Inhibiting PI3K as a therapeutic strategy against cancer

Clinical and Translational Oncology

Volumn 11, Issue 9, 2009, Pages 572-579

  Paz Ares, Luis ^a   Blanco Aparicio, Carmen ^a   García Carbonero, Rocío ^b   Carnero, Amancio ^c  

^a HOSPITAL UNIVERSITARIO VIRGEN DEL ROCÍO   (Spain)

^b Centro Nacional de Investigaciones Oncológicas   (Spain)

^c CSIC   (Spain)

Author keywords

Cancer; FOXO; Inhibitors; PI3K; Therapy

Indexed keywords

ANTINEOPLASTIC AGENT; ENZYME INHIBITOR; PHOSPHATIDYLINOSITOL 3 KINASE;

ANIMAL; BIOLOGICAL MODEL; DRUG ANTAGONISM; DRUG DELIVERY SYSTEM; DRUG EFFECT; EVALUATION; HUMAN; METABOLISM; NEOPLASM; ONCOGENE RAS; PHYSIOLOGY; REVIEW;

1-PHOSPHATIDYLINOSITOL 3-KINASE; ANIMALS; ANTINEOPLASTIC AGENTS; DRUG DELIVERY SYSTEMS; ENZYME INHIBITORS; GENES, RAS; HUMANS; METABOLIC NETWORKS AND PATHWAYS; MODELS, BIOLOGICAL; NEOPLASMS;

EID: 73349117836     PISSN: 1699048X     EISSN: 16993055     Source Type: Journal    
DOI: 10.1007/s12094-009-0407-x     Document Type: Article

References (116)

1. Maehama T, Dixon JE (1998) The tumor suppressor, PTEN/MMAC1, dephosphorylates the lipid second messenger, phosphatidylinositol 3,4,5-trisphosphate. J Biol Chem 273:13375-13378
2. Carnero A, Blanco-Aparicio C, Renner O et al (2008) The PTEN/PI3K/AKT signalling pathway in cancer, therapeutic implications. Curr Cancer Drug Targets 8:187-198
3. Kandel ES, Hay N (1999) The regulation and activities of the multifunctional serine/threonine kinase Akt/PKB. Exp Cell Res 253:210-229
4. Plas DR, Thompson CB (2005) Akt-dependent transformation: There is more to growth than just surviving. Oncogene 24:7435-7442
5. Datta SR, Brunet A, Greenberg ME (1999) Cellular survival: A play in three Akts. Genes Dev 13:2905-2927
6. Downward J (2004) PI 3-kinase, Akt and cell survival. Semin Cell Dev Biol 15:177-182
7. Blanco-Aparicio C, Renner O, Leal JF, Carnero A (2007) Pten, more than the Akt pathway. Carcinogenesis 28:1379-1386
8. Stokoe D (2001) Pten. Curr Biol 11:R502
9. Dahia PL (2000) PTEN, a unique tumor suppressor gene. Endocr Relat Cancer 7:115-129
10. Vivanco I, Sawyers CL (2002) The phosphatidylinositol 3-Kinase AKT pathway in human cancer. Nat Rev Cancer 2:489-501
11. Datta SR, Dudek H, Tao X et al (1997) Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell 91:231-241
12. Dijkers PF, Birkenkamp KU, Lam EW et al (2002) FKHR-L1 can act as a critical effector of cell death induced by cytokine withdrawal: Protein kinase B-enhanced cell survival through maintenance of mitochondrial integrity. J Cell Biol 156:531-542
13. Tran H, Brunet A, Griffith EC, Greenberg ME (2003) The many forks in FOXO's road. Sci STKE 2003:RE5
14. Robey RB, Hay N (2006) Mitochondrial hexokinases, novel mediators of the antiapoptotic effects of growth factors and Akt. Oncogene 25:4683-4696
15. Zhou BP, Hung MC (2002) Novel targets of Akt, p21(Cipl/WAF1), and MDM2. Semin Oncol 29: 62-70
16. Liang J, Zubovitz J, Petrocelli T et al (2002) PKB/Akt phosphorylates p27, impairs nuclear import of p27 and opposes p27-mediated G1 arrest. Nat Med 8:1153-1160
17. Shin I, Yakes FM, Rojo F et al (2002) PKB/Akt mediates cell-cycle progression by phosphorylation of p27(Kip1) at threonine 157 and modulation of its cellular localization. Nat Med 8:1145-1152
18. Viglietto G, Motti ML, Bruni P et al (2002) Cytoplasmic relocalization and inhibition of the cyclin-dependent kinase inhibitor p27(Kip1) by PKB/Akt-mediated phosphorylation in breast cancer. Nat Med 8:1136-1144
19. Lum JJ, Bui T, Gruber M et al (2007) The transcription factor HIF-1alpha plays a critical role in the growth factor-dependent regulation of both aerobic and anaerobic glycolysis. Genes Dev 21:1037-1049
20. Majumder PK, Febbo PG, Bikoff R et al (2004) mTOR inhibition reverses Akt-dependent prostate intraepithelial neoplasia through regulation of apoptotic and HIF-1-dependent pathways. Nat Med 10:594-601
21. Stiles B, Gilman V, Khanzenzon N et al (2002) Essential role of AKT-1/protein kinase B alpha in PTEN-controlled tumorigenesis. Mol Cell Biol 22:3842-3851
22. Link W, Rosado A, Fominaya J et al (2005) Membrane localization of all class I PI 3-kinase isoforms suppresses c-Myc-induced apoptosis in Rat1 fibroblasts via Akt. J Cell Biochem 95:979-989
23. Chen Z, Trotman LC, Shaffer D et al (2005) Crucial role of p53-dependent cellular senescence in suppression of Pten-deficient tumorigenesis. Nature 436:725-730
24. Samuels Y, Ericson K (2006) Oncogenic PI3K and its role in cancer. Curr Opin Oncol 18:77-82
25. Toker A, Yoeli-Lerner M (2006) Akt signaling and cancer: Surviving but not moving on. Cancer Res 66:3963-3966
26. Mayo LD, Donner DB (2002) The PTEN, Mdm2, p53 tumor suppressor- oncoprotein network. Trends Biochem Sci 27:462-467
27. Brazil DP, Hemmings BA (2001) Ten years of protein kinase B signalling: A hard Akt to follow. Trends Biochem Sci 26:657-664
28. Bayascas JR, Leslie NR, Parsons R et al (2005) Hypomorphic mutation of PDK1 suppresses tumorigenesis in PTEN(+/-) mice. Curr Biol 15:1839-1846
29. Blanco-Aparicio C, Renner O, Leal JF, Carnero A (2007) PTEN, more than the AKT pathway. Carcinogenesis 28:1379-1386
30. Stiles B, Groszer M, Wang S et al (2004) PTENless means more. Dev Biol 273:175-184
31. Zhao JJ, Cheng H, Jia S et al (2006) The p110alpha isoform of PI3K is essential for proper growth factor signaling and oncogenic transformation. Proc Natl Acad Sci U S A 103:16296-16300
32. Zhao L, Vogt PK (2008) Class I PI3K in oncogenic cellular transformation. Oncogene 27:5486-5496
33. Jia S, Liu Z, Zhang S et al (2008) Essential roles of PI(3)K-p110beta in cell growth, metabolism and tumorigenesis. Nature 454:776-779
34. Bos JL (1989) ras oncogenes in human cancer: A review. Cancer Res 49:4682-4689
35. Carnero A, Lacal JC (1995) Activation of intracellular kinases in Xenopus oocytes by p21ras and phospholipases: A comparative study. Mol Cell Biol 15:1094-1101
36. Carnero A, Liyanage M, Stabel S, Lacal JC (1995) Evidence for different signalling pathways of PKC zeta and ras-p21 in Xenopus oocytes. Oncogene 11:1541-1547
37. Carnero A, Beach DH (2004) Absence of p21WAF1 cooperates with c-myc in bypassing Ras-induced senescence and enhances oncogenic cooperation. Oncogene 23:6006-6011
38. Shields JM, Pruitt K, McFall A et al (2000) Understanding Ras: 'it ain't over 'til it's over'. Trends Cell Biol 10:147-154
39. Ulku AS, Der CJ (2003) Ras signaling, deregulation of gene expression and oncogenesis. Cancer Treat Res 115:189-208
40. Morrison DK, Cutler RE (1997) The complexity of Raf-1 regulation. Curr Opin Cell Biol 9:174-179
41. Feig LA (2003) Ral-GTPases: Approaching their 15 minutes of fame. Trends Cell Biol 13:419-425
42. Rangarajan A, Hong SJ, Gifford A, Weinberg RA (2004) Species-and cell type-specific requirements for cellular transformation. Cancer Cell 6:171-183
43. McFarlin DR, Lindstrom MJ, Gould MN (2003) Affinity with Raf is sufficient for Ras to efficiently induce rat mammary carcinomas. Carcinogenesis 24:99-105
44. McFarlin DR, Gould MN (2003) Rat mammary carcinogenesis induced by in situ expression of constitutive Raf kinase activity is prevented by tethering Raf to the plasma membrane. Carcinogenesis 24:1149-1153
45. Lim KH, Counter CM (2005) Reduction in the requirement of oncogenic Ras signaling to activation of PI3K/AKT pathway during tumor maintenance. Cancer Cell 8:381-392
46. Gupta S, Ramjaun AR, Haiko P et al (2007) Binding of ras to phosphoinositide 3-kinase p110alpha is required for ras-driven tumorigenesis in mice. Cell 129:957-968
47. Engelman JA, Chen L, Tan X et al (2008) Effective use of PI3K and MEK inhibitors to treat mutant Kras G12D and PIK3CA H1047R murine lung cancers. Nat Med 14:1351-1356
48. Yu K, Toral-Barza L, Shi C et al (2008) Response and determinants of cancer cell susceptibility to PI3K inhibitors: Combined targeting of PI3K and Mek1 as an effective anticancer strategy. Cancer Biol Ther 7:307-315
49. Parsons DW, Wang TL, Samuels Y et al (2005) Colorectal cancer: Mutations in a signalling pathway. Nature 436:792
50. Liaw D, Marsh DJ, Li J et al (1997) Germline mutations of the PTEN gene in Cowden disease, an inherited breast and thyroid cancer syndrome. Nat Genet 16:64-67
51. Marsh DJ, Kum JB, Lunetta KL et al (1999) PTEN mutation spectrum and genotype-phenotype correlations in Bannayan-Riley-Ruvalcaba syndrome suggest a single entity with Cowden syndrome. Hum Mol Genet 8:1461-1472
52. Nelen MR, van Staveren WC, Peeters EA et al (1997) Germline mutations in the PTEN/MMAC1 gene in patients with Cowden disease. Hum Mol Genet 6:1383-1387
53. Schrager CA, Schneider D, Gruener AC et al (1998) Clinical and pathological features of breast disease in Cowden's syndrome: An underrecognized syndrome with an increased risk of breast cancer. Hum Pathol 29:47-53
54. Samuels Y, Wang Z, Bardelli A et al (2004) High frequency of mutations of the PIK3CA gene in human cancers. Science 304:554
55. Kang S, Bader AG, Vogt PK (2005) Phosphatidylinositol 3-kinase mutations identified in human cancer are oncogenic. Proc Natl Acad Sci U S A 102:802-807
56. Zhao JJ, Liu Z, Wang L et al (2005) The oncogenic properties of mutant p110alpha and p110beta phosphatidylinositol 3-kinases in human mammary epithelial cells. Proc Natl Acad Sci U S A 102:18443-18448
57. Bader AG, Kang S, Vogt PK (2006) Cancer-specific mutations in PIK3CA are oncogenic in vivo. Proc Natl Acad Sci U S A 103:1475-1479
58. Renner O, Fominaya J, Alonso S et al (2007) Mst1, RanBP2, and eIF4G are new markers for in vivo PI3K activation in murine and human prostate. Carcinogenesis 28:1418-1425
59. Renner O, Blanco-Aparicio C, Grassow M et al (2008) Activation of phosphatidylinositol 3-kinase by membrane localization of p110 predisposes mammary glands to neoplastic transformation. Cancer Res 68:9643-9653
60. Renner O, Blanco-Aparicio C, Carnero A (2008) Genetic modelling of the PTEN/AKT pathway in cancer research. Clin Transl Oncol 10:618-627
61. Renner O, Blanco-Aparicio C, Grassow M et al (2008) Activation of phosphatidylinositol 3-kinase by membrane localization of p110alpha predisposes mammary glands to neoplastic transformation. Cancer Res 68:9643-9653
62. Bachman KE, Argani P, Samuels Y et al (2004) The PIK3CA gene is mutated with high frequency in human breast cancers. Cancer Biol Ther 3:772-775
63. Campbell IG, Russell SE, Choong DY et al (2004) Mutation of the PIK3CA gene in ovarian and breast cancer. Cancer Res 64:7678-7681
64. Kirkegaard T, Witton CJ, McGlynn LM et al (2005) AKT activation predicts outcome in breast cancer patients treated with tamoxifen. J Pathol 207:139-146
65. Xing D, Orsulic S (2005) A genetically defined mouse ovarian carcinoma model for the molecular characterization of pathway-targeted therapy and tumor resistance. Proc Natl Acad Sci U S A 102:6936-6941
66. Nakayama K, Nakayama N, Kurman RJ et al (2006) Sequence mutations and amplification of PIK3CA and AKT2 genes in purifi ed ovarian serous neoplasms. Cancer Biol Ther 5:779-785
67. Asano T, Yao Y, Zhu J et al (2004) The PI 3-kinase/Akt signaling pathway is activated due to aberrant Pten expression and targets transcription factors NF-kappaB and c-Myc in pancreatic cancer cells. Oncogene 23:8571-8580
68. Okano J, Snyder L, Rustgi AK (2003) Genetic alterations in esophageal cancer. Methods Mol Biol 222:131-145
69. Garcia-Rostan G, Costa AM, Pereira-Castro I et al (2005) Mutation of the PIK3CA gene in anaplastic thyroid cancer. Cancer Res 65:10199-10207
70. Broderick DK, Di C, Parrett TJ et al (2004) Mutations of PIK3CA in anaplastic oligodendrogliomas, high-grade astrocytomas, and medulloblastomas. Cancer Res 64:5048-5050
71. Yuan TL, Cantley LC (2008) PI3K pathway alterations in cancer: Variations on a theme. Oncogene 27:5497-5510
72. Downward J (2003) Targeting RAS signalling pathways in cancer therapy. Nat Rev Cancer 3:11-22
73. Nagata Y, Lan KH, Zhou X et al (2004) PTEN activation contributes to tumor inhibition by trastuzumab, and loss of PTEN predicts trastuzumab resistance in patients. Cancer Cell 6:117-127
74. Fujita T, Doihara H, Kawasaki K et al (2006) PTEN activity could be a predictive marker of trastuzumab efficacy in the treatment of ErbB2-overexpressing breast cancer. Br J Cancer 94:247-252
75. Berns K, Horlings HM, Hennessy BT et al (2007) A functional genetic approach identifi es the PI3K pathway as a major determinant of trastuzumab resistance in breast cancer. Cancer Cell 12:395-402
76. Gharbi SI, Zvelebil MJ, Shuttleworth SJ et al (2007) Exploring the specificity of the PI3K family inhibitor LY294002. Biochem J 404:15-21
77. Garcia-Echeverria C, Sellers WR (2008) Drug discovery approaches targeting the PI3K/Akt pathway in cancer. Oncogene 27:5511-5526
78. Stauffer F, Holzer P, Garcia-Echeverria C (2005) Blocking the PI3K/PKB pathway in tumor cells. Curr Med Chem Anticancer Agents 5:449-462
79. Garlich JR, De P, Dey N et al (2008) A vascular targeted pan phosphoinositide 3-kinase inhibitor prodrug, SF1126, with antitumor and antiangiogenic activity. Cancer Res 68:206-215
80. Chiorean EG, Mahadevan D, Harris WB et al (2009) Phase I evaluation of SF1126, a vascular targeted PI3K inhibitor, administered twice weekly IV in patients with refractory solid tumors. J Clin Oncol 27:15
81. Zhu T, Gu J, Yu K et al (2006) Pegylated wortmannin and 17-hydroxywortmannin conjugates as phosphoinositide 3-kinase inhibitors active in human tumor xenograft models. J Med Chem 49:1373-1378
82. Ihle NT, Williams R, Chow S et al (2004) Molecular pharmacology and antitumor activity of PX-866, a novel inhibitor of phosphoinositide-3- kinase signaling. Mol Cancer Ther 3:763-772
83. Ihle NT, Powis G (2009) Take your PIK: phosphatidylinositol 3-kinase inhibitors race through the clinic and toward cancer therapy. Mol Cancer Ther 8:1-9
84. Carnero A (2009) Novel inhibitors of the PI3K family. Exp Opin Investig New Drugs 18:1-13
85. Knight ZA, Shokat KM (2007) Chemically targeting the PI3K family. Biochem Soc Trans 35:245- 249
86. Maira SM, Stauffer F, Brueggen J et al (2008) Identification and characterization of NVPBEZ235, a new orally available dual phosphatidylinositol 3-kinase/mammalian target of rapamycin inhibitor with potent in vivo antitumor activity. Mol Cancer Ther 7:1851-1863
87. Folkes AJ, Ahmadi K, Alderton WK et al (2008) The identification of 2-(1H-indazol-4-yl)-6-(4- methanesulfonyl-piperazin-1-ylmethyl)-4-morpholin -4-yl-thieno[3,2-d]pyrimidine (GDC-0941) as a potent, selective, orally bioavailable inhibitor of class I PI3 kinase for the treatment of cancer. J Med Chem 51:5522-5532
88. Serra V, Markman B, Scaltriti M et al (2008) NVPBEZ235, a dual PI3K/mTOR inhibitor, prevents PI3K signaling and inhibits the growth of cancer cells with activating PI3K mutations. Cancer Res 68:8022-8030
89. Calvo E, Edelman G, Baselga J et al (2008) A phase 1 dose-escalation study of the safety, pharmacokinetics and pharmacodynamics of XL147, a novel PI3K inhibitor administered orally to patients with advanced solid tumors. EORTCNCI- AACR International Conference on Molecular Targets and Cancer Therapeutics, Geneva, Switzerland
90. Foster, PG (2007) Potentiating the antitumor effects of chemotherapy with the selective PI3K inhibitor XL147. AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics, C199
91. Shapiro G, Kwak E, Baselga J et al (2009) Phase I dose-escalation study of XL147, a PI3K inhibitor administered orally to patients with solid tumors. J Clin Oncol 27[15s]:3500
92. Laird D (2007) XL765 targets tumor growth, survival, and angiogenesis in preclinical models by dual inhibition of PI3K and mTOR. AACR-NCIEORTC International Conference on Molecular Targets and Cancer Therapeutics, B250
93. Markman B, LoRusso PM, Patnaik A et al (2008) A phase 1 dose-escalation study of the safety, pharmacokinetics and pharmacodynamics of XL765, a novel inhibitor of PI3K and mTOR, administered orally to patients with solid tumors. EORTC-NCI-AACR International Conference on Molecular Targets and Cancer Therapeutics, Geneva, Switzerland
94. LoRusso P, Markman B, Tabernero J et al (2009) A phase I dose-escalation study of the safety, pharmacokinetics (PK), and pharmacodynamics of XL765, a PI3K/TORC1/TORC2 inhibitor administered orally to patients (pts) with advanced solid tumors. J Clin Oncol 27[15s]:abstr 3502
95. Sarker D, Kristeleit R, Mazina KE et al (2009) A phase I study evaluating the pharmacokinetics (PK) and pharmacodynamics (PD) of the oral pan-phosphoinositide-3 kinase (PI3K) inhibitor GDC-0941. J Clin Oncol 27[15s]:3538
96. Wagner AJ, Von Hoff DH, LoRusso PM et al (2009) A first-in-human phase I study to evaluate the pan-PI3K inhibitor GDC-0941 administered QD or BID in patients with advanced solid tumors. J Clin Oncol 27[15s]abstr 3501
97. - (2009) A phase I/II study of BEZ235 in patients with advanced solid malignancies enriched by patients with advanced breast cancer. http://clinicaltrials. gov/ct2/show/NCT00620594
98. Billottet C, Grandage VL, Gale RE et al (2006) A selective inhibitor of the p110delta isoform of PI 3-kinase inhibits AML cell proliferation and survival and increases the cytotoxic effects of VP16. Oncogene 25:6648-6659
99. Flinn IW, Byrd JC, Furman RR et al (2009) Preliminary evidence of clinical activity in a phase I study of CAL-101, a selective inhibitor of the p1108 isoform of phosphatidylinositol 3-kinase (P13K), in patients with select hematologic malignancies. J Clin Oncol 27[15s]:abstr 3543
100. Hu L, Hofmann J, Lu Y et al (2002) Inhibition of phosphatidylinositol 3'-kinase increases efficacy of paclitaxel in in vitro and in vivo ovarian cancer models. Cancer Res 62:1087-1092
101. Mabuchi S, Ohmichi M, Kimura A et al (2002) Inhibition of phosphorylation of BAD and Raf-1 by Akt sensitizes human ovarian cancer cells to paclitaxel. J Biol Chem 277:33490-33500
102. Nguyen DM, Chen GA, Reddy R et al (2004) Potentiation of paclitaxel cytotoxicity in lung and esophageal cancer cells by pharmacologic inhibition of the phosphoinositide 3-kinase/protein kinase B (Akt)-mediated signaling pathway. J Thorac Cardiovasc Surg 127:365-375
103. Blanco-Aparicio C, Pequeno B, Moneo V et al (2005) Inhibition of phosphatidylinositol-3-kinase synergizes with gemcitabine in low-passage tumor cell lines correlating with Bax translocation to the mitochondria. Anticancer Drugs 16:977-987
104. Ng SSW, Tsao MS, Chow S, Hedley DW (2000) Inhibition of phosphatidylinositide 3-kinase enhances gemcitabine-induced apoptosis in human pancreatic cancer cells. Cancer Res 60:5451- 5455
105. Wang Q, Li N, Wang X et al (2002) Augmentation of sodium butyrate-induced apoptosis by phosphatidylinositol 3'-kinase inhibition in the KM20 human colon cancer cell line. Clin Cancer Res 8:1940-1947
106. Edwards E, Geng L, Tan J et al (2002) Phosphatidylinositol 3-kinase/Akt signaling in the response of vascular endothelium to ionizing radiation. Cancer Res 62:4671-4677
107. Gupta AK, McKenna WG, Weber CN et al (2002) Local recurrence in head and neck cancer: relationship to radiation resistance and signal transduction. Clin Cancer Res 8:885-892
108. Tortora G, Gelardi T, Ciardiello F, Bianco R (2007) The rationale for the combination of selective EGFR inhibitors with cytotoxic drugs and radiotherapy. Int J Biol Markers 22:S47-52
109. She QB, Solit DB, Ye Q et al (2005) The BAD protein integrates survival signaling by EGFR/ MAPK and PI3K/Akt kinase pathways in PTENdefi cient tumor cells. Cancer Cell 8:287-297
110. Sordella R, Bell DW, Haber DA, Settleman J (2004) Gefitinib-sensitizing EGFR mutations in lung cancer activate anti-apoptotic pathways. Science 305:1163-1167
111. Ihle NT, Paine-Murrieta G, Berggren MI et al (2005) The phosphatidylinositol-3-kinase inhibitor PX-866 overcomes resistance to the epidermal growth factor receptor inhibitor gefitinib in A-549 human non-small cell lung cancer xenografts. Mol Cancer Ther 4:1349-1357
112. Yakes FM, Chinratanalab W, Ritter CA et al (2002) Herceptin-induced inhibition of phosphatidylinositol- 3 kinase and Akt Is required for antibodymediated effects on p27, cyclin D1, and antitumor action. Cancer Res 62:4132-4141
113. Eichhorn PJ, Gili M, Scaltriti M et al (2008) Phosphatidylinositol 3-kinase hyperactivation results in lapatinib resistance that is reversed by the mTOR/phosphatidylinositol 3-kinase inhibitor NVP-BEZ235. Cancer Res 68:9221-9230
114. Rahmani M, Davis EM, Bauer C et al (2005) Apoptosis induced by the kinase inhibitor BAY 43-9006 in human leukemia cells involves downregulation of Mcl-1 through inhibition of translation. J Biol Chem 280:35217-35227
115. Rahmani M, Reese E, Dai Y et al (2005) Coadministration of histone deacetylase inhibitors and perifosine synergistically induces apoptosis in human leukemia cells through Akt and ERK1/2 inactivation and the generation of ceramide and reactive oxygen species. Cancer Res 65:2422-2432
116. Landis-Piwowar KR, Milacic V, Chen D et al (2006) The proteasome as a potential target for novel anticancer drugs and chemosensitizers. Drug Resist Updat 9:263-273