Hasil Pencarian  ::  Simpan CSV :: Kembali

Abstrak :
Protein inti virus hepatitis B memegang peranan penting pada berbagai tahap siklus hidup virus. Dengan mengganggu peran protein inti ini oleh suatu senyawa maka replikasi virus akan terhambat. Emodin dipilih sebagai senyawa penuntun dan dilakukan modifikasi struktur dengan mengubah sifat hidrofobisitas, elektronik dan steriknya. Untuk memprediksi interaksi yang terjadi antara emodin dan senyawa turunan ester emodin dengan protein inti virus hepatitis B sebagai target obat, dilakukan pemodelan molekul dengan program simulasi penambatan dan dinamika molekul. Interaksi senyawa turunan ester emodin dengan reseptor protein inti virus hepatitis B terjadi di daerah antarmuka dimer-dimer reseptor dan membentuk interaksi yang stabil dengan asam amino pada daerah yang akan mengubah konformasi protein sehingga mengganggu perakitan kapsid dan proses replikasi virus. Senyawa turunan ester emodin disintesis dengan pereaksi dan kondisi yang sesuai kemudian dikonfirmasi strukturnya menggunakan spekrofotometri LCMS/MS, FTIR, 1H-NMR dan 13C-NMR. Senyawa yang diperoleh adalah: 3-asetil emodin, 3-benzoil emodin, 3-o-toluil emodin, 3-m-toluil emodin, 3-p-toluil emodin dan 3-dimetilkarbamoil emodin. Aktivitas inhibisi in vitro ditentukan menggunakan sel HepG2 yang ditransfeksi plasmid genom inti virus hepatitis B, immunositokimia dan fluorometri. Hasil menunjukkan bahwa turunan ester emodin aktivitasnya lebih tinggi dan sitotoksitasnya lebih rendah daripada emodin. Senyawa 3-o-toluil emodin, 3-m-toluil emodin dan 3-p-toluil emodin menunjukkan aktivitas yang tinggi dengan IC50 berturut-turut sebesar 0,4; 0,23 dan 0,11 M. Analisa hubungan struktur aktivitas menunjukkan bahwa hidrofobisitas dan sterik berkorelasi non linear dengan aktivitas inhibisi replikasi virus hepatitis B.

---

The core protein of hepatitis B virus plays an important role at different stages of the viral life cycle. By disrupting the role of this core protein by a compound, the viral replication will be inhibited. Emodin was chosen as a lead compound and the structure was modified by altering its hydrophobicity, electronic and steric properties to be emodin ester. Prediction of interaction between emodin and their ester derivatives with hepatitis B virus core protein as drug targets, was performed by molecular modeling simulation program of docking and molecular dynamic. The interaction of emodin ester derivatives with hepatitis B virus core protein receptors occurs in the interface area of receptor dimers and forms a stable interaction with amino acids in areas that will alter protein conformation thus disrupting capsid assembly and viral replication processes. Emodin ester derivatives compound were synthesized with reagents and suitable conditions then confirmed their structures using LCMS MS, FTIR, 1H NMR and 13C NMR spectrophotometry. The compounds obtained were 3 acetyl emodin, 3 benzoyl emodin, 3 o toluyl emodin, 3 m toluyl emodin, 3 p toluyl emodin and 3 dimethylcarbamoyl emodin. In vitro inhibitory activity was determined using HepG2 cells transfected plasmid core genomes of hepatitis B virus, immunocytochemistry and fluorometry. The results showed that the emodin ester derivatives activity is higher and its cytotoxicity is lower than emodin. The 3 o toluyl emodin, 3 m toluyl emodin and 3 p toluyl emodin showed high activity with IC50 of 0.4 0.23 and 0.11 M respectively. Structure activity relationship analysis showed that hydrophobicity and steric properties correlate non linearly with activity.

Abstrak :
Senyawa turunan eugenol diduga dapat menginhibisi Bcl-2 pada sel kanker kolorektal HT29. Penelitian ini bertujuan untuk memperoleh senyawa baru turunan eugenol yang dapat menghambat sel kanker kolorektal HT29 secara in vitro dan menurunkan ekspresi Bcl-2 pada mencit yang mengalami pre-klamsia terhadap kolon secara in vivo. Penelitian ini diawali dengan melakukan desain senyawa turunan secara in silico. Hasil senyawa hit disintesis di laboratorium. Uji secara in vitro, uji apoptosis dan uji in vivo dilakukan berturut-turut pada hasil senyawa sintesis. Hasil in silico, dari skrining secara farmakofor dengan rancangan acak lengkap menggunakan 220 senyawa desain. Berdasarkan fitur farmakofor dengan cut off 5 fitur dihasilkan 23 senyawa. Hasil skrining farmakofor dilakukan docking menghasilkan delapan senyawa yaitu senyawa 4 rsquo;- 2-kloro-3-hidroksipropil -2 rsquo;-metoksifenil 2-hidroksibenzoat 57, 4 rsquo;- 2-kloro-3-hidroksi-propil -2 rsquo;-hidroksifenil 2-hidroksibenzoat 167, S -4 rsquo;- 2,3-dihidroksipropil -2 rsquo;-metoksifenil 2-hidroksibenzoat 59, R -4 rsquo;- 2,3-dihidroksipropil -2 rsquo;-metoksifenil 2-hidroksibenzoat 60, 4 rsquo;-alil-2 rsquo;-metoksifenil 4-amino-2-hidroksibenzoat 71, 4 rsquo;-alil-2 rsquo;-hidroksifenil 4-amino-2-hidroksibenzoat 181, 4 rsquo;-alil-2 rsquo;-metoksifenil 3,4,5-trihidroksibenzoat 86 dan 4 rsquo;-alil-2 rsquo;-metoksifenil 3,5-diihidroksi-4-metoksibenzoat 91 dengan energi ikatan lebih negatif dari standar. Delapan senyawa hasil skrining disintesis melalui reaksi esterifikasi, adisi halogen, hidroksilasi dan demetilasi. Hasil sintesis diuji aktivitas penghambatannya secara in vitro terhadap sel HT29 kanker kolon. Aktivitas penghambatan terhadap sel HT29 menunjukkan nilai IC50 antara 82.98 g/mL - 8.455 g/mL. Nilai IC50 tersebut lebih negatif dibandingkan senyawa penuntun eugenol. Hubungan Kuantitatif Struktur Aktivitas terhadap sel line HT29 menghasilkan persamaan Log 1/IC50 = -0.865-0.210 LogP 2 1.264 logP -0.994CMR n=10; r=0.706; SE:0.21; F=0.497, sig=7.86 . Persamaan menunjukkan variabel log P dan CMR berpengaruh terhadap IC50. Sifat hidrofobisitas log P lebih berperan dibandingkan dengan sifat sterik CMR . Hasil uji in vivo terhadap mencit Mus musculus menunjukkan senyawa turunan 59 memiliki nilai HE dan IHK mendekati kontrol positif. Peningkatan dosis pemberian menyebabkan peningkatan degradasi Bcl-2 pada jaringan mendekati kontrol normal. Hasil penelitian menunjukkan bahwa senyawa baru turunan eugenol 59 yang diperoleh dapat menghambat kanker kolorektal secara in vitro dan in vivo.

---

Compounds derived from eugenol are thought to inhibit Bcl 2 in HT29 colorectal cancer cells. The aim of this study was to obtain new compounds of eugenol derivatives that could inhibit in HT29 cell invitro test and decrease of Bcl 2 expression in mice pre clammed on colon with invivo test. This research begins by designing in silico derivative compounds. The result of the hit compound is synthesized in the laboratory. In vitro tests, apoptotic test and in vivo test were performed successively on the result of the synthesis compound. In silico yield, from a complete randomized pharmacophore screening using 220 design compounds. Based on the pharmacophore features with cut off 5 features produced 23 compounds. The results of pharmacophore screening conducted docking which yielded eight compounds of compound 4 rsquo 2 chloro 3 hydroxypropyl 2 rsquo metohoxyphenyl 2 hydroxybenzoat 57, 4 rsquo 2 chloro 3 hidroxy propyl 2 rsquo hydroxyphenyl 2 hydroxybenzoat 167, S 4 rsquo 2,3 dihydroxypropyl 2 rsquo methoxyphenyl 2 hydroxybenzoat 59, R 4 rsquo 2,3 dihydroxypropyl 2 rsquo methoxyphenyl 2 hydroxybenzoat 60, 4 rsquo allyl 2 rsquo methoxyphenyl 4 amino 2 hydroxybenzoat 71, 4 rsquo allyl 2 rsquo hydroxyphenyl 4 amino 2 hydroxybenzoat 181, 4 rsquo allyl 2 rsquo methoxyphenyl 3,4,5 trihydroxybenzoat 86 dan 4 rsquo allyl 2 rsquo methoxyphenyl 3,5 dihydroxy 4 methoxybenzoat 91 with energy binding more negative than standard. The eight compounds of the screening are synthesized by esterification reaction, addition with halogen, hydroxylation. And demetylation The synthesis results were tested in vitro inhibitory activity against HT29 colon cancer cells. The inhibitory activity against HT29 cells shows an IC50 value between 82.98 g mL 8,455 g mL. The value of IC50 is better than the eugenol guiding compound. Quantitative Relation of Structure Activity against cell line HT29 with equation Log 1 IC50 0.865 0.210 LogP 2 1.264 logP 0.994CMR n 10 r 0.706 SE 0.21 F 0.497, sig 7.86 . This equation showed that log P and CMR have effect with IC50. Hydrophobicity log P more of effect compared than steric parameters CMR . In vivo test of Mus musculus that showed compound derivative 59 based on HE and IHK values approaching positive control. Increased dosage of administration leads to an increase in Bcl 2 degradation in tissues near normal control. The results showed that the new compound derived eugenol 59 obtained can inhibit colorectal cancer in vitro and in vivo.

Abstrak :
Senyawa andrografolida, metabolit sekunder pada tumbuhan Sambiloto (Andrographis paniculata), diketahui memiliki aktivitas antiplasmodia 50% in vitro (IC50) < 10 μM, sehingga layak dijadikan sebagai senyawa pemandu di dalam pencarian dan pengembangan obat antimalaria secara rasional. Enzim farnesil difosfat sintase P. falciparum (PfFPPS) merupakan target yang valid untuk obat antimalaria. Tujuan umum penelitian ini adalah membuktikan membuktikan hipotesis bahwa senyawa turunan andrografolida, yang rancangan strukturnya diperoleh melalui optimasi interaksi maya andrografolida dengan enzim PfFPPS berdasarkan pose penambatan maya substrat dan inhibitornya, memiliki aktivitas antiplasmodia in vitro yang lebih tinggi daripada aktivitas antiplasmodia in vitro andrografolida dan tidak toksik terhadap sel manusia. Melalui optimasi interaksi maya andrografolida dengan enzim PfFPPS berdasarkan pose penambatan maya substrat dan inhibitornya, telah diperoleh rancangan struktur turunan andrografolida yang baru yang berpotensi menghambat enzim PfFPPS. Melalui reaksi esterifikasi terhadap senyawa andrografolida, diperoleh senyawa 3,14-andrografolidabis(4'-kloro)benzoat (18). IC50 senyawa 18 terhadap P. falciparum galur 3D7 adalah: 131,60 μM. Persentase pertumbuhan sel DLD-1 di bawah pengaruh senyawa 18 hingga konsentrasi 63,74 μM adalah lebih dari 100%. Dengan demikian, senyawa turunan andrografolida tersebut memiliki aktivitas antiplasmodia in vitro yang lebih rendah daripada aktivitas antiplasmodia in vitro andrografolida, namun tidak toksik terhadap sel manusia.

---

Andrographolide, a secondary metabolite of Sambiloto (Andrographis paniculata), has a 50% in vitro antiplasmodial activity (IC50) < 10 μM, that it is suitable as a lead compund in an antimalarial drug discovery and development. P. falciparum farnesyl diphosphate synthase (PfFPPS) is a valid target for antimalarial drug. The main objective of this research is to prove the hypothesis that andrographolide derivative, whose design was obtained throughout optimization of virtual interaction between andrographolide and PfFPPS based on the docking pose of its substrates and inhibitor, has a higher in vitro antiplasmodial activity than that of andrographolide and is not toxic towards human cells. Throughout optimization of virtual interaction between andrographolide and PfFPPS based on the docking pose of its substrates and inhibitor, design of novel andrographolide derivatives which are potential to inhibit PfFPPS was obtained. Throughout esterification on andrographolide, an andrographolide derivative was obtained: 3,14-andrographolidebis(4'-chloro)benzoate (18). IC50 of 18 against P. falciparum 3D7 was: 131.60 μM. Percentage of human DLD-1 cell growth under the influence of 18 up to 63.74 μM was more than 100%. Therefore, this andrographolide derivative has lower in vitro antiplasmodial activity than that of andrographolide, but is not toxic towards human cells.

Abstrak :
Latar Belakang: Resistensi insulin pada jaringan otot skelet, hepar, dan adiposit merupakan penyebab utama terjadinya DM tipe 2 serta berbagai penyakit metabolik lain. Resistensi insulin masih sulit diatasi menggunakan obat yang tersedia, sehingga pencarian obat sensitisasi insulin, terutama pada jaringan otot skelet menjadi urgensi dalam riset obat antidiabetes. Salah satu tanaman obat yang berpotensi dikembangkan sebagai obat sensitisasi insulin adalah brotowali. Tujuan: Mengidentifikasi target terapi utama dari brotowali sebagai agen sensitisasi insulin melalui pendekatan in silico, menguji aktivitasnya secara in vitro pada kultur sel otot skelet L6.C11, dan mengidentifikasi senyawa aktifnya menggunakan metode LC-MS/MS metabolomik. Metode: Pencarian target terapi utama resistensi insulin yang ditarget oleh brotowali dilakukan melalui analisis jejaring farmakologi yang diikuti dengan simulasi penambatan molekuler dan dinamika molekuler. Kultur sel otot skelet L6.C11 digunakan sebagai model sel resisten insulin pasca-induksi tinggi glukosa dan tinggi insulin. Hasil fraksinasi terhadap ekstrak metanol brotowali (n=33) diuji aktivitasnya terhadap peningkatan kadar glikogen dan inhibisi fosforilasi serin-312 pada IRS1(metode enzime-linked immunosorbent assay), serta peningkatan GLUT4 tertranslokasi (metode konfokal-imunositokimia). Metode LC-MS/MS metabolomik digunakan untuk menganalisis metabolit dari fraksi-fraksi uji. Analisis statistik komparatif melalui uji ANOVA satu arah dan analisi multivariat melalui PCA dan OPLS untuk mengidentifikasi senyawa penanda bioaktif. Hasil: Analisis jejaring farmakologi memprediksi adanya tiga jalur terapi dari resistensi insulin yang ditarget oleh senyawa kandungan brotowali (jalur persinyalan PI3K, TNF, dan MAPK). Fosforilasi serin-312 pada IRS1 ditentukan menjadi target terapi dalam pengujian in vitro didasarkan pada perannya yang besar pada patogenensis resistensi insulin (degree: 12). Hasil uji in vitro mengidentifikasi fraksi 3 sebagai fraksi dengan aktivitas tertinggi dari seluruh fraksi uji (2,55+0,12 ?g/mL; 45,68+3,20%; 64,07+1,78 AU) dalam aktivitas peningkatan glikogen, inhibisi pIRS1 ser-312, dan peningkatan translokasi GLUT4. LC-MS/MS metabolomik mampu mengidentifikasi senyawa penanda bioaktif batang brotowali berupa tinoskorsida D, higenamin, dan tinoskorsida A. Kesimpulan: Analisis komputasi jejaring farmakologi mampu memprediksi dengan baik target terapi dan senyawa aktif brotowali. Secara in vitro, senyawa kandungan batang brotowali mampu meningkatkan sensitisasi insulin dengan senyawa penanda bioaktif berupa tinoskorsida D, higenamin, dan tinoskorsida A. ......Background: Insulin resistance in skeletal muscle tissue, liver and adipocytes is the main cause of type 2 DM and various other metabolic diseases. Insulin resistance is still difficult to overcome using available drugs, so the search for insulin sensitizing drugs, especially in skeletal muscle tissue, is an urgency in antidiabetic drug research. One of the medicinal plants that has the potential to be developed as an insulin sensitizing drug is brotowali. Objectives: To identify potential therapeutic targets of brotowali as an insulin sensitizing agent through an in silico approach, to test its in vitro activity in L6.C11 skeletal muscle cell culture, and to identify its active compounds using the LC-MS/MS method. Methods: The search for the potential therapeutic target of insulin resistance targeted by brotowali was carried out through network pharmacology analysis followed by molecular docking and molecular dynamics simulations. The fractionation of brotowali methanol extract (n=33) were tested for their activity on increasing glycogen levels and inhibition of serine-312 phosphorylation on IRS1 (enzyme-linked immunosorbent assay method), as well as increasing translocated GLUT4 (confocal-immunocytochemical method). The LC-MS/MS metabolomics method was used to analyze the metabolites of the tested fractions. Comparative statistical analysis through one way ANOVA test and multivariate analysis through PCA and OPLS to identify bioactive marker compounds. Results: Network pharmacology analysis predicted three therapeutic pathways of insulin resistance targeted by brotowali’s compounds (PI3K, TNF, and MAPK signaling pathways) with the PI3K pathway as the main pathway. Sequentially the signaling pathway regulate the glucose homeostasis, anti-inflammation, and cell proliferation. Phosphorylation of serine-312 in IRS1 was determined to be a therapeutic target in in vitro testing based on its major role in the pathogenesis of insulin resistance (degree: 12). In vitro tests identified fraction 3 as the fraction with the highest activity of all tested fractions (2.55+0.12 µg/mL; 45.68+3.20%; 64.07+1.78 AU) in glycogen increasing activity , inhibition of pIRS1 ser-312, and increased GLUT4 translocation sequentially. The bioactive marker compounds of brotowali stems were identified as tinoscorside D, higenamin, and tinoscorside A. Conclusion: Network pharmacology computation was successfully predict the therapeutic targets and active compounds of brotowali. At in vitro test, compounds contained in brotowali stems can increase insulin sensitization with bioactive markers were tinoscorsida D, higenamin, and tinoscorsida A.