Sintesis Senyawa Analog Pirazinamida dan Aktivitas Penghambatannya Terhadap Mycobacterium tuberculosis secara In Vitro dan In Silico

Synthesis of Pyrazinamide Analog and Its Inhibition Activity Evaluation of In Vitro and In Silico Against Mycobacterium tuberculosis

ABSTRAK

SINTESIS SENYAWA ANALOG PIRAZINAMIDA DAN AKTIVITAS PENGHAMBATANNYA TERHADAP Mycobacterium tuberculosis SECARA IN VITRO DAN IN SILICO

Tuberkulosis merupakan salah satu ancaman kesehatan dunia dengan angka kasus yang tinggi. Pengobatan tuberkulosis terkendala dengan munculnya kasus multidrug-resistant tuberculosis (MDR-TB). Dalam beberapa tahun terakhir, dikembangkan berbagai analog pirazinamida yang menunjukkan kemampuan menghambat pertumbuhan M. tuberculosis, salah satunya adalah pirazinamida yang tersubstitusi dengan alisiklik. Sintesis analog pirazinamida dapat dilakukan dengan metode Yamaguchi yang melibatkan reagen 2,4,6-triklorobenzoil klorida (20) sebagai agen kopling. Pada penelitian ini bertujuan untuk mensintesis analog pirazinamida, yaitu N-(sikloheksilmetil)pirazina-2-karboksamida (11) menggunakan metode Yamaguchi. Pada penelitian ini dilakukan karakterisasi senyawa menggunakan spektroskopi ultraviolet (UV), inframerah (FTIR), resonansi magnetik inti atau nuclear magnetic resonance (NMR), dan spektrometri massa. Hasil penelitian menunjukkan bahwa senyawa 11 berhasil disintesis menggunakan metode Yamaguchi dengan rendemen sebesar 59,89%. Uji anti tuberkulosis senyawa 11 secara in silico menunjukkan bahwa senyawa aktif dalam menghambat reseptor InhA M. tuberculosis dengan binding energy dan konstanta inhibisi sebesar -7,48 Kkal/mol dan 3,31 M yang lebih kecil dibandingkan pirazinamida (3) dan isoniazid (1). Senyawa 11 tidak melanggar aturan 5 Lipinski, diprediksi memiliki penyerapan yang baik dalam usus halus, serta tidak berpotensi menimbulkan toksisitas terhadap hati, dan non-mutagenik. Hasil uji anti tuberkulosis senyawa 11 secara in vitro menunjukkan senyawa kurang aktif dalam menghambat pertumbuhan M. tuberculosis dalam konsentrasi kecil dengan konsentrasi penghambatan minimum yang diperoleh sebesar >25 g/mL.

Kata Kunci: Analog pirazinamida, anti tuberkulosis, metode Yamaguchi, sintesis, in silico, in vitro.

ABSTRACT

SYNTHESIS OF PYRAZINAMIDE ANALOG AND ITS INHIBITION ACTIVITY EVALUATION OF IN VITRO AND IN SILICO AGAINST Mycobacterium tuberculosis

Tuberculosis is one of the world's major health threats, with a high number of cases. The treatment of tuberculosis is obstructed by the emergence of multidrug-resistant tuberculosis (MDR-TB) cases. In recent years, various pyrazinamide analogues have been developed that show the ability to inhibit the growth of M. tuberculosis, one of which is pyrazinamide substituted with alicyclic groups. The synthesis of pyrazinamide analogs can be performed using the Yamaguchi method, which involves the use of 2,4,6-trichlorobenzoic chloride (20) as a coupling agent. The aim of this study was to synthesize a pyrazinamide analog, namely N-(cyclohexylmethyl)pyrazine-2-carboxamide (11), using the Yamaguchi method. In this study, the compounds were characterized using ultraviolet (UV) spectroscopy, infrared (FTIR) spectroscopy, nuclear magnetic resonance (NMR) spectroscopy, and mass spectrometry. The results showed that compound 11 was successfully synthesized using the Yamaguchi method with a yield of 59.89%. In silico anti-tuberculosis testing of compound 11 showed that the compound is active in inhibiting the InhA receptor of M. tuberculosis with a binding energy and inhibition constant of -7.48 kcal/mol and 3.31 μM, respectively, which are lower than those of pyrazinamide (3) and isoniazid (1). Compound 11 does not violate Lipinski's 5 rules, is predicted to have good absorption in the small intestine, and is not potentially toxic to the liver or mutagenic. In vitro anti-tuberculosis testing of compound 11 showed that the compound is less active in inhibiting the growth of M. tuberculosis at low concentrations, with a minimum inhibitory concentration (MIC) of >25 μg/mL.

Keyword: Pyrazinamide analog, anti-tuberculosis, Yamaguchi method, synthesis, in silico, in vitro.