontina, osteocalcina, osteonectina, proteína morfogenética ósea 4 (BMP4), proteína morfogenética ósea 2 (BMP2), proteína morfogenética ósea 7 (BMP7), fosfatasa alcalina (ALP) , factor de transcripción relacionado con runt 2 (RUNX2), factor de crecimiento de insulina-1 (IGF-1), osterix, fosfatasa ácida tartrato resistente (TRAP), colágeno alfa-1 (COL1A1) obtenido de RCSB-PDB. Fue analizada la afinidad de unión del complejo único HA, PEEK y HA + PEEK a doce proteínas diana relacionadas con la osteointegración. Los tipos de interacciones químicas producidas por los ligandos en el dominio de la proteína objetivo consistieron en Van der Waals, hidrógeno, hidrofóbico, pi y alquilo.

Resultados: La simulación a ciegas de acoplamiento logró identificar la afinidad de unión más negativa. Esta se encontró en el complejo molecular HA + PEEK en comparación con HA y PEEK de forma individual. El tipo de interacción química formada consistió en hidrógeno, van der Waals, pi y alquilo. HA+PEEK mostró la afinidad de unión más negativa con ALP e IGF-1, con una afinidad de unión de -8,7.

Conclusiones: El acoplamiento molecular de PEEK con HA exhibió una afinidad de unión prominente con marcadores osteogénicos como ALP e IGF-1 in silico, lo que le permite tener un mayor potencial que HA o PEEK como biomaterial único para la osteointegración como la fabricación de mini-implantes que puedan soportar el tratamiento de ortodoncia.

Palabras Clave: buena salud y bienestar; dispositivo de anclaje temporal; in silico; odontología; medicina.

This image has an empty alt attribute; its file name is jppres_pdf_free.png

Citation Format: Ardani IGAW, Nugraha AP, Suryani NM, Pamungkas RH, Vitamamy DG, Susanto RA, Sarno R, Fajar A, Kharisma VD, Nugraha AP, Noor TNEBTA (2022) Molecular docking of polyether ether ketone and nano-hydroxyapatite as biomaterial candidates for orthodontic mini-implant fabrication. J Pharm Pharmacogn Res 10(4): 676–686.

Albrektsson T, Johansson C (2001) Osteoinduction, osteoconduction and osseointegration. Eur Spine J 10: S96–S101.

Alqahtani AR, Gufran K, Silva F, Rocha MG, Chang J (2021) A clinical case report of a potential acute allergic reaction with titanium dental implant. Case Rep Dent 2021: 5592934.

Antolis M, Anggani HS, Marshadianti D, Ayu D (2021) Mini implant orthodontic as an anchorage in skeletal class II malocclusion treatment with severe dental protrusion [Indonesian]. J Ked Gi Unpad 33(1):71–78.

Ardani IG, Rahmawati D, Narmada IB, Nugraha AP, Taftazani H, Kusumawardani MK (2020) Surface Electromyography unveil the relationship between masticatory muscle tone and maloclusion class I & II in Javanese ethnic patient. J Int Dent Med Res 13(4): 1447–1454.

Barkarmo S, Wennerberg A, Hoffman M, Kjellin P, Breding K, Handa P, Stenport V (2013) Nano-hydroxyapatite-coated PEEK implants: A pilot study in rabbit bone. J Biomed Mater Res Part A 101A(2): 465–471.

Cheng Z, Guo C, Dong W, He FM, Zhao SF, Yang GL (2012) Effect of thin nano-hydroxyapatite coating on implant osseointegration in ovariectomized rats. Oral Surg Oral Med Oral Pathol Oral Radiol 113(3): e48-e53.

Choi JW, Shin S, Lee CY, Lee J, Seo HH, Lim S, Lee S, Kim IK, Lee HB, Kim SW, Hwang KC (2017) Rapid induction of osteogenic markers in mesenchymal stem cells by adipose-derived stromal vascular fraction cells. Cell Physiol Biochem 44(1): 53–65.

de Oliveira PGFP, de Melo Soares MS, Silveira E Souza AMM (2021) Influence of nano-hydroxyapatite coating implants on gene expression of osteogenic markers and micro-CT parameters. An in vivo study in diabetic rats. J Biomed Mater Res A 109(5): 682–694.

Elias CN, Oliveira Ruellas AC, Fernandes DJ (2012) Orthodontic implants: Concepts for the orthodontic practitioner. Int J Dent 2012: 549761.

Geng YM, Ren DN, Li SY, Li ZY, Shen XQ, Yuan YY (2020) Hydroxyapatite-incorporation improves bone formation on endosseous PEEK implant in canine tibia. J Appl Biomater Funct Mater 18: 2280800020975172.

Gromolak S, Krawczenko A, Antończyk A, Buczak K, Kiełbowicz Z, Klimczak A (2020) Biological characteristics and osteogenic differentiation of ovine bone marrow derived mesenchymal stem cells stimulated with FGF-2 and BMP-2. Int J Mol Sci 21(24): 9726.

Guo T, Kang W, Xiao D, Duan R, Zhi W, Weng J (2013) Molecular docking characterization of a four-domain segment of human fibronectin encompassing the RGD loop with hydroxyapatite. Molecules 19(1): 149–158.

Gupitasari A, Putri LS (2018) The prevalence of bad habits as the etiology of angle’s class I malocclusion in orthodontic clinic dental hospital Jember in 2015-2016 [Indonesian]. Pus Kes 6(2): 365–370.

Hayman AR (2008) Tartrate-resistant acid phosphatase (TRAP) and the osteoclast/immune cell dichotomy. Autoimmunity 41(3):218-223.

Henry JP, Bordoni B (2021) Histology, Osteoblasts. In: StatPearls. Treasure Island (FL): StatPearls Publishing; Update May 10, 2021.

Herwanda H, Arifin R, Lindawati L (2016) Knowledge of adolescents age 15-17 at SMAN 4 Banda Aceh City on the side effects of using fixed orthodontic appliances  [Indonesian]. J Syiah Kuala Dent Soc 1(1): 79–84.

Jubhari EH, Dammar I, Launardo V, Goan Y (2020) Implant coating materials to increase osseointegration of dental implant: A systematic review. Sys Rev Pharm 11(12): 35–41.

Kazimierczak P, Przekora A (2020) Osteoconductive and osteoinductive surface modifications of biomaterials for bone regeneration: A concise review. Coatings 10(10): 971.

Kharisma VD, Ansori AN, Nugraha AP (2020) Computational study of ginger (Zingiber officinale) as E6 Inhibitor in human papillomavirus type 16 (HPV-16) infection. Biochem Cell Arch 20: 3155–3159.

Khotib J, Lasandara CSC, Samirah S, Budiatin AS (2019) Acceleration of bone fracture healing through the use of natural bovine hydroxyapatite implant on bone defect animal model. Fol Med Indones 55(33): 176-187.

Kim S, Thiessen PA, Bolton EE, Chen J, Fu G, Gindulyte A, Han L, He J, He S, Shoemaker BA, Wang J (2016) PubChem Substance and Compound databases. Nucleic Acids Res 44(D1): D1202–D1213.

Levingstone TJ, Ardhaoui M, Benyounis K, Looney L, Stokes JT (2015) Plasma sprayed hydroxyapatite coatings: Understanding process relationships using design of experiment analysis. Surf Coat Technol 283: 29–36.

Loblobly M, Anindita PS, Leman MA (2015) Gambaran Maloklusi Berdasarkan Indeks Handicapping Malocclusion Assessment Record (HMAR) Pada Siswa SMAN 9 Manado [Indonesian]. e-GiGi 3(2): 625­–633.

Luqman A, Kharisma VD, Ruiz RA, Götz F (2020) In silico and in vitro study of trace amines (TA) and dopamine (DOP) interaction with human alpha 1-adrenergic receptor and the bacterial adrenergic receptor QseC. Cell Physiol Biochem 54(5): 888–898.

Ma Z, Zhao X, Zhao J, Zhao Z, Wang Q, Zhang C (2020) Biologically modified polyether ether ketone as dental implant material. Front Bioeng Biotechnol 8: 620537.

Megat Badarul Hisham PN, Narmada IB, Alida A, Rahmawati D, Nugraha A,  Putranti N (2019) Effects of vitamin D in alveolar bone remodeling on osteoblast numbers and bone alkaline phosphatase expression in pregnant rats during orthodontic tooth movement. J Orofac Sci 11(2): 79–83.

Mešić E, Muratović E, Redžepagić-Vražalica L, Pervan N, Muminović AJ, Delić M, Glušac M (2021) Experimental & fem analysis of orthodontic mini-implant design on primary stability. Appl Sci 11(12): 5461.

Najeeb S, Bds ZK, Bds SZ, Bds MS (2016) Bioactivity and osseointegration of PEEK are inferior to those of titanium: A systematic review. J Oral Implantol 42(6):512–516.

Naini A, Rubianto M, Latief FDE, Gunadi A, Kristiana D, Hendrijantini N (2020) Inflammatory and immunogenic response of the tissue after application of freeze-dried hydroxyapatite gypsum puger scaffold compared to freeze-dried hydroxyapatite bovine scaffold. Contemp Clin Dent 11(4): 371–375.

Nayak UK, Malviya N (2011) Role of mini-implants in orthodontics. Int J Oral Implantol Clin Res 2(3): 126–134.

Ntolou P, Tagkli A, Pepelassi E (2018) Factors related to the clinical application of orthodontic mini-implants. J Int Oral Health 10(3): 110.

Oka S, Li X, Zhang F (2020) MicroRNA-21 facilitates osteoblast activity. Biochem Biophys Rep 25: 100894.

Papathanasiou I, Kamposiora P, Papavasiliou G, Ferrari M (2020) The use of PEEK in digital prosthodontics: A narrative review. BMC Oral Health 20(1): 217.

Peng XB, Zhang Y, Wang YQ, He Q, Yu Q (2019) IGF-1 and BMP-7 synergistically stimulate articular cartilage repairing in the rabbit knees by improving chondrogenic differentiation of bone-marrow mesenchymal stem cells. J Cell Biochem 120(4): 5570–5582.

Prahasanti C, Nugraha AP, Kharisma VD, Ansori AN, Devijanti R, Ridwan TP, Ramadhani NF, Narmada IB, Ardani IG, Noor TN (2021) A bioinformatic approach of hydroxyapatite and polymethylmethacrylate composite exploration as dental implant biomaterial. J Pharm Pharmacogn Res 9(5): 746–754.

Prahasanti C, Nugraha AP, Saskianti T, Suardita K, Riawan W, Ernawat DS (2020) Exfoliated human deciduous tooth stem cells incorporating carbonate apatite scaffold enhance BMP-2, BMP-7 and attenuate MMP-8 expression duringinitial alveolar bone remodeling in Wistar rats (Rattus norvegicus). Clin Cosmet Investig Dent 12: 79–85.

Purnama YHC, Mastutik G, Putra ST (2018) Increased activity of mature osteoblast from rat bone marrow mesenchymal stem cells in osteogenic medium exposed to melatonin. Fol Med Indones 54(4): 282–288.

Ramadhani NF, Nugraha AP, Rahmadani D, Puspitaningrum MS, Rizqianti Y, Kharisma VD, Noor TNEBTA, Ridwan RD, Ernawati DS, Nugraha AP (2022) Anthocyanin, tartaric acid, ascorbic acid of roselle flower (Hibiscus sabdariffa L.) for immunomodulatory adjuvant therapy in oral manifestation coronavirus disease-19: An immunoinformatic approach. J Pharm Pharmacogn Res 10(3): 418–428.

Rose PW, Prlić A, Altunkaya A, Bi C, Bradley AR, Christie CH, Costanzo LD, Duarte JM, Dutta S, Feng Z, Green RK (2017) The RCSB protein data bank: Integrative view of protein, gene and 3D structural information. Nucleic Acids Res 45(D1): D271–D281.

Samirah, Budiatin AS, Mahyudin F, Khotib J (2021) Fabrication and characterization of bovine hydroxyapatite-gelatin-alendronate scaffold cross-linked by glutaraldehyde for bone regeneration. J Basic Clin Physiol Pharmacol 32(4): 555–560.

Sheoran L, Kumar P, Kumar S, Ulla ST, Hussain F (2021) Implants in orthodontics: A brief review. IP Indian J Orthod Dentofacial Res 7(1): 45–48.

Simangunsong S, Muttaqin Z, Tampubolon IA (2018) Description of malocclusion in Batak students based on dental aesthetic index [Indonesian]. Prima JODS 1(1): 40–48.

Sitasari PI, Narmada IB, Hamid T, Triwardhani A, Nugraha AP, Rahmawati D (2020) East Java green tea methanolic extractcan enhance RUNX2 and osterixexpression during orthodontic tooth movement in vivo. J Pharm Pharmacogn Res 8(4): 290–298.

Syada AN, Kurniawan FK, Wibowo D (2017) Comparison of severity levels and levels of orthodontic treatment needs using the malalignment index overview in junior high schools that have school health units and junior high schools that do not have UKS [Indonesian]. Jur Ked Gigi 2(1): 78–83.

Wang X, Mei L, Jiang X, Jin M, Xu Y, Li J, Li X, Meng Z, Zhu J, Wu F (2021) Hydroxyapatite-coated titanium by micro-arc oxidation and steam-hydrothermal treatment promotes osseointegration. Front Bioeng Biotechnol 9: 625877.

Valenti MT, Dalle Carbonare L, Mottes M (2016) Osteogenic differentiation in healthy and pathological conditions. Int J Mol Sci 18(1): 41.

Vieira-Andrade RG, Paiva S, Marques LS (2015) Impact of malocclusions on quality of life from childhood to adulthood. Iss Contemp Orthod 3: 39–55.

Zhou M, Geng YM, Li SY, Yang XB, Che YJ, Pathak JL, Wu G (2019) Nanocrystalline hydroxyapatite-based scaffold adsorbs and gives sustained release of osteoinductive growth factor and facilitates bone regeneration in mice ectopic model. Journal of Nanomaterials 2019: 1202159.

Zogheib T, Walter-Solana A, De la Iglesia F, Espinar E, Gil J, Puigdollers A (2021) Do titanium mini-implants have the same quality of finishing and degree of contamination before and after different manipulations? An in vitro study. Metals 11(2): 245.

© 2022 Journal of Pharmacy & Pharmacognosy Research (JPPRes)

Virtual screening of lead flavonoids against DENV2

J. Pharm. Pharmacogn. Res., vol. 10, no. 4, pp. 660-675, July-August 2022.

Original Article

Flavonoids as potential inhibitors of dengue virus 2 (DENV2) envelope protein

[Flavonoides como posibles inhibidores de la proteína de la cubierta del virus del dengue 2 (DENV2)]

Rachel Raditya Renantha1, Alvin Richardo Liga1, Christy Bianca Tanugroho1, Lovine Xaviera Denovian1, Siti Lateefa Az Zahra Budiyanto2, Arli Aditya Parikesit2*

1Department of Biomedicine, School of Life Sciences, Indonesia International Institute for Life Sciences, Jl. Pulomas Barat Kav.88 Jakarta 13210 Indonesia.

2Department of Bioinformatics, School of Life Sciences, Indonesia International Institute for Life Sciences, Jl. Pulomas Barat Kav.88 Jakarta 13210 Indonesia.



Context: Dengue viruses (DENVs) are the cause of dengue disease, which is one of the most frequent diseases caused by mosquito-borne viral infections. Currently, no specific treatment is available for dengue.

Aims: To identify the most promising inhibitors of dengue virus 2 (DENV2) envelope protein of DENV2 envelope protein from flavonoids compounds through computational methods.

Methods: Structures of 54 flavonoids were collected, then the compounds were screened based on Lipinski’s rules, and there were only 34 compounds that passed the screening. Then QSAR analysis was performed, followed by molecular docking analysis, ADMET evaluation, and molecular dynamics simulations to assess the stability of the protein.

Results: Based on the QSAR analysis, only 32 compounds were subjected to molecular docking analysis. Silymarin had the highest docking score, while juglanin had the lowest ACE score compared to positive controls. The ADMET evaluation showed silymarin and juglanin had good absorption and could not penetrate the blood-brain barrier. In contrast to silymarin which had negative results for the Ames test, carcinogenicity, skin sensitization, and eye irritation, juglanin was positive for Ames test and skin sensitization. Even though the molecular dynamic simulation of both ligands with DENV2 envelope protein showed unstable confirmation, it did not necessarily mean that the ligands cannot be used as inhibitors since the molecular docking results provide evidence of the ligands binding to the DENV2 envelope protein.

Conclusions: Based on the favorable results of QSAR analysis, molecular docking, and ADMET evaluation, juglanin and silymarin were chosen as the candidate with the most potential for DENV2 envelope protein inhibitors. However, further analyses such as in vitro and in vivo analyses are necessary to validate the result of this study.

Keywords: DENV-2; envelope protein; flavonoids; molecular docking; virtual screening.


Contexto: Los virus del dengue (DENV) son los causantes de la enfermedad del dengue, que es una de las enfermedades más frecuentes causada por infecciones virales transmitidas por mosquitos. Actualmente, no se dispone de un tratamiento específico para el dengue.

Objetivos: Identificar los inhibidores más prometedores de la proteína de la envoltura del virus del dengue 2 (DENV2) de la proteína de la envoltura del DENV2 a partir de compuestos de flavonoides a través de métodos computacionales.

Métodos: Las estructuras de 54 flavonoides fueron recolectadas. Los compuestos se seleccionaron según las reglas de Lipinski y solo 34 compuestos pasaron la selección. Luego se realizó el análisis QSAR, seguido de análisis de acoplamiento molecular, evaluación ADMET y simulaciones de dinámica molecular para evaluar la estabilidad de la proteína.

Resultados: Según el análisis QSAR, solo 32 compuestos se sometieron a análisis de acoplamiento molecular. La silimarina obtuvo la puntuación de acoplamiento más alta, mientras que juglanina obtuvo la puntuación ACE más baja en comparación con los controles positivos. La evaluación ADMET mostró que la silimarina y la juglanina tenían una buena absorción y no podían penetrar la barrera hematoencefálica. En contraste con la silimarina que tuvo resultados negativos para la prueba de Ames, carcinogenicidad, sensibilización de la piel e irritación de los ojos, la juglanina fue positiva para la prueba de Ames y la sensibilización de la piel. Aunque la simulación de la dinámica molecular de ambos ligandos con la proteína de la cubierta de DENV2 mostró una confirmación inestable, no significa necesariamente que los ligandos no puedan usarse como inhibidores, ya que los resultados del acoplamiento molecular proporcionan evidencia de que los ligandos se unen a la proteína de la cubierta de DENV2.

Conclusiones: En base a los resultados favorables del análisis QSAR, el acoplamiento molecular y la evaluación ADMET, la juglanina y la silimarina fueron elegidas como las candidatas con mayor potencial para los inhibidores de la proteína de la envoltura de DENV2. Sin embargo, se necesitan más análisis, como análisis in vitro e in vivo, para validar el resultado de este estudio.

Palabras Clave: acoplamiento molecular; DENV-2; flavonoides; proteína de envoltura; proyección virtual.

This image has an empty alt attribute; its file name is jppres_pdf_free.png
Citation Format: Renantha RR, Liga AR, Tanugroho CB, Denovian LX, Budiyanto SLAZ, Parikesit AA (2022) Flavonoids as potential inhibitors of dengue virus 2 (DENV2) envelope protein. J Pharm Pharmacogn Res 10(4): 660–675.

Badshah S, Faisal S, Muhammad A, Poulson B, Emwas A, Jaremko M (2021) Antiviral activities of flavonoids. Biomed Pharmacother 140(9): 111596.

Bekhit A, Bekhit A (2014) Natural antiviral compounds. Stud Nat Prod Chem 42(1): 195–228.

Boonyasuppayakorn S, Reichert E, Manzano M, Nagarajan K, Padmanabhan R (2014) Amodiaquine, an antimalarial drug, inhibits dengue virus type 2 replication and infectivity. Antivir Res 106(6): 125–134.

Dong ZW, Yuan YF (2018) Juglanin suppresses fibrosis and inflammation response caused by LPS in acute lung injury. Int J Mol Med 41(6): 3353–3365.

Duhovny D, Nussinov R, Wolfson H (2002) Efficient unbound docking of rigid molecules. Lect Notes Comput Sci 2452(1):185–200.

Filimonov D, Lagunin A, Gloriozova T, Rudik A, Druzhilovskii D, Pogodin P, Poroikov V (2014) Prediction of the biological activity spectra of organic compounds using the pass online web resource. Chem Heterocy Comp 50(3): 444–457.

Guan L, Yang H, Cai Y, Sun L, Di P, Li W, Liu G, Tang Y (2019) ADMET-score – a comprehensive scoring function for evaluation of chemical drug-likeness. Medchemcomm 10(1): 148–157.

Hanwell M, Curtis D, Lonie D, Vandermeersch T, Zurek E, Hutchison G (2012) Avogadro: an advanced semantic chemical editor, visualization, and analysis platform. J Cheminform 4(1): 17.

Hengstler J, Oesch F (2001) Ames Test. Ency Gen 1(1): 51–54.

Ho L, Liu P, Wang C, Wu J (2007) The development of a drug discovery virtual screening application on Taiwan. Unigrid Adv Grid Perv Comp 2(3): 38–47.

Hollingsworth SA, Dror RO (2018) Molecular dynamics simulation for all. Neuron 99(6): 1129–1143.

Hou G, Zeng K, Lan H, Wang Q (2018) Juglanin ameliorates UVB‑induced skin carcinogenesis via anti‑inflammatory and proapoptotic effects in vivo and in vitro. Int J Mol Med 42(3): 41–52.

Ismail N, Jusoh S (2016) Molecular docking and molecular dynamics simulation studies to predict flavonoid binding on the surface of DENV2 E protein. Interdiscip Sci 9(4): 499–511.

Karim A, Riahi V, Mishra A, Newton M, Dehzangi A, Balle T, Sattar A (2021) Quantitative toxicity prediction via meta ensembling of multitask deep learning models. ACS Omega 6(18): 12306–12317.

Krug RM, Aramini JM (2009) Emerging antiviral targets for influenza A virus. Trends Pharm Sci 30(6): 269–277.

Kurcinski M, Oleniecki T, Ciemny M, Kuriata A, Kolinski A, Kmiecik S (2018) CABS-flex standalone: a simulation environment for fast modeling of protein flexibility. Bioinformatics 35(4): 694–695.

Lagunin A, Stepanchikova A, Filimonov D, Poroikov V (2000) PASS: prediction of activity spectra for biologically active substances. Bioinformatics 16(8): 747–748.

Loaiza-Cano V, Monsalve-Escudero L, Filho C, Martinez-Gutierrez M, Sousa D (2020) Antiviral role of phenolic compounds against dengue virus: A review. Biomolecules 11(1): 11.

Low Z, OuYong B, Hassandarvish P, Poh C, Ramanathan B (2021) Antiviral activity of silymarin and baicalein against dengue virus. Sci Rep 11(1): 21221.

Mangas-Sanjuan V, González-Alvarez M, Gonzalez-Alvarez I, Bermejo M (2010) Drug penetration across the blood–brain barrier: an overview. Ther Del 1(4): 535–562.

Modrow S, Falke D, Truyen U, Schätzl H (2013) Viruses with single-stranded, positive-sense RNA genomes. Mol Vir 9(12): 185–349.

Morris G, Lim-Wilby M (2008) Molecular docking. Met Mol Bio 443(8): 365–382.

Ninfali P, Antonelli A, Magnani M, Scarpa E (2020) Antiviral properties of flavonoids and delivery strategies. Nutrients 12(9): 2534.

Parikesit AA (2018) Introductory Chapter: The Contribution of Bioinformatics as Blueprint Lead for Drug Design. In Ivana Glavic (Ed.), Molecular Insight of Drug Design (p. 7). InTech.

Parikesit AA, Nurdiansyah R (2021) Natural products repurposing of the H5N1-based lead compounds for the most fit inhibitors against 3C-like protease of SARS-CoV-2. J Pharm Pharmacogn Res 9(5): 730–745.

Poh MK, Yip A, Zhang S, Priestle JP, Ma NL, Smit JM, Schul W (2009) A small molecule fusion inhibitor of dengue virus. Antivir Res 84(3): 260–266.

Qamar M, Ashfaq U, Tusleem K, Mumtaz A, Tariq Q, Goheer A, Ahmed B (2017) In-silico identification and evaluation of plant flavonoids as dengue NS2B/NS3 protease inhibitors using molecular docking and simulation approach. Pak J Pharm Sci 30(6): 2119–2137.

Rajapakse S, Rodrigo C, Rajapakse A (2012) Treatment of dengue fever. Infect Drug Resist 5(1): 103–112.

Schaefer T, Panda P, Wolford R (2021) Dengue Fever. Treasure Island (FL): StatPearls Publishing.

Schneidman-Duhovny D, Inbar Y, Nussinov R, Wolfson H (2005) PatchDock and SymmDock: servers for rigid and symmetric docking. Nucleic Acids Res 33(7): 363–367.

Schwarz S, Sauter D, Wang K, Zhang R, Sun B, Karioti A, Bilia A R, Efferth T, Schwarz W (2014) Kaempferol derivatives as antiviral drugs against the 3a channel protein of coronavirus. Planta Med 80(2-3): 177–182.

Shiloputra AF, Parikesit AA, Darmawan JT, Pricillia V, Turista DDR, Ansori, ANM (2021) An overview of the curcumin-based and allicin bioactive compounds as potential treatment to SARS-CoV-2 with structural bioinformatics tools. J Tek Lab 10(2): 59–67.

Surai P (2015) Silymarin as a natural antioxidant: An overview of the current evidence and perspectives. Antioxidants 4(1): 204–247.

Tantawichien T, Thisayakorn U (2017) Dengue. Negl Trop Dis- South Asia 5(10): 329–348.

Tian W, Chen C, Lei X, Zhao J, Liang J (2018a) CASTp 3.0: computed atlas of surface topography of proteins. Nucleic Acids Res 46(1): 363–367.

Tian Y, Zhou Y, Takagi T, Kameoka M, Kawashita N (2018b) Dengue virus and its inhibitors: A brief review. Chem Pharm Bull (Tokyo) 66(3): 191–206.

Verma R, Jatav V, Sharma S (2015) Identification of inhibitors of dengue virus (DENV1, DENV2 and DENV3) NS2b/NS3 serine protease: A molecular docking and simulation approach. Asian J Pharm Clin Res 8(1): 287–292.

Vicente C, Herbinger K, Fröschl G, Malta Romano C, de Souza Areias Cabidelle A, Cerutti Junior C (2016) Serotype influences on dengue severity: a cross-sectional study on 485 confirmed dengue cases in Vitória, Brazil. BMC Infect Dis 16(1): 320.

Vora J, Patel S, Athar M, Sinha S, Chhabria MT, Jha PC, Shrivastava N (2019) Pharmacophore modeling, molecular docking and molecular dynamics simulation for screening and identifying anti-dengue phytocompounds. J Biomol Struct Dyn 38(6): 1726–1740.

Wang L, Song J, Liu A, Xiao B, Li S, Wen Z, Lu Y, Du G (2020) Research progress of the antiviral bioactivities of natural flavonoids. Nat Prod Bioprospect 10(5): 271–283.

Wang N, Huang C, Dong J, Yao Z, Zhu M, Deng Z (2017) Predicting human intestinal absorption with modified random forest approach: a comprehensive evaluation of molecular representation, unbalanced data, and applicability domain issues. RSC Adv 7(31): 19007–19018.

Weber C, Sliva K, von Rhein C, Kümmerer B, Schnierle B (2015) The green tea catechin, epigallocatechin gallate, inhibits chikungunya virus infection. Antiviral Res 113(1): 1–3.

Wessel M, Mente S (2001) Chapter 25. ADME by computer. Annu Rep Med Chem 36(1): 257–266.

WHO – World Health Organization (2009) Dengue Guidelines for Diagnosis, Treatment, Prevention and Control, pp. 10–11.

WHO – World Health Organization (2022) Dengue and severe dengue. [Consulted 24 February 2022].

Xiong G, Wu Z, Yi J, Fu L, Yang Z, Hsieh C (2021) ADMETlab 2.0: an integrated online platform for accurate and comprehensive predictions of ADMET properties. Nucleic Acids Res 49(1): 5–14.

Yan A, Wang Z, Cai Z (2008) Prediction of human intestinal absorption by GA feature selection and support vector machine regression. Int J Mol Sci 9(10): 1961–1976.

Yennamalli R, Subbarao N, Kampmann T, McGeary R, Young P, Kobe, B (2009) Identification of novel target sites and an inhibitor of the dengue virus E protein. J Comput Aided Mol Des 23(6): 333–341.

Zandi K, Teoh BT, Sam SS, Wong PF, Mustafa M, AbuBakar S (2011) Antiviral activity of four types of bioflavonoid against dengue virus type-2. Virol J 8(1): 560.

Zarei M, Abidin N, Auwal S, Chay S, Abdul Haiyee Z, Md Sikin A, Saar N (2019) Angiotensin converting enzyme (ACE)-peptide interactions: Inhibition kinetics, in silico molecular docking and stability study of three novel peptides generated from palm kernel cake proteins. Biomolecules 9(10): 569

© 2022 Journal of Pharmacy & Pharmacognosy Research (JPPRes)

Lysozyme and early childhood caries

J. Pharm. Pharmacogn. Res., vol. 10, no. 4, pp. 652-659, July-August 2022.

Original Article

Lysozyme quantity and quality in relation with early childhood caries: A longitudinal study

[Cantidad y calidad de lisozima en relación con las caries de la primera infancia: Un estudio longitudinal]

Essie Octiara1*, Heriandi Sutadi2,Yahwardiah Siregar3, Ameta Primasari4

1Department of Pediatric Dentistry, Faculty of Dentistry, Universitas Sumatera Utara, Medan, Indonesia.

2Departement of Pediatric Dentistry, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia.

3Departement of Biochemistry, Faculty of Medicine, Universitas Sumatera Utara, Medan, Indonesia,

4Departement of Oral Biology, Faculty of Dentistry, Universitas Sumatera Utara, Medan, Indonesia.



Context: Several studies have reported the relationship between lysozyme quantity (concentration) and quality (activity) with early childhood caries (ECC), but the results remain controversial. These are controversial, probably due to the cross-sectional design used in previous studies.

Aims: To analyze the relation between lysozyme quantity and quality with ECC in two years old children.

Methods: An observational analytic with a cohort study for nine months, with a total sample of 68 caries-free children aged 9-24 months old, selected by purposive sampling from Integrated Healthcare Center in Medan, Indonesia. The subject must have at least two primary upper incisors that have erupted. Lysozyme examination was carried out three times, at the beginning of the month, the third and the ninth month. ECC examination was assessed using the American Association of Pediatric Dentistry criteria. Lysozyme concentration was measured using a competitive ELISA method with human lysozyme C (Fine Test). Then lysozyme reading was done at an absorbance of 450 nm using a microplate reader. Lysozyme activity was assessed using the Lysozyme Detection Kit (Sigma-Aldrich and ready by spectrophotometer. Data analysis was processed with an unpaired t-test, Mann-Whitney test, and longitudinal analysis using a generalized estimating equation (linear) test with a significant value, p<0.05.

Results: This showed that there was not any relationship found between lysozyme concentration and ECC (p>0.05). However, there was a relationship between lysozyme activity and ECC (p=o.oo8).

Conclusions: ECC children had higher lysozyme activity when compared to caries-free children.

Keywords: early childhood caries; longitudinal study; lysozyme activity; lysozyme concentration.

This image has an empty alt attribute; its file name is jppres_pdf_free.png


Contexto: Varios estudios han informado la relación de la cantidad (concentración) y la calidad (actividad) de lisozima con las caries de la primera infancia (ECC), pero los resultados siguen siendo controvertidos. Estos son controvertidos probablemente debido al diseño transversal utilizado en estudios previos.

Objetivos: Analizar longitudinalmente la relación entre la cantidad y calidad de lisozima con la ECC en niños de dos años.

Métodos: Un análisis observacional con un estudio de cohorte durante nueve meses, con una muestra total de 68 niños sin caries de 9 a 24 meses de edad, seleccionados mediante un muestreo intencional del Centro de Salud Integrado en Medan, Indonesia. Los sujetos debieron tener al menos dos incisivos superiores primarios que hayan erupcionado. Examen de lisozima fue realizado tres veces, al mes inicial, al tercero y al noveno mes. El examen de ECC se evaluó utilizando los criterios de la Asociación Estadounidense de Odontología Pediátrica. La concentración de lisozima se midió utilizando un método ELISA competitivo con lisozima C humana (prueba fina). Luego se realizó la lectura de lisozima a una absorbancia de 450 nm utilizando un lector de microplacas. La actividad de la lisozima se evaluó utilizando el kit de detección de lisozima (Sigma-Aldrich y leido por espectrofotómetro. El análisis de datos se procesó con la prueba t no pareada, la prueba de Mann-Whitney y el análisis longitudinal utilizando una prueba de ecuación de estimación generalizada (lineal) con valor significativo, p<0.05.

Resultados: Esto mostró que no se encontró relación entre la concentración de lisozima y la ECC (p>0.05). Sin embargo, hubo una relación entre la actividad de la lisozima y la ECC (p=o.oo8).

Conclusiones: Los niños con ECC presentaron mayor actividad de lisozima en comparación con los niños sin caries.

Palabras Clave: actividad de lisozima; caries de la primera infancia; concentración de lisozima; estudio longitudinal.

This image has an empty alt attribute; its file name is jppres_pdf_free.png

Citation Format: Octiara E, Heriandi S, Yahwardiah S, Ameta P (2022) Lysozyme quantity and quality in relation with early childhood caries: A longitudinal study. J Pharm Pharmacogn Res 10(4): 652–659.

AAPD – American Academy of Pediatric Dentistry (2016) Policy on early childhood caries (ECC): Classifications, consequences, and preventive strategies. [Consulted January 10, 2020].

Amerongen AVN, Bolscher JGM, Veerman ECI (2004) Salivary protein: protective and diagnostic value in cariology? Caries Res 38: 247-253.

Amerongen AVN, Veerman ECI (2002) Saliva – the defender of the oral cavity. Oral Dis 8: 12-22.

Bai J, Zhou Q, Bao ZY, Li XX, Qin M (2007) Comparison of salivary proteins between children with early childhood caries and children without caries. Zhonghua Kouqiang Yixue Zazhi 42: 21-23.

Bhalla S, Tandon S, Satyamoorthy K (2010) Salivary proteins and early childhood caries: A gel electrophoretic analysis. Contemp Clin Dent 1: 17-22.

Dawes C, Pedersen AM, Villa A, Ekström J, Proctor GB, Vissink A, Aframian D, McGowan R, Aliko A, Narayana N, Sia YW, Joshi RK, Jensen SB, Kerr AR, Wolff A (2015) The functions of human saliva: A review sponsored by the World Workshop on Oral Medicine VI. Arch Oral Biol 60(6): 863-874.

de Andrade FB, de Oliveira JC, Yoshie MT, Guimarães BM, Gonçalves RB, Schwarcz WD (2014) Antimicrobial activity and synergism of lactoferrin and lysozyme against cariogenic microorganisms. Braz Dent J 25: 165-169.

Fejerskov O, Kidd E (2008) Dental caries: the disease and its clinical management, 2nd edn Oxford: Blackwell Munksgraard Ltd.

Felizardo KR, Goncalves RB, Schwarcz WD, Frederico RCP, Maciel SM, de Andrade FB (2010) An evaluation of the expression profiles of salivary proteins lactoferrin and lysozyme and their association with caries experience and activity. Rev Odonto Ciênc 25(4): 344-349.

Grahn E, Tenovuo J, Lehtonen OP, Eerola E, Vija P (1988) Antimicrobial system of human whole saliva in relation to dental caries, cariogenic bacteria and gingival inflammation in young adults. Acta Odontol Scan 46: 67-74.

Hao GF, Lin HC (2009) Relashionship of concentration of lactoferrin and lysozyme in saliva and dental caries in primary dentition. Zhonghua Kouqiang Yixue Zazhi 44: 82-84.

Hu S, Sim Y, Toh JY, Saw SM, Goodfrey KM, Chong YS, Yap F, Lee YS, Shek LPC, Tan KH, Chong MFF, Hsu CYS (2019) Infant dietary patterns and early childhood caries in a multi-ethnic Asian cohort. Sci Rep 9(1): 852.

Jenzano JW, Hogan SL, Lundblad RL (1986) Factors influencing measurement of human salivary lysozyme in lysoplate and turbidimetric assays. J Clin Microbiol 24(6): 963-967.

Kebijakan Kesehatan Indonesia (2019) Indonesia diharapkan bebas karies pada 2030. [Consulted August 8, 2020].

Leone CW, Oppenheim FG (2001) Physical and chemical aspects of saliva as indicators of risk for dental caries in humans. J Dent Educ 65(10): 1054-1062.

Lertsirivorakul J, Petsongkram B, Chaiyarit P, Klaynongsruang S, Pitiphat W (2015) Salivary lysozyme in relation to dental caries among Thai preschoolers. Pediatr Dent 39(4): 343-347.

Li Y, Wulaerhan J, Liu Y, Abudureyimu A, Zhao J (2017) Prevalence of severe early childhood caries and associated socioeconomic and behavioral factors in Xinjiang, China: a cross-sectional study. BMC Oral Health 17: 144.

Ministry of Health Republic Indonesia (2019) Basic Health Survey, Jakarta- Ministry of Health Republic Indonesia. [Consulted January 5, 2020].

Moslemi M, Sattari M, Kooshki F, Fotuhi F, Modarresi N, Sadrabad ZK, Shadkar MS (2015) Relationship of salivary lactoferrin and lysozyme concentrations with early childhood caries. J Dent Res Dent Clin Dent Prospects 9(2): 109-114.

Nakayama Y, Mori M (2017) Risk factors associated with early childhood caries in 18 to 23-month-old children in a Japanese city. J Natl Inst Public Health 66(5): 545-552.

Nguyen YHT, Ueno M, Zaitsu T, Nguyen T, Kawaguchi Y (2018) Early childhood caries and risk factors in Vietnam. J Clin Pediatr Dent 42(3): 173-181.

Octiara E, Sutadi H, Siregar Y, Primasari A (2018) sIgA and lysozyme as biomarker of early childhood caries. Adv Health Sci Educ Theory Pract 8: 96-101.

Octiara E, Tamba EA (2012) Relationship of family economic and mother education with early childhood caries (ECC) among children 12-36 months aged in district Medan Denai. Dentika Dent J 17(1): 78-82.

Twetman S, Lindner A, Modeer T (1985) Lysozyme and salivary immunoglobulin A in caries-free and caries-susceptible preschool children. Swed Dent J 5: 9-14.

Vasilescu A, Wang Q, Li M, Boukherroub R, Szunerits S (2016) Aptamer-based electrochemical sensing of lysozyme. Chemosensors 4(10): 10.

© 2022 Journal of Pharmacy & Pharmacognosy Research (JPPRes)

Modified pectins with activity against colon cancer

J. Pharm. Pharmacogn. Res., vol. 10, no. 4, pp. 616-651, July-August 2022.


Pectinas modificadas con actividad contra el cáncer de colon: Una revisión sistemática de 2010-2021

[Modified pectins with activity against colon cancer: A systematic review from 2010-2021]

Matías Pérez-Loyola, Marisela Valdés-González, Gabino Garrido*

Departamento de Ciencias Farmacéuticas, Facultad de Ciencias, Universidad Católica del Norte, Angamos 0610, Antofagasta, Chile.



Context: Colon cancer is a serious disease that can be hereditary or arise from exposure to carcinogens in unhealthy food. Current chemotherapy treatment has serious cardiovascular adverse effects, is not always effective, and there may be resistance to treatment. Recently, studies have found an optimal anticancer activity of modified pectins on colon cancer.

Aims: To systematically review the scientific literature, with PRISMA guidelines, of the articles found in the last 11 years, on the potential anticancer effects of pectins on colon cancer.

Methods: PRISMA and Rayyan were used for the selection of studies and a standardized search was followed in four databases with the terms: modified pectins, cancer, therapy. Data extraction was performed using Excel and relevant information on anticancer activity was collected.

Results: In total, 16 articles were included, finding seven in vitro, three mixed and six in vivo studies. The anticancer activity and optimization of the formulation, encapsulation, release of pectin-based drugs were evaluated by different methods in 12 and six articles, respectively. Most of the studies were classified as high quality with the AXIS tool. Furthermore, 11 out of 12 articles showed positive effects of modified pectins on colon cancer both in vivo and in vitro and six out of six articles found positive effects on pectin-based formulations.

Conclusions: The evidence found suggests a beneficial effect of modified pectins on colon cancer, however, more in vivo studies are required.

Keywords: colon cancer; modified pectin; systematic review; therapy.

This image has an empty alt attribute; its file name is jppres_pdf_free.png


Contexto: El cáncer de colon es una patología grave que puede ser hereditaria o surgir por exposición a carcinógenos en la alimentación no saludable. El tratamiento mediante quimioterapia actual posee efectos adversos cardiovasculares graves, no siempre es efectivo y puede existir resistencia al tratamiento. Recientemente, estudios han encontrado una óptima actividad anticancerígena de las pectinas modificadas sobre cáncer de colon.

Objetivos: Revisar sistemáticamente la literatura científica, con directrices PRISMA, de los artículos encontrados en los últimos 11 años, sobre los potenciales efectos anticancerígenos de las pectinas sobre el cáncer de colon.

Métodos: Se utilizó PRISMA y Rayyan para la selección de estudios y se siguió una búsqueda estandarizada en cuatro bases de datos con los términos: modified pectins, cancer, therapy. La extracción de datos se realizó mediante Excel y se recopilo información relevante sobre la actividad anticancerígena.

Resultados: En total, se incluyeron 16 artículos, encontrándose siete estudios in vitro, tres mixtos y seis in vivo. Se evaluó por diferentes métodos la actividad anticancerígena y la optimización de la formulación, encapsulación, liberación de fármacos a base de pectina, en 12 y seis artículos, respectivamente. Mayoritariamente, los estudios fueron clasificados de alta calidad con la herramienta AXIS. Además, 11 de 12 artículos mostraron efectos positivos de las pectinas modificadas sobre el cáncer de colon tanto in vivo como in vitro y seis de seis artículos obtuvieron efectos positivos en las formulaciones a base de pectina.

Conclusiones: La evidencia encontrada sugiere un efecto beneficioso de las pectinas modificadas sobre el cáncer de colon, sin embargo, se requieren más estudios in vivo.

Palabras Clave: cáncer de colon; pectina modificada; revisión sistemática; terapia.

This image has an empty alt attribute; its file name is jppres_pdf_free.png

Citation Format: Pérez-Loyola M, Valdés-González M, Garrido G (2022) Pectinas modificadas con actividad contra el cáncer de colon: Una revisión sistemática de 2010-2021. [Modified pectins with activity against colon cancer: A systematic review from 2010-2021]. J Pharm Pharmacogn Res 10(4): 616–651.

Aldulaymi R, Al Meslamani AZ (2022) Systematic review of the safety and efficacy of antazoline in the treatment of atrial fibrillation. J Pharm Pharmacogn Res 10(1): 147–157.

Almeida EAMS, Facchi SP, Martins AF, Nocchi S, Schuquel ITA, Nakamura CV, Rubira AF, Muniz EC (2015) Synthesis and characterization of pectin derivative with antitumor property against Caco-2 colon cancer cells. Carbohydr Polym 115: 139–145.

American Cancer Society [Internet]. Tratamiento. 2018. Disponible en: [Consultado 7 noviembre 2021].

Arya M, Singh P, Tripathi CB, Parashar P, Singh M, Kanoujia J, Guleria A, Kaithwas G, Gupta KP, Saraf SA (2019) Pectin-encrusted gold nanocomposites containing phytic acid and jacalin: 1,2-dimethylhydrazine-induced colon carcinogenesis in Wistar rats, PI3K/Akt, COX-2, and serum metabolomics as potential targets. Drug Deliv Transl Res 9(1): 53–65.

Basak S, Annapure US (2022) Trends in “green” and novel methods of pectin modification – A review. Carbohydr Polym 278: 118967.

BeMiller JN (1967) Acid-catalyzed hydrolysis of glycosides. Adv Carbohydr Chem 22: 25–108.

Biagi G, Cipollini I, Grandi M, Zaghini G (2010) Influence of some potential prebiotics and fibre-rich foodstuffs on composition and activity of canine intestinal microbiota. Animal Feed Sci Technol 159(1): 50–58.

Blachier F, Beaumont M, Andriamihaja M, Davila AM, Lan A, Grauso M, Armand L, Benamouzig R, Tomé D (2017) Changes in the luminal environment of the colonic epithelial cells and physiopathological consequences. Am J Pathol 187(3): 476–486.

Cao J, Yang J, Wang Z, Lu M, Yue K (2020) Modified citrus pectins by UV/H2O2 oxidation at acidic and basic conditions: Structures and in vitro anti-inflammatory, anti-proliferative activities. Carbohydr Polym 247: 116742.

Cervantes A (2018) Resistencia a la quimioterapia: mecanismos y vías de modulación. Investigación sobre el cáncer en España: de la Biología Molecular a la clínica, pp. 93–99.

Chang C, Wang ZC, Quan CY, Cheng H, Cheng SX, Zhang XZ, Zhuo RX (2007) Fabrication of a novel pH-sensitive glutaraldehyde cross-linked pectin nanogel for drug delivery. J Biomater Sci Polym Ed 18(12): 1591–1599.

Chang WC, Chapkin RS, Lupton JR (1997) Predictive value of proliferation, differentiation and apoptosis as intermediate markers for colon tumorigenesis. Carcinogenesis 18(4): 721–730.

Charles River (2021) Modelos de Investigación y Servicios. Charles River Acquires Vigene Biosciences. Disponible en: [Consultado 18 de diciembre del 2021].

Chasquibol-Silva N, Arroyo-Benites E, Morales-Gomero J (2008) Extracción y caracterización de pectinas obtenidas a partir de frutos de la biodiversidad peruana. Ing Ind 26: 175–199.

Cheewatanakornkool K, Niratisai S, Manchun S, Dass CR, Sriamornsak P (2017) Characterization and in vitro release studies of oral microbeads containing thiolated pectin–doxorubicin conjugates for colorectal cancer treatment. Asian J Pharm Sci 12(6): 509–520.

Chen L, Hao M, Yan J, Sun L, Tai G, Cheng H, Zhou Y (2021) Citrus-derived DHCP inhibits mitochondrial complex II to enhance TRAIL sensitivity via ROS-induced DR5 upregulation. J Biol Chem 296: 100515.

Cheng H, Li S, Fan Y, Gao X, Hao M, Wang J, Zhang X, Tai G, Zhou Y (2011) Comparative studies of the antiproliferative effects of ginseng polysaccharides on HT-29 human colon cancer cells. Med Oncol 28(1): 175–181.

Cho Y, Turner ND, Davidson LA, Chapkin RS, Carroll RJ, Lupton JR (2012) A chemoprotective fish oil/pectin diet enhances apoptosis via Bcl-2 promoter methylation in rat azoxymethane-induced carcinomas. Exp Biol Med 237(12): 1387–1393.

Clapper ML, Chang WL, Cooper HS (2020) Dysplastic aberrant crypt foci: biomarkers of early colorectal neoplasia and response to preventive intervention. Cancer Prev Res 13(3): 229–240.

Comoglu T, Savaşer A, Ozkan Y, Gönül N, Baykara T (2007) Enhancement of ketoprofen bioavailability by formation of microsponge tablets. Pharmazie 62(1): 51–54.

Courts FL (2013) Profiling of modified citrus pectin oligosaccharide transport across Caco-2 cell monolayers. PharmaNutrition 1(1): 22–31.

Dahlin AM, Palmqvist R, Henriksson ML, Jacobsson M, Eklöf V, Rutegård J, Oberg A, Van Guelpen BR (2010) The role of the CpG island methylator phenotype in colorectal cancer prognosis depends on microsatellite instability screening status. Clin Cancer Res 16(6): 1845–1855.

Das S, Ng KY, Ho PC (2011) Design of a pectin-based microparticle formulation using zinc ions as the cross-linking agent and glutaraldehyde as the hardening agent for colonic-specific delivery of resveratrol: In vitro and in vivo evaluations. J Drug Target 19(6): 446–457.

Das S, Ng KY, Ho PC (2010) Formulation and optimization of zinc pectinate beads for the controlled delivery of resveratrol. AAPS PharmSciTech 11(2): 729–742.

do Prado SBR, Shiga TM, Harazono Y, Hogan VA, Raz A, Carpita NC, Fabi JP (2019) Migration and proliferation of cancer cells in culture are differentially affected by molecular size of modified citrus pectin. Carbohydr Polym 211: 141–151.

Downes MJ, Brennan ML, Williams HC, Dean RS (2016) Development of a critical appraisal tool to assess the quality of cross-sectional studies (AXIS). BMJ Open 6: e011458.

Eliaz I, Raz A (2019) Pleiotropic effects of modified citrus pectin. Nutrients 11(11): 2619.

Escobar L, Rivera A, Aristizabal FA (2010) Comparison of resazurin and MTT methods on studies of citotoxicity in human tumor cell lines. Vitae 17(1): 67–74.

Escobar ML (2007) Estudio comparativo de los métodos de rezasurina y MTT empleados en la evaluación de citotoxicidad sobre tres líneas celulares tumorales [tesis de pregrado]. Pontificia Universidad Javeriana, Bogotá, Colombia.

Escobar ML, Aristizabal FA, Alfonso P (2009) Valoración de dos métodos de tinción en ensayos de citotoxicidad sobre líneas celulares tumorales. Rev Col de Biotecnol 11(2): 49–56.

Fan L, Zuo S, Tan H, Hu J, Cheng J, Wu Q, Nie SP (2020) Preventive effects of pectin with various degrees of esterification on ulcerative colitis in mice. Food Funct 11: 2886–2897.

Ferlay J, Ervik M, Lam F, Colombet M, Mery L, Piñeros M, et al. (2020) Global Cancer Observatory: Cancer Today. Lyon: Centro Internacional de Investigaciones sobre el cáncer. Disponible en: [Consultado el 09 de junio del 2021].

Ferreira-Lazarte A, Fernández J, Gallego-Lobillo P, Villar CJ, Lombó F, Moreno FJ, Villamiel M (2021) Behaviour of citrus pectin and modified citrus pectin in an azoxymethane/dextran sodium sulfate (AOM/DSS)-induced rat colorectal carcinogenesis model. Int J Biol Macromol 167: 1349–1360.

Fröhlich E (2012) The role of surface charge in cell uptake and cytotoxicity of medical nanoparticles. Int J Nanomed 7: 5577–5591.

Gao X, Zhi Y, Sun L, Peng X, Zhang T, Xue H, Tai G, Zhou Y (2013) The inhibitory effects of a rhamnogalacturonan I (RG-I) domain from ginseng Pectin on galectin-3 and its structure-activity relationship. J Biol Chem 288(47): 33953–33965.

Glinsky VV, Raz A (2009) Anti-metastatic properties of modified citrus pectin: one bullet, multiple targets. Carbohydr Res 344(14): 1788–1791.

Global Cancer Observatory (2020) New Global Cancer Data: GLOBOCAN 2020, UICC.

Grau de Castro JJ (2005) Inhibidores de la ciclooxigenasa-2 en la prevención del cáncer. Rev Clin Esp 205(9): 446–456.

Habermann N, Schön A, Lund EK, Glei M (2010) Fish fatty acids alter markers of apoptosis in colorectal adenoma and adenocarcinoma cell lines but fish consumption has no impact on apoptosis-induction ex vivo. Apoptosis 15(5): 621–630.

Hansen LT, Allan-Wojtas PM, Jin YL, Paulson AT (2002) Survival of Ca-alginate microencapsulated Bifidobacterium spp. in milk and simulated gastrointestinal conditions. Food Microbiol 19(1): 35–45.

Holze C, Michaudel C, Mackowiak C, Haas DA, Benda C, Hubel P (2018) Oxeiptosis, a ROS-induced caspase-independent apoptosis-like cell-death pathway. Nat Immunol 19(2): 130–140.

Hu YL, Wang XB, Chen DD, Guo XJ, Yang QJ, Dong LH, Cheng L (2016) Germanicol induces selective growth inhibitory effects in human colon HCT-116 and HT29 cancer cells through induction of apoptosis, cell cycle arrest and inhibition of cell migration. J BUON 21(3): 626–632.

Inohara H, Raz A (1994) Effects of natural complex carbohydrate (citrus pectin) on murine melanoma cell properties related to galectin-3 functions. Glycoconj J 11(6): 527–532.

Instituto Nacional del Cáncer (2021) ¿Que es el cáncer?», NIH, Estados Unidos. Disponible en: [Consultado el 9 de junio del 2021].

Itriago GL, Silva NI, Cortes GF (2013) Cáncer en Chile y el mundo: Una mirada epidemiológica, presente y futuro. Rev Méd Clín Las Condes 24(4): 531–552.

Jackson CL, Dreaden TM, Theobald LK, Tran NM, Beal TL, Eid M, Gao MY, Shirley RB, Stoffel MT, Kumar MV, Mohnen D (2007) Pectin induces apoptosis in human prostate cancer cells: Correlation of apoptotic function with pectin structure. Glycobiology 17(8): 805–819.

Jacobasch G, Dongowski G, Florian S, Müller-Schmehl K, Raab B, Schmiedl D (2008) Pectin does not inhibit intestinal carcinogenesis in APC-deficient min/+ mice. J Agric Food Chem 56(4): 1501–1510.

Jacobs LR, Lupton JR (1986) Relationship between colonic luminal pH, cell proliferation, and colon carcinogenesis in 1,2-dimethylhydrazine treated rats fed high fiber diets. Cancer Res 46: 1727–1734.

Jar AM (2014) Bienestar animal y el uso de animales de laboratorio en la experimentación científica. Animal welfare and the use of laboratory animals in scientific research. Rev Argent Microbiol 46(2): 77–79.

Khazir J, Mir BA, Pilcher L, Riley DL (2013) Role of plants in anticancer drug discovery. Phytochem Lett 7(1): 173–181.

Kim WY, Kim JM, Han SB, Lee SK, Kim ND, Park MK, Kim CK, Park JH (2000) Steaming of ginseng at high temperature enhances biological activity. J Nat Prod 63(2):1702–1704.

Leclere L, Fransolet M, Cambier P, Bkassiny SEL, Tikad A, Dieu M, Vincent SP, Van Cutsem P, Michiels C (2016) Identification of a cytotoxic molecule in heat-modified citrus pectin. Carbohydr Polym 137: 39–51.

Leclere L, Van Cutsem P, Michiels C (2013) Anti-cancer activities of pH-or heat-modified pectin. Front Pharmacol 4: 128.

Li D-Q, Li J, Dong H-L, Li X, Zhang J-Q, Ramaswamy S, Xu F (2021) Pectin in biomedical and drug delivery applications: A review. Int J Biol Macromol 185: 49–65.

Li Y, Fan L, Niu Y, Mian W, Zhang F, Xie M, Mei Q (2017) An apple oligogalactan enhances the growth inhibitory effect of 5-fluorouracil on colorectal cancer. Eur J Pharmacol 804: 13–20.

Li Y, Niu Y, Sun Y, Mei L, Zhang B, Li Q, Liu L, Zhang R, Chen J, Mei Q (2014) An apple oligogalactan potentiates the growth inhibitory effect of celecoxib on colorectal cancer. Nutr Cancer 66(1): 29–37.

Liu L, Fishman ML, Kost J, Hicks KB (2003) Sistemas basados en pectina para la administración de fármacos específicos al colon por vía oral. Biomateriales 24(19): 3333–3343.

Liu L, Li YH, Niu YB, Sun Y, Guo ZJ, Li Q, Li C, Feng J, Cao SS, Mei QB (2010) An apple oligogalactan prevents against inflammation and carcinogenesis by targeting LPS/TLR4/NF-κB pathway in a mouse model of colitis-associated colon cancer. Carcinogenesis 31(10): 1822–1832.

Lizano Soberon M, Barrera Rodriquez R (1993) Resistencia múltiple a drogas: un problema en la quimioterapia de cáncer. Rev Invest Clin 45(5): 481–492.

López I, Balderas-Renteríac I, Gómeza I, González-Santiagoc O, González-Barrancoa P, Garza-Ocañas L, Ramírez-Cabrera MA (2020) MTT Vs WST-1, efficiency, cost, time, and waste generation: evaluating the silver nanoparticles cytotoxicity. Rev Cienc Farm Biomed 3: 3–12.

Marangoni VS, Paino IM, Zucolotto V (2013) Synthesis and characterization of jacalin-gold nanoparticles conjugates as specific markers for cancer cells. Colloids Surf B Biointerfaces 112: 380–386.

Martins AF, de Oliveira DM, Pereira AGB, Rubira AF, Muniz EC (2012) Chitosan/TPP microparticles obtained by microemulsion method applied in controlled release of heparin. Int J Biol Macromol 51(5): 1127–1133.

Mat Sharil AT, Ezzat MB, Widya L, Nurhakim Amri HM, Nor Hikmah AR, Nabilah Zafira Z, Haris MS (2022) Systematic review of flaxseed (Linum usitatissimum L.) extract and formulation in wound healing. J Pharm Pharmacogn Res 10(1): 1–12.

Maxwell EG, Belshaw NJ, Waldron KW, Morris VJ (2012) Pectin – an emerging bioactive food polysaccharide. Trends Food Sci Technol 24: 64–73.

Moghaddam FA, Atyabi F, Dinarvand R (2009) Preparation and in vitro evaluation of mucoadhesion and permeation enhancement of thiolated chitosan-pHEMA core-shell nanoparticles. Nanomed Nanotechnol Biol Med 5: 208–215.

Moher D, Liberati A, Tetzlaff J, Altman DG (2009) Preferred reporting items for systematic reviews and meta-analyses: The PRISMA statement. BMJ 339: b2535.

Morales Yera R, Sierra Pérez L, Triana Díaz A (2018) Cardiotoxicidad inducida por quimioterapia. CorSalud 10(1): 68–77.

Morris ER, Powell DA, Gidley MJ, Rees DA (1982) Conformations and interactions of pectins. I. Polymorphism between gel and solid states of calcium polygalacturonate. J Mol Biol 155(4): 507–516.

Nangia-Makker P, Hogan V, Honjo Y, Baccarini S, Tait L, Bresalier R, Raz A (2002) Inhibition of human cancer cell growth and metastasis in nude mice by oral intake of modified citrus pectin. J Natl Cancer Inst 94(24): 1854–1862.

Odun-Ayo F, Mellem J, Naicker T, Reddy L (2015) Chemoprevention of azoxymethane-induced colonic carcinogenesis in Balb/c mice using a modified pectin alginate probiotic. Anticancer Res 35(9): 4765–4776.

Odun-Ayo F, Mellem J, Reddy L (2016) Improving the survival of probiotic in simulated conditions and azoxymethane induced colon tumor bearing mice using modified citrus pectin-alginate microencapsulation. Universidad Tecnológica, Cape Town, South. Afr J Tradit Complement Altern Med 13(2): 101–109.

Organización Mundial de la Salud (2021) Cáncer, WHO. Disponible en: [Consultado 3 de marzo 2021].

Orlandi P, Gentile D, Banchi M, Cucchiara F, Di Desidero T, Cremolini C, Moretto R, Falcone A, Bocci G (2020) Pharmacological effects of the simultaneous and sequential combinations of trifluridine/tipiracil (TAS-102) and 5-fluorouracil in fluoropyrimidine-sensitive colon cancer cells. Investig New Drugs 38(1): 92–98.

Ouzzani M, Hammady H, Fedorowicz Z, Elmagarmid A (2016) Rayyan-a web and mobile app for systematic reviews. Syst Rev 5(1): 210.

Öztürk K, Mashal AR, Yegin BA, Çalış S (2017) Preparation and in vitro evaluation of 5-fluorouracil-loaded PCL nanoparticles for colon cancer treatment. Pharm Dev Technol 22(5): 635–641.

Pai R, Soreghan B, Szabo IL, Pavelka M, Baatar D, Tarnawski AS (2002) Prostaglandin E2, transactivates EGF receptor: A novel mechanism for promoting colon cancer growth and gastrointestinal hypertrophy. Nat Med 8(3): 289–293.

Perše M, Anton C (2011) Morphological and molecular alterations of 1,2-dimethylhydrazine and azoxymethane induced colon carcinogenesis in rats. J Biomed Biotechnol 2011: 473964.

Popov SV, Ovodov YS (2013) Polypotency of the immunomodulatory effect of pectins. Biochemistry 78(7): 823–835.

RevMan (2020) Review Manager [Computer program]. Version 5.4. The Cochrane Collaboration.

Rojas Castillo N, Zuñiga Corvalan C (2020) Potenciales Efectos Anticaries de Fibras Prebióticas: Una Revisión Sistemática de la Literatura [Tesis de Pregrado]. Universidad de Talca, Talca, Chile.

Romero-Fernandez W, Batista-Castro Z, De Lucca M, Ruano A, García-Barceló M, Rivera-Cervantes M, García-Rodríguez J, Sánchez-Mateo S (2016) El 1, 2, 3 de la experimentación con animales de laboratorio. Rev Perú Med Exp Salud Publica 33(2): 288–299.

Sabra R, Billa N, Roberts CJ (2018) An augmented delivery of the anticancer agent, curcumin, to the colon. React Funct Polym 123: 54–60.

Sabra R, Billa N, Roberts CJ (2019) Cetuximab-conjugated chitosan-pectinate (modified) composite nanoparticles for targeting colon cancer. Int J Pharm 572: 118775.

Salama L, Pastor ER, Stone T, Mousa SA (2020) Emerging nanopharmaceuticals and nanonutraceuticals in cancer management. Biomedicines 8(9): 347.

Sánchez C (2013) Conociendo y comprendiendo la célula cancerosa: Fisiopatología Del Cáncer. Rev Méd Clín Las Condes 24(4): 553–562.

Santana-Gálvez J, Villela-Castrejón J, Serna-Saldívar SO, Cisneros-Zevallos L, Jacobo-Velázquez DA (2020) Synergistic combinations of curcumin, sulforaphane, and dihydrocaffeic acid against human colon cancer cells. Int J Mol Sci 21(9): 3108.

Saz-Peiró P, Tejero-Lainez MC (2016) Fitoterapia en la prevención y tratamiento del cáncer. Med Natur 10(2): 88–99.

Sriamornsak P, Nunthanid J (1998) Calcium pectinate gel beads for controlled release drug delivery: I. Preparation and in vitro release studies. Int J Pharm 160: 207–212.

Srivastava R, Kumar D, Pathak K (2012) Colonic luminal surface retention of meloxicam microsponges delivered by erosion based colon-targeted matrix tablet. Int J Pharm 427(2):153–162.

Tremaroli V, Bäckhed F (2012) Interacciones funcionales entre la microbiota intestinal y el metabolismo del huésped. Nature 489 (7415): 242–249.

Tyagi AK, Singh RP, Agarwal C, Chan DC, Agarwal R (2002) Silibinin strongly synergizes human prostate carcinoma DU145 cells to doxorubicin-induced growth Inhibition, G2-M arrest, and apoptosis. Clin Cancer Res 8(11): 3512–3519.

Valdespino-Gómez VM, Valdespino-Castillo PM, Valdespino-Castillo VE (2015) Interacción de las vías de señalización intracelulares participantes en la proliferación celular: potencial blanco de intervencionismo terapéutico. Cir Cir 83(2): 165–174.

Wang H, Gao Z, Liu X, Agarwal P, Zhao S, Conroy DW, Ji G, Yu J, Jaroniec CP, Liu Z, Lu X, Li X, He X (2018) Targeted production of reactive oxygen species in mitochondria to overcome resistance to anticancer drugs. Nat Commun 9(1): 562.

Wang Y, Nangia-Makker P, Balan V, Hogan V, Raz A (2010) Activation of calpain through galectin-3 inhibition sensitizes prostate cancer cells to cisplatin treatment. Cell Death Dis 1(11): 101–110.

Weber K (2017) Differences in types and incidence of neoplasms in Wistar Han and Sprague-Dawley rats. Toxicol Pathol 45(1): 64–75.

Wirtz S, Neufert C, Weigmann B, Neurath MF (2007) Chemically induced mouse models of intestinal inflammation. Nat Protocol 2(3): 541–546.

Wu KL, Kuo CM, Huang EY, Pan HM, Huang CC, Chen YF, Hsiao CC, Yang KD (2018) Extracellular galectin-3 facilitates the migration of colon cancer cells and is related to the epidermal growth factor receptor. Am J Transl Res 10(8): 2402–2412.

Zhang W, Xu P, Zhang H (2015) Pectin in cancer therapy: A review. Trends Food Sci Technol 44(2): 258–271.

Zhao B, Wang L, Qiu H, Zhang M, Sun L, Peng P, Yu Q, Yuan X (2017a) Mechanisms of resistance to anti-EGFR therapy in colorectal cancer. Oncotarget 8 (3): 3980–4000.

Zhao X, Pan J, Li W, Yang W, Qin L, Pan Y (2018) Gold nanoparticles enhance cisplatin delivery and potentiate chemotherapy by decompressing colorectal cancer vessels. Int J Nanomedicine 13: 6207–6221.

Zhao Y, Hu X, Liu Y, Dong S, Wen Z, He W, Zhang S, Huang Q, Shi M (2017b) ROS signaling under metabolic stress: cross-talk between AMPK and AKT pathway. Mol Cancer 16(1): 79.

© 2022 Journal of Pharmacy & Pharmacognosy Research (JPPRes)

Phytochemistry and tyrosinase inhibition of Salvia officinalis extracts

J. Pharm. Pharmacogn. Res., vol. 10, no. 4, pp. 605-615, July-August 2022.

Original Article

Phytochemical characterization and mushroom tyrosinase inhibition of different extracts from Salvia officinalis L. leaves

[Caracterización fitoquímica e inhibición de la tirosinasa de hongos de diferentes extractos de hojas de Salvia officinalis L.]

Lana Y.M. Juee

Department of Pharmacognosy, Pharmacy College, Hawler Medical University, Erbil, Kurdistan Region, Iraq.



Context: Sage (Salvia officinalis) is an ancient valuable plant used in the treatment of variant health issues.

Aims: To evaluate the depigmentation activity of S. officinalis leaf chloroformic (SOCF) and ethanolic (SOMF) extracts via its efficacy to inhibit tyrosinase enzyme using in vitro model and bioassay-guided identification and quantification of the main active constituents.

Methods: Plant extracts efficacy as a depigmentation agent has been studied via mushroom tyrosinase inhibition using in vitro model at two concentrations (100 and 200 µg/mL). Extracts were analyzed for phenolic compounds that could be responsible for the biological activity using LC-MS/MS analysis.

Results: Significant potency at a high concentration of 200 µg/mL for the methanolic extract were recorded (p≤0.05). The LC-MS/MS analysis of S. officinalis leaf extracts revealed the presence of eight and fourteen analytes of origin of thirty-seven analytes in both SOCF and SOMF, respectively. Analytes’ quantification recorded the highest amount for rosmarinic acid (46 016 µg/g) in SOMF and the lowest was hesperidin (0.6 µg/g) in SOCF.

Conclusions: S. officinalis extracts recorded significant tyrosinase inhibition potency could control the melanin synthesis process and exhibit beneficiary effect in hyperpigmentation issues.

Keywords: hesperidin; hyperpigmentation; LC-MS/MS spectroscopy; rosmarinic acid; sage; tyrosinase inhibitors.

This image has an empty alt attribute; its file name is jppres_pdf_free.png


Contexto: La salvia (Salvia officinalis) es una planta antigua y valiosa utilizada en el tratamiento de problemas de salud variantes.

Objetivos: Evaluar la actividad despigmentante de los extractos chorofórmico (SOCF) y etanólico (SOMF) de hojas de S. officinalis a través de su eficacia para inhibir la enzima tirosinasa utilizando un modelo in vitro y la identificación y cuantificación guiada por bioensayos de los principales componentes activos.

Métodos: La eficacia de los extractos de plantas como agente despigmentante se ha estudiado mediante la inhibición de la tirosinasa de hongos utilizando un modelo in vitro a dos concentraciones (100 y 200 µg/mL). Los extractos se analizaron en busca de compuestos fenólicos que pudieran ser responsables de la actividad biológica mediante análisis LC-MS/MS.

Resultados: Se registró potencia significativa a alta concentración 200 µg/mL para el extracto metanólico (p≤0.05). El análisis LC-MS/MS de extractos de hojas de salvia reveló la presencia de ocho y catorce analitos de origen de treinta y siete analitos tanto en SOCF como en SOMF, respectivamente. La cuantificación de los analitos registró la mayor cantidad de ácido rosmarínico (46 016 µg/g) en SOMF y la menor de hesperidina (0,6 µg/g) en SOCF.

Conclusiones: Los extractos de S. officinalis registraron una potencia significativa de inhibición de la tirosinasa que podría controlar el proceso de síntesis de melanina y exhibir un efecto beneficioso en los problemas de hiperpigmentación.

Palabras Clave: ácido rosmarínico; espectroscopia LC-MS/MS; hesperidina; hiperpigmentación; inhibidores de tirosinasa; salvia.

This image has an empty alt attribute; its file name is jppres_pdf_free.png

Citation Format: Juee LYM (2022) Phytochemical characterization and mushroom tyrosinase inhibition of different extracts from Salvia officinalis L. leaves. J Pharm Pharmacogn Res 10(4): 605–615.

Adhikari A, Devotka HP, Takano K, Nakane T, Basnet P, Skalko-Basnet N (2008) Screening of Nepalese crude drugs traditionally used to treat hyperpigmentation: in vitro tyrosinase inhibition. Int J Cosm Sci 30: 353–360.

Akdeniz M (2018) Screening of Chemical Content Specific to Hypericum Species Growing in Different Parts of Turkey by LC-MS/MS and Method Validation; Investigation of Their Biological Activities and Chemometric Evaluation. Dicle University Institute of Science Department of Chemistry, Ph.D. Thesis, Diyarbakır.

Bauer J, Kuehnl S, Rollinger JM, Scherer O, Northoff H, Stuppner H, Werz O, Koeberle A (2012). Carnosol and carnosic acids from Salvia officinalis inhibit microsomal prostaglandin E-2 synthase-1. J Pharmacol Exp Ther 342: 169–176.

Ben Taarit M, Msaada K, Hosni K, Marzouk B (2012) Physiological changes, phenolic content and antioxidant activity of Salvia officinalis L. grown under saline conditions. J Sci Food Agric 92: 1614–1619.

Bisset NG, Wichtl M (2001) Herbal Drugs and Phytopharmaceuticals: A Handbook for Practice on a Scientific Basis with Reference to German Commision E Monographs. 2nd Edition, Boca Raton FI: CRC Press, pp. 440–443.

Biswas R, Mukherjee PK, Dalai MK, Mandal PK, Nag M (2015) Tyrosinase inhibitory potential of purpurin in Rubia cordifolia-A bioactivity guided approach. Ind Crops Prod 74: 319–326.

Boonsiripiphat K, Theerakulkait C (2009) Extraction of rice bran extract and some factors affecting its inhibition of polyphenol oxidase activity and browning in potato. Prep Biochem Biotechnol 39: 147–158.

Burnett CL, Bergfeld WF, Donald VB, Ronald AH, Curtis DK, Daniel CL, James GM, Ronald CS, Thomas JS, Paul WS, Andersen FA (2010) Final report of the safety assessment of kojic acid as used in cosmetics. Int J Toxicol 29: 6.

Chang TS (2009) An updated review of tyrosinase inhibitors. Int J Mol Sci 10: 2440–2475.

Costa RS, Carneiro TCB, Cerqueira-Lima AT, Queiroz NV, Alcantara-Neves NM, Pontes-de-Carvalho LC, Velozo ED, Oliveira EJ, Figueiredo CA (2012) Ocimum gratissimum Linn. and rosmarinic acid, attenuate eosinophilic airway inflammation in an experimental model of respiratory allergy to Blomia tropicalis. Int Immunopharmacol 13: 126–134.

Crespo MI, Chaban MF, Lanza PA, Joray MB, Palacios SM, Vera DMA, Carpinella MC (2019) Inhibitory effects of compounds isolated from Lepechinia meyenii on tyrosinase. Food Chem Toxicol 125: 383–391.

De Oliveira NCD, Sarmento MS, Nunes EA, Porto CM, Rosa DP, Bona SR, Rodrigues G, Marroni NP, Pereira P, Picada JN, Ferraz ABF, Thiesen FV, Da Silva J (2012) Rosmarinic acid as a protective agent against genotoxicity of ethanol in mice. Food Chem Toxicol 50: 1208–1214.

Dinç M, Pinar NM, Dogu S, Yildirimli S (2009) Micromorphological studies of Lallemantia L. (Lamiaceae) species growing in Turkey. Acta Biol Crac Ser Bot 51: 45–54.

Fatiha B, Didier H, Naima G, Khodir M, Martin K, Léocadie K, Caroline S, Mohamed C, Pierre D (2015) Phenolic composition, in vitro antioxidant effects and tyrosinase inhibitory activity of three Algerian Mentha species: M. spicata (L.), M. pulegium (L.) and M. rotundifolia (L.) Huds (Lamiaceae). Ind Crops Prod 74: 722–730.

Fiocco D, Fiorentino D, Frabboni L, Benvenuti S, Orlandini G, Pellati F, Gallone A (2011) Lavender and peppermint essential oils as effective mushroom tyrosinase inhibitors: a basic study. Flavour Fragr J 26: 6, 441–446.

Gunia-Krzy˙zak A, Popiol J, Marona H (2016) Melanogenesis inhibitors: strategies for searching for and evaluation of active compounds. Curr Med Chem 23: 3548–3574.

Ha SK, Koketsu M, Lee K, Choi SY, Park JH, Ishihara H, Kim SY (2005) Inhibition of tyrosinase activity by N, N-unsubstituted selenourea derivatives. Biol Pharm Bull 28: 838–840.

Iuvone T, De Filippis D, Esposito G, D’Amico A, Izzo AA (2006) The spice sage and its active ingredient rosmarinic acid protect PC12 cells from amyloid-beta peptide-induced neurotoxicity. J Pharmacol Exp Ther 317: 1143–1149.

Jakovljević M, Jokić S, Molnar M, Jašić M, Babić J, Jukić H, Banjari I (2019) Bioactive profile of various Salvia officinalis L. preparations. Plants (Basel) 8: 55.

Kang MA, Yun SY, Won J (2003) Rosmarinic acid inhibits Ca2+-dependent pathways of T-cell antigen receptor-mediated signaling by inhibiting the PLC-gamma 1 and Itk activity. Blood 101: 3534–3542.

Karina BO, Érika P, Almeriane M, Weffort-Santos Oliveira BH (2013) Influence of rosmarinic acid and Salvia officinalis extracts on melanogenesis of B16F10 cells. Rev Bras Farmacogn 23: 249–258.

Karmokar A, Marczylo TH, Cai H, Steward WP, Gescher AJ, Brown K (2012) Dietary intake of rosmarinic acid by ApcMin mice, a model of colorectal carcinogenesis: levels of parent agent in the target tissue and effect on adenoma development. Mol Nutr Food Res 56: 775– 783.

Kim DH, Lee JH (2019) Comparative evaluation of phenolic phytochemicals from perilla seeds of diverse species and screening for their tyrosinase inhibitory and antioxidant properties. S Afr J Bot 123: 341–350.

Kim YJ, Uyama H (2005) Tyrosinase inhibitors from natural and synthetic sources: structure, inhibition mechanism and perspective for the future. Cell Mol Life Sci 62: 1707–1723.

Kumari S, Tien GTS, Kumar VN, Gautam HK (2018) Melanogenesis inhibitors. Acta Dermatol Venereol 98: 924–931.

Lee SY, Baek N, Nam TG (2016) Natural, semisynthetic and synthetic tyrosinase inhibitors. J Enzyme Inhib Med Chem 31: 1–13.

Lin VC, Ding H, Kuo S, Chin L, Wu J, Chang T (2011) Evaluation of in vitro and in vivo depigmenting activity of raspberry ketone from Rheum officinale. Int J Mol Sci 12: 4819–4835.

Lu YR, Foo LY (2002) Polyphenolics of Salvia – a review. Phytochemistry 59: 117–140.

Macrini DJ, Suffredini IB, Varella AD, Younes RN, Ohara MT (2009) Extracts from Amazonian plants have inhibitory activity against tyrosinase: an in vitro evaluation. Braz J Pharm Sci 45: 715–21.

Martina J, Stela J, Maja M, Midhat J, Jurislav B, Huska J, Ines B (2019) Bioactive profile of various Salvia officinalis L. preparations. Plants 8: 55.

Nagarani G, Abirami A, Siddhuraju P (2014) A comparative study on antioxidant potentials, inhibitory activities against key enzymes related to metabolic syndrome, and anti-inflammatory activity of leaf extract from different Momordica species. Food Sci Hum 3: 36–46.

Omar SH, Scott CJ, Hamlin AS, Obied HK (2018) Biophenols: enzymes (β-secretase, cholinesterases, histone deacetylase and tyrosinase) inhibitors from olive (Olea europaea L.). Fitoterapia 128: 118–129.

Panzella L, Napolitano A (2019) Natural and bioinspired phenolic compounds as tyrosinase inhibitors for the treatment of skin hyperpigmentation: recent advances. Cosmetics 6: 57.

Pillaiyar T, Namasivayam V, Manickam M, Jung SH (2018) Inhibitors of melanogenesis: an updated review. J Med Chem 61: 7395–7418.

Psotova J, Svobodova A, Kolarova H, Walterova D (2006) Photoprotective properties of Prunella vulgaris and rosmarinic acid on human keratinocytes. J Photochem Photobiol B 84: 167–174.

Sarikurkcu C, Zengin G, Oskay M, Uysal S, Ceylan R, Aktumsek A (2015) Composition, antioxidant, antimicrobial and enzyme inhibition activities of two Origanum vulgare subspecies (subsp. vulgare and subsp. hirtum) essential oils. Ind Crops Prod 70: 178–184.

Sharmila R, Manoharan S (2012) Anti-tumor activity of rosmarinic acid in 7,12-dimethylbenz(a)anthracene (DMBA) induced skin carcinogenesis in Swiss albino mice. Indian J Exp Biol 50: 187–194.

Siriangkhawut W, Kaewboo I (2013) Ultrasonic extraction method for alizarin from roots of Morinda citrifolia. Anal Chem Indian J 12: 65–69.

Souza PM, Elias ST, Simeoni LA, De Paula JE, Gomes SM, Guerra ENS, Fonseca YM, Silva EC, Silveira D, Magalhãe PO (2012) Plants from Brazilian Cerrado with potent tyrosinase inhibitory activity. PLoS ONE 7: 11.

Teixeira RS, Rocha PR, Polonini HC, Brandão, MAF, Chaves MGA, Raposo NRB (2012) Mushroom tyrosinase inhibitory activity and major fatty acid constituents of Amazonian native flora oils. Braz J Pharm Sci 48: 399–404.

Thanigaimalai P, Manoj M, Vigneshwaran N (2017) Skin whitening agents: medicinal chemistry perspective of tyrosinase inhibitors. J Enzyme Inhib Med Chem 32: 403–425.

Virador VM, Kobayashi N, Matsunaga J, Hearing VJ (1999) A standardized protocol for assessing regulators of pigmentation. Anal Biochem 270: 207–219.

Walker JB, Sytsma KJ (2007) Staminal evolution in the genus Salvia (Lamiaceae): molecular phylogenetic evidence for multiple origins of the staminal lever. Ann Bot 100: 375–391.

Wang KH, Lin R, Hsu FL, Huang YH, Chang HC, Huang CY, Lee MH (2006) Cosmetic applications of selected traditional Chinese herbal medicines. J Ethnopharmacol 106: 353–362.

Zolghadri S, Bahrami A, Hassan Khan MT, Munoz-Munoz J, Garcia-Molina F, Garcia-Canovas F, Saboury AA (2019) A comprehensive review on tyrosinase inhibitors. J Enzyme Inhib Med Chem 34: 279–309.

© 2022 Journal of Pharmacy & Pharmacognosy Research (JPPRes)