Aijijiyah, Nur Pasca (2026) Desain, Sintesis, Dan Uji Bioaktivitas Senyawa-Senyawa Sinamamida Dan Hidrazida-Hidrazon Sebagai Antituberkulosis, α-Glukosidase, Antiinflamasi. Doctoral thesis, Institut Teknologi Sepuluh Nopember.
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Abstract
Tuberkulosis akibat bakteri Mycobacterium tuberculosis (Mtb) merupakan penyakit menular yang dapat meningkatkan risiko terinfeksi Human Immunodeficiency Virus. Isoniazid merupakan senyawa aromatik heterosiklik yang menjadi salah satu obat paling efektif untuk pengobatan tuberkulosis. Meskipun demikian, resistensi terhadap obat tuberkulosis dan belum adanya obat tuberkulosis baru menjadi desakan global dalam upaya melawan tuberkulosis. Diabetes tipe 2 sebagai komorbiditas tuberkulosis meningkatkan angka kematian akibat penyakit ini. Studi literatur mengungkap bahwa asam sinamat dan turunannya memiliki aktivitas antituberkulosis serta mampu menghambat enzim α-glukosidase. Molekul-molekul hibrida isoniazid dan asam sinamat berupa senyawa sinamamida menunjukkan potensi penghambatan terhadap bakteri Mtb. Sintesis sinamamida dapat dilakukan menggunakan pereaksi 2-(1H-benzotriazol-1-il)-1,1,3,3-tetrametil-uronium heksafluorofosfat (HBTU). Namun, senyawa ini tergolong sebagai bahan peledak kelas satu dan menunjukkan dekomposisi autokatalitik selama proses penyimpanan dan pengangkutan. Senyawa 2-metil-6-nitrobenzoat anhidrida (MNBA) (14) selanjutnya dimanfaatkan untuk menggantikan HBTU dalam sintesis senyawa ester. Eksperimen in silico melalui evaluasi sifat farmakokinetika dan toksisitas (ADMET), sifat kemiripan obat, dan penambatan molekuler berhasil mendapatkan senyawa hibrida sinamamida (29a-d). Senyawa ini memenuhi kriteria obat oral sesuai Lipinski, aman berdasarkan prediksi toksisitasnya, dan menunjukkan energi pengikatan yang lebih rendah daripada standar asam 6-kloropirazin-2-karboksilat (POA) (30) dan akarbosa (6) masing-masing terhadap reseptor Mtb dan α-glukosidase. Senyawa sinamamida 29a-d selanjutnya berhasil disintesis melalui reaksi amidasi yang melibatkan asam sinamat (32a-d), isoniazid (2), MNBA (14) dan 4-dimetilaminopiridina (DMAP) (16) dengan rendemen 38-75%. Hasil pengujian aktivitas penghambatan α-glukosidase secara in vitro menunjukkan bahwa keempat senyawa memiliki aktivitas penghambatan moderat dengan nilai konsentrasi penghambatan 50% (inhibitory concentration, IC50) 44,43 ± 5,7 hingga 60,86 ± 6,8 µM yang lebih baik daripada akarbosa (6) (IC50 = 185,00 ± 9,4 μM). Aktivitas penghambatan α-glukosidase senyawa sinamamida sangat bergantung pada jenis substituen pada strukturnya. Gugus pensubstitusi penarik elektron pada kerangka sinamat dapat meningkatkan aktivitas penghambatan. Hasil pengujian menunjukkan bahwa senyawa 29a dengan gugus pensubstitusi dimetilamino pada posisi para pada kerangka sinamat memiliki potensi penghambatan terbaik. Studi penambatan molekuler menunjukkan bahwa senyawa 29a dengan energi pengikatan -8,33 kcal/mol berinteraksi dengan berbagai residu katalitik pada sisi aktif α-glukosidase (kode PDB: 3W37), yakni His626, Ile358, Asp357, Asp568, Asp469, dan Phe601. Lebih lanjut, senyawa sinamamida 29a-d juga mampu menghambat pertumbuhan bakteri Mtb H37Rv dengan konsentrasi penghambatan minimum (minimum inhibitory concentration, MIC) 1,56-12,5 μg/mL yang lebih baik atau sebanding dengan pirazinamida (3) (6,25-12,5 µg./mL). Berbeda dengan hubungan struktur-aktivitas penghambatan α-glukosidase, aktivitas antituberkulosis sinamamida dapat diuntungkan oleh substitusi gugus penarik maupun pendorong elektron pada posisi para pada kerangka sinamat. Hasil pengujian menunjukkan bahwa senyawa 29b dengan gugus pensubstitusi bromin memiliki potensi penghambatan terbaik (MIC > 1,56 μMg/mL). Studi penambatan molekuler menunjukkan bahwa senyawa 29b terikat pada subkantung pengikatan dan kantong hidrofobik potensial pada sisi aktif enoil-acyl carrier protein (ACP) reduktase (InhA) dalam konformasi memanjang. Senyawa sinamamida 29a-d menunjukkan karakteristik fisikokimia dan farmakokinetika dengan toksisitas minimum. Secara keseluruhan, hasil penelitian ini menunjukkan potensi sinamamida 29a-d sebagai lead compound untuk dioptimasi lebih lanjut sebagai agen penghambat α-glukosidase. Inflamasi atau peradangan akut yang tidak terkontrol dapat berubah menjadi peradangan kronis yang menjadi mediator penyakit dengan prevalensi tinggi seperti diabetes dan osteoartritis (OA). Pengobatan untuk mengurangi rasa sakit dan peradangan dilakukan dengan terapi obat analgesik dan obat antiinflamasi non-steroid (OAINS). Meskipun demikian, swamedikasi yang dilakukan sering kali menimbulkan efek samping akibat dosis yang tidak sesuai dengan gejala klinis. Terapi alternatif dalam pengobatan inflamasi dapat dilakukan dengan menggunakan inhibitor enzim siklooksigenase-2 (COX-2), seperti senyawa golongan coxib, di antaranya celecoxib dan rofecoxib. Namun, obat-obat ini menunjukkan efek pada sistem kardiovaskular dan sistem ginjal. Oksindola merupakan senyawa aromatik heterosiklik yang digunakan sebagai lead compound dalam pengembangan senyawa bioaktif karena memiliki beragam aktivitas biologis, seperti analgesik dan antiinflamasi. Selanjutnya, senyawa turunan furan hidrazida-hidrazon menunjukkan aktivitas antiinflamasi yang lebih baik daripada nimesulida. Pembentukan hidrazon pada kondisi netral relatif lambat sehingga menjadi hambatan dalam pemanfaatannya. Sintesis hidrazon dapat berjalan cepat pada pH 5 atau lebih rendah. Senyawa hidrazida-hidrazon (31a-d) berhasil didapatkan melalui studi in silico, yaitu evaluasi ADMET, sifat kemiripan obat, dan studi penambatan molekuler. Senyawa 31a-d memenuhi aturan Lipinski, tidak toksik, dan memiliki energi pengikatan yang lebih rendah dibandingkan dengan tenidap terhadap reseptor COX-2. Senyawa hidrazida-hidrazon 31a-d berhasil disintesis melalui reaksi adisi nukleofilik yang melibatkan isatin (33a-d) dan 5-bromofuran-2-karbohidrazida (34) dalam kondisi asam, dengan rendemen 79-88%. Hasil pengujian antiinflamasi melalui penghambatan denaturasi bovine serum albumin (BSA), proteinase, hemolisis yang diinduksi panas, dan hemolisis yang diinduksi hipotonisitas secara in vitro menunjukkan bahwa senyawa 31a-d memiliki potensi penghambatan yang tergolong sangat baik, dengan nilai IC50 berkisar antara 1,07 ± 1,13 (3,20 ± 3,38 µM) hingga 19,43 ± 5,6 (55,18 ± 15,90 µM) µg/mL, yang sebanding atau lebih baik dibandingkan dengan natrium diklofenak (67) dengan IC50 berkisar antara 1,97 ± 4,19 (6,19 ± 13,17 µM) dan 12,20 ± 2,99 (38,35 ± 9,40 µM) µg/mL. Hasil pengujian penghambatan enzim siklooksigenase-2 (COX-2) menunjukkan bahwa (E)-5-bromo-N'-(5,7-dibromo-2-oksoindolin-3-iliden)furan-2-karbohidrazida (31c) memiliki potensi penghambatan yang lebih baik, dengan nilai IC50 sebesar 0,61 ± 2,77 µg/mL (1,24 ± 5,63 µM). Hasil studi penambatan molekuler menunjukkan bahwa senyawa 31c (energi pengikatan -9,81 kcal/mol) menempati saluran spesifik yang hanya terdapat pada sisi aktif reseptor COX-2 (kode PDB: 6COX) dan kantong yang sama dengan kantong pengikatan gugus trifluorometil pada struktur SC-558. Lebih lanjut, hidrazida-hidrazon 31a-d menunjukkan karakteristik fisikokimia dan farmakokinetika dengan toksisitas minimum. Secara keseluruhan, hasil penelitian ini menunjukkan potensi hidrazida-hidrazon 31a-d sebagai lead compound untuk dioptimasi lebih lanjut sebagai agen penghambat siklooksigenase-2.
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Tuberculosis, caused by Mycobacterium tuberculosis (Mtb), is an infectious disease that increases the risk of infection with the human immunodeficiency virus. Isoniazid is a heterocyclic aromatic compound, one of the most effective drugs for treating tuberculosis. However, resistance to tuberculosis drugs and the lack of new tuberculosis drugs have become a global urgency in the fight against tuberculosis. Type 2 diabetes as a comorbidity of tuberculosis increases the mortality rate from this disease. Literature studies have shown that cinnamic acid and its derivatives exhibit antituberculosis activity and inhibit the enzyme α-glucosidase. Isoniazid and cinnamic acid hybrid molecules in the form of cinnamamide show the potential to inhibit Mtb. Hybrid molecules of isoniazid and cinnamic acid, namely cinnamamide compounds, have shown potential for inhibiting Mtb. Cinnamamide synthesis can be carried out using the reagent 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU). However, this compound is classified as a class I explosive and exhibits autocatalytic decomposition during storage and transportation. The compound 2-methyl-6-nitrobenzoic anhydride (MNBA) (14) was then utilized to replace HBTU in the synthesis of ester compounds. In silico experiments, including evaluation of pharmacokinetic and toxicological properties (ADMET), drug-like properties, and molecular docking, yielded a hybrid series of cinnamamides (29a-d). This compound meets the criteria for oral drugs according to Lipinski, is safe based on its predicted toxicity, and shows lower binding energy than the standards 6-chloropyrazine-2-carboxylic acid (POA) (30) and acarbose (6) to the Mtb receptor and α-glucosidase, respectively. Cinnamamides 29a-d were successfully synthesized via amidation of cinnamic acid (32a-d) with isoniazid (2), MNBA (14), and 4-dimethylaminopyridine (DMAP) (16), with a yield of 38-75%. The results of in vitro α-glucosidase inhibitory activity testing showed that the four compounds had moderate inhibitory activity with IC50 values of 44.43 ± 5.7 to 60.86 ± 6.8 μM, which were superior to acarbose (6) (IC50 = 185.00 ± 9.4 μM). The α-glucosidase inhibitory activity of cinnamamide compounds is highly dependent on the type of substituent in their structure, where electron-withdrawing substitution groups on the cinnamate skeleton increase the inhibitory activity. The test results showed that compound 29a, with a dimethylamino substitution group at the para position on the cinnamate skeleton, had the best inhibitory potential. Molecular docking studies showed that compound 29a, with a binding energy of -8.33 kcal/mol, interacted with various catalytic residues on the active site of α-glucosidase (PDB ID: 3W37), namely His626, Ile358, Asp357, Asp568, Asp469, and Phe601. Furthermore, cinnamamides 29a-d were also able to inhibit the growth of Mtb H37Rv bacteria with a minimum inhibitory concentration (MIC) of 1.56-12.5 μg/mL, which was better than pyrazinamide (3) (6.25-12.5 μg/mL). In contrast to the structure-activity relationship of α-glucosidase inhibition, the antituberculosis activity of cinnamamide can be benefited by the substitution of electron-withdrawing or electron-donating groups at the para position of the cinnamate skeleton. The test results showed that compound 29b, bearing a bromine substitution group, had the best inhibitory potential (MIC > 1.56 μg/mL). Molecular docking studies indicated that compound 29b bound to the binding subpocket and potential hydrophobic pocket in the active site of enoyl-acyl carrier protein (ACP) reductase (InhA) in an elongated conformation. Furthermore, cinnamamides 29a-d showed physicochemical and pharmacokinetic characteristics with minimal toxicity. Overall, the results of this study indicate the potential of cinnamamides 29a-d as lead compounds for further optimization as α-glucosidase inhibitors. Uncontrolled acute inflammation can progress to chronic inflammation, mediating highly prevalent diseases such as diabetes and osteoarthritis (OA). Treatment to reduce pain and inflammation involves analgesics and non-steroidal anti-inflammatory drugs (NSAIDs). However, self-medication often results in side effects due to inappropriate dosages for clinical symptoms. Alternative therapies for the treatment of inflammation include the use of cyclooxygenase-2 (COX-2) enzyme inhibitors, such as coxib compounds, including celecoxib and rofecoxib. However, these drugs have been shown to affect the cardiovascular and renal systems. Oxindole, a heterocyclic aromatic compound, is used as a lead compound in the development of bioactive compounds due to its diverse biological activities, such as analgesic and anti-inflammatory properties. Furthermore, furan hydrazide-hydrazone derivatives showed better anti-inflammatory activity than nimesulide. The formation of hydrazone is relatively sluggish under neutral conditions, which impedes its utilization. Hydrazone synthesis can proceed rapidly at a pH of 5 or lower. Hydrazide-hydrazone compounds (31a-d) were successfully identified through in silico ADMET, drug-like properties, and molecular docking evaluations. Compounds 31a-d fulfill Lipinski's rule, are non-toxic, and have lower binding energy compared to tenidap to the COX-2 receptor. Hydrazides-hydrazones 31a-d were successfully synthesized through a nucleophilic addition reaction involving isatin (33a-d) and 5-bromofuran-2-carbohydrazide (34) under acidic conditions, with a yield of 79-88%. The results of anti-inflammatory testing through inhibition of denaturation of bovine serum albumin (BSA) denaturation, proteinase, heat-induced hemolysis, and hypotonicity-induced hemolysis in vitro showed that compound 31a-d had very good inhibitory potential with IC50 values ranging from 1.07 ± 1.13 (3.20 ± 3.38 µM) to 19.43 ± 5.6 (55.18 ± 15.90 µM) µg/mL, which were comparable or better than diclofenac sodium (67), with IC50 values ranging from 1.97 ± 4.19 (6.19 ± 13.17 µM) to 12.20 ± 2.99 (38.35 ± 9.40 µM) µg/mL. Meanwhile, the results of the inhibition test of cyclooxygenase-2 (COX-2) enzymes showed that (E)-5-bromo-N'-(5,7-dibromo-2-oxoindoline-3-ylidene)furan-2-carbohydrazide (31c) had better inhibitory potential, with an IC50 value of 0.61 ± 2.77 µg/mL (1.24 ± 5.63 µM). The results of molecular docking studies showed that compound 31c (binding energy -9.81 kcal/mol) occupies a specific channel that is only found on the active site of the COX-2 receptor and the same pocket as the trifluoromethyl group binding pocket in the SC-558 structure. Furthermore, hydrazide-hydrazones 31a-d exhibited physicochemical and pharmacokinetic characteristics with minimal toxicity. Overall, the results of this study demonstrate the potential of hydrazide-hydrazones 31a-d as lead compounds for further optimization as a cyclooxygenase-2 inhibitor.
| Item Type: | Thesis (Doctoral) |
|---|---|
| Uncontrolled Keywords: | antituberkulosis, inhibitor α-glukosidase, antiinflamasi, sinamamida, hidrazida-hidrazon, studi in silico, antituberculosis, α-glucosidase inhibitor, anti-inflammatory, cinnamamide, hydrazide-hydrazone, in silico studies |
| Subjects: | Q Science Q Science > QD Chemistry Q Science > QD Chemistry > QD251.2 Chemistry, Organic. Biochemistry Q Science > QD Chemistry > QD471 Chemical compounds - Structure and formulas Q Science > QD Chemistry > QD481 Chemical structure. |
| Divisions: | Faculty of Science and Data Analytics (SCIENTICS) > Chemistry > 47001-(S3) PhD Thesis |
| Depositing User: | Nur Pasca Aijijiyah |
| Date Deposited: | 06 Aug 2026 02:53 |
| Last Modified: | 06 Aug 2026 02:53 |
| URI: | http://repository.its.ac.id/id/eprint/144134 |
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