Cell Chemical Biology, Volume 24

Supplemental Information

Proteome-wide Map of Targets

of T790M-EGFR-Directed Covalent Inhibitors

Sherry Niessen, Melissa M. Dix, Sabrina Barbas, Zachary E. Potter, Shuyan Lu, OlegBrodsky, Simon Planken, Douglas Behenna, Chau Almaden, Ketan S. Gajiwala, KevinRyan, RoseAnn Ferre, Michael R. Lazear, Matthew M. Hayward, John C.Kath, and Benjamin F. Cravatt

1

Supplemental Information

Proteome-wide map of targets of T790M-EGFR-directed covalent inhibitors

Sherry Niessen, Melissa M. Dix, Sabrina Barbas, Zachary E. Potter, Shuyan Lu, Chau Almaden, Oleg Brodsky,

Simon Planken, Douglas Behenna, Ketan S. Gajiwala, Kevin Ryan, RoseAnn Ferre, Michael R Lazear, Matthew M.

Hayward, John C. Kath, Benjamin F. Cravatt

I. Supplemental Figures

II. Supplemental Tables

III. Methods S1

IV. References

2

I. Supplemental Figures

Figure S1. Co-Crystal Structures of Inhibitor 2 and 3 with L858R/T790M, Related to Figure 1.

(A) 1.58 Å co-crystal structure of inhibitor 2 with L858R/T790M EGFR (PDBID: 5UGC) (Planken et al., 2017).

The pyrazole-N1 position, high-lighted with a red arrow, is solvent exposed and was selected as the site to install the

alkyne group.

(B) 3.1 Å co-crystal structure of inhibitor 3 with L858R/T790M EGFR (PDBID: 5UWD). The piperazine amide,

high-lighted with a red arrow, is solvent exposed and was selected as the site to install the alkyne group.

3

Figure S2. Characterizing Probes 4-6, Related to Figure 1.

A) Competitive ABPP assessing blockade of probe 4-6 reactivity by corresponding parent inhibitors 1-3. A431 cells

were pre-incubated with increasing concentrations of inhibitor (1-3; 1 hr) followed by treatment with the

corresponding probe (4-6; 1 hr) and total cellular proteome was analyzed by gel-based ABPP. Probe reactivity of the

170 kDa band corresponding to WT-EGFR is shown. IC50 values are calculated and displayed below (n = 2 per

group, + SD).

(B) In situ proteome reactivity of second- and third-generation EGFR probes in A431 cells. A431 cells, were treated

with designated probes (1 µM, 1 hr) and total cellular proteome was analyzed by gel-based ABPP.

(C) Coommassie stain for SDS-PAGE gels showing in situ proteome reactivity of second- and third-generation

EGFR probes in human cancer cells (Figure 1B) and competitive blockade of probes 4-6 reactivity by

corresponding parent inhibitors 1-3 (Figure 1C).

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5

Figure S3. In Situ Proteome Reactivity of Third-Generation EGFR Inhibitors, Related to Figure 1.

(A) Evaluation of time-dependence of proteomic reactivity of probes 4-6. H1975 cells were treated with designated

probes (4-6; 1 µM) for indicated lengths of time and analyzed by gel-based ABPP.

(B) Evaluation of proteomic reactivity of probe 4-6 in cancer cell lines expressing WT-EGFR (A431) and clinically

relevant EGFR mutants including; EGFR-Del19 (PC9), EGFR-L858R (H3255) and L858R/T790M-EGFR (H1975).

Cells were treated in situ with either 100 nM or 1 µM of the indicated probes (4-6; 24 hr or 4 hr, respectively) and

analyzed by gel-based ABPP.

(C) In vitro proteomic reactivity profiles of probes 4-6. H1975 soluble and insoluble cell lysates at 1 mg/mL were

treated in vitro with probes 4-6 (1 µM, 1 hr) and analyzed by gel-based ABPP.

(D) Competition of probe 7 labeling of WT-EGFR expressed in A431 cells with second and third- generation EGFR

inhibitors. A431 cells were treated with indicated inhibitors (100 nM, 1 hr) or DMSO followed by probe 7 treatment

(1 µM, 1 hr) and analyzed by gel-based ABPP. PF-06459988 is a potent EGFRT790M inhibitor (Cheng et al.,

2016), PF-06274484 is a pan-ERBB family inhibitor (Lanning et al., 2014) and AZD5104 is a metabolite of

AZD9291 with increased potency for WT-EGFR (designated as compound 27 in (Finlay et al., 2014).

(E) Competitive ABPP analysis of probes 4 and 6 by corresponding parent inhibitors 1 and 3. H1975 cells were pre-

incubated with increasing concentrations of inhibitor (1 or 3; 2 hr) followed by treatment with the corresponding

probe (4 or 6; 4 hr) and analyzed by gel-based ABPP. Specifically competed targets of inhibitors 1 and 3 are

marked with “<1” and “<3”, respectively.

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Figure S4. Characterization of NT5DC1 and CHEK2 as Specific Off-Targets of Inhibitor 3, Related to Figure

3.

(A) Full-length labeled gel of the characterization of NT5DC1 as a specific off-target of inhibitor 3/probe 6, Figure

3A. Mock transfected lanes are marked with the letter M and probe-labeled NT5DC1 is marked with an arrow.

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(B) Full-length labeled gel of the characterization of CHEK2 as a specific off-target of inhibitor 3/probe 6, Figure

3B. Mock transfected lanes are marked with the letter M, and probe-labeled CHEK2 is marked with an arrow.

(C) Concentration-dependent competitive blockade of probe 6 labeling of NT5DC1 by inhibitor 3. HEK293T cells

recombinantly expressing NT5DC1 were treated with increasing concentrations of inhibitor 3 (2 hr) followed by

probe 6 (1 μM, 4 hr) and total cellular lysate was analyzed by gel-based ABPP (left, upper panel). Anti-NT5DC1

western blotting was used to confirm equivalent protein expression in each sample (left, bottom panel). Right graph,

in situ IC50 values for inhibitor 3, as measured by gel-based competitive ABPP. Data represent average values +

SEM for three independent experiments.

(D) Concentration-dependent competitive blockade of probe 6 labeling of CHEK2 by inhibitor 3. HEK293T cells

recombinantly expressing CHEK2 were treated with increasing concentrations of inhibitor 3 (2 hr) followed by

probe 6 (1 μM, 4 hr) and analyzed by gel-based ABPP (left, upper panel). Anti-Flag western blot was used to

confirm protein expression (left, bottom panel). Right graph, in situ IC50 values for inhibitor 3, as measured by gel-

based competitive ABPP. Data represent average values + SEM for three independent experiments.

(E) Concentration-dependent competitive blockade of probe 6 labeling of CHEK2 by inhibitor 2. HEK293T cells

recombinantly expressing CHEK2 were treated with increasing concentrations of inhibitor 2 (2 hr) followed by

probe 6 (1 μM, 4 hr) and analyzed by gel-based ABPP (upper panel). Anti-Flag western blot was used to confirm

protein expression (bottom panel) (n=2). Right graph, estimated IC50 value for inhibitor 2, as measured by gel-based

competitive ABPP. Data represent average values + SD for two independent experiments.

(F) Competitive ABPP analysis of purified CHEK2 with probe 6 pre-incubated with inhibitors 1-3. Purified CHEK2

protein was incubated with inhibitors 1-3 (1 hr) before the addition probe 6 (1 µM, 1 hr) and analyzed by gel-based

ABPP (upper panel). Equivalent protein loading was confirmed by coommassie staining (bottom panel).

(G) Competitive ABPP analysis of purified CHEK2 with probes 4-6 pre-incubated with inhibitors 1-3. Purified

CHEK2 protein was incubated with 1 µM inhibitors 1-3 (1 hr) before the addition of probe 4-6 (1 µM, 1 hr) and

analyzed by gel-based ABPP.

(H) Identification of the residue of inhibitor 3 reactivity on purified CHEK2. Inhibitor 3 treated CHEK2 was

digested with trypsin and analyzed by LC-MS. Shown is a representative MS2 spectrum of an inhibitor-modified

peptide containing C231 (highlighted in red), which was identified as a site of covalent modification. The y-axis

(relative intensity) was scaled to highlight fragment ions; unidentified fragment ion shown in grey.

(I) Inhibitor 3 modeled into the CHEK2 kinase crystal structure. Docking of inhibitor 3 into the CHEK2 structure

(PDBID:4BCD) highlights the potential for covalent engagement of C231 on the G-loop of CHEK2 by the

acrylamide of 3. Most known structures of CHEK2 kinase show a disordered G-loop, consistent with the knowledge

that the G-loop is a dynamic structural feature of kinases. The flexibility of the G-loop combined with the proximal

placement of the acrylamide of 3 is consistent with mass spectrometry data designating C231 as the site of covalent

modification by 3.

(J) HEK293T cells recombinantly expressing FLAG-tagged WT or C231A mutant CHEK2 were treated in situ with

probe 5 (1 μM, 4 hr) and analyzed by gel-based ABPP (upper panel). Anti-Flag western blotting was used to

confirm equivalent protein expression in the samples (lower panel).

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Figure S5. Characterization of Cathepsin Off-Targets and In Vitro Versus In Situ Reactivity Profiles for

NT5DC1 and CHEK2, Related to Figure 4.

(A) Full-length gel of the characterization of CTSC as a specific off-target of inhibitor 1/probe 4, Figure 4A. Mock

transfected lanes are marked with the letter M, and probe-labeled CTSC is marked with an arrow.

(B) Concentration-dependent competitive blockade of probe 4 labeling of CTSC labeling by inhibitor 1. HEK293T

cells recombinantly expressing CTSC were treated in situ with increasing concentrations of inhibitor 1 (2 hr)

followed by probe 4 (1 μM, 4 hr) and the soluble lysate was analyzed by gel-based ABPP (upper panel). Probe-

labeled CTSC is marked with an arrow. Anti-CTSC western blotting was used to confirm equivalent protein

expression in each sample (bottom panel).

(C) Comparing in vitro versus in situ reactivity of probe 4 with CTSC. HEK293T cells recombinantly expressing

CTSC were treated in situ (probe 4: 1 µM, 4 hr) or in vitro (probe 4: 5, 10, or 50 µM, 3 hr) and analyzed by gel-

based ABPP (upper panel). Probe-labeled CTSC is marked with an arrow. Anti-CTSC western blotting was used to

confirm equivalent protein expression in the samples (lower panel). Mock lanes are marked with the letter M.

(D) Comparing in vitro versus in situ labeling of NT5DC1 by probe 6. HEK293T cells recombinantly expressing

NT5DC1 were treated in situ (10 µM, 2 hr for inhibitor 3 and/or 1 µM, 4 hr for probe 6) or in vitro (10 µM, 1 hr for

inhibitor 3 followed by 5 µM, 1 hr or 1 µM, 3 hr for probe 6) and analyzed by gel-based ABPP (upper panel). Anti-

NT5DC1 western blotting was used to confirm equivalent protein expression in each sample (lower panel). Mock

lanes are marked with the letter M.

(E) Comparing in vitro versus in situ labeling of CHEK2 by probe 6. HEK293T cells recombinantly expressing

CHEK2 were treated in situ (10 µM, 2 hr for inhibitor 3 and/or 1 µM, 4 hr for probe 6) or in vitro (10 µM, 1 hr for

inhibitor 3 followed by 5 µM, 1 hr or 1 µM, 3 hr for probe 6) and analyzed by gel-based ABPP (upper panel).

Probe-labeled CHEK2 is marked with an arrow. Anti-Flag western blotting was used to confirm equivalent protein

expression in each sample (lower panel). Mock lanes are marked with the letter M.

(F) In situ reactivity of CTSC with probes 4 and 6 post-lysosomal neutralization. HEK293T cells recombinantly

expressing CTSC were treated with NH4Cl (10 mM, 15 min) or bafilomycin A1 (10 nM, 15 min) prior to treatment

with probe 4 or 6 (1 μM, 4 hr), after which cells were lysed and analyzed by gel-based ABPP (upper panel). Probe-

labeled CTSC is marked with an arrow. Anti-CTSC western blotting was used to confirm equivalent protein

expression in the samples (lower panel). Mock lanes are marked with the letter M.

(G) ABPP-SILAC experiments evaluating probe 4 reactivity in bafilomycin- versus DMSO-treated H1975 cells.

ABPP-SILAC experiments in the soluble proteome of H1975 cells treated with probe 4 (1 μM, 4 hr) and either

DMSO or 10 nM bafilomycin. Data represent the mean SILAC ratio value for proteins across two biological

replicates. CTSC and CTSL1 are highlight by the red dots.

(H) In situ reactivity of probe 4 post-lysosomal neutralization in H1975 cells and western blot analysis of cathepsins.

H1975 cells were treated with NH4Cl (10 mM, 15 min) or bafilomycin A1 (10 nM, 15 min) prior to probe 4 labeling

in situ (1 μM, 4 hr) and the soluble lysate analyzed by gel-based ABPP (upper panel). Lysates were transferred and

western blotted for CTSC (middle panel) and CTSL1 (bottom panel).

(I) LTR (LysoTracker Red) staining of ARPE-19 cells treated with inhibitors 1-3. ARPE-19 cells were treated with

indicated concentrations of inhibitors 1-3 (2 hr) and LTR staining was quantified. Data represents average values +

SEM for three independent experiments.

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II. Supplemental Tables

H1975 (IC50)

EGFR-pY1068 (nM)A

GSH T½

(canertinib/inhibitor)B

Inhibitor 1 12 ± 1 3 ± 0.4

Probe 4 76 ± 23 5 ± 0.4

Inhibitor 2 17 ± 2 15 ± 2.2

Probe 5 20 ± 2 >17

Inhibitor 3 14 ± 2 8 ± 0.6

Probe 6 11 ± 1

6 ± 0.3

Table S1, Related to Figure 1. Characterizing third-generation EGFR inhibitor/probe pairs. (A) Cellular IC50

values for blockade of Y1068 autophosphorylation of EGFR in H1975; data represent average values ± SD; n = 5

for inhibitors 1-3 and n = 4 for probes 4-6. (B) Chemical reactivity with glutathione relative to the second-

generation EGFR inhibitor canertinib (half-life in glutathione buffer at pH 7.4). Data represent average values ± SD;

n = 2 per group.

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A431 (IC50)

EGFR-pY1068 (nM)A

A431 (IC50)

Competition assay (nM)B

Inhibitor 1 550 ± 240 540 ± 12

Inhibitor 2 190 ± 46 440 ± 39

Inhibitor 3 3400 ± 1200 3000 ± 1900

Table S2, Related to Figure 1. Characterizing third-generation EGFR inhibitors for: (A) Cellular IC50 values for

blockade of Y1068 autophosphorylation of WT-EGFR in A431 (n = 10-30 per group, ± SD), and, (B) Competitive

blockade of probe labeling of WT-EGFR in A431 cells for the following inhibitor/probe pairs (n = 2 per group, ±

SD): inhibitor 1/probe 4, inhibitor 2/probe 5, and inhibitor 3/probe 6. Also see Figure S2A for representative ABPP

gels from which IC50 values were derived.

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Probe 4 Probe 4 Probe 5 Probe 6

(DMSO/10 µM

inhibitor 1) A

(DMSO/1 µM

inhibitor 1) B

(DMSO/10 µM

inhibitor 2) C

(DMSO/10 µM

inhibitor 3) D

CHEK2 ND ND 2.5 5.4

CLN3 ND ND ND 10.7

CTSC 17 6.0 ND 2.6

CTSL1 20 3.5 ND ND

CTSH 5.7 1.1 ND 0.4

EGFR 17 10 13 6.6

ERBB2 19 13 12 4.4

IFI30 5.4 1.1 ND ND

NT5DC1 ND ND 0.6 20

RFTN1 11 6.0 ND 1.1

RTN4 0.8 0.6 ND 6.3

SCARB1 7.2 2.0 ND 0.9

TEC 20 20 20 20

TEX264 10.1 2.1 ND 0.9

TNK1 ND ND ND 5.3

TRMT61A 1.3 0.9 0.6 4.9

Table S3, Related to Figure 2 and Table 1. High-occupancy protein targets of inhibitors 1-3 identified by

competitive ABPP-SILAC analysis with probes 4-6. Competition ratios are displayed as DMSO/inhibitor-treated

cells with increasing ratio values reflecting greater inhibitor competition. A value of 20 was defined as maximal

competition that could be quantified. (A) 10 µM inhibitor 1 (2 hr) (B) 1 µM inhibitor 1 (2 hr), (C) 10 µM inhibitor 2

(2 hr), and (D) 10 µM inhibitor 3 (2 hr). ND = Not detected. Competed proteins were required to have at least 8

quantified peptides, SEM < 5.2 and identified in two of the three biological replicates with a minimum of three-fold

competition. TNK1 for probe 6 is uniquely designated as an enriched and competed target of inhibitor 3 due to the

high quality of competition data even though it was found to have an enrichment SEM > 5.2.

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III. Methods S1. Synthetic details for chemical probes, Related to STAR Methods.

Information regarding the purification, NMR and exact mass measurements can be found in STAR methods.

Synthesis of Probe 4.

N-(2-((2-(But-3-yn-1-yl(methyl)amino)ethyl)(methyl)amino)-4-methoxy-5-((4-(1-methyl-1H-indol-3-yl)pyrimidin-

2-yl)amino)phenyl)acrylamide) (4). Synthesized in analogy to published methods: (Butterworth et al., 2013).

https://www.google.com/patents/WO2013014448A1?cl=en

1H NMR (400 MHz, DMSO-d6) δ ppm 9.74 (s, 1H), 9.08 (s, 1H), 8.65 (s, 1H), 8.32 (d, J=4.8 Hz, 1H), 8.25 (d,

J=8.0 Hz, 1H), 7.91 (s, 1H), 7.52 (d, J=8.3 Hz, 1H), 7.32 - 7.19 (m, 2H), 7.19 - 7.10 (m, 1H), 7.01 (s, 1H), 6.53 (dd,

J=9.9, 16.9 Hz, 1H), 6.26 (d, J=17.1 Hz, 1H), 5.77 (d, J=9.3 Hz, 1H), 3.91 (s, 3H), 3.86 (s, 3H), 2.93 - 2.84 (m, 2H),

2.82 - 2.77 (m, 1H), 2.71 (s, 3H), 2.63 - 2.57 (m, 2H), 2.44-2.41 (m, 2H), 2.35 - 2.29 (m, 2H), 2.21 (s, 3H).

14

13C NMR (101MHz, DMSO-d6) d 162.5, 161.6, 159.9, 157.7, 146.1, 137.9, 137.7, 133.8, 132.5, 126.9, 126.1, 125.3,

125.1, 122.0, 121.3, 120.9, 113.9, 112.4, 110.5, 107.1, 104.9, 83.2, 71.8, 56.0, 56.0, 55.4, 54.0, 42.1, 41.9, 32.9,

15.8.

m/z (APCI+) for C31H36N7O2 538.2939 (calculated 538.2925 Δ ppm -2.6) (M+H)+.

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Synthesis of Probe 5

N-[(3R,4R)-1-(6-([1-(But-3-yn-1-yl)-3-methoxy-1H-pyrazol-4-yl]amino)-9-methyl-9H-purin-2-yl)-4-

fluoropyrrolidin-3-yl]prop-2-enamide (5). Synthesized in analogy to submitted methods: (Planken et al., 2017).

1H NMR (400 MHz, DMSO-d6) δ ppm 8.43 (d, J=6.6 Hz, 1H), 7.95 (s, 2H), 7.79 (s, 1H), 6.09 - 6.31 (m, 2H), 5.62

(dd, J=9.7, 2.6 Hz, 1H), 5.03 - 5.23 (m, 1H), 4.46 (dt, J=11.9, 6.0 Hz, 1H), 4.10 (t, J=6.9 Hz, 2H), 3.84 (s, 4H), 3.73

- 3.83 (m, 2H), 3.67 (d, J=12.1 Hz, 1H), 3.62 (s, 3H), 2.83 (t, J=2.6 Hz, 1H), 2.64 (td, J=6.8, 2.6 Hz, 2H).

13C NMR (101 MHz, DMSO-d6) d 164.7, 157.3, 154.9, 151.8, 151.5, 138.8, 131.1, 126.1, 124.9, 113.2, 105.4, 94.3

(d, JC-F=179.0 Hz), 81.5, 72.8, 56.0, 53.1 (d, JC-F=29.0 Hz), 51.1 (d, JC-F=20.0 Hz), 50.1, 49.7, 29.0, 19.7.

19F NMR (377 MHz, DMSO-d6) δ ppm -177.8 (s, 1F).

m/z (APCI+) for C21H25FN9O2 454.2117 (calculated 454.2110 Δ ppm -1.5) (M+H)+.

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Synthesis of Probe 6

N-(3-((2-((2-Methoxy-4-(4-(pent-4-ynoyl)piperazin-1-yl)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-

yl)amino)phenyl)acrylamide (6). Synthesized in analogy to published methods: (Lee et al., 2015)

https://www.google.com/patents/US8975249.

1H NMR (400 MHz, DMSO-d6) δ ppm 10.13 (s, 1H), 8.61 (br. s., 1H), 8.28 (s, 1H), 8.06 (s, 1H), 7.76 (br. s., 1H),

7.56 - 7.48 (m, 2H), 7.26 (t, J=8.0 Hz, 1H), 7.17 (br. s., 1H), 6.60 (d, J=2.3 Hz, 1H), 6.44 (dd, J=10.0, 17.0 Hz, 1H),

17

6.26 (dd, J=2.0, 17.0 Hz, 1H), 6.22 (br. s, 1H), 5.75 (dd, J=2.0, 10.0 Hz, 1H), 3.77 (s, 3H), 3.62 - 3.55 (m, 4H), 3.10

- 2.97 (m, 4H), 2.76 (t, J=2.6 Hz, 1H), 2.63 - 2.56 (m, 2H), 2.39 (dt, J=2.4, 7.3 Hz, 2H).

13C NMR (101 MHz, DMSO-d6) d 168.9, 163.1, 160.9, 157.2, 155.8, 155.7, 148.1, 139.0, 138.4, 131.9, 128.8,

128.5, 124.8 (q, JC-F=269.0 Hz), 126.9, 120.8, 120.4, 120.1, 116.5, 115.9, 107.0, 100.6, 84.1, 71.2, 55.6, 49.4, 49.0,

44.6, 41.0, 31.3, 13.9.

19F NMR (377 MHz, DMSO-d6) δ ppm -59.02 (s, 3F).

m/z (APCI+) for C30H31F3N7O3 594.2418 (calculated 594.2435, Δ ppm 2.9) (M+H)+.

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Synthesis of Probe DCG-04 - Rhodamine

Ethyl (2S,3S)-3-(((14S,24S,27S)-1-(3',6'-bis(dimethylamino)-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthen]-5-yl)-14-

carbamoyl-24-(4-hydroxybenzyl)-29-methyl-1,8,16,23,26-pentaoxo-2,9,15,22,25-pentaazatriacontan-27-

yl)carbamoyl)oxirane-2-carboxylate (Rh-DCG-04). Synthesized in analogy to published methods: (Bogyo et al.,

2000, Greenbaum et al., 2000).

1H NMR (500 MHz, MeOD) δ ppm 8.87 (app. t, J=4.8, 0.5H), 8.77 (d. J=1.5, 1H), 8.26 (dd, J=1.5, 8.0, 1H), 7.80

(app. t, J=5.5, 0.5H), 7.52 (d, J=8.0, 1H), 7.13 (dd, J=0.5, 9.5, 2H), 7.05 (d, J=9.5, 2H), 7.01-6.98 (m, 4H), 6.67 (d,

J=8.5, 2H), 4.44 (m, 1H), 4.37 (m, 1H), 4.30 (m, 1H), 4.28-4.18 (m, 2H), 3.65 (d, J=2.0, 1H), 3.56 (d, J=2.0, 1H),

3.47 (m, 2H), 3.20-3.10 (m, 4H), 3.05 (m, 1H), 2.95 (m,1H), 2.85-2.75 (m, 1H), 2.22 (app. t, J=7.5, 4H), 1.85-1.75

(m, 1H), 1.75-1.60 (m, 5H), 1.60-1.35 (m, 14H), 1.30 (app. t, J=7.5, 4H), 1.25-1.15 (m, 2H), 0.92 (d, J=6.5, 3H),

0.89 (d, J=6.5, 3H).

m/z (ESI+) for C64H83N9O14 1202.6113 (calculated 1202.6132, Δ ppm 1.6) (M+H)+.

19

IV. References

BUTTERWORTH, S., FINLAY, M. R. V., WARD, R. A., KADAMBAR, V. K., CHANDRASHEKAR, R. C., MURUGAN, A. & REDFEARN, H. M. (2013). 2 - (2, 4, 5 - substituted -anilino) pyrimidine derivatives as egfr modulators useful for treating cancer. Google Patents.

LEE, K., NIU, D., PETTER, R. C., BAEVSKY, M. F. & SINGH, J. (2015). Heterocyclic compounds and uses thereof. Google Patents.