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`NOVARTIS EXHIBIT 2077
`Par v. Novartis, IPR 2016-01479
`Page 1 of 2
`
`

`

`Clinical Cancer Research 0 Volume 6 0 November 2000 (Supplement)
`
`Poster Session 17
`
`4549s
`
`observed also. Other adverse events, generally mild-moderate in severity, oc-
`curred over a broad range of doses. Toxicities include asthenia, cutaneous
`toxicity, mucositis, and hypertriglyceridemia. In 17 pts receiving 075 to 3.12
`mg/mZ/d, CCl-779 exhibited little accumulation from cycle to cycle, preferential
`binding to R803, dose-related increase in AUC, and mean tv: of 32.6 h.
`Preliminary evidence of antitumor activity has been observed, with 1 PR (non—
`small cell lung cancer) and minor responses in other tumor types. The safety
`profile and antitumor activity observed to date, associated with plasma con—
`centrations at which biological activity was observed in vitro, are encouraging.
`
`415 Effect of the proteasome inhibitor PS-341 on cell cycle progression
`and bcl-2: A potentially unique mechanism of action. Fl. Perez-SolerT, YH
`Ling-r, B NgT, J Adams‘, P Elliott*, L Liebest. Kap/an Cancer Center, TNew York
`University School of Medicine, New York, NY, and “Millennium Pharmaceuticals,
`Cambridge, MA.
`PS—341 is a proteasome inhibitor currently in Phase I clinical evaluation. We
`studied the effects of PS-841 on cell cycle progression and related events in
`human NSCLC H460 (p53 wild type) and H358 (p53 null) cells. Exposure to 0.1
`pM PS-341 for 6 h resulted in a marked accumulation of cells at G2/M. This
`blockade was associated with a 6—10 fold time-dependent accumulation of
`cyclins A and B and a 10—fold elevation of cyclins A and B kinase activities as
`assessed by 32 P-y-ATP incorporation into histone-Hi. in addition. bcl—2 phos-
`phorylation, a marker of mitotic arrest, was detected as early as 3 h after
`exposure to P8341. More importantly, a 25 kDa bcI-2 degradation product was
`detected as early as 12 h after exposure to P8341. This degradation product
`appeared specific for proteasome inhibition since it was observed with the
`proteasome inhibitors MG152 and PSI but not with the chemotherapeutic
`agents paclitaxel, vinblastine, camptothecin, etoposide, and cisplatin, or the
`PKC inhibitor staurosporine. In addition, it was not caspase-dependent since it
`was observed in the presence of caspase inhibitors and appeared to localize in
`the triton X»100 insoluble cellular fraction.
`In view of the ability of P8341 to
`induce arrest at G2/M we then studied in vitro cytotoxicity of the combination of
`P8341 and the antitubulin agent docetaxel against H460 and H358 cells. Cells
`were treated concomitantly with P3341 (0.1pM or 0.5pM) and dccetaxel (0.1 to
`4|).M) for 48 h. An additive cytotoxic effect was observed with the combination
`0.5pM P8341 and 0.5 and 1pM docetaxel. in conclusion, our results indicate
`that P5341 induces unique changes in bcl-2 that appear to be specific for
`proteasome inhibition. The functional consequences of these bcl-2 changes
`and their potential relationship with the demonstrated ability of this agent to
`retain its cytotoxicity against bcI-2 transfected cells is being investigated.
`
`416 A phase I pharmacodynamic study of the proteasome inhibitor
`PS-341. J. P. Thomas, A. Adjei, C. Ehrlichman, P. Geiger, A. Haas, R. Arzooma-
`nian, D. Alberti, R. Marnocha, K. Binger, J. Volkman, C. Feierabend. K. Tutsch,
`J. Adams, P. Eliot and G. Wilding. University of Wisconsin Comprehensive
`Cancer Center, Madison, Wl, Mayo Clinic, Rochester, MN and Mil/enium Phar-
`maceuticals, Cambridge, MA.
`The ubiquitin-proteasome pathway is the principal enzymatic degradation path-
`way for most intracellular proteins including those involved in cell cycle regulation,
`apoptosis and angiogenesis. PS—341 is a dipeptlde boronic acid compound that is
`a potent inhibitor of the 208/268 proteasome. Proteasome inhibitiepigftpggsigqhfiasmp,“
`_——_—aflmfilflflm¥hb_———-——’—fi
`
`414 CCl-779, an ester analogue of rapamycin that interacts with PTEN/
`Pl3 kinase pathways: A phase I study utilizing a weekly intravenous sched-
`ule. E. Raymond, J. Alexandre, H. Depenbrock, N. Ady Vago, S. Faivre, A.
`Lahr-Randak, E. Materman, J. Boni, S. Abbas, E. Angevin, B. Escudier, J.P.
`Armand. Inst/tut Gustave Roussy, Villejuif, France; Onkologische Tagesk/inrk &
`Wyeth Ayerst Research / Genetics Institute, Munich, Germany.
`Background. CCl-779 inhibits mTOR, thus the phosphorylation of elF4E-
`BP1 and p70SE kinases, prevents elF4E to initiate protein synthesis and the
`phosphorylation of the ribosomal protein 86 requrred for the translation of
`mRNAs. Patients and Methods. CCl-779 was given as a weekly 30—min
`infusion in patients (pts) with advanced tumors using the modified CRM. Re-
`sults. 18pts (M/F: 12/6) received: 7.5 (1 pt), 15 (2pts), 22.5 (1 pt), 34 (3pts), 45
`(4pts), 60 (1 pt), 80 (1 pt), 110 (1pt), 165 (1pt) and 220 mg/mzlweek (3pts). DLT
`was observed in only 1pt; MTD has not been reached. No prolonged immuno-
`suppression has been induced. Grade (Gr) 1—2 skin toxicity was observed:
`dryness with mild itching (Bpts), eczema-like lesions (2pts), sub—acute urticaria
`(2pts), and aseptic folliculitis (11pts). Gr1—2 and Gr-3 mucositls/stomatitis were
`observed in 10pts and 1pt, respectively. All pts receiving 2 8 doses experi—
`enced Gr-1 nail changes. Thrombocytopenia was observed in ths; 2pts with
`G-3 at 34 and 45 mg/mz/week. Leukopenia was reported In 4pts and anemia in
`7pts. Asymptomatic increases of triglyceride and cholesterol levels were ob-
`served in 9pts and 5pts, respectively. A reversible decrease in testosterone
`concentrations with increased levels of LH/FSH were observed in 5/9 men
`receiving 24 doses at dosages 215mgim2/week. Pharmacokinetic analysis
`from 12pts (doses: 7.5—60 mg/mZ/week) indicates that CCl-779 me increased
`linearly but AUC increased sub-proportionately. Clearance and volume of dis-
`tribution at steady state increased with increasing dose. Mean half-life was
`about 20hrs. Of the 16pts evaluable for anti-tumor activity, Spts had a partial
`response (renal cell carcinoma with lung metastases; neuro-endocrine tumor
`with hepatic metastases and breast cancer with liver, lymph node and peri-
`orbital metastases]. Conclusion. Current data show that 001—779 has promis-
`ing activity and mild-moderate toxicity over a broad range of doses.
`
`Proceedings estimombtoshaowsc 0 AACR Symposium
`
`diverse metabolic processes including stabilization of cell cycle regulatory proteins
`and inhibition of NF-KB activation. PS—341 has broad activity including MDR and
`Bcl-2 overexpressing cancer cell
`lines. in vivo PS—341 inhibits the growth of a
`number of tumors including the HT—29, NCl-H23 and PC-3 models. Toxicity was
`seen in preclinical models when the proteasome was inhibited by greater than
`80%. We are conducting a phase I
`trial of PS—341 in patients with advanced
`refracton/ cancers. PS—341 is administered intravenously twice weekly for 4 weeks
`followed by a two week break. Dose levels of 0.5, 0.9, 1.25 and 1.50 mg/m2 have
`been explored. A total of 9 patients have been treated at the UW. Toxicities seen
`have included rash, fatigue and thrombocytopenia. A MTD has not yet been
`reached. Proteasome inhibition by PS—341 has been monitored by measuring 208
`proteasome activity in whole blood samples using a flourogenic peptide substrate.
`203 proteasome inhibition in this study measured 1 hour after PS—341 administra-
`tion correlates highly with PS—341 dose. We are achieving levels of proteasome
`inhibition (> 60%) associated with anti-tumor activity in the preclinical models. We
`have also examined patient peripheral blood mononuclear cells to determine
`whether levels of proteasome inhibition achieved in this study may be associated
`with accumulation of ubiquitinated proteins. Cell lysates were analyzed by Western
`blot for ubiquitin protein conjugates. Up to a 3 fold increase in ubiquitinated
`proteins were seen in some patients, peaking at 5 hours after F’s-341 admini—
`stration.
`
`417 Proteasome inhibition by PS-341: A phase I study. A Hamilton‘, JP
`Eder2, A Pavlick‘, JW Clark“, A Chachoua‘, DP Ryans, K Farrell1 , H Wasserstrom‘,
`L Liebes1 , J Wright“, P Elliott5, J Adams5 and F Muggia‘. ’NYU Sch. ofMed., zDana
`Farber Can. Inst, 3Mass. General Hosp, 4CTEP NCI,5MiI/ennium Pharm.
`The proteasome is a multimeric protease complex that regulates cellular
`proteins by degrading ubiquinated proteins. Proteasome inhibition results in
`increased levels of a variety of key cellular proteins that may contribute to
`anti-tumor activity; iKB inhibits nuclear factor KB (NF-KB) mediated transcrip-
`tion, p53 inhibits apoptosis, and p21 inhibits cyclin—dependent kinase (CDK)
`activity. PS—341 is a dipeptide boronic acid derivative that inhibits the protea-
`some by stabilization of its active site. Animal models predicted dose limiting
`gastrointestinal toxicity at 280% proteasome inhibition (Pl). PS-341 was ad-
`ministered as an IV bolus on D1&4 of a 2-week cycle. Five dose levels have
`been studied to date: 0.25mg/m2, 0.8mg/m2, 1mg/m2, 1.2mg/m2 and 1.45mg/
`m2. 19 pts have been treated: 11M / 8F. Age: median 57, range 25—78. Primary
`tumors: colorectal (3), renal
`(3), NSCLC (3), melanoma (2), ST sarcoma (2).
`osteosarcoma (1), lymphoma (1), prostate (1), endometrial (1), esophagus (1),
`hepatoma (1). Prior therapies: chemotherapy (17), radiotherapy (13). Toxicities
`have been mild and non-specific. 1/6 pts treated at 1.2mg/m2 experienced
`self-limiting GS diarrhea. No objective responses have been documented. one
`pt with melanoma treated at 1mg/m2 maintained a PR in lung with SD in skin fer
`6 months. Pl was measured at 1, 4 and 24hrs after dosing. At all dose levels,
`peak Pl was seen at 1hr, and recovery to approximately 50% of peak Pl was
`seen at 24h. Peak mean Pl were 21%, 54%, 48% and 59% at dose levels 1, 2'
`3 and 4 respectively. Tumor PI at 24h in one pt was 87% and averaged 54% in
`2 biopsies at 2—3 h in another pt. Accrual is ongomg at 1.9mg/n12, and phase "
`studies are planned. Supported by U01 CA76642, M01 RR00096 and the Lynne
`Cohen Foundation (NY), and U01 62490 (Boston).
`
`418 Pharmacodynamic evaluation of the protein kinase C (PKC) inhib-
`itor CGP41251 (PKC412) in patients with metastatic melanoma. M. Mill-
`ward‘, C. House‘, L. Webster‘, B. Linahan‘, l. Olver2, G. Toner‘, J. ZaiCbergt
`D. Bowtell‘. ’Peter MacCa/lum Cancer Institute, Melbourne, 2Floya/ Adelaidé
`Hospital, Australia.
`.
`_
`PKC412 selectively inhibits PKC (lC50 <1piM) and has preclinical activity as a
`cytostatic and modulator of MDR. The recommended Phase ll dose is 75mg (d3
`which produces potentially active trough plasma levels (10pmol/l), and suppresses
`cytokine release and lymphocyte ERKZ levels (Thavasu 1999). Patients (pts) with
`measurable metastatic melanoma and 2 2 superfrcral lesrons received 75mg tds.
`tumor biopsies and plasma were collected prior to and after 28 days treatment:
`lntra-tumoral total PKC activity was measured in cytosolic and particulate fractions
`using protamine sulphate as the substrate. lnitial experiments showed addition of
`10pM PKC412 to melanoma biopsies inhibited phosphorylatron. Abrlity of plasma
`to modulate ex vivo intracellular daunorubicin accumulatro'nlrn MDR cells was
`measured with activity of 20pg/ml valspodar (PSC833) defining 100% reversaL
`Compared to the pretreatment biopsy, cytosolic PKC actrvrty was reduced by 7%
`to 91% in 7/9 pts. Particulate PKC activity was reduced by 11% to 79% in 4/9 p13.
`Only 1 pt had >50% inhibition in both fractions. Tumor PKC rsoform profile in 1 pt
`resistant to PKC412 (unchanged cytosolic activrty and >200% increase particulate
`activity) showed an abundance of PKCg, an isoform refractory to inhibition b
`PKC412 (IC50 >1000pM). Addition of 20pg/ml PKC412 to pretreatment plasma
`produced 14%—64% (mean 40%) reversal of MDR. Plasma taken following 28
`days PKC412 showed <10% reversal in 8/8 patients. Alllpatrents had progressiVe
`disease. This Phase IIA trial did not demonstrate consrstent target inhibition or
`pharmacodynamic efficacy of PKC412 in melanoma patients.
`
`NOVARTIS EXHIBIT 2077
`Par v. Novartis, IPR 2016-01479
`Page 2 of 2
`
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