Pentagamavunon-0 Research Articles

EFFECT OF VANILLIN QUALITY ON THE SYNTHESIS AND SOLID DISPERSION SYSTEMS ON PGV-0 ANTIOXIDANT CAPACITY

Objective: Pentagamavunon-0 (PGV-0) was a curcumin analog with antioxidant properties and has been synthesizing with improvement techniques using vanillins with different qualities. PGV-0 has a low bioavailability but could be increased solubility by making a solid dispersion system. This study aimed to determine the antioxidant properties of PGV-0 that were synthesized with different quality starting materials of vanillin and also to obtain the effect of PEG 6000 and maltodextrin solid dispersion systems.
 Methods: PGV-0 was synthesizing by mixing cyclopentanone into two different groups of vanillins (vanillin pro analysis and vanillin “Kapal Layar”) using an HCl catalyst. The purity of products was identified with TLC (Thin Layer Chromatography) and spectrophotometrically compared with PGV-0 standard. Solid dispersion system consisting of two treatment groups: PEG 6000 and maltodextrin groups. The measurement of antioxidant properties was examined by Cupric Ion Reducing Antioxidant Capacity (CUPRAC).
 Results: PGV-0 of vanillin pro analysis group (EC50= 13.62 ppm) was found higher antioxidant capacity as compared to the vanillin “Kapal Layar” group (EC50= 15.35 ppm). The solid dispersion system of PGV-0-PEG 6000 at ratio 1:10 has shown powerful antioxidant capacity (EC50= 9.00 ppm) than the PGV-0-PEG 6000 at ratio 1:5 group (EC50= 10.65 ppm). PGV-0-maltodextrin at ratio 1:10 (EC50= 11.96 ppm) also has powerful antioxidant activity when compared with PGV-0-maltodextrin at ratio 1:5 (EC50= 13.63 ppm).
 Conclusion: The results of the study showed that the use of vanillin with different qualities in PGV-0 synthesis and also solid dispersion systems can affect the antioxidant activity of PGV-0.

Curcumin Analogs Induce Apoptosis and G2/M Arrest In 4T1 Murine Triple-Negative Breast Cancer Cells

Chemotherapy is the first-line treatment for triple-negative breast cancer (TNBC), yet toxicity and resistance effects have been the current problems. Curcumin,a natural compound, has been reported to exert anti-proliferative effects on various cancer cells, including breast carcinoma cells. However, the β-diketone moiety influences the stability of curcumin. Curcumin analogs, pentagamavunon-0 (PGV-0), and pentagamavunon-1 (PGV-1) were synthesized to improve the stability and activity of curcumin by modified the β-diketone moiety into mono-ketone pentanone. In this study, we evaluated the cytotoxicity, inhibition of cell cycle progression, and induction of apoptosis of curcumin and its analogs (PGV-0 and PGV-1) in murine triple-negative breast cancer 4T1 cell line. The cytotoxic evaluation was done by MTT assay, while apoptosis induction and cell cycle evaluation was performed by annexin V staining and detected by flow cytometry. Curcumin and its analogs, PGV-0, and PGV-1, significantly inhibit the viability of 4T1 breast cancer cells with an IC50 value of 34.34µg/mL, 13.76µg/mL and 38.21μg/mL, respectively. Apoptosis analysis with a dose of 10µg/mL and 15µg/mL in 4T1 breast cancer cells showed that curcumin and its analogs effectively induce apoptotic in a dose-dependent manner. In cell cycle analysis using a dose of 15µg/mL, curcumin inhibited the cell cycle progression in the S phase, whereas PGV-0 and PGV-1 inhibited the cell cycle in the G2/M phase. It could be concluded that curcumin analogs, PGV-0 and PGV-1, have higher potential to be developed as anti-cancer agents by inducing cell cycle arrest and apoptosis in triple-negative breast cancer.

Pentagamavunon-0 (PGV-0) Enhance Cytotoxic Effect of Doxorubicin through Increasing of Apoptosis, Senescence and ROS Level on Triple Negative Breast Cancer 4T1

TNBC, one of the sub type of breast cancers was widely known with high tumorigenic and poor prognosis than others. The development of combination agent (co-chemotherapy) with doxorubicin for chemotherapy of TNBC were carried out to decrease doxorubicin side effect and resistance in cancer. This present study aims to explore the co-chemotherapeutic properties of PGV-0 and investigate induction of doxorubicin on apoptosis, senescence and ROS against TNBC. 4T1 Cell line were used as a TNBC in vitro model. Cytotoxic measurement was performed using MTT assay resulting in IC50 values of 52 μM. Meanwhile, the combination of doxorubicin and PGV-0 showed synergistic effect which decreased cell viability of 4T1 better than single treatment of doxorubicin. Apoptosis analysis was performed using annexin V/PI assay indicated that the combination treatment of PGV-0 and doxorubicin increased apoptosis evidence. Senescence detection was carried out using senescence-associated-β galactosidase (SA-β-gal) assay. The results showed that a single treatment of PGV-0 induced cellular senescence and increased senescence cells in combination treatment. Moreover, DCFDA staining showed that PGV-0 increased ROS level at single treatment, whereas combination treatment increased ROS intracellular compared to the positive control of doxorubicin. Based on these results, PGV-0 has potential as a co-chemotherapeutic candidate on TNBC.Keyword: 4T1, PGV-0, Co-chemotherapy, Cytotoxic, Senescence, Apoptosis, ROS

Analog Curcumin, Pentagamavunon-0 (PGV-0), Induces Senescence and Increases Cytotoxic Effect of Doxorubicin on HCC 1954 Cells

Senescence defined as an irreversible cell cycle arrest. Senescence can inhibit cancer growth and suppress the progression of cancer. Some anticancer compounds are known to have the potential to induce senescence. Senescence defence against tumor development by preventing proliferation of cells with DNA damage. The study aimed to determine the cytotoxic effects and senescence induction of Pentagamavunon-0 (PGV-0) on Human Epidermal Growth Factor Receptor 2-positive (HER2-positive) breast cancer cells, HCC 1954. Cytotoxic tests carried out with 3- (4.5-dimethylthiazzol-2yl) -2.5-tidiphenyltetrazolium (MTT) assay showed that PGV-0 exhibited a strong cytotoxic effect with a the half maximal inhibitory concentration (IC50) value of 39 μM. Treatment with IC50 in sub-doses combined with doxorubicin showed cytotoxic enhancement effects. The senescence assay using SA-β-Galactosidase showed that the PGV-0 in a single treatment was able to induce senescence with a percentage of cell senescent of 15%. The combination treatment of PGV-0 at the half dose of IC50 with doxorubicin 100 nM was able to induce senescence with the percentage of senescent cells of 25%. Moreover, PGV-0 also increased intracellular reactive oxygen species (ROS). The results of this study indicate that PGV-0 exhibits cytotoxic effect, increases cytotoxic effect of doxorubicin and induces senescence that may correlate to the increasing of intracellular ROS in 1954 HCC cells.Keywords: Pentagamavunon (PGV-0), HCC 1954, Cytotoxic, Senescence

Effects of pentagamavunon-0 (PGV-0) as alternative analgesics on orthodontic tooth movement in rats

Introduction: Some analgesic drugs may have adverse effects on bone remodelling and, thus, on orthodontic tooth movement rate (OTM). GV-0 is synthesized by reacting vanillin and cyclopentanone catalyzed in acidic condition, and it has been revealed as a selective COX-2 inhibitor. This study was aimed to investigate the effect of pentagamavunon-0 (PGV-0), one of the curcumin analogues, on OTM. Methods: This study was conducted on 50 male Wistar rats (350-450 g) which were randomly divided into five groups (n = 10 each): 1) no treatment group (NT), 2) orthodontic treatment only (ORT), 3) ORT plus 0.4% sodium carboxymethyl cellulose (Na-CMC) analgesic carrier, 4) ORT plus 200 mg/kg BW Paracetamol (PCT) as the positive control, and 5) ORT plus PGV-0 (50 mg/kg BW (PGV-0). Results: Drug and day interaction was statistically significant on two-way ANOVA. Post-hoc analyses showed that OTM increased from day 3 to 7 in all orthodontic groups over the same distance (p>0.05). Maximum OTM was found on day 6, which was significantly farther than the distance on day 4. On day 7, OTM was less than on day 6. OTM in all orthodontic groups, including in the PGV-0 group, was higher than in the NT group (p<0.05). No differences was seen in OTM between PGV-0 group and other orthodontic groups (p>0.05). Post-hoc analysis (intra days) revealed that OTM in PGV-0 and other orthodontic treatment groups increased. Conclusion: After a single orthodontic force, PGV-0 does not inhibit tooth movement in rats from day 1 to day 7. Therefore, it is possible to develop PGV-0 as an alternative analgesics during orthodontic therapy.Keywords: Analgesic drug, orthodontics, tooth movement, curcumin.

Cytotoxicity of Tetrahydropentagamavunon-0 (THPGV)-0 and Tetrahydropentagamavunon-1 (THPGV-1) in Several Cancer Cell Lines

Tetrahydropentagamavunon-0 (THPGV-0) and Tetrahydro-pentagamavunon-1 (THPGV-1), are analogs of a curcumin metabolite, tetrahydrocurcumin, and a derivate of Pentagama-vunon-0 (PGV-0) and Pentagamavunon-1 (PGV-1), respectively. THPGV-0 and THPGV-1 have been successfully synthesized and are investigated for their anticancer potency. Cytotoxic assays were performed toward several cancer cell lines to determine values of the IC 50 against those cell lines. Assessing cytotoxicity in Vero normal cell line showed the selectivity of those compound. THPGV-1 showed highest cytotoxic activity in lymphoma Raji cells, a suspension cell line, with an IC 50 of 180mM. Both THPGV-0 and THPGV-1 showed similar potencies in T47D breast cancer cell line with IC 50 values of 250-270mM. Regardless their high selectivity, however, cytotoxic activities of THPGV-0 and THPGV-1 were lower compared to PGV-0 and PGV-1 in HeLa cervical, T47D breast, and WiDr colon cancer cell lines. Further study using different types of cancer cell lines and confirmation of cell viability by another assays and apoptosis detection may give more benefit.

Molecular Modeling of An Analog Of Curcumin Compounds Pentagamavunon-0 (PGV-0) And Pentagamavunon-1 (PGV-1) Through Computational Chemistry Methods Ab-Initio HF/4-31G

Breast cancer is the second largest number of cancer cases in Indonesia, after cervical cancer. The growth of these cancer cells can be prevented with compounds Pentagamavunon-0 (PGV-0) and Pentagamavunon-1 (PGV-1). This compound is an analog of curcumin compounds that have anti breast cancer activity. Modeling the structure of compound PGV-0 and PGV-1 through computational chemistry methods Ab-initio HF/4-31G could be used to predict the geometry and structure elucidation spectra associated with pharmacological activity such as anticancer compounds theoretically.This research involves modeling the structures and spectra prediction calculation compounds PGV-0 and PGV-1 by computational chemistry methods Ab-initio HF/4-31G, using Gaussian03W. The result using Ab-initio HF/4-31G method then compared with data from experimental geometry and the results of calculations with AM1.The results showed that computational chemistry methods Ab-initio HF/4-31G calculations give better results for modeling the structure compared semiempirik method AM1.

APOPTOSIS INDUCTION EFFECT OF CURCUMIN AND ITS ANALOGS PENTAGAMAVUNON-0 AND PENTAGAMAVUNON-1 ON CANCER CELL LINES

ABSTRACTObjectives: This experiment aims to investigate the apoptosis effect of curcumin and its analogs pentagamavunon-0 (PGV-0) and PGV-1 on normaland other cancer cell lines.Methods: Growth inhibition effect was investigated using the MTT method. Double staining used acridine orange, 2-(4-aminodiphenyl)-6-indolcarbamidine dihydrochloride and ethidium bromide was performed to determine morphological changes of cells. Detection of PARP, caspase-3,PUMA and BAX using a western blot method was conducted to elucidate the apoptosis effect of the compounds.Results: PGV-1 (2.5 μM) and PGV-0 (5.0 μM) could inhibit T47D-cell growth on 72 h observation, but not for curcumin. DNA staining showed PGV-1has the strongest apoptosis induction effect on T47D-cells compared to PGV-0 and curcumin as well. Western blot analysis resulted in cleavage PARP(83 kD) on HeLa, T47D, and MCF-7 cells treated with PGV-1 (2.5 μM), PGV-0 (5.0 μM). Curcumin (10.0 μM) just induced apoptosis on T47D-cell andMCF-7 cell, but not HeLa cell. Cleavage PARP resulted by apoptosis process in the cell. PGV-1 (2.5 μM) had a stronger apoptosis effect compared toPGV-0 (5.0 μM) and curcumin (10.0 μM) based on cleaved PARP result qualitatively. On the normal cell (NH3T3), cells that were treated with thecompounds resulted in a negative cleavage PARP. This result indicated that the compounds were part of a selectively induced cancer cell line apoptosisprocess.Conclusion: Curcumin, PGV-0 and PGV-1 could inhibit cell growth by induce apoptosis on cancer cells but not on normal cells, which PGV-1 hasstrongest apoptosis induction effect on cancer cell lines.Keywords: Curcumin and analogs, Apoptosis, Cancer cell lines.

Konsentrasi Aman Kurkumin dan PGV-0 terhadap Sel Vero Berdasarkan Hasil Uji Sitotoksik

Curcumin (1,7-bis(3-methoxyphenyl 4'hidroksi) -1.6 heptadien, 3,5-dione) is the yellow pigment of Curcuma longa. Curcumin has various biological activities such as antioxidant, antibacterial, antifungal, antiprotozoa l and antivirus. Various benefits of curcumin can not be separated from the weakness which is not stable to pH and light. Pentagamavunon-0 (PGV-0) were made by changing the β diketone group on cluster analog of curcumin into monoketon while eliminating active methylene group so it is more stable to pH and light. PGV-0 also has potential as a stronger antioxidant and antiinflammatory agent than other curcumin analogues. The objective of this research was to determine the safe concentrations of curcumin and PGV-0 on vero cells due to the increased use of the two compounds through the cytotoxic test. This research includes experimental research. Cytotoxic test performed with microculture tetrazolium technique ( MTT ) method. Use of MTT to evaluate the cytotoxic is based on changes of tetrazolium salt into formazan crystals by mitochondrial enzyme succinate dehydrogenase with the help of cellular NADH. The results showed that the safe concentrations of curcumin and PGV-0 on vero cells respectively are 6.25 and 1.5625 ppm. Based on the cytotoxic test the secure concentration of curcumin was higher than PGV-0.

Hepatoprotective and antioxidant activity of pentagamavunon-0 against carbon tetrachloride-induced hepatic injury in rats

ObjectiveTo investigate the hepatoprotective and antioxidant activity of pentagamavunon-0(PGV-0) against CCl4-induced hepatic injury in rats. MethodsThe groups of animals were administered with PGV-0 at the doses 2.5, 5, 10, and 20 mg/kgb.w., p.o. once in a day for 6 days and at day 7 the animals were administrated with carbon tetrachloride (CCl) (20%, 2 mL/kgb.w. in liquid paraffin (i.p.). The effect of PGV-0 on serum transaminase (SGPT), alkaline phosphates (ALP) and total bilirubin were determined in CCl4-induced hepatotoxicity in rats. Further, the effects of PGV-0 on glutathione (GSH) content, catalase (CAT) and NO free radical scavenging activity also were investigated. ResultsThe results demonstrated that PGV-0 significantly reduced the activity of SGPT, serum ALP and total bilirubin in CCl4induced rat hepatotoxicity. PGV-0 has effect on the antioxidant and free radical defense system. It prevented the depletion level of GSH and decrease activity of CAT in CCl4-induced liver injury in rats. PGV-0 also demonstrated the free radical scavenger effects on NO free radical scavenging activity with ES value of 32.32μM. ConculsionAll of our findings suggests that PGV-0 could protect the liver cells from CCl4-induced liver damages and the mechanism may through the antioxidative effect of PGV-0 to prevent the accumulation of free radicals and protect the liver damage.

SYNTHESIS NEW POTENTIAL ANTI-INFLAMMATORY AGENT SODIUM SALT OF PENTAGAMAVUNON-0

Inflammation is the response of living tissues to injury. The process affects physiological changes such as erythema, edema, asthma and fever. Non-steroid Anti-inflammatory Drugs (NSAIDs) have been developed since they could inhibit inflammation process because of its ability to inhibit biosynthesis of prostaglandin, one of inflammation mediators, through inhibition of cyclooxigenase (COX) enzymes. Molecules, which have been reported having anti-inflammatory activity, for example, are curcumin, some curcumin derivatives and curcumin analogues. One of curcumin analogues that has been developed is pentagamavunon-0 (PGV-0) whose IUPAC name is 2,5-bis(4'-hidroxy-3'-methoxy-benzylidene)cyclo-pentanone. But PGV-0, which is like curcumin, practically insoluble in water, so it causes problems in the development. The aim of this research is to synthesize a derivative of PGV-0, a natrium salt of PGV-0 (natrium pentagamavunonate-0/Na-pentagamavunonate-0), which is hoped to have a better anti-inflammatory activity and solubility in water than PGV-0. PGV-0 was synthesized by reacting vanillin and cyclopentanone catalized by acid. Na-pentagamavunonate-0 was synthesized with PGV-0 as a starting material using an appropriate method. This research was able to synthesize new compound that was estimated as a natrium salt of PGV-0 (natrium pentagamavunonate-0/Na-pentagamavunonate-0). Keywords: Curcumin, PGV-0, Na-pentagamavunonate-0, anti-inflammation

Synthesis of potassium pentagamavunonat-0 using potassium hydroxyde and solvent tertrahydrofuranethanol

Pentagamavunon-0 (PGV-0) (2,5-bis(4’-hydroxy-3’-methoxy benzy-lidene) cyclopentanone) is a curcumin analogue. To improve its bioavailability it has been succesfully synthesized the Na-salt of this compound. However, sodium may disturb the electrolyte liquid-balance in the body which in turn hypertension. Therefore, the synthesis of potassium pentagamavunonat-0 is attempted. Synthesis has been done using potassium hydroxide as a potassium source, in tetrahydrofuran-ethanol solvent, with molar variations. The result is dipotassium pentagamavunonate-0 (elucidated with IR, 1H-NMR, 13C-NMR and MS spectra), which is more soluble in water and has better stability at high pHs. Molar variations did not show correlation with its yield. The yields are more than 100 %, may be caused by water crystals that contaminate the compound. Key words : pentagamavunon-0, dikalium pentagamavunonat-0, synthesis

IDENTIFICATION OF PENTAGAMAVUNON-0 METABOLITE IN FAECES AFTER INTRAVENOUS INJECTION OF WHITE-MALE SPRAGUE DAWLEY-DERIVED RATS

The compound 2,5-bis-(4’-hydroxy-3’-methoxybenzilidin) cyclopentanone or Pentagamavunon-0 (PGV-0) as the new anti-inflamatory drug candidate, is one of curcumin structure modification. Identification of metabolites in faeces could be strengthen the whole information about biotransformation of drug and could be use for studying on PGV-0 metabolites which are excreted through bile. Male Sprague Dawley derived rats were used in all experiments. PGV-0 was given intravenously using dose at 40 mg/kg BB. Then the faeces callected after 24 and 48 hours. The metabolite investigation devided into two parts, etil acetate fraction and water fraction, of faeces. The faeces which is excreted after 24 hours, found was more-polar metabolite according to PGV-0; it has shoft-yellow flourencese and having Rf value = 0,48 on TLC, using silica gel F254 as the stationary phase and etil acetate as the mobile phase. No PGV-0 glucuronide and sulfate metabolites were detectable in faeces, which are excreted on 24 or 48 hours. Key words : Pentagamavunon-0, biotransformation, intravenous, faeces

The stability of PGV-0 (Pentagamavunon-0) as an Antiinflammatory drug in liquid dosage forms

PGV-0 is a curcumin derivate, a synthetic compound which may be a candidate of a new drug. This substance has a potent inflammatory effect with a very low toxicity. One of the first step which must be searched for a candidate of drug, is to perform a stability study. There are many degraded drugs causing the adverse reactions. Most of them could be the initiator in forming an antigen at anaphylactic reaction or allergic reaction. Further more some of the degraded drugs are very toxic. So, the study of degradation or the stability investigations of a new drug should be carried out. The PGV-0 stability in a buffer solution at pH 10,0 had been studied by an accelerated temperature method. The temperatures were held at 50°, 55° and 60° C. The intact PGV-0 has been analysed by HPLC. The results then were used to define the shelf-life, the half life and the activation energy of the degradation of PGV-0. It was evident that PGV-0 was unstable in aqueous solution, the shelf-life was only 45.3 hours, the half-life was 299 hours and the activation energy was 14,2 kkal mol-1. Because the PGV-0 was not stable in aqueous solution, it is suggested that this substance should be made into solid dosage forms instead of the liquid dosage forms. Key word : PGV-0, stability, solution

BIOTRANSFORMATION OF PENTAGAMAVUNON-0 (PGV-O) : IN VITRO AND IN VIVO STUDIES

Pentagamavunon-0 (PGV-0) or 2,5-bis-(4-hydroxy-3-methoxy benzyilidin) cyclopentanon is a synthetic compound which has been proved to have an anti-inflamatory effect by animal experiments. The study was performed in order to investigate the possible route of biotransformation of PGV-0, which in turn can assist to elucidate the mechanism of its action. The in vivo biotransformation was predicted by identification of all metabolites of PGV-0 presence in urine and faeces after administration orally and intravenously. The in vitro studies were performed using microsomal or sitosolic fraction of the hepar. The metabolites were determined by thin layer chromatography and spectrophotometry. The results indicated that PGV-0 underwent oxydative biotransformation (phase I) and at least produced one metabolite. In vitro studies, PGV-0 also underwent glucuronidation and sulfatation. However, in vivo studies indicated that only metabolites of PGV-0 which have hydroxyl groups underwent conjugation process but PGV-0. Key words : Pentagamavunon-0, biotransformation, in vivo and in vitro.

PENGARUH PEMBERIAN PENTAGAMAVUNON-0 PADA AKTIVITAS GLUTATION-S-TRANSFERASE PADA HATI TIKUS SECARA IN VIVO

ABSTRACT Glutathione S-transferase is amount of enzymes catalyze conjugation reac-tion of glutathione (GSH) with electrofilic endogen compound or electrofilic xenobiotic. At the particular cancer there is increasing GST, especially on mu and phi class that it cause therapy on cancer cell by citostatic drug (commonly as electrofilic compound) not effectively. Pentagamavunon-0 (PGV-0) is antiinflammation compound. Base on in vivo research denoted this compound has five times strengthened than curcumin to inhibit GST enzyme activity. But using PGV-0 by in vivo to GST activity never report or publish yet. This research was done by measuring isolated GST enzyme activity from rat liver was given in vivo PGV-0, with nine groups treatment. Each group consist of control group without treatment, control group CMC-Na 0.5 % per oral and intraperitoneal DMSO, group with doses treatment of PGV-0 20, 40, 80 and 160 mg/Kg BB per oral, rat group was given benzo(a)pirene (BP) 1 mg/Kg BB i.p. and group was given BP 1 mg/Kg BB i. p. and PGV-0 80 mg/Kg BB per oral. GST enzyme activity is measured from reaction of conjugate GSH and DCNB that it’s absorption/minute. Base on rate (absorption/minute) GST enzyme activity can be accounted. Then inhibition percentage can be accounted from each rat treatment compared with control without treatment and solvent control. Analyze use statistic test (Kruskal-Wallis then continued by Mann-Whitney test), to know specific different of GST class mu enzyme activity in rat liver at each treatments group. Result research denoted that PGV-0 give inhibition effect to GST class mu activity of rat liver by in vivo. Doses level did not give proportional result to GST class mu activity of rat liver. Doses level of PGV-0 give optimum result to GST class mu activity of rat liver inhibition at 80 mg/Kg BB with inhibition value 19,99 % compared with CMC-Na as solvent. Key word: pentagamavunon-0, in vivo, glutathione S-transferase, inhibition