SİNYAL MOLEKÜLLERİNİN KLİNİKTE HEDEFLENMESİ

Transkript

1 Hacettepe Üniversitesi Onkoloji Enstitüsü SİNYAL MOLEKÜLLERİNİN KLİNİKTE HEDEFLENMESİ Emin Kansu

2 NUCLEUS

3 HÜCREDE HEDEF YAPILAR?

4 HÜCREDE HEDEF MOLEKÜLLER ve YOLLAR

5 HÜCRE BİYOLOJİSİ GENLER mrna PROTEİNLER HÜCRENİN KADERİ

6 HÜCRE BİYOLOJİSİNDE ROL ALAN AKTÖRLER RESEPTÖRLER SİNYAL MOLEKÜLLERİ TRANSKRİPSİYON FAKTÖRLERİ HÜCRE SİKLUSU DÜZENLEYİCİLERİ ADEZYON MOLEKÜLLERİ KROMATİN DÜZENLEYİCİLERİ

7 KEMOTERAPİ Kanser Hücreleri Chemotherapy drugs target cancer cells because they are rapidly dividing Chemotherapy drugs enter the nucleus where they interfere with the DNA İLAÇ Chemotherapy drug Cells destroyed Leukaemic cells may survive chemotherapy and resist further doses of chemotherapy drugs. This is known as being refractory to chemotherapy

8 Targets of Cancer Therapy Plasma Membrane Growth factors 2. Growth factor receptors 3. Adaptor proteins 4. Docking proteins/ binding proteins Guanine nucleotide exchange factors 6. Phosphatases and phospholipases 7 Growth Factor Signaling 8 RNA Translation 11 Nuclear Membrane Microtubule Dynamics Signaling kinases 8. Ribosomes 9. Transcription factors 10. Histones 11. Molecular chaperones 12. DNA 13. Microtubules Cyclins 15. Cyclin-dependent kinases 10 Gene Transcription DNA Replication and Repair 16. Cell death receptors 17. Apoptosis-effector caspases 18. Caspase inhibitors 19. CD40-CD40L Cell Growth Motility Survival Proliferation Angiogenesis 14 15

9 HÜCREDE HEDEF MOLEKÜLER YAPILAR

10 HÜCRE Hücre Membranı TİROZİN KİNAZ SİNYAL MOLEKÜLLERİ TF Nukleus PROTEOZOM

11 RESEPTÖR SİNYAL*** TF GENLER

12 HÜCRE-İÇİ SİNYAL YOLLARI TRANSKRİPSİYON FAKTÖRLERİ YAŞAM PROLİFERASYON YÖNELİM FARKLILAŞMA GEN EKSPRESYONU

13 SİNYAL İLETİMİ SONUCU... SİTOPLAZMİK MOLEKÜLLERİN AKTİVASYONU NUKLEUS TA GEN TRANSKRİPSİYONU PROLİFERASYON AKTİVASYON FARKLILAŞMA APOPTOZİS

14 HÜCRE Hücre Membranı TİROZİN KİNAZ SİNYAL MOLEKÜLLERİ TF Nukleus PROTEOZOM

15 HÜCRE YÜZEY RESEPTÖRLERİNİN LİGAND BAĞLANMASI LİGAND RESEPTÖR SİNYAL YOLLARI

16 HÜCRE YÜZEY RESEPTÖRLERİ ve SİNYAL LİGAND RESEPTÖR SİTOPLAZMA SİNYAL UÇLARI TAILS

17 KİNAZLARIN YERLEŞİM YERLERİ

18 TİROZİN KİNAZ ETKİSİ FOSFORİLASYON SİNYALİN BAŞLAMASI

19 PROTEİN KİNAZLAR Tirozin Kinaz Serin Kinaz Treonin Kinaz

20 LİGAND RESEPTÖR BAĞLANMASI VE TİROZİN KİNAZ LİGAND RESEPTÖR RESEPTÖR TİROZİN KİNAZ

21 Reseptör Tirozin Kinase (RTK) Aileleri TGF-a EGF HER (erbb) Family None Heregulin Epiregulin Heregulin Epiregulin VEGF-A VEGF-B VEGFR Family VEGF-A VEGF-C VEGF-D VEGF-C VEGF-D PDGFR Family FL PDGF SCF EGFR HER2 HER3 HER4 VEGFR-1 VEGFR-2 VEGFR-3 Flt-3 PDGFR KIT Receptor Dimerization and Activation FGF-1 FGF-14 FGF-16 FGF-23 IGF-1 IGF-2 P P P P P P P P P P P P P P P P FGFR1 FGFR2 FGFR3 FGFR4 IGF-1R VEGFRs, PDGFR, KIT, Flt-3 EGFR/HER2 Mutant Flt-3

22 HEDEF : TİROZİN KİNAZ SİNYAL SİSTEMLERİ TİROZİN KİNAZ İNHİBİTÖRLERİ

23 NORMAL TİROZİN KİNAZLARIN AKTİVASYONLARI İNAKTİF DURUM

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25 Kronik Myeloid Lösemi (KML) FÜZYON GENİ

26 TİROZİN KİNAZ İNHİBİTÖRLERİ Bcr-Abl KRONİK MİYELOİD LÖSEMİ t ( 9;22 ) TRANSLOKASYONU Bcr-Abl Füzyon Geni GLIVEC TİROZİN KİNAZ Tirozin Kinaz HÜCRE İÇİ SİNYALLER Aktivasyonu SİNYAL İLETİMİ

27 IMATINIB MESYLATE - Gleevec

28 IMATINIB İN ETKİ MEKANİZMASI

29 KANSERLERDE TİROZİN KİNAZ DİSREGÜLASYONU TK-PROTEİN FÜZYONLARI

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33 DASATINIB DASATİNİB!

34 HEDEFLENMİŞ TEDAVİLER : SİNYAL İLETİMİ İNHİBİTÖRLERİ IMATINIB MESYLATE (Tirozin Kinaz İnhibitörü) GENEFITINIB (Epidermal Büyüme Faktör Reseptörü Tirozin Kinaz İnhibitörü EGFR-TK) CETUXIMAB (Epidermal Büyüme Faktör Reseptörü İnhibitörü) BORTEZOMİB (Proteozom İnhibitörü) TRASTUZUMAB (HERCEPTİN) (Epidermal Büyüme Faktörü Reseptör TK İnhibitörü)

35 HEDEF : HÜCRE YÜZEY RESEPTÖRLERİ EGFR VEGFR HER-2 Flt- 3

36 KANSERLERDE TİROZİN KİNAZ DİSREGÜLASYONU

37 RESEPTÖR MONOKLONAL ANTİKOR SİNYAL İNHİBE OLUR RESEPTÖRLERİN MONOKLONAL ANTİKORLAR İLE BLOKE EDİLMESİ

38 Trastuzumab HERCEPTİN

39 HER-2 / neu C-erbB-2 Human Epidermal büyüme faktörü Reseptorü-2 HER 2, p185 HER2 olarak da bilinir HER2/neu = HER2 reseptörünün, onkojenin kodladığı üretimi

40 HER-2 nin AŞIRI ARTIMI (OVEREXPRESSION ) HER-2 meme kanserinde yaklaşık % oranında eksprese olur. Penault-Llorca J Clin Oncol (Meeting Abstarct ),2005

41 HER-2 reseptör dimer transmembran sinyal iletim yolu Büyüme faktörü Bağlanan bölge Plazma membranı Nükleusa sinyal iletimi Tirosin kinaz aktivitesi Sitoplazma Nükleus Gen aktivitesi HÜCRE BÖLÜNMESİ

42 TRASTUZUMAB

43 HER2 reseptörüne trastuzumab bağlanması :

44 ANJİOGENEZ İNHİBİTÖRLERİ EGFR / HER-2 VEGFR

45 TÜMÖRLERDE DAMAR AĞ YAPILARI ANORMALDİR Sarkoma doğru büyüyen kan damarları SARKOM NORMAL DAMARLAR ANORMAL DAMARLAR Uyarlanmıştır, Marx İzin alınarak basılmıştır, University of Louisville School of Medicine. Copyright Reference: Marx J. Science. 2003;301:

46 Yeni damarların büyümesinin inhibisyonunun vasküler pencere fare modelinde gözlenmesi Tümör hücrelerinin ekilmesi ve iskemik ortam oluşturulmasından sonraki neovaskülarizasyon Kontrol LLC ekiminden önce Ekimden 1 gün sonra Ekimden 6 gün sonra Ekimden 9 gün sonra LLC ekiminden önce Ekimden 1 gün sonra Ekimden 6 gün sonra Ekimden 9 gün sonra Anti-VEGF tedavi* Uyarlanmıştır, Osusky İzin alınarak basılmıştır, Angiogenesis. *Anti-VEGF ajan: SU11248 (VEGF tirozin kinaz inhibitörü). Reference: Osusky KL et al. Angiogenesis. 2004;7:

47 TÜMÖRLERDE ANJİOGENEZ

48 VEGF RESEPTÖR MONOKLONAL ANTİKOR SİNYAL İNHİBE OLUR RESEPTÖRLERİN MONOKLONAL ANTİKORLAR İLE BLOKE EDİLMESİ

49 ANTİ-VEGFR TEDAVİSİ İLE

50 BEVACİZUMAB - ALTUZAN ALTUZAN 400 mg / 16 ml ALTUZAN 100 mg / 4 ml

51 BEVACIZUMAB ( ALTUZAN) ENDİKASYONU 5-florourasil/folinik asit veya 5-florourasil/folinik asit/irinotekan ile kombine olarak METASTATİK KOLON VE METASTATİK REKTUM KANSERLERİNİN BİRİNCİ BASAMAK TEDAVİSİNDE KULLANILIR Referanslar: 1. Altuzan prospektüs.

52 SOLİD TÜMÖRLERDE TİROZİN KİNAZ İNHİBİTÖRLERİ TİROZİN KİNAZ KANSER HEDEF TEDAVİ EGFR (ERBB1) NSCLC GEFINITIB COLORECTAL ERLOTINIB HER-2 (ERBB2) BREAST LUNG TRASTUZUMAB C-KIT GIST IMATINIB

53 SOLİD TÜMÖRLERDE TİROZİN KİNAZ İNHİBİTÖRLERİ TİROZİN KİNAZ KANSER HEDEF TEDAVİ PDGFR GLİOBLASTOM IMATINIB OSTEOSARKOM VEGFR-1,-2 NSCLC-MEME BEVACIZUMAB PROSTAT-RENAL KOLO-REKTAL

54 HEDEFLENMİŞ TEDAVİLER : MONOKLONAL ANTİKORLAR RITUXIMAB ALEMTUZUMAB TRASTUZUMAB GEMTUZUMAB OZOGAMICIN IBRITUMOMAB TIOXETAN

55 ANTİ-KANSER MONOKLONAL ANTİKORLARI Monoklonal Ticari Antikor Şekli Endikasyonları RITUXIMAB MABTHERA NHL - KLL TRASTUZUMAB HERCEPTiN Meme Ca ALEMTUZUMAB CAMPATH-1 NHL - KLL CETUXIMAB ERBITUX Kolon Ca ANTI-VEGF AVASTİN Kolon Ca

56 YENİ HEDEFLER : Flt-3 ve Flt-3 Mutasyonları

57 HEDEFLENMİŞ TEDAVİLERDE YENİ UFUKLAR..

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59 GLEEVEC FİLMİ

60 Sağlıklı bir insanın hücrelerinde yalnızca 2 tane gen kopyası bulunur. Kanserli hücrelerde ise gen kopyası sayısı bu rakamın çok üstündedir. ONKOLOJİDE HER2

61 Anti-VEGF TEDAVİSİ ERKEN ETKİLER DEVAM EDEN ETKİLER 1 Mevcut mikrodamarlanmanın regresyonu Yeni tümör damarlarının inhibisyonu 1,2,9,10 2 Geride kalan tümör damarlarının normalizasyonu 5-8 Referanslar: 1. Baluk P et al. Curr Opin Genet Dev. 2005;15: Inai T et al. Am J Pathol. 2004;165: Erber R et al. FASEB J. 2004;18: Tong RT et al. Cancer Res. 2004;64: Jain RK. Nat Med. 2001;7: Jain RK. Science. 2005;307: Lee CG et al. Cancer Res. 2000;60: Willett CG et al. Nat Med. 2004;10: Gerber HP et al. Cancer Res. 2005;65: Warren RS et al. J Clin Invest. 1995;95:

62 Anti-VEGF tedavi, damar yapısını normalleştirir Over karsinomu* Kontrol Gliom* Anti-VEGF tedavi Uyarlanmıştır, Byrne İzin alınarak basılmıştır, The American Association for Cancer Research. Copyright Kontrol Anti-VEGF tedavi *Fare modeli Anti-VEGF ajan: VEGF Trap. Reference: Byrne AT et al. Clin Cancer Res. 2003;9:

63 HEDEFLENMİŞ TEDAVİLER : SİNYAL İLETİMİ İNHİBİTÖRLERİ IMATINIB MESYLATE (Tirozin Kinaz İnhibitörü) GENEFITINIB (Epidermal Büyüme Faktör Reseptörü Tirozin Kinaz İnhibitörü EGFR-TK) CETUXIMAB (Epidermal Büyüme Faktör Reseptörü İnhibitörü) BORTEZOMİB (Proteozom İnhibitörü) TRASTUZUMAB (HERCEPTİN) (Epidermal Büyüme Faktörü Reseptör TK İnhibitörü)

64 HEDEFLENMİŞ TEDAVİLERİN GELECEĞİ...

65 FIRST MONOCLONAL ANTIBODY INDICATED IN THE TREATMENT OF RELAPSED OR CHEMOTHERAPY-RESISTANT LOW-GRADE AND FOLLICULAR NON-HODGKIN S LYMPHOMA Roche Onkoloji

66 B-LENFOSİT YÜZEY ANTİKORLARI sig CD 19 B LENFOSİT ANTİ CD20 (RITUXIMAB) CD 20

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69 KANSERLERDE TİROZİN KİNAZ DİSREGÜLASYONU MUTASYONLAR Fms-Like TK3

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71 CD20 B-LENFOSİT ANTİJENİ % >95 ten fazla B-Hücreli NHL de Pozitif bulunur Plazma hücrelerinde bulunmaz.

72 İnsan meme kanser hücrelerine HER-2 gen amplifikasyonunun etkisi İnsan meme kanser hücreleri DNA sentezi % Hücre büyüme oranı % HER2 negatif HER2 geniyle transfekt HER2 pozitif Yumuşak agarda büyüme % 225 Tümörojenisite nude farede Metastatik potansiyel % 220 nude farede Değişmiş fenotip Slamon DJ ve ark. yayınlanmamış veri

73 Özetleyecek olursak : 80 % Trastuzumab + Dosetaksel Dosetaksel % 61 Ay Trastuzumab + Dosetaksel ay Dosetaksel % ay Cevap Oranları Ortalama Sağkalım Marty et al JCO Vol 23 No 19:

74 SİNYAL İNHİBİTÖRLERİ

75 B-Cell Life Cycle : CD20 Expression Bone Marrow Blood Lymphatic System Pluripotent Stem Cell Lymphoid Stem Cell Pre-B Cell B Cell Activated B Cell Plasma Cell CD20 Antigen Expression Precursor B-Cell Acute Leukemias B-Cell Lymphomas, CLL Myeloma Seldin DW. J Nuc Technol 2002; 30: Stashenko P, et al. J Immunol 1980; 125: Slide 3

76 B-LENFOSİT YÜZEY ANTİKORLARI sig CD 19 B LENFOSİT ANTİ CD20 (RITUXIMAB) CD 20

77 HÜCRE İÇİ HEDEF MOLEKÜLLER

78 LAÇLARI HANGİ ETKİN HEDEFLERE YÖNLENDİREBİLİRİZ? HEDEFLENMİŞ TEDAVİ = TARGETED THERAPY

79 HEDEFLEMEDE YENİ UFUKLAR APOPTOZİS-UYARICI AJANLARIN GELİŞTİRİLMESİ BORTEZOMIB (VELCADE ) OBLİMERSEN (GENASENSE ) ANTİ-ANJİOGENEZ İNHİBİTÖRLERİ BEVASİZUMAB (ALTUZAN ) EPİDERMAL BÜYÜME-FAKTÖR RESEPTÖR (EGFR) İNHİBİTÖRLERİ GEFITINIB (IRESSA) ERLOTINIB (TARCEVA ) CETUXIMAB (ERBITUX )

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82 HÜCREDE HEDEF MOLEKÜLER YAPILAR

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90 HÜCRE BİYOLOJİSİ GENLER mrna PROTEINLER HÜCRENİN KADERİ

91 HEDEF : ANJİOGENEZ İNHİBİTÖRLERİ

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93 B-LENFOSİT GELİŞİMİ : CD20 EKSPRESYONU KEMİK İLİĞİ KAN Lenfatik Sistem Pluripotent Stem Cell Lymphoid Stem Cell Pre-B Cell B Cell CD20 ANTİJENİ Activated B Cell Plasma Cell Precursor B-Cell Acute Leukemias EKSPRESYONU B-HÜCRELİ LENFOMA KLL MYELOM Seldin DW. J Nuc Technol 2002; 30: Stashenko P, et al. J Immunol 1980; 125:

94 HÜCRE Hücre Membranı TİROZİN KİNAZ SİNYAL MOLEKÜLLERİ TF Nukleus PROTEOZOM

95 * * HÜCRE YÜZEY * ANTİJENLERİ ve RESEPTÖRLERİ

96 1. MabCampath : ETKİ MEKANİZMASI T-CELL CD52 2. LENFOSİT YÜZEYİNE BAĞLANMA T-CELL LENFOSİT YÜZEYİNDE CD 52 ANTİJENİ 3. APOPTOTİK HÜCRE ÖLÜMÜ

97 MONOKLONAL ANTİKORLARIN YÜKLENMESİ STAIN (FITC / TEXAS RED) TOXIN (DIPHTERIA) * DRUG (DAUNOMYCIN) ISOTOPE 131 Iodine, 90 Yttrium

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99 İZOTOP RADYOİZOTOP İLE ETKİN RADYASYON DOZU!

100 Iodine 131- TOSITUMOMAB BEXXAR ANTİ-CD20 IODINE 131* Slide 7

101 RADYOISOTOPE-LABELLED ANTI-CANCER MOABs Monoclonal Antibody Brand Name Indications IBRITUMOMAB* ZEVALIN NHL * Indium-111 * Yttrium-90 TOSITUMOMAB* BEXXAR NHL

102 TOSITUMOMAB (BEXXAR) :Iodine-131 in Crossfire Etkisi Radyoizotop taşımayan soğuk Antikor RADYOİZOTOP İŞARETLİ ANTİKOR Courtesy of Andrew Zelenetz, MD.

103 HEDEF : HÜCRE SİKLUSU MOLEKÜLLERİ

104 YENİ YAKLAŞIMLAR : APOPTOZİSİ İNDÜKLEYİCİ AJANLARIN GELİŞTİRİLMESİ Cytochrome c Caspase Aktivasyonu APOPTOZİS

105 APOPTOSİS DE ROL ALAN MOLEKÜLLER

106 Targeting the Tumor and Its Microenvironment 1. Growth factor signaling 2. Other growth stimulating/ 9 suppressing receptors 3. Microtubule dynamics 4. Histone acetylation/ deacetylation 5. DNA replication, transcription, repair 6. Protein synthesis 7. Protein folding 8. Cell cycle 10 Cancer Cell 9. Activators and inhibitors of apoptosis 10. Metastasis Tumor Cell Death Cancer Cell Endothelial Cell Tumor Cell Growth/Replication Angiogenesis

107 HEDEFLENMİŞ TEDAVİLER : SİNYAL İLETİMİ İNHİBİTÖRLERİ IMATINIB MESYLATE (Tirozin Kinaz İnhibitörü) GENEFITINIB (Epidermal Büyüme Faktör Reseptörü Tirozin Kinaz İnhibitörü EGFR-TK) CETUXIMAB (Epidermal Büyüme Faktör Reseptörü İnhibitörü) BORTEZOMİB (Proteozom İnhibitörü TRASTUZUMAB (HERCEPTİN) (Epidermal Büyüme Faktörü Reseptör TK İnhibitörü)

108 VEGF Receptors in Angiogenesis and Lymphangiogenesis TKI PlGF VEGF-A, -B VEGF-A, -C, -D, -E VEGFR-2 / KDR VEGF-C, -D VEGFR-1 / Flt-1 PI(3,4,5)P3 PDK1,2 PI(4,5)P2 PTEN PI3-K P P P P Sck PI(4,5)P2 PLCg DAG + IP3 PKC RAS VEGFR-3 Ca ++ RAC Akt/PKB FAK RAF Permeability Survival Paxillin MEK Migration ERK Angiogenesis Lymphangiogenesis

109 TİROZİN KİNAZ AKTİVİTELERİNİN BİRDEN FAZLA RESEPTÖRDE HEDEFLENEREK BLOKE EDİLMESİ *

110 Signaling Through mtor View Animation Notes: 1. You must be in slideshow mode to view animation. 2. If you receive an ActiveX Message Box, you should allow blocked content to run in Internet Explorer.

111 mtor Inhibition May Enhance the Antitumor Effects of Other Therapies Chemotherapy Radiation mtor Inhibition EGFR Inhibitors Antiestrogens Antiangiogenics

112 2006 in Physiology and Medicine ANDREW FIRE Carnegie Inst of Washington, USA CRAIG MELLO Univ Massachusets Cancer Ctr,USA Fundamental mechanisms by which gene expression and silencing is controlled with Process of RNA Interference (MicroRNA )

113 HEDEFLENMİŞ TEDAVİDE NANOTEKNOLOJİ

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115 Micro RNA - MiRNA - MiR

116 UZAY ve GALAXY LER... Star Wars Episode III

117 ATOM BOMBASI FİLMİ

118 RESEPTÖRLER HÜCRE MEMBRANI

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122 GLEEVEC

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125 HASTAYA YARARI?

126 HÜCRE Hücre Membranı TİROZİN KİNAZ SİNYAL MOLEKÜLLERİ TF Nukleus PROTEOZOM

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128 T-LENFOSİT HÜCRE HÜCRE TEMASI AKTİVASYON MONOSİT SİNYAL İLETİMİ DİĞER...

129 ANTİKOR MOLEKÜLÜ L H H L Fab-Kısmı Antigen Binding region IgG IgA IgM IgD IgE H H Fc-Kısmı PLAZMA HÜCRELERİ TARAFINDAN SENTEZLENİR

130 MONOKLONAL ANTİKORLARIN ETKİ MEKANİZMALARI

131 Antikor Moleküllerinin Hücre Yüzey Antijenlerine Bağlanması

132 MabCampath = Alemtuzumab CAMPATH MabCampath AMPATH T-CELL CD52 antigen MabCampath is the first monoclonal antibody that specifically targets the CD52 antigen CD52 is abundant on the surface of 95% of normal and malignant B- and T-lymphocytes CD52 is not found on stem cells Stem cells are therefore not damaged and differentiate to replace malignant lymphocytes with healthy lymphocytes MabCampath triggers the death of the target cell by binding to CD52

133 CONJUGATED FORMS OF ANTI-CANCER MONOCLONAL ANTIBODIES RADIOIMMUNOCONJUGATE 3 IODINE TOSITUMOMAB :BEXXAR 90 YTTRIUM IBRITUMOMAB:ZEVALIN CHEMOTHERAPEUTIC CONJUGATE Calicheamycin + Anti-CD33 MYELOTARG

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137 ANTİ-KANSER MONOKLONAL ANTİKORLARI Monoklonal Ticari Antikor Şekli Endikasyonları RITUXIMAB MABTHERA NHL - KLL TRASTUZUMAB HERCEPTiN Meme Ca ALEMTUZUMAB CAMPATH-1 NHL - KLL CETUXIMAB ERBITUX Kolon Ca ANTI-VEGF AVASTİN Kolon Ca

138 GELECEKTE YENİ HEDEFLER...

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140 HÜCREDE MİKRO ANATOMİK YAPILAR

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142 İlaçları Başka Hangi Etkin bölgelere Yönlendirebiliriz? HANGİ HEDEFLERİ SEÇELİM?

143 KANSER HÜCRESİNİN BÜYÜME ÇOĞALMA MOLEKÜLLERİ

144 GF VE HER-2 RESEPTÖRLERİNİN SİNYALLERİ : HEDEF KİNAZ İNHİBİTÖRÜ

145 KİNAZ İNHİBİTÖRLERİNİN IGF-RESEPTÖR ÜNÜ BLOKAJI

146 RADYOİZOTOP *** ANTİ-KANSER MONOKLONAL ANTİKORLARI Monoklonal Antikor Ticari İsmi Endikasyonları IBRITUMOMAB* ZEVALİN NHL * Indium-111 * Yttrium-90 TOSITUMOMAB* BEXXAR NHL

147 APOPTOSIS PROGRAMMED CELL DEATH ANTIGEN Antijen CD20 ANTI-CD 20 RITUXIMAB Antikor B-LYMPHOCYTE B hücresi CD20 (+) ACTIVATION OF CASPASES AND PROCESS OF CELL DEATH APOPTOTIC CELL DEATH OF B-LYMPHOCYTE

148 Complement-Dependent-Cellular Cytotoxicity (CDCC) Antigen CD20 Fc region + RITUXIMAB Complement System C1r C1s C1q C1 B-LYMPHOCYTE CD 20 (+) Lysis H 2 0/ Ions Pores formed (C8 - C9s)

149 TOPOİZOMERAZ İNHİBİTÖRLERİ **

150 Growth Factors IGF-1, EGF, TGFα, VEGF, etc Growth Factors and the mtor Pathway Oxygen, energy, and nutrients Ras/Raf pathway kinases S6 PTEN TSC2 TSC1 S6K1 PI3-K Akt/PKB mtor Protein Production Cell Growth and Proliferation Ras/Raf Abl ER Angiogenesis 4E-BP1 elf-4e mtor Intracellular protein Central controller of cell growth and proliferation mtor signaling is often deregulated in cancer Downstream inhibition of mtor has potential for Antiproliferative effects on tumor cells Angiogenesis inhibition Enhancement of the effects of chemotherapy This slide shows only a few of the many kinases involved in these signaling pathways

151 mtor Tumor Cell Growth and Angiogenesis Cancer cell Growth factors Endothelial cell Smooth muscle cell (pericyte) PI3-K TSC2 TSC1 PTEN Akt/ PKB PI3-K Akt/ PKB PI3-K Akt/ PKB mtor mtor mtor Protein production VHL HIF-1a HIF-2a Cell growth and proliferation Angiogenic growth factors Cell growth and proliferation Cell growth and proliferation

152 The Role of mtor in Cancer Cells

153 mtor Inhibition Blocks Cell Cycle Progression at the G1 S Restriction Point M G 2 mtor G 1 S Restriction point From Israels and Israels. Oncologist. 2000;5: , with permission.

154 The mtor Pathway and Cancer mtor is a central controller of cell growth and proliferation in normal cells mtor pathway deregulation causes loss of growth control in cancer mtor controls responses essential to cancer cells mtor regulates cell growth and cell division mtor influences responses to DNA damage mtor regulates response to angiogenic stimuli mtor pathway and responses to antiestrogens

155 mtor Inhibition May Enhance the Cytotoxicity of DNA-Damaging Agents Cisplatin Cell Death DNA Cross-linked DNA Cycle Arrest DNA Repair Survival DNA damage caused by agents such as cisplatin activates p53 p53 triggers DNA repair, which allows the cell to survive, or, failing that, p53 initiates cell death mtor regulates production of p21, a cell cycle inhibitor that allows DNA repair mtor inhibition blocks p21 translation, forcing cell death even when the DNA damage is otherwise nonlethal mtor inhibition can enhance the activity of certain drugs such as cisplatin and other platinum derivatives Beuvink et al. Cell. 2005;120:

156 Teşekkür Ederim...

157 Summary Rationale for Targeting mtor (cont) Aberrant signaling through upstream pathways can activate mtor inappropriately, promoting Abnormal cell growth, proliferation, and angiogenesis Survival of cancer cells in the nutrientand oxygen-depleted tumor environment Targeting deregulated pathways has been a successful clinical strategy in cancer

158 Summary Rationale for Targeting mtor mtor, an intracellular protein, acts as a central regulator of multiple signaling pathways that may mediate abnormal growth and proliferation mtor provides a stable genetic target mtor activity is influenced by nutrient and energy levels and by signaling through pathways often deregulated or overexpressed in cancer, such as Cell surface receptors, such as EGFR/HER, IGFR, VEGFR

159 FGF Receptor Signaling in Tumor Cell Growth and Angiogenesis TKI PKC DAG + IP3 Ca ++ PI(4,5)P2 PLCg SHP-2 Grb2 SOS RAS- GDP GTP P RAS- GTP RAF PLA 2 RAF Platelet-Activating Factor MEK ERK MEK JAK ERK Gene Expression Apoptosis Proliferation Migration Differentiation Angiogenesis

160 Growth Factor Signaling Through EGFR/HER2 EGF, TGF-α EGFR HER2 TKI PI(3,4,5)P3 PDK1,2 PI(4,5)P2 PTEN PI3-K P P P P Grb2 SOS RAS-GDP GTP P RAS-GTP Akt/PKB RAF Glycogen Synthesis Survival MEK ERK Proliferation Migration Cell Growth, Proliferation, Survival, Transformation

161 The IGF-1R Signalling Pathway TKI IGF-1R P P PI(3,4,5)P3 Akt/PKB PTEN PI(4,5)P2 PI3-K SHP-2 SHP-2 P IRSs P Shc Grb2 SOS RAS-GDP GTP P RAS-GTP RAF mtor 4E-BP1 S6K1 MEK ERK elf-4e Elk-1 Ribosomal Synthesis mrna Translation Cell Growth, Proliferation, Survival, Transformation, Metastasis, Angiogenesis

162 Mutant and Wild-Type Flt-3 Activation in Leukemic (AML, ALL) Cells Normal Hematopoietic Cells FL TKI Flt-3 Mutations (AML) Flt-3-ITD Flt-3-D835 Wild-type Flt-3 RAS-GTP TYK2 MAP Kinase Pathway RAF STAT5 JAK/STAT Pathway Cell Proliferation

163 CD40L (CD154) Stimulates Growth and Survival of B Cells and B-Cell Derived Malignant Cells CD40 Antagonist Antibody X CD40L (CD154) TRAFs Akt/PKB PI3-K X X X X NF-kB JNK ERK B Lymphocytes B-Cell Derived Malignancies P38 MAPK TRAF = TNF receptor associated factor. Inhibition of Apoptosis Cell Growth Cell Proliferation

164 Activation of RAS-RAF Oncogenes by Mutation or Growth Factor Stimulation EGF KI EGFR HER2 Grb2 Shc RAS-GTP MEK RAF Kinases eg, Src, PAK ERK SRF Elk-1 Substrates in Nucleus Substrates in Cytosol and Cytoskeleton Cell Growth, Proliferation, Survival, Transformation, Metastasis, Angiogenesis

165 Central Role of BCR-ABL in Leukemic Cells (CML, ALL) IL-3 Receptor TKI PI3-K Gab2 Shc Grb2 SOS RAS-GDP GTP P RAS-GTP JAK Akt/PKB BCR-ABL RAF FAK SHP-1 MEK Antiapoptosis Paxillin Migration ERK Gene Expression Cell Growth, Proliferation, Survival, Transformation, Metastasis

166 Induction of Apoptosis Through Death Receptors Agonist Antibody Death Receptor Pro- Caspase 8 Bid Mitochondrion Cyto C Bax Bad Bax PI3-K RAS-GTP FLIP Caspase 8 Pro- Caspase 3 Cyto C Bcl-2 Bcl-XL Akt/PKB IAP Caspase 3 Caspase 9 APAF1 Pro- Caspase 9 ICAD CAD PARP Cell Death

167 Multispecific TKIs: Targeting Multiple Growth Factor Signaling Pathways EGFR/HER2 TKI VEGFR Cancer Cell Endothelial Cell

168 Molecular Chaperones and Protein Folding HSP90 Inhibitor ADP ADP Protein Function/ Confirmation HSP90 HSP90 Early complex HOP p23 HSP40 Final complex HSP70 X HSP90 Propylisomerase s Protein Degradation HSP40 HSP40 HSP70 HOP HSP90 HSP70 HOP HSP90 p23 Propylisomerase s Intermediate complex R. Jensen modified from Wegele et al. Rev Physiol Biochem Pharmacol. 2004;151:1-44, Springer-Verlag GmbH

169 Cross-Talk Between the PI3-K/Akt and RAS/RAF Signaling Pathways Growth Factor Receptor Cytokine Receptor RAS PI3-K RAF Akt/PKB MEK mtor ER ERK mrna Translation Gene Transcription Growth Survival Division Motility

170 Histone Acetylation in the Regulation of Gene Transcription HDAC Inhibitor DNA HDAC Ac TF Ac Ac Ac Ac Ac Ac Ac Ac TF Ac Ac Ac Ac Ac Ac Ac Ac Ac Ac Ac Ac Ac Ac Histones Transcriptional Repression HAT Ac Ac Ac Transcriptional Activation Ac Ac Oncogene Expression Ac Ac Ac Tumor Suppressor Gene Expression Apoptosis

171 mtor: A Central Controller of Tumor Cell Growth and Angiogenesis Growth factors VEGF VEGFR Integrins PTEN PI3-K LKB1 PI3-K Oxygen, Nutrients, Amino Acids ILK Akt/ PKB TSC1/TSC2 AMPK Energy Akt/ PKB 4E-BP1 elf-4e mtor S6K1 P S6 mtor Inhibitor FKBP-12 mtor Inhibitor mtor Inhibitor FKBP-12 mtor X Gene transcription Protein production X X X Cell growth Cancer Cell X Proliferation VEGF production Cell growth and proliferation Endothelial Cell

172 mtor: A Central Controller of Tumor Cell Growth and Angiogenesis Growth factors VEGF VEGFR Integrins PTEN PI3-K LKB1 PI3-K Oxygen, Nutrients, Amino Acids ILK Akt/ PKB TSC1/TSC2 AMPK Energy Akt/ PKB 4E-BP1 elf-4e mtor S6K1 P S6 mtor Inhibitor FKBP-12 mtor Inhibitor mtor Inhibitor FKBP-12 mtor X Gene transcription Protein production X X X Cell growth Cancer Cell X Proliferation VEGF production Cell growth and proliferation Endothelial Cell

173 Microtubule Dynamics as a Target of Cancer Therapy Microtubules Depolymerization G2 S Mitosis Microtubule Stabilizer Microtubule Dynamics Polymerization Cell Death G1

174 Topoisomerase I Inhibition Induces DNA Breaks and Cell Death Topoisomerase I Inhibitor 5' DNA Polymerase Topoisomerase I 5' 3' 5' 5' 3' Replication Fork TOPO I Replication Fork TOPO I 3' DNA Ligase Replication 3' Replication 5' 5' 3' 5' 5' 3' Replication Fork TOPO I TOPO I 3' Replication fork encounters stabilized Topo I DNA complex Replication stops 3' DNA single-strand breaks Cell death

175 Histone Acetylation in the Regulation of Gene Transcription HDAC Inhibitor DNA HDAC Ac TF Ac Ac Ac Ac Ac Ac Ac Ac TF Ac Ac Ac Ac Ac Ac Ac Ac Ac Ac Ac Ac Ac Ac Histones Transcriptional Repression HAT Ac Ac Ac Transcriptional Activation Ac Ac Oncogene Expression Ac Ac Ac Tumor Suppressor Gene Expression Apoptosis

176 Cell Cycle Regulation by Cyclins and Cyclin-Dependent Kinases (CDKs) Dephosphorylation CDK Inhibitor Cyclin B CDK1 G2 M G0 Restriction Point G1 Protein Synthesis Cyclins D1, D2, D3 CDK4 CDK6 Cyclins Proteasomal Degradation CDKs CDK Inhibitor Cyclin A CDK1 CDK Inhibitor S Cyclin A CDK2 CDK Inhibitor Cyclin E CDK2 p21 p27 others Modified from Schwartz and Shah. J Clin Oncol. 2005;23: , with permission.

177 Central Role of BCR-ABL in Leukemic Cells (CML, ALL) IL-3 Receptor TKI PI3-K Gab2 Shc Grb2 SOS RAS-GDP GTP P RAS-GTP JAK Akt/PKB BCR-ABL RAF FAK SHP-1 MEK Antiapoptosis Paxillin Migration ERK Gene Expression Cell Growth, Proliferation, Survival, Transformation, Metastasis

178 Activation of RAS-RAF Oncogenes by Mutation EGF or Growth Factor Stimulation KI EGFR HER2 Grb2 Shc RAS-GTP MEK RAF Kinases eg, Src, PAK ERK SRF Elk-1 Substrates in Nucleus Substrates in Cytosol and Cytoskeleton Cell Growth, Proliferation, Survival, Transformation, Metastasis, Angiogenesis

179 Rational Combinations: Proven Therapies and New Targets Proven therapies Growth factor receptors and downstream signaling kinases Cytotoxic chemotherapy Radiation therapy Hormonal therapy Multiple new targets Cell metabolism, cell growth, protein translation, protein folding Gene transcription Cell cycling, DNA replication, and cell division Cell death receptors

180 HSP90 Inhibitor Molecular Chaperones ADP ADP and Protein Folding Protein Function/ Confirmation HSP90 HSP90 Early complex HOP p23 HSP40 Final complex HSP70 X HSP90 Propylisomerase s Protein Degradation HSP40 HSP40 HSP70 HOP HSP90 HSP70 HOP HSP90 p23 Propylisomerase s Intermediate complex R. Jensen modified from Wegele et al. Rev Physiol Biochem Pharmacol. 2004;151:1-44, Springer-Verlag GmbH

181 The Role of mtor in Cancer Cells

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184 -LENFOSİT / LENFOMA YÜZEY ANTİJENLERİ sig CD 19 B LENFOMA HÜCRESİ ANTİ CD20 (RITUXIMAB) CD 20

185 odine 131 BEXXAR

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188 MABTHERA= RITUXIMAB CHIMERIC Human-Mouse IgG Antibody CD20 MOUSE Variable Regions Bind to CD20 20 Antigen on B-Lymphocytes on B-Cells HUMAN k-constant region HUMAN IgG1 Fc Region

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190 MONOKLONAL ANTİKORLARIN İZOTOPLA YÜKLENMESİ VE LENFOMA HÜCRESİNE YÖNLENMESİ / BAĞLANMASI B-LENFOMA HÜCRESİ CD 20 *

191 Iodine I 131- Labeled Tositumomab: BEXXAR :Features TOSITUMOMAB : FEATURES Murine IgG2a ANTI-CD20 ANTIBODY B-cell specific Induction of apoptosis Complement-dependent cytotoxicity (CDC) Antibody-dependent cellular cytotoxicity (ADCC) BEXXAR ANTI-CD20 IODINE 131 RADIOISOTOPE Cytotoxic beta emission Physical half-life of 8 days Short path length Gamma emission allows dosimetry IODINE 131 Slide 7

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195 CONJUGATED FORMS OF ANTI-CANCER MONOCLONAL ANTIBODIES RADIOIMMUNOCONJUGATE 3 IODINE TOSITUMOMAB :BEXXAR 90 YTTRIUM IBRITUMOMAB:ZEVALIN CHEMOTHERAPEUTIC CONJUGATE Calicheamycin + Anti-CD33 MYELOTARG

196 MABTHERA= RITUXIMAB

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199 1. MabCampath : Alemtuzumab T-CELL CD52 2. MabCampath binds to the lymphocyte T-CELL Lymphocyte covered with CD52 antigens 3. Apoptotic Cell death

200 MONOKLONAL ANTİKORLARIN RADYOİSOTOPLARLA İŞARETLENMESİ (RADIO-İMMÜNOTERAPİ) L H H L BOYA (FITC / TEXAS RED) TOKSİN (DİFTERİ) İLAÇ (DAUNOMYCIN) İZOTOP : 131 Iodine, 90 Yttrium

201 MONOKLONAL ANTİKORLAR ETKİLERİNİ NASIL GERÇEKLEŞTİRİR? I. DİREKT TÜMÖR HÜCRESİ ÜZERİNE a. Apoptozisin indüklenmesi ya Hücre Ölümü b. Büyüme Faktörü Reseptörlerinin (GFR) bloke edilmesi c. Anti-İdiotip Antikor (Anti-İd) sentezi yoluyla

202 MONOKLONAL ANTİKORLARIN ETKİ MEKANİZMALARI II.İNDİREKT ETKİLERİ a. Antikora-Bağımlı Hücresel Sitotoksisite (ADCC) b. Kompleman-Aracılı Hücresel Sitotoksisite (CMCC)

203 RADYOİZOTOP *** ANTİ-KANSER MONOKLONAL ANTİKORLARI Monoklonal Antikor Ticari İsmi Endikasyonları IBRITUMOMAB* ZEVALİN NHL * Indium-111 * Yttrium-90 TOSITUMOMAB* BEXXAR NHL

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220 Rituximab : Konak İmmün Effektör Hücreleri İle Etkileşimi

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222 B-Cell Life Cycle : CD20 Expression Bone Marrow Blood Lymphatic System Pluripotent Stem Cell Lymphoid Stem Cell Pre-B Cell B Cell Activated B Cell Plasma Cell CD20 Antigen Expression Precursor B-Cell Acute Leukemias B-Cell Lymphomas, CLL Myeloma Seldin DW. J Nuc Technol 2002; 30: Stashenko P, et al. J Immunol 1980; 125: Slide 3

223 GLEEVEC FİLMİ

224 FIRST MONOCLONAL ANTIBODY INDICATED IN THE TREATMENT OF RELAPSED OR CHEMOTHERAPY-RESISTANT LOW-GRADE AND FOLLICULAR NON-HODGKIN S LYMPHOMA Roche Onkoloji

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226 HÜCREDE HEDEF MOLEKÜLER YAPILAR

227 RESEPTÖR SİNYAL*** TF GENLER

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240 HER2 amplifikasyonu overekspresyon ile sonuçlanır

241 HÜCRE İÇİ HEDEF MOLEKÜLLER