(BIOLOGIC DRUG)
ANA INDRAYATI
FAKULTAS FARMASI USB
anaindrayati@setiabudi.ac.id
08156266891
MATERI
1. Protein terapeutik
2. DNA/RNA terapeutik
3. CRISPR-Cas 9
4. Bakteriofaga sebagai obat
OBAT BIOLOGI: obat yang berasal dari makhluk
hidup berupa jaringan, sel, DNA, RNA dan protein
▪ Dunia Farmasi: obat kimia (sintetik) dan obat biologi
▪ Obat biologi: biopharmaceuticals (biotechnology-derived pharmaceuticals),
biologic(al) medical product, biologic
▪ Obat biologi non-rekombinan-> diisolasi langsung dari sumber
penghasil
▪Insulin: diekstraksi dari pankreas hewan (sapi dan babi)
▪Artemisinin: diekstraksi langsung dari Artemisia annua
▪ Obat biologi rekombinan-> rekayasa genetika (kloning)
▪Insulin manusia: diproduksi di bakteri Escherichia coli
▪Artemisinin: diproduksi di sel ragi Sacharomyces cerevisiae
Metabolit Sekunder Fakta-fakta dan Permasalahannya
Paclitaxel (kanker Kadar pada kulit batang 0,01-0,02%, 1 g taxol/3 batang
ovarium, payudara, paru- pohon, pengobatan membutuhkan 2 g, dibutuhkan waktu
paru, pankreas) 100 tahun untuk mendapatkan pohon dewasa yang cukup
besar untuk dipanen, permintaan pasar 250 kg/tahun.
Sintesis kimia sulit dan sangat kompleks
Artemisia annua
• OBAT BIOLOGI/BIOLOGIC
• OBAT INOVATOR/ORIGINATOR/REFERENCE
• BIOSIMILAR/FOLLOW OF BIOLOGIC (US dan Eropa): similar
(keamanan, kemurnian, profil efikasi) dengan originator> diproduksi
setelah masa paten produk innovator berakhir
• BIOBETTER: better (potensi/efikasi, kemurnian protein, waktu paro
lebih panjang, dosis kecil, imunogenesitas dan toksisitas rendah, efek
samping kecil.
▪ Dapat diproduksi tanpa menunggu berakhirnya masa paten
produk innovator
▪ Komponen active biobetter berbeda dengan produk inovator
▪ Targetnya biobetter sama dengan produk innovator
BIOSMILIAR DAN BIOBETTER:
DERIVATIF DARI BIOLOGIC (ORIGINATOR/INNOVATOR/REFERENCE)
INSULIN NON- INSULIN
REKOMBINAN REKOMBINAN BIOSIMILAR BIOBETTER
(pankreas hewan) (Humulin: innovator)
PERBANDINGAN BIOSIMILAR-BIOBETTER
Biosimilar Biobetter
Keterbatasan: struktur (komponen aktif Tidak ada keterbatasan struktur:
mirip dengan produk innovator) modifikasi kimia/molekuler (komponen
aktif berbeda dengan produk innovator)
Similar: keamanan dan profil Improve: keamanan dan profil
efikasi/potensi efikasi/potensi
Approve: data (comparability data) mirip New drug: data klinik dan non-klinik
dengan produk innovator
Yunani: DIAGNOSIS:
▪ Haima: darah o Pemeriksaan kadar faktor
▪ Philia: cinta atau kasih sayang pembekuan VIII, IX dan XI
JENIS HEMOFILIA: o Diagnosis molekular (gen
hemofilia)
o A (defisiensi faktor VIII)
o B (defisiensi faktor IX)
o Penyakit Christmas: nama
keluarga pasien pertama
o C (defisiensi faktor XI)
FAKTOR PEMBEKUAN DARAH:
TERAPI HEMOFILIA
• HEMOFILIA: darah
tidak dapat membeku
• Defisiensi protein
untuk pembekuan
darah
• Protein tersebut
diproduksi oleh sel-sel
sinusoidal di hati
THE ROYAL
DISEASE
Recombinant FVIII protein is currently used to treat bleeding episodes at the cost of
approximately $55 000 per person year
Antibodimonoklonal (monoclonal antibody,
MAB)
HBsAg: Hepatitis B Surface Antigen • Alemtuzumab (Campath): chronic
HPV: Human Papiloma Virus lymphocytic leukemia (CLL)
Protein sebagai vaksin: vaksin 2 • Bevacizumab (Avastin): cervical,
Hbsag, vaksin HPV colorectal, non-small-cell lung, kidney,
and some brain cancers
1 • Rituximab (Rituxan): non-Hodgkin's
lymphoma.
KLASIFIKASI 3
PROTEIN
Protein untuk diagnosis: leptin,
TERAPEUTIK glukagon, N-protein (Covid-19)
Produksi Ekstraksi,
Kloning
protein Pemurnian,
Protein Formulasi
Skala besar Karakterisasi
rekombinan
(Fermentasi) protein
Memenuhi
persyaratan:
mutu, efikasi,
safety
https://courses.lumenlearning.com/wm-nmbiology1/chapter/reading-
prokaryotic-and-eukaryotic-gene-regulation/
DNA/RNA TERAPEUTIK
RNA ANTISENSE
DNA plasmid
dibungkus dengan
partikel berukuran 1–3
mikron (emas atau
tungsten).
• Beberapa terapi gen telah disetujui oleh FDA US, termasuk
Kymriah® untuk leukemia, Luxturna™ untuk penyakit
genetik pada retina, Zolgensma® untuk Spinal Muscular
Atrophy, eteplirsen untuk Duchene Muscular Dystrophy,
Trikafta® untuk cystic fibrosis, dan Zynteglo™ untuk
talasemia yang masih dalam pertimbangan
• https://www.youtube.com/watch?v=CvJNzxvPCzg
CRISPR-Cas9
TEKNOLOGI MENGEDIT GEN
• Zinc-finger nucleases (ZFNs)
• Transcription activator-effector nucleases (TALENs)
• Clustered regularly interspaced short palindromic
repeats-Cas 9 (CRISPR-Cas 9)
Paling populer
faster, more precise, more efficient, and more cost-
effective
Teknologi mengedit genom semakin popular
• Berpotensi digunakan untuk mencegah dan
mengobati berbagai penyakit (monogenic:
hemophilia, anemia bulan sabit, dan sistik
fibrosis), (kompleks: kanker, kardiovakskuler, DM,
HIV/AIDS, penyakit saraf dll)
SEJARAH CRISPR-CAS 9
• Clustered regularly interspaced short palindromic repeats-Cas 9
• 1987 ditemukan pada bakteri E. coli-> 20 tahun berikutnya (2007)-> CRISPR_Cas 9
sistem imun pada bakteri dan archaebakteri
• Sistem imun: faga (bakteriofaga) dan plasmid
• Infeksi faga-> CRISPR memotong bagian kecil dari DNA faga-> menyimpan
potongan DNA faga-> infeksi faga berikutnya-> mengenali dan melawan faga
https://www.civilsdaily.com/news/nobel-prize-in-
chemistry-for-crispr-technology/
2011:Charpentier dan Doudna
mempelajari fenomena sistem
imun pada bakteri->
Mensintesis Cas9 kombinasi
gRNA-> untuk berbagai tujuan
https://www.civilsdaily.com/news/nobel-prize-in-
chemistry-for-crispr-technology/
▪ Pasangan suami istri Grace
dan Mark (HIV positif)
▪ Bayi tabung fertilisasi in-
vitro (IVF)
▪ Dua anak kembar (Lulu dan
Nana-> nama samaran)
▪ Ilmuwan Cina He Jiankui
dan koleganya
▪ Saat masih embrio (sel
tunggal) He Jianku
mengedit DNA Lulu dan
Nana
▪ Memutasi gen CCR5 (co-
reseptor HIV) dengan
CRISPR-Cas g
▪ Tujuan: Lulu dan Nana
terbebas dari HIV
▪ Proses tidak berhasil
▪ Pertentangan: melanggar
kode etik ilmiah
PENGEMBANGAN OBAT:
BAKTERIOFAGA UNTUK MENGATASI
RESISTENSI BAKTERI TERHADAP
ANTIBIOTIK
Phage therapy: An alternative to antibiotics in the age of multi-drug resistance
ANA INDRAYATI
FAKULTAS FARMASI USB
anaindrayati@setiabudi.ac.id
MULTIDRUG-RESISTEN (MDR)
XDR: Extensively Drug Resistant
▪ Metisilin resistant Staphylococcus aureus
XDR TB adalah TB yang
(MRSA)
disebabkan oleh strain yang
▪ Vancomycin-resistant Enterococci (VRE)
▪ Carbapenem-resistant Acinetobacter resistensi terhadap isoniazid dan
baumannii (CRAB) rifampisin, disertai resisten
▪ Multi-drug resistant Pseudomonas aeruginosa terhadap salah satu fluorokuinolon
▪ Enterobacteriaceae that produce extended dan salah satu dari tiga obat
spectrum β-lactamases
injeksi lini kedua (amikasin,
▪ Carbapenemase-producing
kapreomisin atau kanamisin)->
Enterobacteriacea
lebih FATAL dibandingkan MDR
• BAKTERIOFAGA (FAGA) adalah virus yang
menginfeksi bakteri (dan archaebacteria)
• Bakteriofaga ditemukan secara terpisah oleh
Frederick W. Twort (Inggris, 1915) dan Felix
d’Herelle (Prancis, 1917)
• Bacteriophage, meaning “bacteria eater,” :
bakteriosid (membunuh bakteri)
• Bakteriofaga menyebabkan bakteri lisis
(menganggu metabolisme) > bakteri mati
• Non-living biological (tergantung sel inang),
materi genetik DNA atau RNA yang dilindungi
oleh protein kapsid
BIG HERO
RESEPTOR (‘PINTU MASUK’) FAGA KE DALAM SEL BAKTERI
MEKANISME PERTAHANAN
BAKTERI THD INFEKSI FAGA: RESEPTOR FAGA: SPESIFIK (GAMBAR)
▪ Mengubah struktur reseptor
(tidak dikenali faga)
▪ DNA faga dipotong-potong
dengan gunting molekuler
clustered regularly interspaced
palindromic repeats/CRISPR
associated system
(CRISPR/Cas) system
UPAYA FAGA :
▪ Bermutasi sehingga tetap
dapat masuk melewati
reseptor yang telah berubah
strukturnya
▪ Anti CRISPR/Cas system)
▪ Specificity: Bacteriophages are generally very specific in their host range, which promises less harm to
our microbiota. However, this specificity also means that for most clinically relevant infections, a mixture
of different phage will have to be used to address the polymicrobial nature of the infection
▪ Bactericidal: lytic phages infect their target host bacteria and cause cell death, in comparison to certain
bacteriostatic antibiotics
▪ Active on-site propagation: bacteriophages increase in concentration in situ as they propagate in the
presence of bacterial host (infectious agents). Unlike antibiotics, which often require frequent doses to
kill the bacteria efficiently, only one bacteriophage is theoretically needed to target a single
corresponding host bacterium (single-hit kinetics). It is possible to administer a single low-dose of
phage, which will then propagate itself, given the existing bacterial density as an active therapy,
resulting in continued bacterial adsorption and killing
▪ Low inherent toxicity: bacteriophages are primarily composed of nucleic acid and proteins that have
been studied to be non-toxic with the use of highly purified phage preparations [
▪ Enormous diversity: phage is found in diverse abundances across the biosphere and therefore isolating
and purifying new phages, necessary to target a known pathogenic bacteria, is achievable even when
bacteria evolve resistance
▪ Formulation and application versatility: multiple strains of phages can be added together into a
“phage cocktail” to target multiple bacteria of interest with a broader killing spectrum.
▪ Narrow potential for antibiotic cross-resistance: the mechanism of bacterial resistance to antibiotics
and phage are entirely different. Thus, bacteria that are resistant to antibiotics may be targeted and
treated with the use of phage therapy, and vice-versa, presenting combination therapies as an
attractive strategy .
▪ Biofilm clearance: phages have been demonstrated to lyse and penetrate through some biofilms
that have shown resistance to antibiotics. This is partially attributed to the presence of
depolymerases and lysins that can chew through the biofilm extracellular polymeric matrix.
▪ Low environmental impact: bacteriophages are natural components of the environment that can be
naturally evolved (as opposed to genetic modification), thus easing public acceptance of phage
therapy. Furthermore, because this natural product is composed primarily of proteins and nucleic
acids, unused phage materials can easily be inactivated and discarded
▪ Relatively low discovery and production cost: the costs associated with the discovery, isolation, and
purification of bacteriophages are significantly decreasing with the progression of screening and
sequencing technologies
TERAPI FAGA PADA PASIEN CF YANG TERINFEKSI BAKTERI
Burkholderia cepacian PENYEBAB PNEUMONIA
• In 2006, the FDA recognized the use of a cocktail of six bacteriophages in the form of an application spray
as GRAS (Generally Recognized as Safe). The product is marketed as LISTEX P100 and is produced by the
Phage Technology Center in the Netherlands.
• LISTEX is used in the clean rooms of cheese, meat, and poultry processing plants to inhibit the growth of
Listeria on products such as luncheon meats and hot dogs. The phage preparation is not declared as an
ingredient on the label of a treated product. Additional products are being developed to reduce
Campylobacter on chicken and Salmonella on fish, cheese, and meats.
Biochemistry (Mosc). 2022; 87(8): 777–788.
Published online 2022 Aug 15. doi: 10.1134/S0006297922080090
PMCID: PMC9377665
PMID: 36171658
CRISPR–Cas9: A History of Its Discovery and Ethical Considerations of Its
Use in Genome Editing
Irina Gostimskaya