Antikanker

Suryawati, S.Si., Apt., M.Sc.

Didi Nurhadi Illian, S.Farm., M.Si., Apt.
 SUB-CP-MK
(sbg kemampuan akhir yang diharapkan)
Mampu menjelaskan:
 Hubungan struktur kimia dengan aktivitas biologis obat
 Modifikasi molekul
Outline
 Mekanisme kerja
 Penemuan
 Modifikasi molekul

nitrogen mustard & agen alkilasi lainnya: (5-fluorourasil,

taxol, vinka)
 Apa itu Kanker? – akibat (hasil) dari gen yang
rusak
 Kerusakan pada materi genetik (DNA) menyebabkan:
 Kematian sel
 Perbaikan sel
 Mutasi genetik (jarang)
 Translasi dari gen yang bermutasi  protein mutant yang
menyebabkan:
 Sel kehilangan kemampuan untuk ‘berhenti tumbuh atau mati’
 Peningkatan pertumbuhan sel

CHY860
 Mengapa Kanker Mematikan?
 Tumor tumbuh di ruang terbatas dan menghancurkan jaringan
normal
 Tumor  laju metabolisme tinggi  kekurangan nutrisi 
kelelahan
 Menyebabkan defisiensi imun  resiko infeksi ↑ infections
 Tumor memproduksi hormon  mengganggu fungsi metabolik
normal
 Invasi ke jaringan disekitarnya  perdarahan dan infeksi
 Tumor bermetastasis ke organ yang letaknya jauh (e.g. tulang, hati,
otak dan lambung) dan menghancurkan jaringan tersebut.
CHY860
Siklus Sel
…lanjutan (Siklus Sel)
Mekanisme Kerja
…lanjutan (Mekanisme Kerja)
 Agen Alkilasi

Kimia Medisinal
Covalent DNA-binding drugs
Nitrogen mustards Aziridines Alkane sulphonates
Mechloethamine, Thiotepa, altretamine Busulfan
chlorambucil, melphalan, (hexamethylmelamine),
cyclophosphamide, ifosfamide mitomycin C
O O
N H3C O S O (CH2)n O S O CH3
Cl
O O
N N
R N
N N N
Cl
Triethylenemelamine (TEM)

Nitrosoureas Methylating agents Platinum complexes

Carmustine (BCNU), Dacarbazine, procarbazine, Cisplatin (cis-DDP),
lomustine (CCNU), temozolamide carboplatin
semustine (methyl-CCNU),
streptozotocin
N CONH2
Cl NH3
O Pt
Cl NH3
R R' CH3
N
N N H N N N
H N CH3
Dacarbazine
O
 Target DNA
 Basa DNA sel kanker merupakan target agen alkilasi.
 Basa tersebut adalah:
 Guanine (N-7, O-6) > adenine (N-3 > N-1) > cytosine (N-
1) >> thymine

NH2 O6 NH2 O
7
N N CH3
1N HN N HN
1
N N H2N 2 N N O N O N
3 R R R R
adenine guanine cytosine thymine
 Mekanisme umum Agen alkilasi
 Agen Alkilasi → mengganti H dengan gugus alkil (R) pada kondisi
fisiologis under physiological conditions
 Alkilasi basa DNA menyebabkan:
 Miscoding selama proses replikasi (misal: G-C menjadi G-T ) 
mutagenesis
 Depurinasi dan rantai DNA pecah
 Stimulasi proses perbaikan
 Memicu proses penghancuran diri sendiri (apoptosis)

pH 7.4, 37 oC
HNu R X Nu R + HX
aqueous solution
biological alkylating
nucleophile agent
 Mekanisme kimia dari nitrogen mustards
 Pembentukan ion aziridinium ion – dibantu oleh pasangan
elektronbebas dari nitrogen
 Aziridinium bereaksi dengan nucleophile (basa DNA sel kanker)
 Grup kloroetil kedua mengalami reaksi yang sama
Cl Nu1 Nu1
fast SN1 SN2
R N R N R N

Cl Cl Cl

SN1

2
Nu Nu
2

SN2
R N R N

Nu1 Nu
1
 Mekanisme biologi nitrogen mustards
 Nitrogen mustards bereaksi dengan
dua molekul bifunctional)
 Dapat membentuk inter- dan
intrastrand cross-links
 Ikatan silang antar rantai
(Interstrand cross-links) mencegah
pemisahan rantai DNA sel kanker
 salah kode (miscoding of bases) Interstrand Intrastrand
R
O O
 rantai DNA pecah HN
N
N
N
NH

H2N N N N NH2
N

DNA O O DNA
O O

O O

O P O- -O P O

O O
DNA DNA
DNA crosslink formed from reaction at N-7 guanine on both strands
(G-G crosslink)
Clinically used nitrogen mustards
Cl Cl

N N
O Cl NH2 Cl

NaO HOOC
Chlorambucil Melphalan

Me O NH O
N P Cl
O N
Cl Cl Cl
Nitromin
Cyclophosphamide
 Cyclophosphamide – a prodrug
Cl Cl Cl Cl Cl Cl Cl Cl

oxidative
N O metabolism N O N O N O
P P P P
HN O HN O H2N O H2N O
H
HO O O
Cyclophosphamide 4-Hydroxycyclophosphamide Aldophosphamide

SO 3Na retro-Michael
O S O reaction
detoxified product acrolein
SO 3Na (toxic)
HS
Mesna

 Cyclophosphamide is an unreactive Cl Cl
Cl Cl
prodrug – activated by metabolism in
N O
the liver. N
H P
H2N O
 The mustard is deactivated by the Nornitrogen Phosphoramide
mustard mustard
electron withdrawing P=O group.
DNA reactive metabolites
 Landmarks – Tamoxifen
 Discovered from ICI’s oral contraceptive
program in the 1950’s.
 Currently, tamoxifen is the world’s best selling
anti-cancer drug. Used to treat breast cancer.
 30–40% of ER +ve women respond, duration of O

response 1–3 yrs. CH3

N
 Minimal side effects, non-toxic. Tamoxifen CH3
 Acts as an estrogen, antiestrogen, or mixed
agonist/antagonist depending on target organ
and species.
 How does tamoxifen work?
1. Tamoxifen binds competitively to the steroid binding site on the estrogen receptor (ER).
 Binding affinity for Estradiol = 100; tamoxifen = 4
 ER adopts a different conformation with tamoxifen bound than with estrogen

 
2. ER dimerises, losing a stablising protein HSP90
3. ER locates to the nucleus, binds to DNA at the estrogen response element (ERE).
4. The complex formed is only weakly transcriptionally active
 Overall effect is cytostatic i.e. cells stop growing at G1/S transition

                                                                                         
 Landmarks in chemotherapy
 Johnson et al 1963
 Vinca alkaloids → Isolated from Catharanthus roseus (formerly
classified as Vinca rosea)
 Clinically use: in combination for ALL, non-Hodgkin’s lymphoma,
ovarian, advanced testicular, Karposis sarcoma.
OH
N

N
H
N OCOCH3
H O
COOCH3
H3CO
H3CO N OH
R
Vincristine (VCR): R = CHO
Vinblastine (VBL): R = CH3
 Vinca alkaloids and the cell cycle

mitosis G0 – resting phase

X
M
premitotic interval
G2
G1

DNA synthesis

 Vinca alkaloids act during the M-phase of the cell cycle i.e. cell
division
 They act by interfering with the microtubules within the nucleus
Microtubules and the M-phase
G2 Interphase Prophase Metaphase

Telophase

Anaphase

Microtubule
Organising Centre
 Structure of microtubules

• made of dimers of a- and b- tubulin

• 13 units per tubule

http://www.rpi.edu/dept/bcbp/molbiochem/MBWeb/mb2/part1/microtub.htm
 Vinca alkaloids and microtubules
 Microtubules are organised tubes made up of tubulin dimers.
 Vinca alkaloids prevent the formation of microtubules and so stop
chromosome separation.
 They bind to tubulin dimers and form paracrystaline aggregates
preventing microtubule growth.
Vinca alkaloid

Microtubule
Paracrystaline
Free Vinca –bound aggregates
tubulin tubulin dimers
dimers
 Landmarks – Taxol
O
 1990’s NCI
O
 Taxol isolated from the bark of the H3C O
OH CH3 OH
pacific yew (Taxus brevifola). 10 000 H
N
H CH3
CH3
kg of bark yields 1 kg of taxol – O O HO H
enough for 500 patients. O O O
O
 Clinically use: ovarian, and breast O

cancer
Taxol
 Further Reading: Angew. Chem. Int.
Ed. Eng. 1994, 33, 15-44
 Taxol analogues
2
O R
 Taxotere semi-synthetic derivative from 1
O
O
R NH O
10-deacetylbaccatin III isolated from the OH
Ph O
leaves of Taxus baccata (European Yew). OH
HO H
 Twice as active as taxol. O O O
O
O

T a xo l R 1 = P h; R 2 = O A c;
T a x o te re R 1 = tB u O ; R 2 = O H ;

OH
O

OH
HO

HO H
O O O
O
O

1 0 -D e a c e ty lb a c c a tin III
 How do taxoids work?
 Taxol binds to tubulin dimers and acts by stabilising microtubules.
 During G1 causes microtubule bundling and cell death.
 During G2 – M-phase transition causes accumulation of disorganised
filaments around the MTOC. Prevents progression into M-phase and
causes cell death.

X-ray crystal structure of a,b tubulin dimer with taxol (red) bound
Antimetabolites

CHY360
Dr I Hardcastle
 Antimetabolit?
 Antimetabolit – agen yang mencegah proses metabolisme penting
dari sel DNA.
 Terdiri dari:
(1) antagonis folat
 Penghambat enzim dihydrofolate reductase e.g. methotrexate
 Penghambat enzim thymidylate synthase e.g. Tomudex

(2) Analog pyrimidine

 e.g. 5-fluorouracil, gemcitabine

(3) Antagonis purine

 e.g. 6-mercaptopurine

(4) Antagonis asam amino

 e.g. alanosine or azaserine
 Antimetabolites and the cell cycle
mitosis G0 – resting phase

M
premitotic interval
G2
G1

DNA synthesis

 Antimetabolit bekerja selama fase S pada sklus sel pada sintesa DNA
 Mekanisme:
– Penghambatan enzim yang terlibat pada sintesa basa nukleosida
– Penghambatan enzim yang terlibat pada sintesa kofaktor2 penting
– Penghambatan DNA polymerase
– Inkorporasi ke dalam sintesa RNA dan DNA synthesis
– Penghambatan proses umpan balik terhadap sintesa basa nukleosida
 Analog pirimidine: 5-fluorouracil
 Ditemukan oleh Heidelberger – 1965.
 Penelitian menunjukkan bahwa sel tumor lebih membutuhkan urasil
dibandingkan asam orotat.
 Dengan landasan bahwa asam asetat adalah cuka, sedangkan fluoroacetic
acid adalah racun tikus maka:
fluorinasi urasil  dapat menjadi toksik bagi sel tumor
 Substitusi yang mungkin terjadi adalah posisi 5 dan 6.
 Substitusi pada posisi 5 lebih mudah terjadi dan menghasilkan senyawa
yang stabil
 Thymidine adalah 5-metilurasil

O O O O

5 5 CH3 5 F
HN HN HN HN
O 6
O N O N O N O 1N
H H H H
OH
orotic acid uracil thymidine 5-fluorouracil
 5-FU Metabolism
 Aktivasi 5-FU terjadi melalui proses metabolisme melalui reaksi berikut;
i. Menjadi ribonukleosida (basa + gula);
ii. Dengan fosforilasi menjadi ribonukleotida;
iii. Dengan modifikasi menjadi bentuk deoksida
 FUTP di-inkorporasi ke dalam RNA – sitotoksik
 dFUTP di-inkorporasi ke dalam DNA – sitotoksik
O O
F F
HN HN

-
O3PO O N H FUDP FUTP FU-RNA
O N O RNA Polymerase
H
5-FU OH OH
FUMP

O
F
HN

- O N H FdUDP FdUTP FU-DNA

O3PO
O DNA Polymerase

OH
FdUMP
 from Lecture of:
Dr Ian Hardcastle
I.R.Hardcastle@ncl.ac.uk