1 / 35

BASI MOLECOLARI DELL’AZIONE DEL FARMACO BIOTECNOLOGIE FARMACOLOGICHE LEZIONE 11 Ingegneria animale II

CORSO DI LAUREA SPECIALISTICA IN BIOTECNOLOGIE DEL FARMACO Adriana Maggi. BASI MOLECOLARI DELL’AZIONE DEL FARMACO BIOTECNOLOGIE FARMACOLOGICHE LEZIONE 11 Ingegneria animale II. Produzione di biofarmaci. Utilizzo di vettori tessuto specifici per

zora
Download Presentation

BASI MOLECOLARI DELL’AZIONE DEL FARMACO BIOTECNOLOGIE FARMACOLOGICHE LEZIONE 11 Ingegneria animale II

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. CORSO DI LAUREA SPECIALISTICA IN BIOTECNOLOGIE DEL FARMACO Adriana Maggi BASI MOLECOLARI DELL’AZIONE DEL FARMACO BIOTECNOLOGIE FARMACOLOGICHE LEZIONE 11 Ingegneria animale II

  2. Produzione di biofarmaci Utilizzodivettoritessutospecifici per esprimereproteineterapeutichenel latte o nelsierodianimalitransgenici

  3. The cost of making one transgenic animal ranges from $20,000 to $300,000 It has been estimated that one transgenic animal can produce, in its lifetime, $200 to $300 million worth of pharmaceuticals.

  4. AviGenics comparison of production inputs and costs for monoclonal antibodies* using traditional cell culture versus using transgenic poultry or goats *100 kg of raw material per yearSource: AviGenics www.avigenics.com

  5. Nutritional food.

  6. Partiallistofbioactivecomponents in milk withhumanhealthimplications

  7. Pharming products currently in development.

  8. Giugno 2006 APPROVAZIONE DA PARTE DELL’EMEA DEL PRIMO FARMACO IN UN BIOREATTORE ANIMALE: ANTITROMBINA UMANA PRODOTTA IN CAPRA (per la terapia della deficienza ereditaria di antitrombina, una patologia che affligge 3000-5000 soggetti) Nature Biotechnology, 24,8:877

  9. Alcuni farmaci in fase avanzata di studio prodotti in bioreattori animali Ditta prodotto indicazione fase sviluppo GTC BiotherapeuticsantitrombinhereditaryantitrombinApproved EU (Framigham Ma USA) deficiencyphase III USA PharmingC1 inhibitor (rabbit) Hereditaryangiodemaphase III (LeidenNetherlands) h Lactoferrin (rabbit) infection and inflammpreclinical hfibrinogen (rabbit) tissuesealantpreclinical Bioprotein Technologies Rothavirusparticlesrothinfectionpreclinical (Paris) (rabbit) PharmAtheneprotexia (goat) neuraltissuepreclinical

  10. Risposta agli xenobiotici ‘umanizzata’ • SXR e’ un recettore intracellulare che lega • ed induce la trascrizione di CYP3A un citocromo • essenziale nel metabolismo dei farmaci • PXR e’ l’omologo murino del recettore umano • SXR • Diverse specie animali rispondono ad un determinato xenobiotico (inducendo CYP3a) in modo specifico; questa caratteristica e’ insita nelle differenze tra i diversi recettori • (Es SXR e PXR)

  11. Strategia per il Knock out di PXR

  12. Strategia per l’inserzione di SXR

  13. Risposta agli xenobiotici ‘umanizzata’ • L’ablazione di PXR e la sua sostituzione con • SXR ha permesso di creare un modello di topo • ‘umanizzato’ per il metabolismo dei farmaci. • il modello ‘umanizzato’ permette di prevedere • il metabolismo di un determinato farmaco e di • studiare l’interazione tra farmaci diversi

  14. Modelli per lo screening di farmaci Animali reporter

  15. Adriana Maggi Metodologie innnovative in drug discovery: animali reporter Nature Review Drug Discovery March 2005

  16. ER ER ER POL II, TF, co-regulators transcription ER ER ERE Reporter: easily detectable protein REPORTER SYSTEMS TO STUDY GENE EXPRESSION IN VIVO Reporter GFP

  17. REPORTER SYSTEMS: FROM CELLS TO MICE THE REPORTER/ DETECTION SYSTEM THE PROMOTER TRANSGENESIS/Kin

  18. RAPID TURNOVER! Strategia per la generazione del topo reporter ER-luc

  19. “Optical Imaging” after estradiol injection (50mg/Kg s.c.) 100 30 HEAD ABDOMEN CTS/S 50 15 50 3 HOURS TIME 0 CHEST TAIL 2 CTS/S X (10-3) 25 1 TIME 0 6 HOURS 6 HOURS 24 HOURS 3 HOURS 24 HOURS

  20. IS THE ERE-LUC MOUSE A FAITHFULL REPORTER OF THE ACTIVITY OF ESTROGEN RECEPTORS? IS THE ERE-LUC MOUSE A FAITHFULL REPORTER OF THE ACTIVITY OF ESTROGEN RECEPTORS? ANALYSIS OF RESPONSIVE TISSUES CO-LOCALIZATION OF ERs AND LUCIFERASE BY IMMUNOCYTOCHEMISTRY STUDIES TIME-COURSE, DOSE-RESPONSE AFTER ADMINISTRATION OF ESTRADIOL PARALLEL MEASUREMENT OF ENDOGENOUS ESTROGEN- RESPONSIVE GENES (PROGESTERONE RECEPTOR) ANALYSIS OF LUCIFERASE ACTIVITY AFTER TREATMENT WITH ICI 182,780 TO MEASURE THE CONTRIBUTION OF ERRs Ciana et al. Mol. Endocrinol. 2001, Nature Med. 2003

  21. ERE-Luc mouse a model to study ER transcriptional activity Embryo’s development 16.5 17.5 18.5 15.5 12.5 13.5 14.5 19.5 immature adult day10 day1 proestrus estrus diestrus 1 diestrus 2

  22. REPORTER MICE A NEW APPROACH FOR THE DEVELOPMENT OF THERAPEUTICS

  23. DRUG DEVELOPMENT – PRECLINICL PHASES Preclinical studies Pharmacology: pharmacodynamics and organ-specificity of action drug disposition ADME Toxicology: single, repeated-dose toxicity testing special toxicity tests

  24. day 0 day 1 day 2 day 3 day 4 day 5 day 6 day 7 day 8 day 9 day 10 day 11 day 12 day 13 day 14 day 15 day 16 day 17 Bioluminescence analysis of ERE-Luc cycling female mice day 18 day 19 day 20 day 21

  25. 250 Control E2 200 ER activity in LIVER of adult cycling ERE-Luc mice treated with: E2 5.5 ug/kg/day Tamoxifen 5.5 ug/kg/day Tamoxifene 666 ug/kg/day Raloxifene 5.5 ug/kg/day Raloxifene 2111 ugkgday (each data point represents the photon emission mean of a minimum of 5 mice) 150 cts/s 100 50 0 2 4 6 8 14 16 18 10 12 20 22 E2 200 T low T high 150 cts/s 100 50 0 2 4 6 8 10 14 18 20 12 16 E2 200 R low R high 150 cts/s 100 50 0 days 2 6 8 0 4 14 18 10 16 20 12

  26. DRUG DEVELOPMENT – PRECLINICL PHASES Preclinical studies Pharmacology: pharmacodynamics and organ-specificity of action drug disposition ADME Toxicology: single, repeated-dose toxicity testing special toxicity tests

  27. ERE-LUC mouse in preclinical studies Adsorption, Distribution, MEtabolism With reporter mice the study of drug levels in plasma becomes meaningless: drug dosage can be directly based on the pharmacological response even in case of prolonged treatments Drug metabolism may maintain some interest to evaluate the enzymes responsible for the process

  28. DRUG DEVELOPMENT – PRECLINICL PHASES Preclinical studies Pharmacology: pharmacodynamics and organ-specificity of action drug disposition ADME Toxicology: single, repeated-dose toxicity testing special toxicity tests

  29. ERE-LUC mouse in preclinical studies: toxicology the actual knowledge of the entire spectrum of drug targets facilitate the prediction of side effect gender pharmacology effects in embryos effects during lactation differential effects in all of the target organs during development, maturity and aging

  30. LIVER BRAIN 14 10 10 8 * * * 12 * * 7,5 7,5 3 6 * * 5 2 4 5 L.U/mg prot. (x100,000) * L.U./mg prot. (x100,000) L.U./mg prot. (x10,000) * * * 2,5 1 2 2,5 * 0 0 0 0 ADULT ADULT SUCKLING SUCKLING CONTROL p,p’-DDT o,p’-DDT MICE LACTATING FROM MOTHERS EXPOSED TO DDT Di Lorenzo et. al. Endocrinology 2002

  31. 16,5 DPC 18,5 DPC IMAGING OF ERE-LUC PREGNANT MICE

More Related