Download powerpoint

ANTI-TUMOR EFFECTS
OF AN IMIDAZOQUINOLINE
IN RENAL CELL CARCINOMA
Michael Schwartz, MD
3/14/2007
What are
imidazoquinolines?
• Family of immune response modifiers of which imiquimod is
the most commonly used.
• Act on Toll-like receptors which can increase cellular
production of TNF, IFN-, and a variety of other cytokines.
• Shown to improve antigen presentation by dendritic cells
• Imiquimod is FDA approved for topical treatment of
genital condyloma and basal cell carcinoma.
Project Background
• Renal cell carcinoma (RCC) responds to
immunomodulators (IL-2, IFN-)
• Imiquimod has been used as an immune
response modifier in basal cell carcinoma
• We hypothesized that imidazoquinolines
may have direct in vitro and in vivo
biological activity against renal cell
carcinoma.
Study Design
• 4 renal cell carcinoma cell lines used
– 1 mouse (RENCA)
– 3 human (CAKI-1, CAKI-2, A-498)
Experimental drug: 3M-011
• Closely related to Imiquimod (3M
Pharmaceuticals)
• Potential for use as an oral agent
Do renal cell carcinoma cell lines
express Toll-like receptor 7?
TLR-7 Expression in RCC cell lines
300
280
260
RENCA
Band Intensity
240
220
CAKI-2
200
180
160
140
(-) Control
CAKI-1
120
100
Cell Line
A-498
Can 3M-011 affect cell
viability?
• Evaluated by cell counts
• Cells cultured in with media
supplemented with 3M-011at 100 g/ml
• Cells harvested and counted at 24, 48,
and 72 hour time points
3M-011 Affect on Cell Viability
TLR-7 Expression in RCC cell lines
Relative to Control
300
280
120
260
RENCA
Band Intensity
100
80
% decrease
in cell
viability
RENCA
240
220
CAKI-2
200
180
160
140
120
60
100
(-) Control
A-498
CAKI-1
CAKI-2
Cell Line
40
20
CAKI-1
0
Cell Lines
Potential correlation between cell viability
and level of TLR-7 expression
A-498
Apoptosis in RENCA cells induced by 3M-011
1.4
Figure 2
3M-011
Absorbance (405 nm)
1.2
1
0.8
0.6
0.4
Control
0.2
0
Media supplement
In vivo affects of 3M-011 on a murine
metastatic RCC model
Tail vein injection
(2 x 105 RENCA cells)
One week for tumor growth
Group 1: Placebo
Group 2: 3M-011
Two week treatment
(every other day oral gavage)
Sacrifice for pathology
In vivo results
120%
100%
100%
70%
80%
60%
% of mice with tumor
40%
20%
0%
Placebo
3M-011
45
37.2
40
35
30
Mean # of macroscopic tumor nodules
25
20
12.6
15
10
5
0
Placebo
60
3M-011
51.6
50
Mean # of microscopic tumor nodules
40
22.9
30
20
10
0
Placebo
• 30% of mice in 3M-011 group showed
no pathologic evidence of tumor
3M-011
PLACEBO
3M-011
Summary and Conclusions
• TLR-7 is expressed in both murine and human renal cancer
cell lines.
• Imidazoquinolines have direct biological effects on renal
cell carcinoma cell lines by decreasing cell viability and
inducing apoptosis.
• Our results suggest a potential correlation between the
TLR-7 expression level and the effect of 3M-011 on cell
viability.
• Initial results in an immune competent mouse model of
metastatic renal cell carcinoma suggest anti-tumor effects
in vivo.
References
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
Jemal A, Siegel R, Ward E, Murray T Xu J, , Murray T, Smigel C,
Thun M. Cancer statistics, 2006. CA Cancer J Clin 2006;56:106-130.
Lam JS, Leppert JT, Figlin RA, Belldegrun AS. Surveillance
following radical or partial nephrectomy for renal cell carcinoma.
Cur Urol Rep 2005;6:7-18.
Milowsky MI, Nanus DM. Chemotherapeutic strategies for renal cell
carcinoma. Urol Clin North Am 2003;30:601-609.
Yagoda A, Abi-Rached B, Petrylak D. Chemotherapy for advanced
renal-cell carcinoma: 1983-1993. Semin Oncol 1995;22:42-60.
Flanigan RC, Salmon SE, Blumenstein BA et al. Nephrectomy
followed by interferon-2b compared with interferon-2b alone for
metastatic renal-cell cancer. N Engl J Med 2001;345:1655-1659.
Medical Research Council Renal Cancer Collaborators. Interferon-
and survival in metastatic renal carcinoma: early results of a
randomized controlled trial. Lancet 1999;353:14-17.
Pyrhonen S, Salminen E, Ruutu M et al. Prospective randomized
trial of interferon-2a plus vinblastine versus vinblastine alone in
patients with advanced renal cell cancer. J Clin Oncol 1999;17:28592867.
Motzer RJ, Rini BI, Bukowski RM et al. Sunitinib in patients with
metastatic renal cell carcinoma. JAMA 2006;295:2516-24.
Ratain MJ, Eisen T, Stadler WM et al. Phase II placebo-controlled
randomized discontinuation trial of sorafenib in patients with
metastatic renal cell carcinoma. J Clin Oncol 2006;24:2505-12.
Seya T, Akazawa T, Uehori J, Matsumoto M, Azuma I, Toyoshima
K. Role of toll-like receptors and their adaptors in adjuvant
immunotherapy for cancer. Anticancer Res 2003;23:4369-76.
Okamoto M, Sato M. Toll-like receptor signaling in anti-cancer
immunity. J Med Invest 2003;50:9-24.
Cook DN, Pisetcky DS, Schwartz, DA. Toll-like receptors in the
pathogenesis of human disease. Nat Immunol 2004;5:975-9.
Iwasaki A, Medzhitov R. Toll-like receptor control of the adaptive
immune responses. Nat Immunol 2004;5:987-95.
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
24.
25.
26.
Kodner CM, Nasraty S. Management of genital warts. Am Fam Physician.
2004;70:2335-42.
Tyring S, Conant M, Marini M, Van Der Meijden W, Washenik K.
Imiquimod; an international update on therapeutic uses in dermatology. Int J
Dermatol. 2002;41:810-6.
Horiguchi A, Sumitomo M, Asakuma J, Asano T, Asano T, and Hayakawa M.
3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitor, fluvastatin, as a
novel agent for prophylaxis of renal cancer metastasis. Clin Can Res
2004;10:8648-55.
Lebwohl M, Dinehart S, Whiting D, Lee PK, Tawfik N, Jorizzo J, Lee JH,
Fox TL. Imiquimod 5% cream for the treatment of actinic keratosis: results
from two phase III, randomized, double-blind, parallel group, vehiclecontrolled trials. J Am Acad Dermatol 2004;50:714-21.
Geisse J, Caro I, Lindholm J, Golitz L, Stampone P, Owens M. Imiquimod
5% cream for the treatment of superficial basal cell carcinoma: results from
two phase III, randomized, vehicle-controlled studies. J Am Acad Dermatol
2004;505:722-33.
Bong AB, Bonnekoh B, Franke I, Schon MP, Ulrich J, Gollnick H.
Imiquimod, a topical immune response modifier, in the treatment of
cutaneous metastases of malignant melanoma. Dermatology 2002;205:135-8.
Schon M, Bong AB, Drewniok C, Herz J, Geilen CC, Reifenberger J,
Benninghoff B, Slade HB, Gollnick H, Schon MP. Tumor-selective
induction of apoptosis and the small-molecule immune response modifier
imiquimod. J Natl Cancer Inst 2003;95:1138-49.
Schon MP, Wienrich BG, Drewniok C, Bong AB, Gollnick H, Schon M, et al.
Death receptor-independent apoptosis in malignant melanoma induced by the
small-molecule immune response modifier imiquimod. J Invest Dermatol
2004;122:1266-1276.
Albert ML, Sauter B, Bhardwaj N. Dendritic cells acquire antigen from
apoptotic cells and induce class I-restricted CTLs. Nature 1998;392:86-9.
Schon MP, Schon M. Immune modulation and apoptosis induction: two
sides of the antitumoral activity of imiquimod. Apoptosis. 2004;9:291-8.
Gibson SJ, Lindh JM, Riter TR, Gleason RM, Rogers LM, Fuller AE,
Oesterich JL, Gorden KB, Qiu X, McKane, et al. Plasmacytoid dendritic
cells produce cytokines and mature in response to the TLR7 agonists,
imiquimod and resiquimod. Cell Immunol 2002;218:74-86.
Imbertson LM, et al. Cytokine induction in hairless mouse and rat skin after
topical application of the immune response modifiers imiquimod and S28463. J Invest Dermatol. 1998;110:734-9.
Witt PL, Ritch PS, Reding D, McAuliffe TL, Westrick L, Grossberg SE,
Borden EC. Phase I trial of an oral immunomodulator and interferon inducer
in cancer patients. Cancer Res 1993;53:5176-80.