A Single-Dose Study of Orally Administrated Defactinib or VS-6766 in Patients With Glioblastoma
Phase: EARLY_PHASE1
Conditions: Glioblastoma, Recurrent Glioblastoma
Sponsor: Emory University

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 1 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
P R O T O C OL TI TL E: A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n 
p ati e nts wit h gli o bl ast o m a
WI N S HI P P R O T O C OL #: WI N S HI P 5 7 5 1- 2 2
I N D #: 1 6 4 3 0 5
C O O R DI N A TI N G C E N T E R: E m or y Wi ns hi p C a nc er I nstit ut e
P RI N CI P AL I N V E S TI G A T O R:
N a m e: 
J effr e y J. Ols o n, M D
D e p art m e nt: 
N e ur os ur g er y 
T el e p h o n e N u m b er:
4 0 4- 7 7 8- 3 0 9 1 
E m ail A d dr ess:
j ols o n @ e m ory. e d u
C O-I N V E S TI G A T O R S:
E X T E R N AL ( N O N- E M O R Y) C OLL A B O R A T O R S:
N a m e 
Lili a K uc h er y a v y k h
Titl e: 
Ass oci at e Pr of ess or
A d dr ess 
D e pt. of Bi oc h e mistr y, U ni v ersi d a d C e ntr al d el C ari b e, P u ert o Ric o 
T el e p h o n e 
7 8 7- 7 9 8 3 0 0 1
e- m ail a d dr ess:
lili a. k uc h er y a v y k h @ ucc ari b e
I R B:                                    U NI V E R SI D A D C E N T R AL D EL C A RI B E E S C U EL A D E M E DI CI N A C O MI T E D E D E R E C H O S 
H U M A N O S (I R B)  I N S TI T U TI O N AL R E VI E W B O A R D
V E R SI O N: 6. 0, 8/ 3 0/ 2 0 2 4
F U N DI N G S O U R C E : V er ast e m, I nc. a n d U ni v ersi d a d C e ntr al d el C ari b e

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 2 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
R E VI SI O N HI S T O R Y
R e visi o n #
V ersi o n 
D at e
S u m m ar y of C h a n g es
1
9- 2 2- 2 3
P a g e 5: R e m o v al of i m m u n ol o g y ass a ys a n d 
p h ar m ac o ki n etics fr o m scr e e ni n g p eri o d i n D ef acti ni b 
sc h e d ul e of e v e nts
P a g e 6: R e m o v al of i m m u n ol o g y ass a ys a n d 
p h ar m ac o ki n etics fr o m scr e e ni n g p eri o d i n Av ut o m eti ni b 
sc h e d ul e of e v e nts.
2
1- 3 1- 2 4
Gr a m m atic al u p d at es t hr o u g h o ut
S ecti o n 6. 1: U p d ati n g bl o o d s a m pl e a m o u nt a n d t y p e of 
t u b e b ei n g us e d t o o bt ai n t h e s a m pl e
3
3- 1 9- 2 4
Gr a m m atic al u p d at es t hr o u g h o ut
U p d at e d eli gi bilit y t o i ncl u d e acti v e e y e dis or d ers
4
4- 4- 2 4
U p d ati n g accr u al n u m b ers t o acc o u nt f or scr e e n f ail ur es a n d 
gr a m m atic al u p d at es t hr o u g h o ut
5
7- 3 0- 2 4
U p d at es t o sc h e d ul e of ass ess m e nts
6
8- 3 0- 2 4
U p d at e d e nr oll m e nt n u m b ers

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 3 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
T a bl e of C o nt e nts
1.
St u d y S u m m ar y
3
2.
O bj ecti v es
3
3.
B ac k gr o u n d
4
4.
St u d y E n d p oi nts
5
5.
St u d y I nt er v e nti o n/I n v esti g ati o n al A g e nt 
6
6.
Pr oc e d ur es I n v ol v e d 
8
7.
St atistic al A n al ysis Pl a n 
9
8.
D at a S p eci m e n B a n ki n g
1 2
9.
S h ari n g of R es ults wit h P artici p a nts
1 2
1 0. St u d y Ti m eli n es
1 3
1 1.
I ncl usi o n a n d Excl usi o n Crit eri a
1 3
1 2.
V ul n er a bl e P o p ul ati o ns
1 5
1 3.
L oc al N u m b er of P artici p a nts
1 5
1 4.
R ecr uit m e nt M et h o ds
1 5
1 5.
Wit h dr a w al of P artici p a nts
1 6
1 6.
Ris k t o P artici p a nts
1 6
1 7.
P ot e nti al B e n efits t o P artici p a nts
1 7
1 8.
C o m p e ns ati o n t o P artici p a nts
1 7
1 9.
D at a M a n a g e m e nt a n d C o nfi d e nti alit y
1 7
2 0.
Pr o visi o ns t o M o nit or t h e D at a t o E ns ur e t h e S af et y of P artici p a nts
1 8
2 1.
Pr o visi o ns t o Pr ot ect t h e Pri v ac y I nt er est of P artici p a nts
1 9
2 2.
Ec o n o mic B ur d e n t o P artici p a nts
1 9
2 3.
I nf or m e d C o ns e nt
1 9
2 4.
S etti n g
2 0
2 5.
R es o urc es A v ail a bl e 
2 1
2 6.
M ulti- Sit e R es e arc h W h e n E m or y is t h e L e a d Sit e
2 2
2 7.
R ef er e nc es
2 2

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 4 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
1.
St u d y S u m m ar y
Pr oj ect Titl e
I d e ntific ati o n of tr e at m e nt c o nc e ntr ati o ns of 
d ef acti ni b a n d a v ut o m eti ni b f or gli o bl ast o m a 
tr e at m e nt
Pr oj ect D esi g n
L a b or at or y i n v esti g ati o n of p osts ur gic al gli o bl ast o m a 
s p eci m e ns aft er si n gl e d os e or al a d mi nistr ati o n of 
d ef acti ni b or a v ut o m eti ni b
Pri m ar y O bj ecti v e(s)

C h ar act eriz e or d escri b e t h e a bilit y of a n or al 
d os e of d ef acti ni b a n d a v ut o m eti ni b t o 
r e ac h t u m or a n d br ai n ar o u n d t u m or

Ass ess t h e s af et y of t h e a d mi nistr ati o n of a 
si n gl e or al d os e of d ef acti ni b or a v ut o m eti ni b 
i n p ati e nts wit h gli o bl ast o m a
S ec o n d ar y O bj ecti v e(s)

Ass ess t h e a bilit y of d ef acti ni b a n d 
a v ut o m eti ni b t o i n hi biti o n of P y k 2/ F A K a n d 
M E K/ E R K p h os p h or yl ati o n i n t h os e s a m pl es, 
r es p ecti v el y

Ass ess t h e p h ar m ac o d y n a mics of d ef acti ni b 
a n d a v ut o m eti ni b
R es e arc h
I nt er v e nti o n(s)/I nt er acti o ns
Si n gl e d os e or al a d mi nistr ati o n
St u d y P o p ul ati o n
S u bj ects ol d er t h a n 2 1 y e ars of a g e di a g n os e d wit h 
gli o bl ast o m a ( G B M) t h at r e q uir e s ur g er y as p art of 
t h eir m a n a g e m e nt
S a m pl e Siz e
1 2 p ati e nts
St u d y D ur ati o n f or 
i n di vi d u al p artici p a nts
Fr o m ti m e of c o ns e nt u ntil s ut ur e or st a pl e r e m o v al, 
e nc o m p assi n g a p pr oxi m at el y 3 w e e ks.
St u d y S p ecific
A b br e vi ati o ns/ D efi niti o ns
Gli o b ast o m a ( G B M)
F u n di n g S o urc e (if a n y)
V er ast e m, I nc

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 5 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
Pr oc e d ur es f or S u bj ects r ec ei vi n g 
D ef acti ni b
Scr e e ni n g
D a y 1- 1 4
D ef acti ni b
( 2 0 0 m g)
Or  
( 4 0 0 m g) *
S ur gic al 
r es ecti o n 
e v al u ati o n
P ost- S ur gic al 
m o nit ori n g
F or at l e ast t h e 4 
h o urs f oll o wi n g 
s ur g er y
E n d of tr e at m e nt 
visit
2 w e e ks p ost 
s ur g er y
I nf or m e d c o ns e nt
X
D e m o gr a p hics
X
M e dic al hist or y
X
C o n. M e dic ati o n r e vi e w
A d mi nist er st u d y i nt er v e nti o n
X
P h ysic al ex a m (i ncl u di n g h ei g ht a n d 
w ei g ht)
X
X
El ectr oc ar di o gr a m ( E K G) d
X
Ec h oc ar di o gr a m ( E C H O) e
X
St a pl e r e m o v al; p h ysic al ex a m 
(i ncl u di n g s ki n a n d or al)
X
Vit al si g ns
X
X
X
H ei g ht
X
W ei g ht
X
S er u m c h e mistr y a
X
X f
Pr e g n a nc y t est b
X
Tiss u e s a m pl e extr acti o n
X
A Es r evi e w a n d e v al u ati o n
X -----------------------------------------------------------------------
X
Ot h er ass ess m e nts ( e. g., 
i m m u n ol o g y ass a ys, 
p h ar m ac o ki n etic) c
X
C o m pl et e C as e R e p ort F or ms ( C R Fs)
X
X
X
X
X
1. 2 Sc h e d ul e of Ass ess m e nts
*I n D ef acti ni b ar m, t hr e e s u bj ects will r ec ei v e 2 0 0 m g d os e. If n o d os e li miti n g t oxiciti es occ ur, t hr e e s u bj ects will r ec ei v e 4 0 0 m g d os e.

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 6 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
a)  
M et a b olic p a n el, li v er f u ncti o n p a n el i ncl u di n g bilir u bi n l e v el, ki d n e y f u ncti o n p a n el, Q Tc e v al u ati o n, P T/I N R/ P T T
b) 
S er u m pr e g n a nc y t est wit hi n 2 w e e ks of st u d y dr u g a d mi nistr ati o n ( w o m e n of c hil d b e ari n g p ot e nti al)
c)  
W est er n bl ot a n al ysis of r ati o of P y k 2, F A K, M E K 1/ 2 a n d Er k 1/ 2
d) 
E K G, Q Tc t o occ ur wit hi n 2 w e e ks of st u d y dr u g a d mi nistr ati o n
e) 
E C H O t o occ ur wit hi n 2 w e e k of st u d y dr u g a d mi nistr ati o n
f)  
M et a b olic p a n el, li v er f u ncti o n p a n el i ncl u di n g bilir u bi n l e v el, ki d n e y f u ncti o n p a n el, a n d P T/I N R/ P T T

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 7 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
*I n A V U T O M E TI NI B ar m, t hr e e s u bj ects will r ec ei v e 3. 2 m g d os e. If n o d os e li miti n g t oxiciti es occ ur, t hr e e s u bj ects will r ec ei v e 4 m g d os e.
Pr oc e d ur es f or S u bj ects r ec ei vi n g 
A V U T O M E TI NI B
Scr e e ni n g
A V U T O M E TI NI B
( 3. 2 m g)
Or 
( 4 m g) *
S ur gic al 
r es ecti o n 
e v al u ati o n
P ost- S ur gic al 
m o nit ori n g
F or at l e ast t h e 
4 h o urs 
f oll o wi n g 
s ur g er y
E n d of 
tr e at m e nt 
visit
2 w e e ks p ost 
s ur g er y
I nf or m e d c o ns e nt
X
D e m o gr a p hics
X
M e dic al hist or y
X
C o n. M e dic ati o n r e vi e w
A d mi nist er st u d y i nt er v e nti o n
X
P h ysic al ex a m (i ncl u di n g h ei g ht 
a n d w ei g ht)
X
X
El ectr oc ar di o gr a m ( E K G)
X
Ec h oc ar di o gr a m ( E C H O)
X
St a pl e r e m o v al
X
Vit al si g ns
X
X
X
H ei g ht
X
W ei g ht
X
S er u m c h e mistr y a
X
X f
Pr e g n a nc y t est b
X
Tiss u e s a m pl e extr acti o n
X
A Es r e vi e w a n d e v al u ati o n
X -----------------------------------------------------------------------
X
Ot h er ass ess m e nts ( e. g., 
i m m u n ol o g y ass a ys, 
p h ar m ac o ki n etic) c
X
C o m pl et e C as e R e p ort F or ms 
( C R Fs)
X
X
X
X
a)  
M et a b olic p a n el, li v er f u ncti o n p a n el i ncl u di n g bilir u bi n l e v el, ki d n e y f u ncti o n p a n el, Q Tc e v al u ati o n, a n d P T/I N R/ P T T.
b) 
S er u m pr e g n a nc y t est wit hi n 2 w e e ks of st u d y dr u g a d mi nistr ati o n ( w o m e n of c hil d b e ari n g p ot e nti al)
c)  
W est er n bl ot a n al ysis of r ati o of P y k 2, F A K, M E K 1/ 2 a n d Er k 1/ 2.

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 8 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
d) 
E K G, Q Tc t o occ ur wit hi n 2 w e e ks of st u d y dr u g a d mi nistr ati o n
e) 
E C H O t o occ ur wit hi n 2 w e e k of st u d y dr u g a d mi nistr ati o n
f)  
M et a b olic p a n el, li v er f u ncti o n p a n el i ncl u di n g bilir u bi n l e v el, ki d n e y f u ncti o n p a n el, a n d P T/I N R/ P T T
2.
O bj ecti v es
T his st u d y is d esi g n e d as a p h as e 0 cli nic al tri al w h os e p ur p os e is t o esti m at e t h e w or ki n g 
c o nc e ntr ati o ns of P y k 2/ F A K i n hi bit or d ef acti ni b a n d R A F/ M E K cl a m p a v ut o m eti ni b, as w ell as 
t h e i n hi biti o n of P y k 2/ F A K a n d M E K/ E R K p h os p h or yl ati o n i n t u m or tiss u e a n d br ai n ar o u n d 
t u m or p ati e nts wit h gli o bl ast o m a, r es p ecti v el y.
O bj ecti v es: ( 1) esti m at e ( or c h ar act eriz e) t h e c o nc e ntr ati o n of d ef acti ni b or
a v ut o m eti ni b t h at acc u m ul at es i n t h e G B M a n d br ai n ar o u n d t u m or, ( 2) ass ess t h e s af et y of 
t h e a d mi nistr ati o n of a si n gl e or al d os e of d ef acti ni b or a v ut o m eti ni b i n p ati e nts wit h 
gli o bl ast o m a, ( 3) ass ess t h e i n hi biti o n of P y k 2/ F A K or M E K, Er k si g n ali n g i n t u m or a n d br ai n 
ar o u n d t u m or, r es p ecti v el y, a n d ( 4) ass ess t h e p h ar m ac o d y n a mics of d ef acti ni b or 
a v ut o m eti ni b i n p ati e nts wit h gli o bl ast o m a.
H y p ot h esis: A d mi nistr ati o n of d ef acti ni b or a v ut o m eti ni b i n hi bits P y k 2/ F A K a n d R A F/ M E K 
si g n ali n g i n G B Ms, r es p ecti v el y.
3.
B ac k gr o u n d
O ur pr e vi o us st u di es a n d p u blis h e d lit er at ur e r e p orts h a v e i d e ntifi e d d er e g ul ati o n of F A K/ P y k 2 
a n d R A F/ M E K si g n ali n g i n h u m a n G B Ms, r el at e d t o o v er acti v ati o n a n d g ai n of g e n e c o p y 
n u m b er of t h eir u p-str e a m r e g ul at ors E G F R, P D G F R, a n d F G F R r ec e pt or t yr osi n e ki n as es ( N u n ez 
et al., 2 0 2 1; R o bi ns o n et al., 2 0 1 0). Alt h o u g h o nc o g e nic dri v er K R A S a n d B R A F m ut ati o ns ar e 
n ot c o m m o n i n G B Ms ( M a kit o et al., 2 0 2 1), B R A F V 6 0 0 h as b e e n r e p ort e d i n 6 9 % of e pit h eli oi d 
gli o bl ast o m as ( A n dr e ws et al., 2 0 2 2). A d diti o n all y, a si g nific a nt 1 2-f ol d u p-r e g ul ati o n of B R A F 
a n d 2-f ol d u p-r e g ul ati o n of R A F- 1 w er e f o u n d i n t h e m aj orit y of G B M tiss u e s a m pl es c o m p ar e d 
t o n or m al br ai n c ort ex (L y usti k m a n et al., 2 0 0 8). N F 1 m ut ati o ns a n d d el eti o ns, w hic h ar e t h e 
h all m ar k f or G B M wit h m es e nc h y m al m ol ec ul ar si g n at ur es ( P hilli ps et al, 2 0 0 6; V er h a a k et al, 
2 0 1 0), h a v e b e e n s h o w n t o acc el er at e Kr as - dri v e n t u m ori g e n esis ( B e h n a n et al., 2 0 1 9; Xi a oji n g 
et al., 2 0 1 9) a n d ar e als o ass oci at e d wit h F A K h y p er acti v ati o n ( Xi a oji n g et al., 2 0 1 9). T a ki n g i n 
acc o u nt t h e fr e q u e nc y of E G F R m ut ati o ns ( 3 1. 5 4 %, acc or di n g t o M y C a nc er G e n o m e d at a b as e 
a n d S a g e S y n a ps e pl atf or m: htt ps:// w w w. m yc a nc er g e n o m e. or g , htt ps://s y n a ps e. or g/ g e ni e ), 
P D G F R m ut ati o ns ( 1 0 %) a n d N F 1 m ut ati o ns ( 1 1- 1 9 %) i n n e wl y di a g n os e d G B Ms t o g et h er wit h 
t h e o bs er v ati o ns t h at t w o-t hir ds of pri m ar y G B Ms e v e nt u all y d ev el o p a tr a nscri pti o n al pr ofil e 
c o nsist e nt wit h t h e m es e nc h y m al G B M m ol ec ul ar-si g n at ur e u p o n r ec urr e nc e ( P hilli ps et al., 
2 0 0 6), R A F a n d F A K si g n ali n g ar e p ot e nti al t ar g ets i n n e wl y di a g n os e d a n d r ec urr e nt G B Ms, a n d 
m a y b e es p eci all y i m p ort a nt i n e pit h eli oi d gli o bl ast o m as.
As it w as s h o w n i n o ur pr e vi o us st u di es a n d b y ot h ers, F A K/ P y k 2 a n d R A F ex pr essi o n i n 
m ali g n a nt gli o m as ar e ess e nti al f or t h e m ai nt e n a nc e of t u m or pr olif er ati o n a n d dis p ers al
(Li pi ns ki et al., 2 0 0 8; L oft us et al., 2 0 0 9; R ol o n- R e y es et al., 2 0 1 5; P a oli n o et al., 2 0 1 0; N u n ez et 

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 9 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
al., 2 0 2 1). T ar g eti n g of F A K/ P y k 2, as w ell as R A F/ M E K, i n gli o m a a ni m al m o d els r es ult e d i n 
si g nific a nt i n hi biti o n of t u m or gr o wt h (Li pi ns ki et al., 2 0 1 8, S at h or ns u m et e e et al., 2 0 0 6).
H o w e v er, h u m a n cli nic al tri als i d e ntifi e d j ust m o d est pr o gr essi o n-fr e e s urvi v al i n p ati e nts, 
r ec ei vi n g i n hi bit ors of F A K/ P y k 2 or R A F si g n ali n g i n di vi d u all y ( Cl o u g h es y et al., 2 0 0 6; Li u et al., 
2 0 2 1). C o m p e ns at or y u p-r e g ul ati o n of F A K/ P y k 2 si g n ali n g w as o bs er v e d i n c a nc er c ells i n 
r es p o ns e t o R A F i n hi biti o n, dri vi n g t h e tr e at m e nt r esist a nc e ( Hir at a et al., 2 0 1 5). C o-i n hi biti o n 
of B R A F a n d F A K/ P y k 2 a b olis h e d c o m p e ns at or y Er k r e acti v ati o n, l e a di n g t o b ett er c o ntr ol of 
t u m or gr o wt h.
A v ut o m eti ni b is a n o v el R A F/ M E K i n hi bit or ( R A F/ M E K cl a m p) t h at bl oc ks b ot h R A F a n d M E K vi a 
a si n gl e m ol ec ul e a n d h as t h e a d v a nt a g e of i n hi biti n g d o w nstr e a m Er k si g n ali n g m or e 
c o m pl et el y t h a n ot h er t ar g et e d a g e nts. d ef acti ni b is a s m all m ol ec ul e f oc al a d h esi o n ki n as e 
( F A K) i n hi bit or t h at i n hi bits p ar all el p at h w a y si g n ali n g a n d h as d e m o nstr at e d s y n er g y wit h 
R A F/ M E K i n hi bit or A V U T O M E TI NI B ( d at a o bt ai n e d fr o m pr eli mi n ar y cli nic al st u di es, c o n d uct e d 
b y dr u g s p o ns or of t h e st u d y V er ast e m I nc). I n cli nic al o bs erv ati o ns, a v ut o m eti ni b i n d uc e d 
el e v at e d F A K p h os p h or yl ati o n ( p F A K) as a p ot e nti al r esist a nc e m ec h a nis m i n t h e m aj orit y of 
p ati e nts, w hil e t h e c o m bi n ati o n of a v ut o m eti ni b a n d d ef acti ni b r e d uc e d t his c o m p e ns at or y 
p F A K si g n al, l e a di n g t o b ett er tr e at m e nt o utc o m e.
As a n o v er arc hi n g c o nc e pt, w e h y p ot h esiz e t h at wit h pr o p er tr e at m e nt d ev el o p m e nt, d u al 
i n hi biti o n of R A F/ M E K a n d F A K/ P y k 2 si g n ali n g wit h us e of c o m bi n at ori al a d mi nistr ati o n of 
a v ut o m eti ni b a n d d ef acti ni b f or G B M tr e at m e nt will t ar g et a n d i n hi bit t u m or c ell pr olif er ati o n. 
As a n i niti al st e p, p h ar m ac o d y n a mic st u di es of i n di vi d u al a d mi nistr ati o ns of a v ut o m eti ni b a n d 
d ef acti ni b i n G B M tiss u es will pr o vi d e a b asis f or n ext st e p d e v el o p m e nt of c o m bi n at ori al 
t h er a pi es utilizi n g P y k 2/ F A K a n d R A F/ M E K i n hi bit ors t o pr o vi d e si g nific a nt tr e at m e nt b e n efit.
4.
St u d y E n d p oi nts
Pri m ar y E n d p oi nts:
1.
C o nc e ntr ati o n of d ef acti ni b t h at acc u m ul at es i n t h e G B M a n d br ai n ar o u n d t u m or.
2.
C o nc e ntr ati o n of a v ut o m eti ni b t h at acc u m ul at es i n t h e G B M a n d br ai n ar o u n d t u m or.
3.
Ass ess m e nt of s af et y of d ef acti ni b b y q u a ntific ati o n of t h e r ec o g niz e d si d e eff ects of t his 
a g e nt i ncl u di n g f ati g u e, n a us e a, di arr h e a, v o miti n g, h y p er bilir u bi n e mi a, d ecr e as e d 
a p p etit e, p eri p h er al e d e m a, dizzi n ess, a n d h e a d ac h e a n d b y m o nit ori n g f or n e w or 
u n d escri b e d a d v ers e e v e nts.
4.
Ass ess m e nt of s af et y of a v ut o m eti ni b b y q u a ntific ati o n of t h e r ec o g niz e d si d e eff ects of 
t his a g e nt i ncl u di n g r as h, cr e ati n e p h os p h o ki n as e el e v ati o n, vis u al dist ur b a nc es,

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 1 0 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
h y p o al b u mi n e mi a, a n d f ati g u e a n d b y m o nit ori n g f or n e w or u n d escri b e d a d v ers e ev e nts.
S ec o n d ar y E n d p oi nts:
1.
p P y k 2 a n d p F A K p h os p h or yl ati o n i n t u m or, br ai n ar o u n d t u m or, a n d s er u m.
2.
M E K a n d Er k i n t u m or, br ai n ar o u n d t u m or, a n d s er u m.
3.
P h ar m ac o d y n a mics of d ef acti ni b a n d a v ut o m eti ni b
Pr e vi o us cli nic al tri als h a v e i d e ntifi e d d os e of d ef acti ni b wit h bi ol o gic eff ect as 4 0 0 m g t wic e p er 
d a y ( BI D) a n d m axi m u m t ol er at e d d os e as 7 5 0 m g gi v e n or all y (J o n es et al., 2 0 1 5), a n d t h e 
o pti m u m d os e of a v ut o m eti ni b as 3. 2- 4 m g t wic e p er w e e k ( BI W) ( G u o et al., 2 0 2 0). T h e s a m e 
st u di es i d e ntifi e d t h at m axi m u m o bs er v e d s er u m c o nc e ntr ati o n is ac hi e v e d 1- 3 h o urs aft er 
a d mi nistr ati o n f or b ot h dr u gs.
T h e m ost fr e q u e nt tr e at m e nt-r el at e d a d v ers e eff ects d et ect e d f or d ef acti ni b i n pr e vi o us cli nic al 
tri als w er e h y p er bilir u bi n e mi a, f ati g u e, d ecr e as e d a p p etit e, a n d di arr h e a w h e n gi v e n f or 2 1 
d a ys i n t w o ti m es a d a y r e gi m e n (J o n es et al., 2 0 1 5; S hi miz u et al., 2 0 1 5). A d v ers e eff ects 
d et ect e d f or a v ut o m eti ni b i n pr e vi o us cli nic al tri als w er e r as h, vis u al dist ur b a nc es, 
h y p o al b u mi n e mi a, cr e ati ni n e p h os p h o ki n as e el e v ati o n, a n d f ati g u e w h e n gi v e n f or 2 1 d a ys i n 
t w o ti m es a w e e k r e gi m e n ( M arti n ez- G arci a et al., 2 0 1 2; G u o et al., 2 0 2 0). O ur st u d y pr o p os es 
t o us e a si n gl e d os e of d ef acti ni b or a si n gl e d os e of a v ut o m eti ni b o n e t o t w o h o urs pri or t o t h e 
t u m or r es ecti o n s ur g er y i niti ati o n t o all o w m axi m u m l ev els t o occ ur at t h e ti m e of t u m or 
s a m pli n g. As a p orti o n of t his st u d y is t o ass ess s af et y, t h e st u d y will b e st art e d wit h a l o w er 
d os e of 2 0 0 m g P O si n gl e d os e f or d ef acti ni b a n d 3. 2 m g P O si n gl e d os e f or a v ut o m eti ni b f or a 
t ot al of t hr e e p ati e nts e ac h. T h e n, t h e d os e will b e i ncr e as e d t o 4 0 0 m g P O si n gl e d os e a n d 4 m g 
P O si n gl e d os e (t hr e e p ati e nts e ac h) if n e e d e d, t o r e ac h a w or ki n g c o nc e ntr ati o n of 1 3. 3 n M f or 
d ef acti ni b a n d 1 9 0 n M f or a v ut o m eti ni b i n t u m or t o g et h er wit h t h e r e d ucti o n of P y k 2/ F A K a n d 
M E K/ Er k p h os p h or yl ati o n. I n t h e c as e of s e v er e a d v ers e eff ects t h at pl ac e t h e r es e arc h s u bj ect 
at ris k d uri n g st u d y p artici p ati o n, eit h er d uri n g s ur g er y or d uri n g t h e p osts ur gic al c o urs e, t h e 
st u d y of hi g h er c o nc e ntr ati o ns will b e s us p e n d e d u ntil t h e ass ess m e nt of t h e c a us e of t h e 
a d v ers e e v e nt. If t h e a d v ers e e v e nts ar e s h o w n t o n ot b e r el at e d t o t h e d ef acti ni b or 
a v ut o m eti ni b a d mi nistr ati o n, t h e st u d y will b e r es u m e d.  If t h er e is r e as o n a bl e s us pici o n t h e 
a d v ers e e v e nts ar e r el at e d t o t h e a g e nts, d os a g e will eit h er b e m o difi e d, or t h e st u d y c urt ail e d.
5.
St u d y I nt er v e nti o n/I n v esti g ati o n al A g e nt
D escri pti o n: d ef acti ni b a n d a v ut o m eti ni b ar e n o n-c o v al e nt P y k 2/ F A K a n d R A F/ M E K ki n as e 
i n hi bit ors wit h n a n o m ol ar p ot e nc y. B ot h dr u gs w er e i n h u m a n cli nic al tri als b e gi n ni n g i n 2 0 1 2 
a n d I N Ds w er e d e v el o p e d f or b ot h dr u gs.

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 1 1 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
D os e Esc al ati o n D esi g n
T h o u g h t his is n ot a p h as e I st u d y l o o ki n g f or a m axi m u m t ol er at e d or a p h as e 2 d os e, a d os e 
esc al ati o n pl a n is pr o p os e d t o e ns ur e t h at s u bj ects ar e n ot ex p os e d t o exc essi v e t oxicit y. T h e 
i niti al pl a n will b e f or t hr e e s u bj ects t o r ec ei v e t h e l o w er d os e of d ef acti ni b ( 2 0 0 m g) a n d f or 
t hr e e t o r ec ei v e t h e l o w er d os e of A V U T O M E TI NI B ( 3. 2 m g). If n o n e of t h e first 3 s u bj ects f or 
e ac h dr u g ex p eri e nc es d os e li miti n g t oxicit y, t h e d os e will b e i ncr e as e d t o t h e hi g h er pl a n n e d 
d os e f or t h e n ext c o h ort. If o n e of t h e first 3 s u bj ects ex p eri e nc es a d os e li miti n g t oxicit y, t h e 
c o h ort will b e ex p a n d e d t o six s u bj ects. If n o n e of t h e r e m ai ni n g s u bj ects i n t h e ex p a n d e d 
c o h ort ex p eri e nc e d os e li miti n g t oxicit y, t h e n ext c o h ort will b e a d mi nist er e d t h e n ext hi g h er 
pl a n n e d d os e. If a s ec o n d s u bj ect ex p eri e nc es a d os e li miti n g t oxicit y, accr u al at t h e d os e l ev el 
s h o ul d b e h alt e d wit h o ut f urt h er esc al ati o ns. C o ns ult ati o n will t h e n b e h el d wit h t h e D S M C, 
V er ast e m a n d t h e F D A r e g ar di n g alt er n ati v e d osi n g str at e gi es v ers us st u d y cl os ur e. T his s a m e 
3 + 3 pl a n m e nti o n e d f or t h e first d os e l ev el will a p pl y t o t h e s ec o n d d os e l ev el f or d ef acti ni b 
( 4 0 0 m g) a n d f or A V U T O M E TI NI B ( 4 m g).
If n o m e a ni n gf ul a m o u nt of d ef acti ni b or a v ut o m eti ni b is i d e ntifi e d i n t u m or s a m pl es at t h e 
s ec o n d d os e l e v el, t h e st u d y will b e p ut o n h ol d a n d c o ns ult ati o ns will b e h el d wit h t h e D S M C, 
V er ast e m a n d F D A t o d et er mi n e if hi g h er d os es, as r e p ort e d i n t h e i n v esti g at or’s br oc h ur e, 
s h o ul d b e p urs u e d, or if t h e st u d y s h o ul d b e e n d e d. If n o m e as ur a bl e a m o u nts of eit h er 
d ef acti ni b or a v ut o m eti ni b is i d e ntifi e d at t h e t w o d os e l e v els pr o p os e d, a n d a d v ers e e v e nts 
e nc o u nt er e d d o n ot m e et crit eri a f or d os e li miti n g t oxicit y, t h e i n v esti g at ors will c o ns ult wit h 
t h e D S M C, V er ast e m a n d t h e F D A, a n d pr o p os e f urt h er d os e l ev els (t e nt ati v el y 5 0 0 m g a n d 7 5 0 
m g f or d ef acti ni b, a n d 5 m g f or a v ut o m eti ni b, b as e d o n t h e m axi m u m ex pl or e d si n gl e d os es 
t ests n ot e d i n i n v esti g at or’s br oc h ur es f or e ac h) t o t h e D S M C. Pr o vi d e d t h er e is a p pr o v al, t h e 
st u d y will t h e n c o nti n u e usi n g t h e s a m e d os e esc al ati o n pl a n, m e nti o n e d i n t h e pr e vi o us 
p ar a gr a p h, as w as us e d i n l o w er d os e l e v els.  If t h er e is n o a p pr o v al, t h e st u d y will b e st o p p e d. 
If t h er e is still n o m e as ur a bl e dr u g i n t u m or aft er c o m pl eti o n of t h es e a d diti o n al d os e l ev els, t h e 
st u d y will b e st o p p e d, a n d c o ns ult ati o n h el d wit h V er ast e m a n d t h e D S M C r e g ar di n g f urt h er 
d osi n g str at e gi es or cl osi n g t h e st u d y.
Dr u g A d mi nistr ati o n
T h e dr u gs will b e gi v e n s e p ar at el y, or all y i n a si n gl e d os e. D ef acti ni b will b e gi v e n at l o w est 
d os e of 2 0 0 m g a n d esc al ati n g t o 4 0 0 m g, a n d a v ut o m eti ni b will b e gi v e n at 3. 2 m g a n d 
esc al ati n g t o 4 m g 1 t o 2 h o urs b ef or e t h e pl a n n e d G B M t u m or r es ecti o n s ur g er y.
C o nc e ntr ati o ns a n d ti mi n g ar e b as e d o n d at a o bt ai n e d fr o m pr e vi o us cli nic al tri als.
Dr u g h a n dli n g: A v ut o m eti ni b will b e s hi p p e d at 2- 8 C. D ef acti ni b will b e s hi p p e d at a m bi e nt 
t e m p er at ur e ( 1 5- 3 0 C) u nl ess it is s hi p p e d wit h a v ut o m eti ni b i n w hic h c as e it c a n b e s hi p p e d at 
2- 8 C. B ot h c a n b e st or e d at 4 C, b ut f or n o l o n g er t h a n 2 4 m o nt hs aft er m a n uf act uri n g, b ef or e 
b ei n g gi v e n t o t h e s u bj ect. T h e dr u gs will b e us e d o nl y f or c o ns e nti n g s u bj ects a n d b e us e d 
o nl y b y a ut h oriz e d i n v esti g at ors.

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 1 2 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
T h e c o ntr ol of t h e d ef acti ni b a n d a v ut o m eti ni b will b e acc o m plis h e d b y f oll o wi n g a n 
est a blis h e d a p pr o v e d St a n d ar d O p er ati o n Pr oc e d ur e ( S O P) at E m or y U ni v ersit y I n v esti g ati o n al 
P h ar m ac y (I D S).
I D S will b e us e d acc or di n g t o t h e p h ar m acist s u p pli er t o c o ntr ol h a n dli n g, st or a g e, or d eri n g, 
r ec ei vi n g, a n d c o ntr ol of dr u gs a n d ass ur e c o m pli a nc e wit h t h e st a n d ar ds of t h e U. S. F o o d a n d 
Dr u g A d mi nistr ati o n ( F D A) a n d of t h e st u d y s p o ns or.
D ef acti ni b a n d a v ut o m eti ni b ar e n ot c o m m erci all y a v ail a bl e, ar e n ot c o ntr oll e d s u bst a nc es, a n d 
F D A Ris k E v al u ati o n a n d Miti g ati o n Str at e g y ( R E M S) st at us h as n ot b e e n ev al u at e d.
D ef acti ni b a n d a v ut o m eti ni b ar e i n v esti g ati o n al dr u gs. A n E m or y I n v esti g at or h ol ds t h e I N D.
Pr oc e d ur es a n d r e g ul ati o ns t o b e f oll o w e d t o c o m pl y wit h s p o ns or r e q uir e m e nts f or F D A 
r e g ul at e d r es e arc h i ncl u d e:
2 1 C F R 1 1: t o e ns ur e t h at all r ec or ds ar e a ut h e ntic, i nc orr u pti bl e, a n d c o nfi d e nti al, r ec or ds will 
b e k e pt i n a cl os e d c o m p ut er s yst e m s ec ur e d wit h a p ass w or d. O nl y a ut h oriz e d p ers o n n el fr o m 
t h e st u d y gr o u p will h a v e acc ess t o t h e p ass w or d.
2 1 C F R 5 4: n o fi n a nci al i nt er ests of t h e cli nic al i n v esti g at or i n t h e o utc o m e of t h e st u d y b ec a us e 
of t h e p a y m e nt or b ec a us e t h e i n v esti g at or h as a pr o pri et ar y i nt er est i n t h e pr o d uct or b ec a us e 
t h e i n v esti g at or h as a n e q uit y i nt er est i n t h e s p o ns or of t h e c o v er e d st u d y.
2 1 C F R 2 1 0, 2 1 1: t h e dr u g will b e m a n uf act ur e d, pr oc ess e d, a n d p ac k e d i n F D A lic e ns e d 
p h ar m ac ol o gic al f aciliti es a n d c o m pl y wit h t h e r e q uir e m e nts of t h e m a n uf act ur er of m e dic al 
pr o d ucts t o e ns ur e t h at t h er e is n o ris k f or s u bj ects d u e t o i n a d e q u at e s af et y pr oc e d ur es.
2 1 C F R 3 1 2: d ef acti ni b a n d a v ut o m eti ni b ar e i n v esti g ati o n al n e w dr u gs f or w hic h I N Ds ar e i n 
eff ect a n d ar e ex e m pt fr o m t h e pr e m ar k eti n g a p pr o v al r e q uir e m e nts. L a b els wit h t h e 
st at e m e nt " C a uti o n: N e w Dr u g - Li mit e d b y F e d er al ( or U nit e d St at es) l a w t o i n v esti g ati o n al 
us e" will b e us e d. T h e dr u gs will n ot b e r e pr es e nt e d i n a pr o m oti o n al c o nt ext or c o m m erci all y 
distri b ut e d. T h e us e of dr u gs will n ot b e pr ol o n g e d aft er t h e e n d of t h e fi n di n g p eri o d t o 
s u p p ort a m ar k eti n g a p plic ati o n. S u bj ects will n ot b e c h ar g e d f or t h e us e of d ef acti ni b or 
a v ut o m eti ni b.
5. 1. A v ut o m eti ni b Pr o p h yl a xis
Pr o p h yl actic m e dic ati o ns m ust b e us e d f or a mi ni m u m of t w o w e e ks aft er r ec ei vi n g t h e 
si n gl e d os e t o miti g at e a g ai nst d er m at ol o gic t oxiciti es. H y dr oc ortis o n e 1 % cr e a m, 
m oist uriz er, a n d s u nscr e e n (s u n pr ot ecti o n f act or [ S P F] ≥ 5 0) s h o ul d b e a p pli e d t o pic all y 
BI D, al o n g wit h a s yst e mic a nti bi otic ( mi n oc ycli n e 1 0 0 m g d ail y or d ox yc ycli n e 1 0 0 m g 
BI D). A p plic ati o n of t o pic al a g e nts s h o ul d i ncl u d e t h e m ost c o m m o nl y aff ect e d s ki n ar e as 
s uc h as f ac e, sc al p, n ec k, u p p er c h est, a n d u p p er b ac k. I n a d diti o n, p ati e nts s h o ul d b e 
a d vis e d t o a v oi d u n n ec ess ar y ex p os ur e t o s u nli g ht.
F or p ati e nts w h o c a n n ot t ol er at e a c o m p o n e nt of t h e pr o p h yl axis, t h at c o m p o n e nt 
m a y b e eli mi n at e d, or a n ot h er a g e nt c o ul d b e us e d.

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 1 3 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
P ati e nts w h o d e v el o p r as h/s ki n t oxiciti es s h o ul d b e s e e n b y a q u alifi e d d er m at ol o gist a n d 
s h o ul d r ec ei v e e v al u ati o n f or s y m pt o m atic/s u p p orti v e c ar e 
m a n a g e m e nt. G e n er al 
r ec o m m e n d ati o ns f or s y m pt o m atic c ar e i ncl u d e:

Pr uritic l esi o ns: c o ol c o m pr ess es a n d or al a nti hist a mi n e t h er a pi es

Fiss uri n g l esi o ns: M o ns el’s s ol uti o n, sil v er nitr at e, or zi nc oxi d e cr e a m

D es q u a m ati o n: t hic k e m olli e nts a n d mil d s o a p

P ar o n yc hi a: a ntis e ptic b at h, l oc al p ot e nt c ortic ost er oi ds i n a d diti o n t o 
a nti bi otics; if n o i m pr o v e m e nt, c o ns ult d er m at ol o gist or s ur g e o n

I nf ect e d l esi o ns: a p pr o pri at e b act eri al/f u n g al c ult ur e- dri v e n s yst e m atic or 
t o pic al a nti bi otics
6.
Pr oc e d ur es I n v ol v e d
T ot al 1 7 s u bj ects will b e e nr oll e d i n t h e st u d y. D ef acti ni b will b e gi v e n t o 6 s u bj ects a n d 
a v ut o m eti ni b will b e gi v e n t o ot h er 6 s u bj ects or all y wit hi n 1- 2 h o urs pri or t o t h e G B M t u m or 
r es ecti o n i n pl a n n e d esc al ati n g d os es of 2 0 0 m g (t hr e e s u bj ects) a n d 4 0 0 m g (t hr e e s u bj ects) f or 
d ef acti ni b, a n d 3. 2 m g (t hr e e s u bj ects) a n d 4 m g (t hr e e s u bj ects) f or a v ut o m eti ni b. P ati e nts will 
b e gi v e n t h e m e dic ati o ns at t h e l o w er d os es i n alt er n ati n g f or m at.
6. 1 Ass ess m e nt f or D os e Li miti n g T oxicit y
Alt h o u g h t his is n ot a p h as e I st u d y l o o ki n g f or d os e li miti n g t oxiciti es, o bs er v ati o n f or t h es e 
e v e nts will b e u n d ert a k e n, a n d w h e n t h e y occ ur, us e d as a g ui d e f or d os e esc al ati o n. P ati e nts 
will b e gi v e n t h e m e dic ati o ns at t h e l o w er d os es i n alt er n ati n g f or m at.
D os e-li miti n g t oxiciti es will i ncl u d e t h e f oll o wi n g e v e nts t h at ar e n ot cl e arl y d u e t o t h e 
u n d erl yi n g dis e as e extr a n e o us c a us es: a n y d e at h, a n y n o n- h e m at ol o gic t oxicit y of gr a d e 3 or 
hi g h er, gr a d e 3 or hi g h er t hr o m b oc yt o p e ni a wit h cli nic all y si g nific a nt bl e e di n g. Excl u d e d will b e 
gr a d e 3 n a us e a a n d v o miti n g, or di arr h e a f or l ess t h a n 7 2 h o urs wit h a d e q u at e a nti- e m etic a n d 
ot h er s u p p orti v e c ar e; gr a d e 3 f ati g u e f or m or e t h a n o n e w e e k; gr a d e 3 or hi g h er el ectr ol yt e 
a b n or m alit y t h at l asts f or u p t o 7 2 h o urs, is n ot c o m plic at e d, a n d r es ol v es s p o nt a n e o usl y or 
r es p o n ds t o c o n v e nti o n al m e dic al i nt er v e nti o ns; a n d gr a d e 3 or hi g h er a m yl as e or li p as e t h at is 
n ot ass oci at e d wit h s y m pt o ms or cli nic al m a nif est ati o ns of p a ncr e atitis. If t h er e ar e r e p orts of 
a d v ers e eff ects q u alif yi n g as d os e li miti n g t oxiciti es at l o w er or u p p er d os es, t h e n d os a g e will 
eit h er b e m o difi e d, or t h e st u d y c urt ail e d.
S u bj ects will u n d er g o t h eir pl a n n e d br ai n s ur g er y i n t h e s a m e w a y as if t h e y di d n ot p artici p at e 
i n t h e st u d y. N o a d diti o n al tr e at m e nts or pr oc e d ur es will b e p erf or m e d d uri n g t h e s ur g er y or 
aft er.
Tiss u e s a m pl e extr acti o n: 
T w o s a m pl es will b e t a k e n:

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 1 4 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
●
T u m or s a m pl e: Tiss u e s a m pl es 1. 5- 2. 0 c m 3 i n a p pr oxi m at e siz e will b e s e p ar at e d fr o m 
t h e m aj or t u m or m ass r e m o v e d fr o m t h e p ati e nt a n d pl ac e d i nt o a t u b e c o nt ai ni n g ic e- 
c ol d p h os p h at e b uff er e d s ali n e s ol uti o n ( P B S) i m m e di at el y aft er s ur gic al diss ecti o n. T h e 
ti m e of t h e s a m pl e r es ecti o n will b e r ec or d e d. T h e s a m pl es will b e t a k e n fr o m t h e n o n- 
n ecr otic ar e a of t h e t u m or. T his s p eci m e n will b e pl ac e d o n dr y ic e i n t h e O R p at h ol o g y 
l a b wit hi n 3 0 mi n ut es of h ar v est a n d br o u g ht t o t h e i n v esti g at or’s l a b or at or y o n dr y ic e 
at t h e c o m pl eti o n of t h e s ur g er y.
●
Br ai n ar o u n d t u m or: Tiss u e s a m pl es 0. 2 5- 0. 5 c m 3 a p pr oxi m at e siz e will b e t a k e n fr o m 
n or m al br ai n n ot r el at e d t o el o q u e nt f u ncti o n ar o u n d t h e t u m or a n d pl ac e d i nt o a t u b e 
c o nt ai ni n g ic e-c ol d p h os p h at e b uff er e d s ali n e s ol uti o n ( P B S) i m m e di at el y aft er s ur gic al 
diss ecti o n. T h e ti m e of t h e s a m pl e c oll ecti o n will b e r ec or d e d. T his s p eci m e n will b e 
pl ac e d o n dr y ic e i n t h e O R p at h ol o g y l a b wit hi n 3 0 mi n ut es of h ar v est a n d br o u g ht t o 
t h e i n v esti g at or’s l a b or at or y o n dr y ic e at t h e c o m pl eti o n of t h e s ur g er y.

Bl o o d S a m pl es: a 1 0 ml s a m pl e i n a p ur pl e t o p t u b e will b e o bt ai n e d d uri n g t h e t u m or 
r es ecti o n. T h e ti m e of t h e bl o o d c oll ecti o n will b e r ec or d e d. T h e bl o o d s a m pl e will b e 
br o u g ht t o t h e i n v esti g at or’s l a b or at or y at t h e s a m e ti m e as t h e tiss u e s p eci m e ns.
L a b or at or y pr oc e d ur es:
O nc e i n t h e l a b or at or y, e ac h tiss u e s a m pl e will b e c ut i nt o t w o p arts. O n e p art of t h e t u m or 
s a m pl e, n or m al br ai n, a n d t h e pl as m a fr o m t h e bl o o d s a m pl e will b e s e nt t o E m or y Li pi d o mics 
c or e f or p h ar m ac o d y n a mic st u di es f or i d e ntific ati o n of d ef acti ni b a n d a v ut o m eti ni b 
c o nc e ntr ati o ns i n t h e tiss u e a n d s er u m. T h e ot h er p art of t h e tiss u e s a m pl e will b e us e d f or 
pr ot ei n extr acti o n a n d p h ar m ac o d y n a mic st u di es. Extr act e d pr ot ei ns will b e s hi p p e d i n dr y ic e 
t o P u ert o Ric o, C e ntr al C ari b b e a n U ni v ersit y ( U C C) f or w est er n bl ot a n al ysis of r ati o of t ot al a n d 
p h os p h or yl at e d f or ms of P y k 2 a n d F A K as w ell as t ot al a n d p h os p h or yl at e d M E K 1/ 2 a n d Er k 1/ 2. 
Pr ot ei n s a m pl es will b e s hi p p e d t o t h e U ni v ersi d a d C e ntr al d el C ari b e b as e d o n a M at eri al 
Tr a nsf er A gr e e m e nt.
P artici p a nts will b e m o nit or e d f or s af et y b e gi n ni n g i m m e di at el y aft er t h e dr u g c o ns u m pti o n 
a n d d uri n g pr e- a n d p ost-s ur gic al p eri o d. I n pr e vi o us cli nic al tri als n o r e acti o ns t h at c o ul d 
p ot e nti all y i nt erf er e wit h t u m or r es ecti o n s ur g er y w er e r e p ort e d. H o w ev er, if s uc h r e acti o ns 
ar e o bs er v e d d uri n g t h e pr e-s ur gic al p eri o d ( e. g., f ev er, n a us e a, r as h, bl e e di n g, p ai n) t h e 
s ur g er y will b e d el a y e d u ntil s y m pt o ms ar e r es ol v e d.
P ost-s ur g er y pr oc e d ur es:
F or u p t o 4 h o urs (t h e l e n gt h of t his will b e i n di vi d u aliz e d t o t h e s u bj ects r ec o v er y n e e ds p er 
st a n d ar d of c ar e) f oll o wi n g s ur g er y t h e f oll o wi n g p osts ur gic al o bs er v ati o ns will b e c oll ect e d:

H R, R R, S p O 2, B P a n d T e m p er at ur e

N e ur ol o gic al Ass ess m e nt

P ai n Sc or e

Ass ess m e nt of W o u n d Sit es / Dr essi n gs

Pr es e nc e of dr ai ns a n d p at e nc y of s a m e w h e n utiliz e d

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 1 5 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4

Ot h er c o m plic ati o ns/ ass ess m e nt fi n di n gs as p ati e nts c o n diti o n dict at es
Fr e q u e nc y of t h es e r o uti n e p ost-s ur g er y o bs er v ati o ns will b e as p er st a n d ar d of c ar e i n di vi d u aliz e d t o t h e 
gi v e n s u bj ect.
U p o n st a pl e r e m o v al a n d wit hi n t w o w e e ks p ost-s ur g er y wi n d o w, t h e f oll o wi n g m o nit ori n g 
t ests m ust b e c o m pl et e d: m et a b olic p a n el, li v er f u ncti o n p a n el i ncl u di n g bilir u bi n l ev el, ki d n e y 
f u ncti o n p a n el, a n d w h e n cli nic all y i n dic at e d P T/I N R/ P T T.
Gi v e n t h e ris k of d er m atitis a n d st o m atitis, a f ull p h ysic al ex a m i ncl u di n g s ki n a n d or al ex a ms 
m ust b e p erf or m e d at t h e ti m e of st a pl e r e m o v al.
T o l ess e n t h e pr o b a bilit y or m a g nit u d e of ris ks, t h e st u d y will b e p erf or m e d b e gi n ni n g wit h t h e 
s m all er pl a n n e d d os e of d ef acti ni b ( 2 0 0 m g) or s m all est d os e of a v ut o m eti ni b ( 3. 2 m g). If 
si g nific a nt a d v ers e r e acti o ns will b e d et ect e d i n pr e-s ur gic al p eri o d, d uri n g t h e s ur g er y, or i n 
p ost-s ur gic al p eri o d t hr o u g h t h e ti m e of s ut ur e or st a pl e r e m o v al ( us u all y a b o ut 1 4 d a ys), t h e 
st u d y will b e p a us e d f or t h e e v al u ati o n of a d v ers e r e acti o ns. T h e st u d y will pr oc e e d t o hi g h er 
d os es o nl y w h e n it is asc ert ai n e d t h at t h e l o w er d os e d o es n ot c a us e si g nific a nt h e alt h 
c o m plic ati o ns or ris ks f or s u bj ects.
N o a d diti o n al dr u gs a n d d e vic es o utsi d e of t h e st a n d ar d tr e at m e nt pr actic e, exc e pt f or 
d ef acti ni b or a v ut o m eti ni b, will b e us e d i n t h e m a n a g e m e nt of t h e p artici p a nts.
S u bj ect d at a will b e a n al yz e d a n d r e p ort e d i n a n a n o n y m o us m a n n er. O nl y i nf or m ati o n r el at e d 
t o a g e, s ex, r ac e, ass oci at e d di a g n osis, a n d m ol ec ul ar m ar k ers will b e c oll ect e d.
Dr u g s p ecific i n v esti g ati o n al d at a c oll ect e d d uri n g t h e st u d y: c o nc e ntr ati o n of d ef acti ni b or 
a v ut o m eti ni b i n t h e s er u m, r es ect e d G B M tiss u e s a m pl es, a n d br ai n ar o u n d t u m or s a m pl es will 
b e m e as ur e d aft er c o ns u m pti o n of i n dic at e d d os es of d ef acti ni b or a v ut o m et a ni b r es p ecti v el y. 
T his will i d e ntif y t h e n or m al tiss u e a n d t u m or u pt a k e of d ef acti ni b a n d a v ut o m eti ni b. W est er n 
bl ot a n al ysis of t ot al a n d p h os p h or yl at e d v ersi o ns of t h e P y k 2, F A K, M E K 1/ 2 a n d Er k 1/ 2 i n t h e 
n or m al tiss u e a n d t u m or s p eci m e n will b e c orr el at e d wit h t h e d os a g e a n d s er u m c o nc e ntr ati o n 
of d ef acti ni b a n d a v ut o m et a ni b a d mi nist er e d a n d i n t his m a n n er will i d e ntif y t h e a m o u nt of 
d ef acti ni b a n d a v ut o m et a ni b n e e d e d f or m e a ni n gf ul bi ol o gic eff ect.
N o l o n g-t er m f oll o w- u p is pr o p os e d f or t h e st u d y.
7.
St atistic al A n al ysis Pl a n

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 1 6 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
T his is a t w o- ar m st u d y t o ass ess t h e a bilit y of d ef acti ni b a n d a v ut o m et a ni b t o r e ac h 
Gli o bl ast o m as ( G B M) a n d t h e br ai n tiss u e ar o u n d t h e m. D ef acti ni b will b e gi v e n t o 6 
s u bj ects, a n d a v ut o m et a ni b will b e gi v e n t o 6 s u bj ects or all y 1- 2 h o urs pri or t o t h e G B M 
t u m or r es ecti o n i n pl a n n e d esc al ati n g d os es of 2 0 0 m g (t hr e e s u bj ects) a n d 4 0 0 m g 
(t hr e e s u bj ects) f or d ef acti ni b, a n d 3. 2 m g (t hr e e s u bj ects) a n d 4 m g (t hr e e s u bj ects) f or 
a v ut o m et a ni b. P ati e nts will b e gi v e n t h e m e dic ati o ns i n a n alt er n ati n g f or m at. A 
m o nit ori n g p eri o d of 2 1 d a ys will b e all o w e d t o el a ps e aft er t h e l ast s u bj ect r ec ei v es t h e 
l o w er d os e t o all o w f or o bs er v ati o n f or d os e li miti n g t oxiciti es.
S u m m ar y st atistics of dr u g c o nc e ntr ati o ns i n t u m or, br ai n ar o u n d t u m or, a n d i n 
c o nc urr e nt s er u m s a m pl e, as w ell as P y k 2/ F A K, M E K/ Er k p h os p h or yl ati o n i n t u m or a n d 
br ai n ar o u n d t u m or will b e r e p ort e d usi n g d escri pti v e st atistics i ncl u di n g m e a n, m e di a n, 
i nt er q u artil e r a n g e, mi ni m u m/ m axi m u m, a n d st a n d ar d d evi ati o n. P ati e nt d e m o gr a p hics 
will b e s u m m ariz e d d escri pti v el y. 9 5 % c o nfi d e nc e i nt erv als will b e r e p ort e d, u nl ess 
ot h er wis e s p ecifi e d. All t ests will b e t w o-si d e d, a n d a si g nific a nc e l e v el of 0. 0 5 will b e 
c o nsi d er e d.
7. 1 
S a m pl e siz e c o nsi d er ati o ns:
N u m b er of P artici p a nts E nr oll e d: I n t his i niti al st u d y of t h es e a g e nts i n h u m a ns wit h 
gli o bl ast o m a, w e will b e r ecr uiti n g u p t o 1 7 p artici p a nts o v er a p eri o d of a p pr oxi m at el y 
1 2 m o nt hs. D ef acti ni b will b e gi v e n t o 6 s u bj ects a n d a v ut o m et a ni b will b e gi v e n t o 
ot h er 6 s u bj ects or all y 1- 2 h o urs pri or t o t h e G B M t u m or r es ecti o n i n pl a n n e d esc al ati n g 
d os es of 2 0 0 m g (t hr e e s u bj ects) a n d 4 0 0 m g (t hr e e s u bj ects) f or d ef acti ni b, a n d 3. 2 m g 
(t hr e e s u bj ects) a n d 4 m g (t hr e e s u bj ects) f or a v ut o m et a ni b. W e h a v e f o u n d a s a m pl e 
siz e of 3 ( p er d os e a n d p er dr u g) t o b e s uffici e nt f or c h ar act erizi n g c o nc e ntr ati o n of 
st u d y dr u gs i n pri or P h as e 0 st u di es [ Q uilli n et al., 2 0 2 0]. T h e n u m b er of s u bj ects 
a v ail a bl e f or r ecr uit m e nt is b as e d o n t h e pri nci p al i n v esti g at or’s esti m at e fr o m his 
n e ur os ur gic al o nc ol o g y pr actic e. T his n u m b er ( 1 7) is a n esti m at e of t h e n u m b er of c as es 
wit h ( 1) i m a gi n g c o nsist e nt q u alif yi n g hist ol o g y ( or pr e vi o us di a g n osis of gli o bl ast o m a) 
t h at ( 2) r e q uir e cr a ni ot o mi es f or s ur gic al r es ecti o n as t h e n ext st e p i n t h eir t h er a p y t h at 
c a n b e accr u e d o v er a p pr oxi m at el y a y e ar at o ur i nstit uti o n.  B as e d o n o ur accr u al 
ex p eri e nc e, it is a ntici p at e d t h at s o m e s u bj ects will n ot h a v e fi n al p at h ol o g y s h o wi n g 
gli o bl ast o m a or will f ail scr e e ni n g.  T h us, it is ex p ect e d t h at t o o bt ai n 1 2 i n di vi d u als 
m e eti n g scr e e ni n g crit eri a, act u all y h a vi n g a gli o bl ast o m a a n d w h os e tiss u e a n al ysis is 
m e a ni n gf ul, as m a n y as 1 7 s u bj ects m a y n e e d t o b e e nr oll e d.
7. 2 
Pri m ar y e n d p oi nts:

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 1 7 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
M e as ur e m e nt of d ef acti ni b a n d a v ut o m et a ni b L e v els i n T u m or, Br ai n Ar o u n d T u m or, 
a n d S er u m: D ef acti ni b c o nc e ntr ati o n will b e m e as ur e d i n a s a m pl e of t h e gli o bl ast o m a, 
br ai n ar o u n d t h e gli o bl ast o m a, a n d i n a s er u m s a m pl e fr o m e ac h s u bj ect r ec ei vi n g

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 1 8 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
d ef acti ni b. A v ut o m et a ni b c o nc e ntr ati o n will b e m e as ur e d i n a s a m pl e of t h e 
gli o bl ast o m a, br ai n ar o u n d t h e gli o bl ast o m a, a n d i n a s er u m s a m pl e fr o m e ac h s u bj ect 
r ec ei vi n g a v ut o m et a ni b. D escri pti v e st atistics, s uc h as m e a n a n d st a n d ar d d evi ati o n, 
will b e g e n er at e d wit h t h es e r es ults. C o nc e ntr ati o n will b e c o m p ar e d b et w e e n d os e 
l e v els wit hi n st u d y dr u g usi n g t w o-s a m pl e t-t ests or n o n- p ar a m etric e q ui v al e nts s uc h as 
M a n n W hit n e y U t ests.
S af et y: As t his is t h e first st u d y of t h es e a g e nts i n s u bj ects wit h br ai n t u m ors, t h e n at ur e 
a n d i nci d e nc e of a d v ers e e v e nts a n d ris k is n ot k n o w n i n a q u a ntit ati v e m a n n er. I n 
i n di vi d u als wit h ot h er f or ms of c a nc er, r e g ul ar us e o v er ti m e h as b e e n ass oci at e d wit h 
a d v ers e e v e nts s uc h as f ati g u e, n a us e a, di arr h e a, v o miti n g, h y p er bilir u bi n e mi a, 
d ecr e as e d a p p etit e, p eri p h er al e d e m a, dizzi n ess, a n d h e a d ac h e f or d ef acti ni b [ G er b er 
et al., 2 0 1 9] a n d r as h, cr e ati n e p h os p h o ki n as e el ev ati o n, h y p o al b u mi n e mi a, a n d f ati g u e 
f or a v ut o m et a ni b [ G u o, 2 0 2 0]. As t his st u d y will o nl y b e usi n g o n e d os e of t h e dr u g, t h e 
li k eli h o o d of t h es e a d v ers e e v e nts is s m all er t h a n r e p ort e d i n t h es e ot h er dis e as e t y p es. 
T h e i nci d e nc e of t h es e e v e nts is esti m at e d t o b e i n t h e r a n g e of 0. 5 % or l ess f or t h es e 
e v e nts c o m bi n e d. S af et y st o p pi n g r ul es ar e d escri b e d b el o w. S u m m ar y st atistics will 
i ncl u d e fr e q u e nci es a n d p erc e nt a g es of t h es e a d v ers e ev e nts.
7. 3 
S ec o n d ar y e n d p oi nts
P y k 2 a n d F A K p h os p h or yl ati o n: T o ex pl or e t h e eff ect o n its pr o p os e d t ar g ets, s a m pl es 
fr o m p ati e nts r ec ei vi n g d ef acti ni b will b e pr oc ess e d t o m e as ur e p P y k 2 a n d p F A K 
p h os p h or yl ati o n. T h es e r es ults will b e s u m m ariz e d wit h d escri pti v e st atistics. T h es e 
r es ults will b e c o m p ar e d t o c urr e nt a n d hist oric al c o ntr ol s a m pl es of gli o bl ast o m a 
(c oll ect e d i n o ur pr e vi o us h u m a n G B M s p eci m e ns’ st u di es) a n d a n al yz e d i n p ar all el wit h 
st u d y s u bj ect s a m pl es a n d r u n o n o n e w est er n bl ot m e m br a n e t o r e d uc e t ec h ni q u e 
v ari ati o n ( N u ñ ez R E, et al., 2 0 2 1). As m e nti o n e d els e w h er e i n t his pr ot oc ol w e 
a ntici p at e t h er e will b e 1 2 s ets of s u bj ect s p eci m e ns. T h e m e a n p h os p h or yl ati o n will b e 
c o m p ar e d b et w e e n gr o u ps usi n g t w o-s a m pl e t-t est. s or n o n- p ar a m etric e q ui v al e nts 
s uc h as M a n n W hit n e y U t ests. D os e l ev els will b e c o m p ar e d t o a hist oric al c o ntr ol, 
b ot h al o n e a n d c o m bi n e d.
M E K 1/ 2 a n d E R K p h os p h or yl ati o n: T o ex pl or e t h e eff ect o n its pr o p os e d t ar g ets, 
s a m pl es fr o m p ati e nts r ec ei vi n g a v ut o m et a ni b will b e pr oc ess e d t o m e as ur e M E K 1/ 2 
a n d Er k 1/ 2 p h os p h or yl ati o n. T h es e r es ults will b e s u m m ariz e d wit h d escri pti v e 
st atistics. T h e r es ults will b e c o m p ar e d t o hist oric al gli o bl ast o m a s p eci m e ns as 
d escri b e d a b o v e f or d ef acti ni b. P h os p h or yl ati o n will b e c o m p ar e d b et w e e n d os e l e v els 
wit hi n st u d y dr u g usi n g t w o-s a m pl e t-t ests or n o n- p ar a m etric e q ui v al e nts s uc h as M a n n 
W hit n e y U t ests.

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 1 9 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
P h ar m ac o d y n a mics: Pr o vi d e d s a m pl es of t h e t u m or, br ai n ar o u n d t u m or, a n d s er u m 
pr o vi d e t ec h nic all y v ali d d at a, t h e dr u g c o nc e ntr ati o n a n d p h ar m ac o d y n a mic d at a 
n or m alit y di a g n ostics will b e d o n e wit h S h a pir o- Wil k esti m ati o ns t o ev al u at e t h e r a w 
st atistic al distri b uti o n of o ur d at as ets. Fr e q u e nc y a n d S E M gr a p hs will b e c alc ul at e d f or 
u ni v ari at e p ur p os es. T w o-s a m pl e t-t ests or M a n n- W hit n e y U t ests will b e us e d t o 
c o m p ar e t h e m e a n ( or m e di a n) diff er e nc es b et w e e n d os a g e gr o u ps f or e ac h dr u g.
7. 4 
A n al ysis p o p ul ati o ns
P h ar m ac o d y n a mics/s af et y: All p ati e nts u n d er g oi n g a d mi nistr ati o n of d ef acti ni b or 
a v ut o m eti ni b pri or t o s ur gic al r es ecti o n of t h eir gli o bl ast o m a.
7. 5 
S af et y st o p pi n g r ul es
T o all o w f or t h e a bilit y t o t a k e st e ps t o ass ess a n d mi ni miz e a d v ers e e v e nts, t h e i n v esti g at ors 
will st o p accr u al if o n e p ati e nt ex p eri e nc es at l e ast o n e s ev er e a d v ers e ev e nt ( as d efi n e d b y 
st a n d ar d C T C A E 5. 0 crit eri a) occ urr e nc e p ossi bl y or pr o b a bl y r el at e d t o t h e dr u g o v er a s eri es of 
3 c o ns ec uti v e s u bj ects f or a gi v e n a g e nt f or a gi v e n d os e. A d v ers e ev e nts a n d s ev er e a d v ers e 
e v e nts attri b ut a bl e t o t h e t u m or, t h e s ur g er y f or it, or t h e st a n d ar d of c ar e dr u gs us e d b ef or e 
a n d aft er s ur g er y will n ot b e i ncl u d e d i n t his c o nsi d er ati o n. T h e s e v er e a d v ers e e v e nts will t h e n 
b e st u di e d b y t h e i n v esti g at ors a n d st e ps t o miti g at e t h e m f or f ut ur e c as es will b e o utli n e d f or 
t h e W CI D at a S af et y M o nit ori n g C o m mitt e e ( D S M C) a n d V er ast e m. O nc e t h es e st e ps ar e 
a p pr o v e d b y t h e W CI D at a S af et y M o nit ori n g C o m mitt e e a n d V er ast e m accr u al will b e 
r es u m e d. If a r e m e d y c a n n ot b e d et er mi n e d t o miti g at e t h e t oxicit y, accr u al will st o p. T h e 
i niti al pl a n is f or o nl y 3 p ati e nts m axi m u m will b e tr e at e d at a gi v e n d os e l ev el f or a st u d y dr u g. 
H o w e v er, p er t h e D os e Esc al ati o n D esi g n, ex p a nsi o n t o 6 p ati e nts i n o n e d os e gr o u p m a y occ ur 
if s e v er e t oxicit y t h at is s ucc essf ull y tr e at e d a n d w h os e f urt h er occ urr e nc e c a n b e miti g at e d p er 
a n a gr e e d u p o n pl a n wit h t h e D S M C a n d V er ast e m. I n t h e c as e t h at a l o w er d os e l ev el pr o vi d es 
s atisf act or y p h ar m ac o ki n etic a n d p h ar m ac o d y n a mic d at a b ef or e r e ac hi n g 6 p ati e nts, a pl a n t o 
m o v e t o a hi g h er d os e l e v el aft er f e w er t h a n 6 p ati e nts t h e st u d y will b e p ut o n h ol d a n d a pl a n 
will b e pr es e nt e d t o t h e D S M C a n d V er ast e m t o ex p e dit e st u d y c o m pl eti o n.
T h e st u d y s af et y wi n d o w will b e fr o m t h e ti m e of dr u g a d mi nistr ati o n j ust pri or t o s ur g er y u ntil 
s ut ur e or st a pl e r e m o v al, w hic h is t h e st at e d off st u d y ti m e p oi nt.
I n or d er t o l o o k f or t h e c o m m o nl y e nc o u nt er e d t oxiciti es list e d b el o w, t h e s u bj ects will u n d er g o 
t h e f oll o wi n g l a b or at or y st u di es at t h e ti m e of s ut ur e or st a pl e r e m o v al: m et a b olic p a n el, l e v er

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 2 0 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
f u ncti o n p a n el i ncl u di n g bilir u bi n l e v el, ki d n e y f u ncti o n p a n el, a n d w h e n cli nic all y i n dic at e d 
P T/I N R/ P T T. I n a d diti o n, at t h e ti m e of s ut ur e or st a pl e r e m o v al, t h e s u bj ect will u n d er g o f ull 
p h ysic al ex a m i ncl u di n g s ki n a n d or al ex a m, wit h e m p h asis o n l o o ki n g f or d er m atitis a n d 
st o m atitis.
C o m m o nl y e nc o u nt er e d t oxiciti es f or d ef acti ni b t h at will b e m o nit or e d f or s eri o us a d v ers e 
e v e nts ( Gr a d e 3 t oxicit y or a b o v e b y C T C A E 5. 0 crit eri a) will b e f ati g u e, n a us e a, di arr h e a, 
v o miti n g, h y p er bilir u bi n e mi a, d ecr e as e d a p p etit e, p eri p h er al e d e m a, dizzi n ess, a n d h e a d ac h e 
( n ot attri b ut a bl e t o t h e br ai n t u m or or it’s tr e at m e nt). T h o u g h t h e t h es e ar e t h e m ost li k el y 
t oxiciti es b as e d o n t h e i n v esti g at or’s br oc h ur e, ot h er t oxiciti es will als o b e r ec or d e d a n d 
a d dr ess e d wit h t h er a p y as a p pr o pri at e f or t h e e v e nt. As m e nti o n e d a b o v e, if a r e m e d y c a n n ot 
b e d et er mi n e d t o miti g at e t h e t oxicit y, accr u al will st o p.
L ess c o m m o nl y e nc o u nt er e d t oxiciti es f or d ef acti ni b t h at will b e m o nit or e d f or t h at ar e n ot 
cl e arl y d u e t o t h e u n d erl yi n g dis e as e or extr a n e o us c a us es: a n y d e at h; a n y n o n- h e m at ol o gic 
t oxicit y of gr a d e 3 or hi g h er; gr a d e 3 or hi g h er t hr o m b oc yt o p e ni a wit h cli nic all y si g nific a nt 
bl e e di n g; a n d n e utr o p e nic f e v er.
T h o u g h t h e t h es e ar e t h e m ost li k el y t oxiciti es b as e d o n t h e i n v esti g at or’s br oc h ur e, ot h er 
t oxiciti es will als o b e r ec or d e d a n d a d dr ess e d wit h t h er a p y as a p pr o pri at e f or t h e e v e nt. As 
m e nti o n e d a b o v e, if a r e m e d y c a n n ot b e d et er mi n e d t o miti g at e t h e t oxicit y, accr u al will st o p.
8.
D at a S p eci m e n B a n ki n g
N o d at a or s p eci m e ns will b e b a n k e d f or f ut ur e us e.
9.
S h ari n g of R es ults wit h P artici p a nts
I n di vi d u al s u bj ect r es ults will n ot aff ect t h e st a n d ar d tr e at m e nt m et h o ds f or t h e p ati e nt. F or 
t his r e as o n, t h e i n di vi d u al r es ults will b e s h ar e d wit h t h e s u bj ect u p o n r e q u est o nl y.
I nf or m ati o n g ai n e d d uri n g t h e r es e arc h pr oj ect m a y b e p u blis h e d i n t h e f or m of r e p orts or 
j o ur n al articl es. H o w e v er, t h e p ers o n al i nf or m ati o n will n ot b e i d e ntifi e d i n a n y w a y i n t h os e 
p u blic ati o ns.
N o i nci d e nt al fi n di n gs ar e a ntici p at e d.
1 0. St u d y Ti m eli n es
• T h e d ur ati o n of a n i n di vi d u al p artici p a nt’s p artici p ati o n i n t h e st u d y is wit hi n a st a n d ar d 
ti m eli n e of pr e- a n d p ost-s ur gic al o bs er v ati o n, wit h t h e m c o mi n g off st u d y at t h e ti m e of 
s ut ur e/st a pl e r e m o v al.
• T h e d ur ati o n a ntici p at e d f or e nr olli n g all st u d y p artici p a nts is o n e y e ar.

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 2 1 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
• T h e esti m at e d d at e f or t h e i n v esti g at ors t o c o m pl et e t his st u d y a n d pri m ar y a n al ys es is 1 8 
m o nt hs fr o m t h e b e gi n ni n g of t h e st u d y
1 1. I ncl usi o n a n d E xcl usi o n Crit eri a
I ncl usi o n crit eri a:
1. 
N e w or r ec urr e nt gli o bl ast o m a di a g n os e d b y n e ur oi m a gi n g t ec h ni q u es f or w hic h 
s ur gic al r es ecti o n is i n dic at e d.
2. 
A g e ol d er t h a n 2 1 y e ars
3. 
A n E ast er n C o o p er ati v e Gr o u p ( E C O G) p erf or m a nc e st at us ≤ 1.
4.  
M ust h a v e a d e q u at e or g a n f u ncti o n d efi n e d b y t h e f oll o wi n g l a b or at or y p ar a m et ers:
a.
A d e q u at e h e m at ol o gic f u ncti o n i ncl u di n g h e m o gl o bi n ( H b) ≥ 9. 0 g/ dL; pl at el ets
≥1 0 0, 0 0 0/ m m 3 ; a n d a bs ol ut e n e utr o p hil c o u nt ( A N C) ≥ 1 5 0 0/ m m3 ). If a r e d 
bl o o d c ell tr a nsf usi o n h as b e e n a d mi nist er e d t h e H b m ust r e m ai n st a bl e a n d ≥
9. 0 g/ dL f or at l e ast 1 w e e k pri or t o first d os e of st u d y t h er a p y.
b.
A d e q u at e h e p atic f u ncti o n: (i) t ot al bilir u bi n ≤ 1. 5 × u p p er li mit of n or m al ( UL N) 
p er t h e i nstit uti o n; p ati e nts wit h Gil b ert s y n dr o m e m a y e nr oll if t ot al bilir u bi n <
3. 0 m g/ dL ( 5 1 μ m ol e/L);  (ii) al a ni n e a mi n otr a nsf er as e ( AL T) a n d as p art at e 
a mi n otr a nsf er as e ( A S T) ≤ 2. 5 × UL N ( or < 5x UL N i n p ati e nts wit h li v er 
m et ast as es).
c.
A d e q u at e r e n al f u ncti o n wit h a cr e ati ni n e cl e ar a nc e r at e of ≥ 5 0 mL/ mi n as 
c alc ul at e d b y t h e C oc kcr oft- G a ult f or m ul a or s er u m cr e ati ni n e of ≤ 1. 5 × UL N.
d. I nt er n ati o n al n or m aliz e d r ati o (I N R) ≤ 1. 5 a n d p arti al t hr o m b o pl asti n ti m e ( P T T) ≤
1. 5 x UL N i n t h e a bs e nc e of a ntic o a g ul ati o n or t h er a p e utic l e v els i n t h e pr es e nc e 
of a ntic o a g ul ati o n.
e.
Al b u mi n ≥ 3. 0 g/ dL ( 4 5 1 μ m ol e/L).
f.
Cr e ati n e p h os p h o ki n as e ( C P K) ≤ 2. 5 x UL N.
g.
A d e q u at e c ar di ac f u ncti o n wit h l eft v e ntric ul ar ej ecti o n fr acti o n ≥ 5 0 % b y 
ec h oc ar di o gr a p h y ( E C H O) or m ulti pl e- g at e d ac q uisiti o n ( M U G A) sc a n.
5. 
B as eli n e Q Tc i nt er v al < 4 6 0 ms f or w o m e n a n d ≤4 5 0  ms f or m e n ( a v er a g e of 
tri plic at e r e a di n gs) ( C T C A E Gr a d e 1) usi n g Fr e d erici a’s Q T c orr ecti o n f or m ul a. N O T E: 
T his crit eri o n d o es n ot a p pl y t o s u bj ects wit h a ri g ht or l eft b u n dl e br a nc h bl oc k.
6. 
A d e q u at e r ec o v er y fr o m t oxiciti es r el at e d t o pri or tr e at m e nts t o at l e ast Gr a d e 1 b y 
C T C A E v 5. 0. Exc e pti o ns i ncl u d e al o p eci a a n d p eri p h er al n e ur o p at h y gr a d e ≤ 2.
7.  
M al e a n d f e m al e p ati e nts wit h r e pr o d ucti v e p ot e nti al a gr e e t o us e hi g hl y eff ecti v e 
m et h o d of c o ntr ac e pti v e ( p er Cli nic al Tri al F acilit ati o n Gr o u p [ C F T G] 
r ec o m m e n d ati o ns) d uri n g t h e tri al a n d f or 3 m o nt hs f oll o wi n g t h e l ast d os e of 6 7 6 6 
f or m al e p ati e nts, a n d 1 m o nt h f oll o wi n g t h e l ast d os e of 6 7 6 6 f or f e m al e p ati e nts.

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 2 2 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
A d ults u n a bl e t o c o ns e nt will b e i ncl u d e d i n t h e st u d y wit h t h e c o n diti o n of t h eir L e g al 
r e pr es e nt ati v e pr o vi di n g i nf or m e d c o ns e nt.
Excl usi o n crit eri a:
1.
Cli nic all y si g nific a nt acti v e g astr oi nt esti n al a b n or m aliti es, r e q uir e m e nt f or s yst e mic 
a ntic o a g ul ati o n or p ot e nt C Y P 2 C 8 i n hi bit ors, a n d hist or y of cli nic all y si g nific a nt c ar di ac 
or p ul m o n ar y dis or d ers
2.
Mi n ors will b e excl u d e d fr o m t h e i n v esti g ati o n. Gli o bl ast o m a is t h e m aj or f or m of br ai n 
c a nc er i n p e o pl e o v er 5 0 y e ars ol d. P e di atric c as es of gli o bl ast o m a ar e r el ati v el y r ar e. 
B esi d es t his, t h er e ar e cr uci al m ol ec ul ar diff er e nc es b et w e e n a d ult a n d p e di atric 
gli o m as. O ur pr eli mi n ar y d at a f or pr o p os e d i n v esti g ati o n w er e o bt ai n e d o n G B M 
s p eci m e ns a n d c ult ur es d e v el o p e d fr o m G B M tiss u es d o n at e d b y a d ult s u bj ects. R es ults 
of i n v esti g ati o n of a d ult gli o m a tiss u e c a n n ot si m pl y b e extr a p ol at e d t o c hil dr e n. 
T h er ef or e, o ur pri m ar y r es e arc h f oc us is t h e i n v esti g ati o n of G B M i n a d ults. If 
a p pr o pri at e, a s e p ar at e, a g e-s p ecific st u d y i n c hil dr e n will b e p erf or m e d.
3.
Pr e g n a nt w o m e n will b e excl u d e d fr o m t h e st u d y as alt er e d h or m o n al a n d 
i m m u n ol o gic al st at us c a n aff ect t h e st u d y r es ults.
4.
Pris o n ers will b e excl u d e d fr o m t h e st u d y.
5.
S yst e mic a nti-c a nc er t h er a p y wit hi n 4 w e e ks of t h e first d os e of st u d y t h er a p y.
6.
Hist or y of pri or m ali g n a nc y, wit h t h e exc e pti o n of c ur ati v el y tr e at e d m ali g n a nci es or 
m ali g n a nci es wit h v er y l o w p ot e nti al f or r ec urr e nc e or pr o gr essi o n.
7.
M aj or s ur g er y wit hi n 4 w e e ks ( excl u di n g pl ac e m e nt of v asc ul ar acc ess), mi n or s ur g er y 
wit hi n 2 w e e ks, or p alli ati v e r a di ot h er a p y wit hi n 1 w e e k of t h e first d os e of t h e 
i n v esti g ati o n al a g e nt ( d ef acti ni b or a v ut o m eti ni b).
8.
Ex p os ur e t o m e dic ati o ns ( wit h or wit h o ut pr escri pti o n), s u p pl e m e nts, h er b al r e m e di es, 
or f o o ds wit h p ot e nti al f or dr u g- dr u g i nt er acti o ns wit h t h e i n v esti g ati o n al a g e nt 
( D ef acti ni b or A v ut o m eti ni b) wit hi n 1 4 d a ys pri or t o t h e first d os e of t h e 
i n v esti g ati o n al a g e nt ( D ef acti ni b or Av ut o m eti ni b) a n d d uri n g t h e c o urs e of t h er a p y, 
i ncl u di n g:
a.
A v ut o m eti ni b : str o n g C Y P 3 A 4, i n hi bit ors or i n d uc ers, d u e t o p ot e nti al dr u g- 
dr u g i nt er acti o ns wit h A v ut o m eti ni b. F or a d diti o n al g ui d a nc e s e e  
htt ps:// w w w.f d a. g o v/ dr u gs/ dr u g-i nt er acti o ns-l a b eli n g/ dr u g- d e v el o p m e nt-
a n d- dr u g-i nt er acti o ns-t a bl e-s u bstr at es-i n hi bit ors- a n d-i n d uc ers
b.
D ef acti ni b: str o n g C Y P 3 A 4, C Y P 2 C 9, a n d P- gl yc o pr ot ei n ( P- g p) i n hi bit ors or 
i n d uc ers, d u e t o p ot e nti al dr u g- dr u g i nt er acti o ns wit h D ef acti ni b. F or a d diti o n al 
g ui d a nc e s e e htt ps:// w w w.f d a. g o v/ dr u gs/ dr u g- i nt er acti o ns-l a b eli n g/ dr u g-
d ev el o p m e nt- a n d- dr u g-i nt er acti o ns-t a bl e-s u bstr at es- i n hi bit ors- a n d-i n d uc ers
9.
K n o w n h e p atitis B, h e p atitis C or h u m a n i m m u n o d efici e nc y vir us ( HI V) i nf ecti o n t h at is 
acti v e a n d/ or r e q uir es t h er a p y.
1 0. Acti v e s ki n dis or d er t h at h as r e q uir e d s yst e mic t h er a p y wit hi n t h e p ast 1 y e ar.
1 1. Hist or y of r h a b d o m y ol ysis.
1 2. C o nc urr e nt oc ul ar dis or d ers:

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 2 3 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
a.
P ati e nts wit h acti v el y tr e at e d gl a uc o m a, r eti n al v ei n occl usi o n ( R V O), i ncl u di n g 
pr e dis p osi n g f act ors f or R V O, i ncl u di n g u nc o ntr oll e d h y p ert e nsi o n, a n d 
u nc o ntr oll e d di a b et es.
b.
P ati e nts wit h c urr e nt e vi d e nc e of visi bl e r eti n al p at h ol o g y t h at is c o nsi d er e d a 
ris k f act or f or R V O, i ntr a oc ul ar pr ess ur e > 2 1 m m H g as m e as ur e d b y 
t o n o m etr y, or ot h er si g nific a nt oc ul ar p at h ol o g y a n d a n at o mic al a b n or m aliti es 
t h at i ncr e as e t h e ris k f or R V O.
c.
P ati e nts wit h acti v e c or n e al er osi o n (i nst a bilit y of c or n e al e pit h eli u m), 
c or n e al d e g e n er ati o n, acti v e or r ec urr e nt k er atitis, a n d ot h er f or ms of s eri o us 
oc ul ar s urf ac e i nfl a m m at or y c o n diti o ns.
1 3. C o nc urr e nt c o n g esti v e h e art f ail ur e, pri or hist or y of cl ass III/ I V c ar di ac dis e as e ( N e w 
Y or k H e art Ass oci ati o n [ N Y H A]), m y oc ar di al i nf arcti o n wit hi n t h e l ast 6 m o nt hs, u nst a bl e 
arr h yt h mi as, u nst a bl e a n gi n a, or s ev er e o bstr ucti v e p ul m o n ar y dis e as e.
1 4. P ati e nts wit h t h e i n a bilit y t o s w all o w or al m e dic ati o ns or i m p air e d g astr oi nt esti n al 
a bs or pti o n d u e t o g astr ect o m y or acti v e i nfl a m m at or y b o w el dis e as e.
1 5. P ati e nts wit h a hist or y of h y p ers e nsiti vit y t o a n y of t h e i n acti v e i n gr e di e nts 
( h y dr ox y pr o p yl m et h ylc ell ul os e, m a n nit ol, m a g n esi u m st e ar at e) of t h e i n v esti g ati o n al 
pr o d uct.
1 2. V ul n er a bl e P o p ul ati o ns
T his r es e arc h d o es n ot i n v ol v e mi n ors, pr e g n a nt w o m e n, b as eli n e c o g niti v el y i m p air e d a d ults, 
or pris o n ers.
1 3. L oc al N u m b er of P artici p a nts
A t ot al of 1 7 p artici p a nts will b e e nr oll e d (c o ns e nt e d) t o r e ac h o ur r ecr uit m e nt g o al of 1 2 p ati e nts 
st u d y wi d e: accr u al will c o nti n u e u ntil t h er e ar e 3 p artici p a nts c o m pl eti n g t h e st u d y i n e ac h 
ex p eri m e nt al gr o u p of d ef acti ni b 2 0 0 m g, d ef acti ni b 4 0 0 m g, a v ut o m et a ni b 3. 2 m g a n d 
a v ut o m et a ni b 4 m g. T his n u m b er of p artici p a nts is n e e d e d t o pr o vi d e m e a ni n gf ul d escri pti v e d at a 
f or t h e r es e arc h pr oc e d ur es. S ex a n d r ac e will n ot b e t a k e n i n c o nsi d er ati o n d uri n g t h e scr e e ni n g 
a n d e nr oll m e nt. D u e t o t his w e a ntici p at e t h at p erc e nt a g e of t h e p artici p a nts acc or di n g t o s ex a n d 
r ac e will r efl ect d e m o gr a p h y i n t h e ar e a of e nr oll m e nt.
1 4. R ecr uit m e nt M et h o ds
P ati e nts will b e i nf or m e d a b o ut t h e st u d y b y t h e i n v esti g at ors a n d/ or a d esi g n at e d c ertifi e d 
R es e arc h C o or di n at or at E m or y Cli nic, t h e Wi ns hi p C a nc er I nstit ut e, or E m or y H os pit al b y 
p ers o n al c o m m u nic ati o n a n d b y t h e pr o vi d e d I nf or m e d C o ns e nt F or m. P ati e nts sc h e d ul e d f or 
G B M t u m or r es ecti o n s ur g er y at E m or y U ni v ersit y H os pit al will b e a p pr o ac h e d. Eli gi bilit y will b e

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 2 4 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
d et er mi n e d b y t h e i n v esti g at ors a n d R es e arc h C o or di n at or b as e d o n t h e s p ecifi e d 
i ncl usi o n/ excl usi o n crit eri a.

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 2 5 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
L a b or at or y r es e arc h ers will n ot h a v e a n y p ers o n al i nf or m ati o n a b o ut t h e p ati e nts d uri n g 
r ecr uit m e nt a n d st u d y.
P ati e nts will h a v e t h e p ossi bilit y t o c o nt act t h e Pri nci p al I n v esti g at or t o cl arif y all p ossi bl e 
q u esti o ns b ef or e a n d d uri n g t h e i n v esti g ati o n.
1 5.
Wit h dr a w al of P artici p a nts
P artici p a nts c a n b e wit h dr a w n fr o m t h e st u d y if t h e br ai n t u m or s ur g er y is c a nc el e d or if 
s ur g er y is p ost p o n e d f or l o n g er t h a n 6- 7 h o urs aft er D E F A C TI NI B or A V U T O M E TI NI B 
a d mi nistr ati o n. I n t h es e c as es, t h e bl o o d s a m pl es a n d s a m pl es of t h e r es ect e d t u m or a n d br ai n 
will n ot b e t a k e n f or a n al ysis.
1 6. Ris k t o P artici p a nts
Pr e vi o us cli nic al tri als h a v e i d e ntifi e d t h e o pti m u m d os e of d ef acti ni b as 4 0 0 m g (J o n es et al., 
2 0 1 5) a n d t h e o pti m u m d os e f or a v ut o m et a ni b as 3. 2 m g ( G u o et al., 2 0 2 0) f or n o n- br ai n t u m or 
m ali g n a nci es. T h e m ost fr e q u e nt tr e at m e nt-r el at e d a d v ers e eff ects d et ect e d i n pr e vi o us 
cli nic al tri als w er e h y p er bilir u bi n e mi a, f ati g u e, d ecr e as e d a p p etit e, a n d di arr h e a f or d ef acti ni b 
w h e n gi v e n f or 2 1 d a ys (t w o ti m es a d a y) (J o n es et al., 2 0 1 5; S hi miz u et al., 2 0 1 5) a n d r as h, 
cr e ati ni n e p h os p h o ki n as e el e v ati o n, h y p o al b u mi n e mi a, a n d f ati g u e f or a v ut o m eti ni b w h e n 
gi v e n f or 2 1 d a ys 2 ti m es a w e e k ( G u o et al., 2 0 2 0). H o w e v er, t h es e r e acti o ns w er e m ostl y n ot 
o bs er v e d at l o w er c o nc e ntr ati o ns, s uc h as 2 0 0 m g a n d 3. 2 r es p ecti v el y f or e ac h dr u g. O ur st u d y 
pr o p os es a si n gl e d os e of d ef acti ni b or a v ut o m eti ni b. F or s af et y, t h e st u d y will b e st art e d wit h 
t h e l o w est d os e of 2 0 0 m g, a n d t h e d os e will b e i ncr e as e d till 4 0 0 m g if t h e i d e ntifi e d 
c o nc e ntr ati o n of d ef acti ni b i n t h e t u m or t o g et h er wit h t h e r e d ucti o n of P y k 2 a n d F A K 
p h os p h or yl ati o n is n ot ac hi e v e d at l o w er d os es. T h e s a m e d os e esc al ati o n fr o m 3. 2 t o 4 m g is 
pr o p os e d f or A v ut o m et a ni b is pr o p os e d if i d e ntifi e d dr u g c o nc e ntr ati o n i n t h e t u m or t o g et h er 
wit h t h e r e d ucti o n of M E K 1/ 2 a n d Er k 1/ 2 p h os p h or yl ati o n is n ot ac hi e v e d at 3. 2 m g d os e. T o 
mi ni miz e ris k t o s u bj ects a n d c o nsi d eri n g a d v ers e r e acti o ns o bs er v e d i n pr evi o us cli nic al tri als, 
s u bj ects wit h cli nic all y si g nific a nt g astr oi nt esti n al a b n or m aliti es, a ntic o a g ul ati o n dis or d ers, a n d 
hist or y of cli nic all y si g nific a nt c ar di ac or p ul m o n ar y dis or d ers will b e excl u d e d fr o m t h e st u d y. 
I n c as e of s e v er e a d v ers e eff ects, t h at pl ac e s u bj ects at ris k r el at e d t o d ef acti ni b or 
a v ut o m eti ni b a d mi nistr ati o n, a p p e ar d uri n g s ur g er y or p ost-s ur gic al r e h a bilit ati o n, t h e st u d y 
wit h gi v e n a n d hi g h er c o nc e ntr ati o ns will b e s us p e n d e d u ntil t h e c a us e of d ef acti ni b or 
a v ut o m eti ni b c o m plic ati o n is u n d erst o o d a n d

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 2 6 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
a d dr ess e d. W e ex p ect t h e l o w ris k of i nj ur y or s ev er e si d e eff ects fr o m all st u d y pr oc e d ur es. 
T h e p ati e nts will b e h a vi n g s ur g er y r e g ar dl ess of t h eir p artici p ati o n i n t his st u d y.
L a b or at or y r es e arc h a n d d at a r e p orti n g will i n v ol v e t h e us e of c o d e d pri v at e i nf or m ati o n a n d 
bi ol o gic al s p eci m e ns o nl y. Ri g hts a n d w elf ar e of s u bj ects ar e ex p ect e d t o b e at mi ni m al ris k.
1 7. P ot e nti al B e n efits t o P artici p a nts
P artici p a nts will n ot r ec ei v e a n y fi n a nci al or m e dic al b e n efits f or t h eir p artici p ati o n.
1 8. C o m p e ns ati o n t o P artici p a nts
N o m o n et ar y c o m p e ns ati o n will b e pr o vi d e d t o p artici p a nts
1 9. D at a M a n a g e m e nt a n d C o nfi d e nti alit y
T h e cli nic al r es e arc h ers will h a v e acc ess t o p ers o n al i nf or m ati o n a b o ut t h e p ati e nts. T h e 
d esi g n at e d c ertifi e d R es e arc h C o or di n at or at E m or y H os pit al will pr o vi d e t h e l a b or at or y 
r es e arc h ers wit h t h e u ni q u e I d e ntific ati o n N u m b er of e ac h s a m pl e. All d oc u m e nts c o nt ai ni n g 
t h e i nf or m ati o n a b o ut t h e n a m es of t h e p ati e nts a n d t h eir I d e ntific ati o n N u m b er will b e k e pt at 
t h e Cli nic al Tri als Offic e at t h e Wi ns hi p C a nc er I nstit ut e i n a p ass w or d pr ot ect e d c o m p ut er i n a 
s af el y l oc k e d offic e.
All m e m b ers of t h e r es e arc h gr o u p, w h o will w or k wit h bi os p eci m e ns, h a v e c ertific ati o n i n 
Bi o m e dic al R es e arc h wit h H u m a ns s u bj ects a n d H e alt h I nf or m ati o n Pri v ac y a n d S ec urit y ( HI P S) 
f or Cli nic al I n v esti g at ors tr ai ni n g. M Ds, t h at ar e pr of essi o n all y i n v ol v e d i n b ot h, p ati e nt’s pr e- 
a n d p ost-s ur gic al m o nit ori n g, s ur g er y pr oc e d ur e, a n d b y t h es e h a v e acc ess t o p ati e nt’s 
i d e ntifi a bl e i nf or m ati o n, a n d w h o ar e als o i n v ol v e d i n t h e st u d y, will m ai nt ai n p ati e nt 
c o nfi d e nti alit y. I n t his c ertific at e t h e y will i n dic at e t h at t h e y n e v er p ass or m a k e it a v ail a bl e b y 
a n y w a y t h e i d e ntifi a bl e p ers o n al i nf or m ati o n r el at e d t o bi os p eci m e ns t o t h e r es e arc h gr o u p.
S p eci m e ns will b e pr o vi d e d t o t h e r es e arc h gr o u p t o g et h er wit h t h e C oll ecti o n I nstr u m e nts o n 
b asic p ati e nt d at a, w h er e g e n d er, a g e, et h nicit y, di a g n osis, a n d ti m e of d ef acti ni b 
i ntr o d ucti o n, a n d t u m or r es ecti o n s ur g er y will b e i n dic at e d.
Bi os p eci m e ns will b e pr oc ess e d i m m e di at el y f or t h e r es e arc h pr oc e d ur es a n d ar e n ot i nt e n d e d 
t o b e st or e d. T h e acc ess t o bi os p eci m e ns will h a v e a Cli nic al R es e arc h C o or di n at or ( w h o will 
p ass s p eci m e ns t o t h e r es e arc h t e a m) a n d t h e m e m b er of t h e r es e arc h gr o u p, w h o will pr oc ess 
s p eci m e ns f or pr ot ei n extr acti o n a n d s e n d t h e m t o E m or y Li pi d o mics C or e. T his m e m b er of t h e

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 2 7 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
r es e arc h gr o u p is t o b e n a m e d l at er a n d will b e r e q uir e d t o h a v e H u m a n S u bj ects R es e arc h a n d 
HI P A A tr ai ni n g a n d c ertific ati o ns. Extr act e d pr ot ei ns f or P Y K 2, F A K, M E K a n d Er k a n al ysis, w hic h 
will o nl y h a v e tiss u e i d e ntific ati o n n u m b er, will b e s hi p p e d fr oz e n t o P u ert o Ric o, C e ntr al 
C ari b b e a n U ni v ersit y ( U C C) o v er ni g ht U P S. T h e tiss u e i nt e n d e d f or p h ar m ac o ki n etic a n d 
p h ar m ac o d y n a mic a n al ysis will b e s e nt t o E m or y Li pi d o mics c or e f or M ass- S p ectr o m etr y
If a p artici p a nt d ecli n es t o p artici p at e i n d ef acti ni b or a v ut o m et a ni b tr e at m e nt a n d f oll o wi n g 
s ur g er y tiss u e c oll ecti o n, t h e p artici p a nt will n ot b e assi g n e d a st u d y I D n u m b er a n d t h e st u d y 
c o or di n at ors/ d at a c oll ect ors will r efr ai n fr o m c oll ecti n g a n y d at a o n t h e p artici p a nt. If t h e 
p artici p a nt a gr e es t o p artici p at e i n d ef acti ni b or a v ut o m et a ni b tr e at m e nt a n d f oll o wi n g t h e 
s ur g er y tiss u e c oll ecti o n, b ut r ef us e d t o pr o vi d e a n y r el at e d i nf or m ati o n, as a g e, g e n d er, 
et h nicit y, t h e p artici p a nt will b e assi g n e d a st u d y I D n u m b er a n d t h e st u d y
c o or di n at ors/ d at a c oll ect ors will b e i nstr uct e d t o c oll ect d at a o nl y o n t h os e as p ects of t h e st u d y 
t o w hic h t h e p artici p a nt h as a gr e e d t o p artici p at e. T h es e pr oc e d ur es will h el p pr ev e nt 
u n a ut h oriz e d i ncl usi o n of t h e p ati e nt’s d at a i n t h e d at a b as e.
Sci e ntific d at a o bt ai n e d i n t h e pr o p os e d i n v esti g ati o n will b e s u bj ect t o p u blic ati o n i n m e dic al 
a n d sci e ntific j o ur n als. T h e p ers o n al d at a r e g ar di n g t h e p ati e nts pr o vi di n g t h e s p eci m e ns will 
n ot b e p u blis h e d.
2 0. Pr o visi o ns t o M o nit or t h e D at a t o E ns ur e t h e S af et y of P artici p a nts
☒ M or e t h a n mi ni m al ris k – C o nti n u e b el o w.
M ar k t h e ris k c at e g oriz ati o n, as d et er mi n e d b y t h e D at a a n d S af et y M o nit ori n g Q u esti o n n air e, 
w hic h a p pli es t o y o ur st u d y b el o w:
S el ect o n e of t h e f oll o wi n g ( d o n ot d el et e t his t a bl e; r e vi e w t h e g ui d a nc e d oc u m e nt 
f or d efi niti o ns):
☐M e di u m C o m pl exit y
☒ Hi g h C o m pl exit y C at e g or y A
☐Hi g h C o m pl exit y C at e g or y B
If c h o osi n g t his c at e g ory f or a st u dy u n d er a n I N D or I D E b ec a us e y o u b eli ev e t h e 
st u dy i nt erv e nti o n d o es n ot si g nific a ntly i m p act m or bi dity or m ort ality, pl e as e 
pr ovi d e y o ur r ati o n al e:

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 2 8 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
D S M P R e q uir e m e nt
H o w t his R e q uir e m e nt is M et
Fr e q u e nc y
R es p o nsi bl e P art y(i es)
R e al-ti m e r e vi e w of 
p artici p a nt d at a d uri n g 
i niti al d at a c oll ecti o n.
T his r e q uir e m e nt will b e m et 
p er Wi ns hi p’s N CI a p pr o v e d 
D S M P
T his will occ ur 
e v er y ti m e n e w 
i nf or m ati o n is 
o bt ai n e d.
SI/st u d y t e a m
Sit e M o nit ori n g at pr e- 
d et er mi n e d i nt er v als: 
T h e Pri nci p al I n v esti g at or 
h as a r es p o nsi bilit y t o 
e ns ur e t h at t h e st u d y is 
f oll o wi n g all as p ects of 
t h e pr ot oc ol.
T his r e q uir e m e nt will b e m et 
p er Wi ns hi p’s N CI a p pr o v e d 
D S M P
bi a n n u all y
D S M C
1 0 0 % r e vi e w of 
r e g ul at or y fil es
D S M C m o nit ors will r e vi e w t h e 
pr ot oc ol, a m e n d m e nts, 
i nf or m e d c o ns e nt d oc u m e nts, 
I R B s u b missi o ns a n d m e et wit h 
t h e pri nci p al i n v esti g at or f or 
cl arific ati o n of st u d y o bj ecti v es
R e vi e w e d at first 
a n d cl os e- o ut visits
D S M C
1 0 0 % r e vi e w of c o ns e nt 
f or ms
M o nt hl y Q A c h ec k of 5- 1 0 
r a n d o ml y s el ect e d c o ns e nts t o 
v ali d at e C e ntr al S u bj ect 
R e gistr ati o n ( C S R) a n d P R M S t o 
c o n d uct Q A c o ns e nt c h ec ks i n 
r e al ti m e as s u bj ects ar e 
r e gist er e d i n O n C or e vis C S R 
pr oc ess
bi a n n u all y
P R M S, Q M
R e vi e w of cr e d e nti als, 
tr ai ni n g r ec or ds, t h e 
d el e g ati o n of 
r es p o nsi bilit y l o gs (if 
a p plic a bl e)
Cli nic al tri als m o nit or will r e vi e w 
t h e el ectr o nic r e g ul at or y bi n d er 
a n d c o m p ar e t h e m a g ai nst t h e 
st aff list e d o n t h e D O A l o g. T h e 
m o nit or als o r evi e ws t h e sit e’s 
s o urc e d oc u m e nts t o e ns ur e 
t h at all st u d y st aff h a v e b e e n 
pr o p erl y list e d o n t h e D O A a n d 
h a v e c orr es p o n di n g 
d oc u m e nt ati o n ( C V, ML, G C P 
c erts, tr ai ni n g l o g) fil e d i n t h e
el ectr o nic r e g ul at or y st u d y 
bi n d er
bi a n n u all y
D S M C
C o m p aris o n of c as e 
r e p ort f or ms ( C R F) t o
s o urc e d oc u m e nt ati o n
T h e PI is r es p o nsi bl e f or 
e ns uri n g t h at i nst a nc es of
e gr e gi o us d at a i ns uffici e nci es
bi a n n u all y
D S M C

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 2 9 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
f or acc ur ac y a n d 
c o m pl eti o n
t h at m a y i m p act t h e sci e ntific 
i nt e grit y of t h e tri al
R e vi e w of 
d oc u m e nt ati o n of all 
a d v ers e e v e nts
D uri n g t h e m o nit ori n g pr oc ess, 
t h e D S M B r e vi e ws tri al s af et y 
d at a f or st o p pi n g r ul es, 
d e vi ati o ns, st u d y a m e n d m e nts, 
accr u al r at es a n d m o nit ori n g 
r e p orts f or t h er a p e utic 
i n v esti g at or-i niti at e d cli nic al 
tri als a n d a n y ot h er tri al as 
d e e m e d n ec ess ar y
bi a n n u all y
D S M C
M o nit ori n g of critic al 
d at a p oi nts ( eli gi bilit y, 
st u d y e n d p oi nts, etc.)
T h e assi g n e d m o nit or will 
r a n d o ml y s el ect s u bj ect(s) f or 
r e vi e w b as e d o n p ar a m et ers i n 
T a bl e 1 or 2 as n ot e d a b o v e.
Alt h o u g h t h e pri nci p al 
i n v esti g at or a n d a p plic a bl e 
st u d y t e a m m e m b ers will 
r ec ei v e n otific ati o n of tri al 
m o nit ori n g i n a d v a nc e, t h e 
s u bj ect s el ecti o n will n ot b e 
r e v e al e d i n a d v a nc e of t h e 
m o nit ori n g visit.
bi a n n u all y
D S M C
L a b or at or y r e vi e w of 
pr oc essi n g a n d st or a g e 
of s p eci m e ns
T h e assi g n e d m o nit or will 
r a n d o ml y s el ect s u bj ect(s) f or
r e vi e w b as e d o n p ar a m et ers i n 
T a bl e 1 or 2 as n ot e d a b o v e
R e vi e w e d at first 
a n d cl os e- o ut visits
a n d at l e ast 
bi a n n u all y
D S M C
Ass ess m e nt of l a b or at or y 
s p eci m e ns st or e d l oc all y
If accr u al at ti m e of i niti al 
m o nit ori n g is > I O b ut : S 2 0, I 0 % 
of s u bj ects will b e m o nit or e d at 
mi ni m u m. T h er e aft er, 
m o nit ori n g will n ot occ ur u nl ess
accr u al r e ac h es 3 0
R e vi e w e d at first 
a n d cl os e- o ut visits 
a n d at l e ast 
bi a n n u all y
D S M C
T est articl e 
acc o u nt a bilit y r e vi e w
I n a d diti o n t o a c o m pr e h e nsi v e 
r e vi e w of a v ail a bl e t oxicit y d at a, 
t h e D S M C r evi e ws all i nt er n al 
m o nit ori n g r e p orts of tri als 
u n d er its p urvi e w
R e vi e w e d at first 
a n d cl os e- o ut visits 
a n d at l e ast 
bi a n n u all y
D S M C
Acc o u nt a bilit y l o gs, 
dis p e nsi n g r ec or ds, a n d 
ot h er p artici p a nt r ec or ds
If accr u al at ti m e of i niti al 
m o nit ori n g is > I O b ut : S 2 0, I 0 % 
of s u bj ects will b e m o nit or e d at 
mi ni m u m. T h er e aft er,
At l e ast bi a n n u all y
D S M C

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 3 0 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
m o nit ori n g will n ot occ ur u nl ess 
accr u al r e ac h es 3 0.
F or F D A r e g ul at e d 
st u di es, t h e f oll o wi n g 
r e q uir e m e nts a p pl y:
m o nit ori n g acti viti es m e et t h e 
F D A's r e q uir e m e nts as 
d eli n e at e d i n 2 1 C F R 5 0, 2 1 C F R
5 6, 2 1 C F R 8 1 2 f or st u di es 
c o n d uct e d u n d er a n I D E a n d 2 1 
C F R 3 1 2 f or st u di es c o n d uct e d 
u n d er a n I N D.
bi a n n u all y
D S M C
M o nit ori n g m et h o ds 
( m a y i ncl u d e c e ntr aliz e d, 
o n-sit e, a n d s elf- 
m o nit ori n g)
T h e i nt er n al m o nit ori n g t e a m is 
i n d e p e n d e nt fr o m a n y st u d y 
pr ot oc ol a n d d o es n ot p erf or m 
a n y tri al-r el at e d s p ecific d uti es 
i n or d er t o u p h ol d a n u n bi as e d 
a p pr o ac h t o st u d y m o nit ori n g. 
O v ersi g ht of t h e m o nit ori n g 
pr oc ess a n d 
i d e ntific ati o n/ assi g n m e nt of 
st u di es f or m o nit ori n g is 
pr o vi d e d b y t h e M a n a g er of t h e 
I nt er n al M o nit ors.
bi a n n u all y
D S M C
* F or i nt er n ati o n al st u di es, y o u ar e r e q uir e d t o e n g a g e a C R O t h at is w or ki n g i n t h e sit e c o u ntr y a n d/ or t o 
c o ns ult wit h E m or y’s l e g al c o u ns el r e g ar di n g c o m pli a nc e wit h t h e c o u ntr y’s cli nic al r es e arc h r e g ul ati o ns.
T h e D at a a n d S af et y M o nit ori n g C o m mitt e e ( D S M C) of t h e Wi ns hi p C a nc er I nstit ut e will pr o vi d e 
o v ersi g ht f or t h e c o n d uct of t his st u d y. T h e D S M C f u ncti o ns i n d e p e n d e ntl y wit hi n Wi ns hi p 
C a nc er I nstit ut e t o c o n d uct i nt er n al m o nit ori n g f u ncti o ns t o e ns ur e t h at r es e arc h b ei n g 
c o n d uct e d b y Wi ns hi p C a nc er I nstit ut e I n v esti g at ors pr o d uc es hi g h- q u alit y sci e ntific d at a i n a 
m a n n er c o nsist e nt wit h g o o d cli nic al pr actic e ( G C P) a n d a p pr o pri at e r e g ul ati o ns t h at g o v er n 
cli nic al r es e arc h.
D e p e n di n g o n t h e ris k l ev el of t h e pr ot oc ol, t h e D S M C r e vi e w m a y occ ur 
e v er y 6 m o nt hs or a n n u all y. F or st u di es d e e m e d Hi g h Ris k, i niti al st u d y m o nit ori n g will occ ur 
wit hi n 6 m o nt hs fr o m t h e d at e of t h e first s u bj ect accr u e d, wit h 2 of t h e first 5 s u bj ects b ei n g 
r e vi e w e d. F or st u di es d e e m e d M o d er at e Ris k, i niti al st u d y m o nit ori n g will occ ur wit hi n 1 y e ar 
fr o m t h e d at e of t h e first s u bj ect accr u e d, wit h 2 of t h e first 5 s u bj ects b ei n g r e vi e w e d.
S u bs e q u e nt m o nit ori n g will occ ur i n r o uti n e i nt er v als p er t h e Wi ns hi p D at a a n d S af et y 
M o nit ori n g Pl a n ( D S M P) .
T h e D S M C will r e vi e w p erti n e nt as p ects of t h e st u d y t o ass ess s u bj ect s af et y, c o m pli a nc e wit h 
t h e pr ot oc ol, d at a c oll ecti o n, a n d ris k- b e n efit r ati o. S p ecific all y, t h e Wi ns hi p C a nc er I nstit ut e 
I nt er n al M o nit ors assi g n e d t o t h e D S M C m a y v erif y i nf or m e d c o ns e nt, eli gi bilit y, d at a e ntr y, 
acc ur ac y a n d a v ail a bilit y of s o urc e d oc u m e nts, A Es/ S A Es, a n d ess e nti al r e g ul at or y d oc u m e nts. 
F oll o wi n g t h e m o nit ori n g r e vi e w, m o nit ors will pr o vi d e a pr eli mi n ar y r e p ort of m o nit ori n g 
fi n di n gs t o t h e PI a n d ot h er p erti n e nt i n di vi d u als i n v ol v e d i n t h e c o n d uct of t h e st u d y. T h e PI is

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 3 1 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
r e q uir e d t o a d dr ess a n d r es p o n d t o all t h e d efici e nci es n ot e d i n t h e pr eli mi n ar y r e p ort. Pri or t o 
t h e c o m pl eti o n of t h e fi n al s u m m ar y r e p ort, m o nit ors will disc uss t h e pr eli mi n ar y r e p ort 
r es p o ns es wit h t h e PI a n d ot h er t e a m m e m b ers ( w h e n a p pr o pri at e). A fi n al m o nit ori n g 
s u m m ar y r e p ort will t h e n b e pr e p ar e d b y t h e m o nit or. Fi n al D S M C r evi e w will i ncl u d e t h e fi n al 
m o nit ori n g s u m m ar y r e p ort wit h c orr es p o n di n g PI r es p o ns e, s u b mitt e d C A P A ( w h e n 
a p plic a bl e), PI S u m m ar y st at e m e nt, a n d a v ail a bl e a g gr e g at e t oxicit y a n d s af et y d at a.
T h e D S M C will r e n d er a r ec o m m e n d ati o n a n d r ati n g b as e d o n t h e o v er all tri al c o n d uct. T h e PI 
is r es p o nsi bl e f or e ns uri n g t h at i nst a nc es of e gr e gi o us d at a i ns uffici e nci es ar e r e p ort e d t o t h e 
I R B. C o nti n ui n g R e vi e w s u b missi o ns will i ncl u d e t h e D S M C r ec o m m e n d ati o n l ett er. S h o ul d a n y 
r e visi o ns b e m a d e t o t h e pr ot oc ol-s p ecific m o nit ori n g pl a n aft er i niti al D S M C a p pr o v al, t h e PI 
will b e r es p o nsi bl e f or n otif yi n g t h e D S M C of s uc h c h a n g es. T h e C o m mitt e e r es erv es t h e ri g ht 
t o c o n d uct a d diti o n al a u dits if n ec ess ar y.
T h e r es e arc h i n v ol v es m or e t h a n Mi ni m al Ris k t o p artici p a nts, as or al a d mi nistr ati o n of a 
d ef acti ni b or a v ut o m eti ni b si n gl e d os e is pr o p os e d. T h e Wi ns hi p C a nc er I nstit ut e D at a a n d S af et y 
M o nit ori n g C o m mitt e e will m o nit or t his P h as e 0 cli nic al tri al. T h e D S M C f u ncti o ns i n d e p e n d e ntl y 
wit hi n Wi ns hi p C a nc er I nstit ut e t o c o n d uct i nt er n al m o nit ori n g f u ncti o ns t o e ns ur e t h at r es e arc h 
b ei n g c o n d uct e d b y Wi ns hi p C a nc er I nstit ut e I n v esti g at ors pr o d uc es hi g h- q u alit y sci e ntific d at a 
i n a m a n n er c o nsist e nt wit h g o o d cli nic al pr actic e ( G C P) a n d a p pr o pri at e r e g ul ati o ns t h at g o v er n 
cli nic al r es e arc h. I niti al st u d y m o nit ori n g will occ ur wit hi n 6 m o nt hs fr o m t h e d at e of t h e first 
s u bj ect accr u e d, wit h 2 of t h e first 5 s u bj ects b ei n g r e vi e w e d. T h e D S M C will r e vi e w p erti n e nt 
as p ects of t h e st u d y t o ass ess s u bj ect s af et y, c o m pli a nc e wit h t h e pr ot oc ol, d at a c oll ecti o n, a n d 
ris k- b e n efit r ati o. S p ecific all y, t h e Wi ns hi p C a nc er I nstit ut e I nt er n al M o nit ors assi g n e d t o t h e 
D S M C m a y v erif y i nf or m e d c o ns e nt, eli gi bilit y, d at a e ntr y, acc ur ac y a n d a v ail a bilit y of s o urc e 
d oc u m e nts, A Es/ S A Es, a n d ess e nti al r e g ul at or y d oc u m e nts. F oll o wi n g t h e m o nit ori n g r evi e w, 
m o nit ors will pr o vi d e a pr eli mi n ar y r e p ort of m o nit ori n g fi n di n gs t o t h e PI a n d ot h er p erti n e nt 
i n di vi d u als i n v ol v e d i n t h e c o n d uct of t h e st u d y. T h e PI is r e q uir e d t o a d dr ess a n d r es p o n d t o all 
t h e d efici e nci es n ot e d i n t h e pr eli mi n ar y r e p ort. Pri or t o t h e c o m pl eti o n of t h e fi n al s u m m ar y 
r e p ort, m o nit ors will disc uss t h e pr eli mi n ar y r e p ort r es p o ns es wit h t h e PI a n d ot h er t e a m 
m e m b ers ( w h e n a p pr o pri at e). A fi n al m o nit ori n g s u m m ar y r e p ort will t h e n b e pr e p ar e d b y t h e 
m o nit or. Fi n al D S M C r e vi e w will i ncl u d e t h e fi n al m o nit ori n g s u m m ar y r e p ort wit h c orr es p o n di n g 
PI r es p o ns e, s u b mitt e d C A P A ( w h e n a p plic a bl e), PI S u m m ar y st at e m e nt, a n d a v ail a bl e a g gr e g at e 
t oxicit y a n d s af et y d at a. T h e D S M C will r e n d er a r ec o m m e n d ati o n a n d r ati n g b as e d o n t h e o v er all 
tri al c o n d uct. T h e PI is r es p o nsi bl e f or e ns uri n g t h at i nst a nc es of e gr e gi o us d at a i ns uffici e nci es 
ar e r e p ort e d t o t h e I R B. C o nti n ui n g R e vi e w s u b missi o ns will i ncl u d e t h e D S M C r ec o m m e n d ati o n 
l ett er. S h o ul d a n y r e visi o ns b e m a d e t o t h e pr ot oc ol-s p ecific m o nit ori n g pl a n aft er i niti al D S M C 
a p pr o v al, t h e PI will b e r es p o nsi bl e f or n otif yi n g t h e D S M C of s uc h c h a n g es. T h e C o m mitt e e 
r es er v es t h e ri g ht t o c o n d uct a d diti o n al a u dits if n ec ess ar y.
D at a a n d S af et y M o nit ori n g pl a n:

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 3 2 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
-
S u bj ects will b e m o nit or e d h o url y aft er or al a d mi nistr ati o n of d ef acti ni b or 
a v ut o m et a ni b f or st o m ac h p ai n, n a us e a, dizzi n ess, f ati g u e, di arr h e a as t h es e eff ects 
w er e r e p ort e d i n pr e vi o us cli nic al tri al st u di es wit h us e of d ef acti ni b a n d 
a v ut o m et a ni b (J o n es et al., 2 0 1 5; G u o et al., 2 0 2 0). If a d diti o n al s y m pt o ms a p p e ar, 
t h e y will b e r ec or d e d a n d a d dr ess e d b y PI a n d Cli nic al R es e arc h c o or di n ati o n st aff.
-
D uri n g t h e s ur g er y, p ossi bl e r el at e d a d v ers e eff ects (s uc h as exc ess bl e e di n g, h e art, a n d 
p ul m o n ar y c o m plic ati o ns) will b e r ec or d e d a n d r e p ort e d b y t h e PI or t h e o p er ati n g 
s ur g e o n.
-
I n t h e p ost-s ur gic al p eri o d s u bj ects will b e m o nit or e d b y t h e h os pit al p ers o n n el a n d 
Cli nic al R es e arc h c o or di n at or acc or di n g t o r o uti n e p osts ur gic al c ar e pr ot oc ol. All 
p ossi bl e r el at e d a d v ers e eff ects will b e r e p ort e d t o PI.
-
A s u bj ect will b e r e m o v e d fr o m st u d y p artici p ati o n i n c as e of a d v ers e r e acti o ns l e a di n g 
t o d el a y of s ur g er y f or l o n g er t h a n 6- 7 h o urs. T h e d ecisi o n will b e m a d e b y t h e PI.
-
If t h er e ar e a d v ers e r e acti o ns, as ev al u at e d b y t h e pri m ar y p h ysici a n ar e m or e t h a n mil d 
( gr a d e 2) a n d ar e s us p ect e d t o b e r el at e d t o d ef acti ni b or a v ut o m eti ni b a d mi nistr ati o n, 
t h e PI will p ost p o n e f urt h er accr u al t o t h e st u d y f or t h e e v al u ati o n of t h e ris k f or f ut ur e 
s u bj ects. If ris k is f o u n d t o b e si g nific a nt, t h e a d mi nist er e d d os e will b e r e d uc e d i n 
c o ns ult ati o n wit h t h e D at a S af et y M o nit ori n g C o m mitt e e f or ot h er p artici p a nts a n d 
st u d y will b e r es u m e d. If ris k is ev al u at e d as n ot si g nific a nt, t h e st u d y will b e c o nti n u e d 
as pl a n n e d. If a s e v er e a d v ers e eff ect is r e p e at e d i n a n ot h er s u bj ect ev e n aft er d os e 
m o dific ati o n, t h e PI will st o p t h e st u d y f or f urt h er e v al u ati o n of m ec h a nis ms of t h e 
s e v er e a d v ers e r e acti o n r est arti n g or c urt aili n g t h e st u d y will b e c arri e d o ut i n 
c o ns ult ati o n wit h t h e D at a S af et y M o nit ori n g C o m mitt e e.
If n o m e as ur a bl e a m o u nts of eit h er d ef acti ni b or a v ut o m et a ni b is i d e ntifi e d at t h e t w o 
d os e l e v els pr o p os e d, a n d a d v ers e e v e nts e nc o u nt er e d h a v e b e e n gr a d e 3 or l ess, t h e 
i n v esti g at ors will c o ns ult wit h V er ast e m a n d pr o p os e f urt h er d os e l ev els (t e nt ati v el y 5 0 0 
m g a n d 7 5 0 m g f or d ef acti ni b, a n d 5 m g f or a v ut o m eti ni b, b as e d o n t h e m axi m u m 
ex pl or e d si n gl e d os es t ests n ot e d i n i n v esti g at or’s br oc h ur es f or e ac h) t o t h e D S M C. 
Pr o vi d e d t h er e is a p pr o v al, t h e st u d y will t h e n c o nti n u e usi n g t h e s a m e sc h e m e as w as 
us e d i n l o w er d os e l e v els. If t h er e is still n o m e as ur a bl e dr u g i n t u m or aft er c o m pl eti o n 
of t h es e a d diti o n al d os e l e v els, t h e st u d y will b e st o p p e d, a n d c o ns ult ati o n h el d wit h 
V er ast e m a n d t h e D S M C r e g ar di n g f urt h er esc al ati o n str at e gi es or cl osi n g t h e st u d y.
R e p orti n g R e q uir e m e nts f or I N D h ol d er
F or I n v esti g at or-s p o ns or e d I N D st u di es, r e p orti n g r e q uir e m e nts f or t h e F D A a p pl y i n acc or d a nc e 
wit h t h e g ui d a nc e s et f ort h i n 2 1 C F R, P art 3 1 2. 3 2. E v e nts m e eti n g t h e f oll o wi n g crit eri a n e e d t o 
b e s u b mitt e d t o t h e F D A as Ex p e dit e d I N D S af et y R e p orts.
7 C al e n d ar- D a y T el e p h o n e or F ax R e p ort
T h e S p o ns or-I n v esti g at or is r e q uir e d t o n otif y t h e F D A of a f at al or lif e-t hr e at e ni n g a d v ers e e v e nt 
t h at is u n ex p ect e d a n d ass ess e d b y t h e i n v esti g at or t o b e p ossi bl y r el at e d t o t h e us e of 
i nv esti g ati o n al a g e nts. A n u n ex p ect e d a d v ers e e v e nt is o n e t h at is n ot alr e a d y d escri b e d i n t h e

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 3 3 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
m ost r ec e nt G ui d a nc e f or I n v esti g at or s ecti o n of t h e I n v esti g at or’s Br oc h ur e. S uc h r e p orts ar e t o 
b e t el e p h o n e d or f ax e d t o t h e F D A, wit hi n 7 c al e n d ar d a ys of t h e first l e ar ni n g of t h e e v e nt.
1 5 C al e n d ar- D a y Writt e n R e p ort
T h e S p o ns or-I n v esti g at or is als o r e q uir e d t o n otif y t h e F D A a n d all p artici p ati n g i n v esti g at ors, i n 
a writt e n I N D S af et y R e p ort, of a n y s eri o us u n ex p ect e d a d v ers e ev e nt t h at is c o nsi d er e d 
r e as o n a bl y or p ossi bl y r el at e d t o t h e us e of i n v esti g ati o n al a g e nt.
Writt e n I N D S af et y R e p orts wit h a n al ysis of si mil ar e v e nts ar e t o b e s u b mitt e d t o t h e F D A, wit hi n 
1 5 c al e n d ar d a ys of first l e ar ni n g of t h e ev e nt. T h e F D A pr ef ers t h es e r e p orts o n a M e d W atc h 
3 5 0 0 F or m b ut alt er n ati v e f or m ats ( e. g., s u m m ar y l ett er) ar e acc e pt a bl e.
F D A F ax n u m b er of I N D S af et y R e p orts: 1-( 8 0 0)- F D A- 1 0 7 8.
T h e I N D s p o ns or will als o ass ess w h et h er t h e ev e nt c o nstit ut es a n u n a ntici p at e d pr o bl e m p osi n g 
ris ks t o s u bj ects or ot h ers ( U P). T his ass ess m e nt will b e pr o vi d e d t o t h e E m or y U ni v ersit y I R B, 
w hic h, i n t ur n will m a k e a fi n al d et er mi n ati o n. If t h e E m or y I R B d et er mi n es a n ev e nt is a U P it will 
n otif y t h e a p pr o pri at e r e g ul at or y a g e nci es a n d i nstit uti o n al offici als.
All A d v ers e E v e nts will b e r e p ort e d t o r e g ul at or y a ut h oriti es, I R B/I E Cs a n d i n v esti g at ors i n 
acc or d a nc e wit h all a p plic a bl e gl o b al l a ws a n d r e g ul ati o ns.
2 1. Pr o visi o ns t o Pr ot ect t h e Pri v ac y I nt er est of P artici p a nts
T h e Cli nic al R es e arc h C o or di n at or will h a v e acc ess t o s u bj ects a n d t h eir p ers o n al i nf or m ati o n. 
T h e Cli nic al R es e arc h C o or di n at or will pr o vi d e s u bj ect i nf or m ati o n r el at e d t o t h e st u d y, a ns w er 
q u esti o ns, a n d cl arif y pr oc e d ur es t o b e p erf or m e d. If a s u bj ect w a nts t o c o nt act t h e PI dir ectl y 
f or d e e p er cl arific ati o ns, t h e y c a n d o it b y t h e p ati e nt p ort al, p h o n e or i n a p ers o n al m e eti n g.
T h e l a b or at or y r es e arc h ers will n ot h a v e acc ess t o a n y s o urc es of p ers o n al i d e ntif yi n g 
i nf or m ati o n a b o ut t h e p artici p a nts. T h e R es e arc h C o or di n at or will pr o vi d e r es e arc h ers wit h t h e 
u ni q u e I d e ntific ati o n N u m b er of e ac h s a m pl e b ut n ot wit h t h e n a m e or p ers o n al i d e ntif yi n g 
i nf or m ati o n of t h e p ati e nt. All d oc u m e nts c o nt ai ni n g t h e i nf or m ati o n a b o ut t h e n a m es of t h e 
p ati e nts a n d t h eir I d e ntific ati o n N u m b er will b e k e pt at t h e Cli nic al Tri als Offic e at t h e Wi ns hi p 
C a nc er I nstit ut e i n a p ass w or d pr ot ect e d c o m p ut er.
2 2. Ec o n o mic B ur d e n t o P artici p a nts
P artici p a nts will n ot r ec ei v e a n y p a y m e nts i n r el ati o n t o t h eir p artici p ati o n i n t h e st u d y.
2 3. I nf or m e d C o ns e nt

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 3 4 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
T h e i niti al i nf or m e d c o ns e nt disc ussi o n will occ ur i n Wi ns hi p C a nc er I nstit ut e or t h e E m or y 
Cli nic.
At Wi ns hi p C a nc er I nstit ut e, t h e i nf or m e d c o ns e nt is a n o n g oi n g, i nt er acti v e pr oc ess r at h er t h a n 
a o n e-ti m e i nf or m ati o n s essi o n. T h e c o ns e nt f or m d oc u m e nt is d esi g n e d t o b e gi n t h e i nf or m e d 
c o ns e nt pr oc ess, w hic h pr o vi d es t h e p ati e nt wit h o n g oi n g ex pl a n ati o ns t h at will h el p t h e m 
m a k e e d uc ati o n al d ecisi o ns a b o ut w h et h er t o b e gi n or c o nti n u e p artici p ati n g i n t h e tri al. T h e 
r es e arc h t e a m k n o ws t h at a writt e n d oc u m e nt al o n e m a y n ot e ns ur e t h at t h e p ati e nt f ull y 
u n d erst a n ds w h at p artici p ati o n m e a ns. T h er ef or e, t h e r es e arc h t e a m will disc uss wit h t h e 
p ati e nt t h e tri al's p ur p os e, pr oc e d ur es, ris ks a n d p ot e nti al b e n efits, a n d t h eir ri g hts as a 
p artici p a nt. T h e t e a m will c o nti n u e t o u p d at e t h e p ati e nt o n a n y n e w i nf or m ati o n t h at m a y 
aff ect t h eir sit u ati o n.
C o ns e nt will b e o bt ai n e d pri or t o a n y r es e arc h- dri v e n pr oc e d ur es. T h e i n v esti g at or will ass ess 
t h e p ati e nt’s c a p acit y d uri n g his/ h er e nc o u nt ers wit h hi m or h er. T h e i n v esti g at or will gi v e t h e 
p ers o n pr o vi di n g c o ns e nt a d e q u at e o p p ort u nit y t o r e a d t h e c o ns e nt d oc u m e nt b ef or e it is 
si g n e d a n d d at e d.
It will b e ex pl ai n e d t o pr os p ecti v e p artici p a nts t h at t h e st u d y i n v ol v es r es e arc h, t h e p ur p os e of 
t h e r es e arc h, t h e ex p ect e d d ur ati o n of p artici p ati o n, as w ell as t h e a p pr oxi m at e n u m b er of 
p artici p a nts t o b e e nr oll e d. T h e st u d y pr oc e d ur es, a n d i d e ntific ati o n of r es e arc h pr oc e d ur es v. 
n o n-r es e arc h will als o b e t h or o u g hl y disc uss e d. It will b e ex pl ai n e d t o p artici p a nts t h at 
p artici p ati o n is v ol u nt ar y a n d t h at t h e s u bj ect m a y disc o nti n u e at a n y ti m e.
R ef us al t o p artici p at e or wit h dr a w will n ot i n v ol v e a p e n alt y or l oss of b e n efits t o w hic h t h e 
p artici p a nt is ot h er wis e e ntitl e d. R ef us al will i n n o w a y aff ect t h e p artici p a nt's f ut ur e ar e. T h e 
p artici p a nt will als o b e t ol d of t h e p ossi bl e c o ns e q u e nc es of t h e d ecisi o n t o wit h dr a w fr o m t h e 
r es e arc h, a n d pr oc e d ur es f or or d erl y t er mi n ati o n of p artici p ati o n.
A n y si g nific a nt n e w fi n di n gs d e v el o p e d d uri n g t h e c o urs e of t h e r es e arc h t h at m a y aff ect t h e 
p artici p a nt's willi n g n ess t o c o nti n u e t o p artici p at e will b e pr o vi d e d. Als o ex pl ai n e d will b e 
a ntici p at e d circ u mst a nc es u n d er w hic h t h e s u bj ect's p artici p ati o n m a y b e t er mi n at e d b y t h e 
i n v esti g at or wit h o ut t h e p artici p a nt's c o ns e nt.
Pr os p ecti v e p artici p a nts will b e pr o vi d e d wit h a d escri pti o n of a n y r e as o n a bl y f or es e e a bl e ris ks 
or disc o mf orts as w ell as a d escri pti o n of a n y b e n efits t o t h e p artici p a nt or t o ot h ers t h at mi g ht 
b e r e as o ns ex p ect e d fr o m t h e r es e arc h. Alt er n ati v e pr oc e d ur es or c o urs es of tr e at m e nt will 
als o b e t h or o u g hl y disc uss e d.
Pr os p ecti v e p artici p a nts will als o b e gi v e n d et ail e d i nf or m ati o n d escri bi n g t h e ext e nt t o w hic h 
c o nfi d e nti alit y of r ec or ds i d e ntif yi n g t h e p artici p a nt will b e m ai nt ai n e d a n d w h at r ec or ds m a y 
b e ex a mi n e d b y t h e r es e arc h st aff, I R Bs, s p o ns or, t h eir r e pr es e nt ati v es, a n d p ossi bl y t h e F D A or 
O H R P.

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 3 5 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
Als o c o m m u nic at e d t o t h e p artici p a nt will b e a n ex pl a n ati o n t h at e m er g e nc y m e dic al c ar e will 
b e arr a n g e d f or a st u d y-r el at e d ill n ess or i nj ur y, a n d a n ex pl a n ati o n of w h et h er f u n ds ar e s et 
asi d e t o p a y f or t his c ar e a n d/ or c o m p e ns ati o n, a n d if s o b y w h o m ( e. g., s p o ns or, s u bj ect, 
i ns ur er). T h e p artici p a nt is t ol d t h e s o urc e of t h e st u d y's f u n di n g.
All p artici p a nts will b e t ol d of a n y a d diti o n al c osts t h at m a y r es ult fr o m p artici p ati o n i n t h e 
r es e arc h.
C o ns e nt will b e d o n e i n p ers o n or r e m ot el y t hr o u g h s ec ur e d e m ail, p h o n e or b y el ectr o nic 
c o ns e nti n g usi n g o n e of t h e m et h o ds t h at is E m or y LI T S a p pr o v e d ( e. g., D oc u Si g n) w h e n 
a v ail a bl e. W e will f oll o w E m or y’s g ui d a nc e o n us e of el ectr o nic i nf or m e d c o ns e nt.
N o n- E n glis h- S p e a ki n g P artici p a nts
A c ertifi e d tr a nsl at or/i nt er pr et er will b e pr es e nt d uri n g t h e c o ns e nti n g pr oc ess a n d all 
q u esti o ns a n d c o nc er ns will b e a ns w er e d b y t h e tr e ati n g p h ysici a n.
A S h ort F or m i n t h at s p ecific l a n g u a g e will b e us e d. A c ertifi e d tr a nsl at or/i nt er pr et er will b e 
pr es e nt d uri n g t h e c o ns e nti n g pr oc ess, a n d t his will b e d oc u m e nt e d. W e will us e w h at's 
a v ail a bl e o n E m or y I R B w e bsit e. F or t h e l a n g u a g es t h at ar e n ot a v ail a bl e, w e will h a v e t h e s h ort 
f or m tr a nsl at e d t o t h at l a n g u a g e a n d s u b mit t h e I R B f or r evi e w a n d a p pr o v al pri or t o us e.
Pr oc ess t o D oc u m e nt C o ns e nt i n Writi n g: Wi ns hi p S O P 2. 1:” O bt ai ni n g I nf or m e d c o ns e nt f or 
I nt er v e nti o n al cli nic al tri al” will b e f oll o w e d.
P artici p a nts w h o ar e n ot y et a d ults (i nf a nts, c hil dr e n, t e e n a g ers) : N/ A
C o g niti v el y I m p air e d A d ults : will b e i d e ntifi e d b as e d o n pr of essi o n al o pi ni o n of 
t h eir pri m ar y p h ysici a n. I n t his c as e t h eir L e g all y A ut h oriz e d R e pr es e nt ati v e will 
u n d er g o a n i nf or m e d c o ns e nt pr oc e d ur e as p er E m or y I R B p olic y.
A d ults U n a bl e t o C o ns e nt : p er missi o n will b e o bt ai n e d, vi a i nf or m e d c o ns e nt, fr o m 
t h e L e g all y A ut h oriz e d R e pr es e nt ati v e i n t h e n ext or d er of pri orit y: s p o us e, a d ult 
c hil d, c o urt- a p p oi nt e d g u ar di a n, or att or n e y f or h e alt h c ar e.
W ai v er or Alt er ati o n of C o ns e nt Pr oc ess (c o ns e nt will n ot b e o bt ai n e d, r e q uir e d 
i nf or m ati o n will n ot b e discl os e d, or t h e r es e arc h i n v ol v es d ec e pti o n): N/ A
2 4. S etti n g
T h e r es e arc h t e a m will c o n d uct t h e r es e arc h i n t w o sit es: E m or y U ni v ersit y H os pit al a n d 
U ni v ersi d a d C e ntr al d el C ari b e.
I d e ntific ati o n a n d r ecr uit m e nt of p ot e nti al p artici p a nts will b e c o n d uct e d at E m or y U ni v ersit y 
H os pit al a n d Cli nic, a n d t h e Wi ns hi p C a nc er I nstit ut e.

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 3 6 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
T h e r es e arc h pr oc e d ur es will b e p erf or m e d at E m or y U ni v ersit y ( pr e p ar ati o n of pr ot ei n s a m pl es 
& p h ar m ac o ki n etics/ p h ar m ac o d y n a mics), a n d at C e ntr al C ari b b e a n U ni v ersit y ( U ni v ersi d a d 
C e ntr al d el C ari b e) ( w est er n bl ot a n al ysis of pr ot ei n s a m pl es i n or d er t o e v al u at e t h e l ev el of 
P y k 2, F A K, M E K a n d Er k si g n ali n g acti v ati o n).
R es e arc h pr oc e d ur es at t h e U ni v ersi d a d C e ntr al d el C ari b e will b e c o n d uct e d b as e d o n t h e 
U ni v ersi d a d C e ntr al d el C ari b e I R B a p pr o v al, w hic h is c o nti n g e nt u p o n a n d b as e d u p o n t h e 
E m or y I R B a p pr o v al.
T h e U ni v ersi d a d C e ntr al d el C ari b e H u m a n R es e arc h S u bj ects Pr ot ecti o n Offic e a n d I nstit uti o n al 
R e vi e w B o ar d will r e vi e w E m or y I R B a p pr o v al d oc u m e nt ati o n a n d c ertif y t h eir r es e arc h 
pr oc e d ur es ar e i n li n e wit h t h e st a n d ar ds at t h e U ni v ersi d a d C e ntr al d el C ari b e.
2 5. R es o urc es A v ail a bl e
A p pr oxi m at el y 1 0 0 G B M s ur g eri es ar e p erf or m e d at E m or y U ni v ersit y H os pit al a n n u all y. B as e d 
o n o ur pr e vi o us ex p eri e nc e, ar o u n d 2 0 % of p ot e nti al p artici p a nts si g n c o ns e nt a n d p artici p at e 
i n st u di es. B as e d o n t his a p pr oxi m ati o n, w e will n e e d t o a p pr o ac h a p pr oxi m at el y 6 0 p ot e nti al 
p artici p a nts t o e nr oll 1 2 p artici p a nts t h at c o m pl et e t h e st u d y as w e pl a n. T h us, E m or y 
U ni v ersit y H os pit al h as r es o urc es f or e nr oll m e nt of t h e n ec ess ar y n u m b er of p artici p a nts 
d uri n g t h e y e ar of accr u al. T h e i n v esti g at ors will d e v ot e a t ot al of a p pr oxi m at el y o n e c al e n d ar 
y e ar t o accr ui n g c as es a n d 1 8 m o nt hs c o m pl eti n g t h e r es e arc h.
E m or y H os pit al h as all m e dic al or ps yc h ol o gic al r es o urc es t h at p artici p a nts mi g ht n e e d as a 
r es ult of a ntici p at e d c o ns e q u e nc es of t h e h u m a n r es e arc h.
All p ers o ns assisti n g wit h t h e r es e arc h will b e m a d e f a mili ar wit h t h e pr ot oc ols a n d d escri b e d 
r es e arc h pr oc e d ur es, t h eir d uti es a n d f u ncti o ns r el at e d t o t h e m.
F aciliti es
All n ec ess ar y f aciliti es t o c o m pl et e t h e pr o p os e d st u di es ar e a v ail a bl e t o Dr. Ols o n ( PI) a n d Dr. 
M a y ol D el V all e ( C O- PI) at t h e E m or y U ni v ersit y H os pit al i n Atl a nt a a n d Dr. K uc h er y a v y k h 
(c oll a b or at or) at t h e U ni v ersi d a d C e ntr al d el C ari b e Sc h o ol of M e dici n e ( U C C) i n B a y a m ó n, 
P u ert o Ric o.
T h e E m or y Li pi d o mic c or e h as t h e m ass-s p ectr o m etr y r es o urc es r e q uir e d f or t h e st u d y a n d will 
d esi g n at e a p pr o pri at e p ers o n n el aft er c o ns ulti n g wit h t h e dir ect or of E m or y i nt e gr at e d c or e 
f aciliti es.

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 3 7 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
Dr. Ols o n h ol ds a n d a p p oi nt m e nt of Pr of ess or of N e ur os ur g er y at E m or y U ni v ersit y 
Dr. M a y ol d el V all e h ol ds a n a p p oi nt m e nt of S e ni or Ass oci at e at E m or y U ni v ersit y
Wi ns hi p C a nc er C e nt er N e ur os ur g er y L a b or at or y will b e us e d f or tiss u e s a m pl e pr ot ei n 
extr acti o n wit h a d d e d p h os p h at as e i n hi bit or, a n d fr oz e n at - 8 0 t o b e s hi p p e d o v er ni g ht t o 
P u ert o Ric o i n dr y ic e.
Dr. K uc h er y a v y k h h ol ds a n a p p oi nt m e nt i n t h e D e p art m e nt of Bi oc h e mistr y, U ni v ersi d a d 
C e ntr al d el C ari b e Sc h o ol of M e dici n e.
L a b or at or y: T h e l a b or at or y is l oc at e d at t h e d e p art m e nt of Bi oc h e mistr y of t h e M e dic al Sc h o ol’s 
M ai n b uil di n g w hic h h o us es m a n y of t h e s h ar e d e q ui p m e nt c e nt ers i ncl u di n g t h e C o nf oc al 
Micr osc o p e a n d t h e O ptic al I m a gi n g f acilit y. T h e l a b or at or y h as t h e b asic l a b or at or y e q ui p m e nt 
t h at i ncl u d es c e ntrif u g es, s h a k ers, r efri g er at ors, – 2 0 o C fr e ez ers, fl u or esc e nt micr osc o p e 
Ol y m p us 1 X 7 1, N aris hi g e S R- 5 M m o us e st er e ot axic s yst e m f or br ai n i m pl a nt ati o n s ur g eri es, 
A msc o p e s ur g er y micr osc o p e. I n t h e c or e f aciliti es, t h er e ar e als o a n Ol y m p us F V 1 0 0 0 c o nf oc al 
micr osc o p e, a Bi or a d V ers a d oc M P 5 0 0 0 m ol ec ul ar i m a g er, a n a p pli e d Bi os yst e ms 7 5 0 0 R e al 
Ti m e P C R S yst e m, f u m e h o o ds, a n d t h e c ell c ult ur e r o o m all a v ail a bl e f or c o m m o n us e.
T h e l a b or at or y at U C C ari b e is c ertifi e d a n d i ns p ect e d y e arl y b y t h e offic e of t h e D e a n of 
A d mi nistr ati o n w h o arr a n g es visits wit h t h e D e p art m e nt of H e alt h a n d S af et y a n d t h e N ati o n al 
Fir e D e p art m e nt.
C o m p ut ers: T h er e ar e m ulti pl e c o m p ut ers a v ail a bl e f or t his pr oj ect. All h a v e I nt er n et acc ess a n d 
ar e c o nfi g ur e d wit h st a n d ar d a p plic ati o n s oft w ar e f or Wi n d o ws s yst e ms, i ncl u di n g Micr os oft 
Offic e 1 1, Gr a p h P a d Pris m 8, A d o b e P h ot os h o p C S 6. T h e g el d oc u m e nt ati o n s yst e m, 
fl u or esc e nt micr osc o p e a n d ot h er e q ui p m e nt h a v e att ac h e d c o m p ut ers wit h a p pr o pri at e 
s oft w ar e. M or e o v er, t h e i nstit uti o n pr o vi d es a c o m pl et e c o m p ut er c o m m u nic ati o ns n et w or k 
e n vir o n m e nt wit h r el at e d t ec h n ol o g y a n d r es o urc es, i ncl u di n g a n e nt er pris e st or a g e a n d 
b ac k u p s yst e m f or m a n a g e m e nt of critic al r es e arc h d at a, e m ail, a n d vir us/s p a m pr ot ecti o n.
Offic e: Dr. K uc h er y a v y k h h as h er offic e at t h e D e p art m e nt of Bi oc h e mistr y. It is e q ui p p e d wit h a 
c o m p ut er f or d at a a n al ysis a n d m a n uscri pt a n d gr a nt pr e p ar ati o n. F ax, c o p y, a n d pri nti n g 
s u p p ort a n d c o n v e ni e nt acc ess t o a n a d mi nistr ati v e assist a nt f or g e n er al cl eric al assist a nc e a n d 
gr a nts m a n a g e m e nt s u p p ort ar e als o a v ail a bl e i n t h e Bi oc h e mistr y D e p art m e nt.
2 6. M ulti- Sit e R es e arc h W h e n E m or y is t h e L e a d Sit e
N/ A
T h e st u d y is a si n gl e-sit e r ecr uit m e nt r es e arc h st u d y.

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 3 8 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
2 7. R ef er e nc es
N u ñ ez R E, D el V all e M M, Ortiz K, Al m o d o v ar L, K uc h er y a v y k h L. Micr o gli al C yt o ki n es I n d uc e 
I n v asi v e n ess a n d Pr olif er ati o n of H u m a n Gli o bl ast o m a t hr o u g h P y k 2 a n d F A K Acti v ati o n.
C a nc ers ( B as el). 2 0 2 1 D ec 7; 1 3( 2 4): 6 1 6 0. d oi: 1 0. 3 3 9 0/c a nc ers 1 3 2 4 6 1 6 0. P MI D: 3 4 9 4 4 7 7 9; 
P M CI D: P M C 8 6 9 9 2 2 8.
R o bi ns o n J P, V a n Br oc kli n M W, G uil b e a ult A R, Si g n or elli DL, Br a n d n er S, H ol m e n SL. Acti v at e d 
B R A F i n d uc es gli o m as i n mic e w h e n c o m bi n e d wit h I n k 4 a/ Arf l oss or A kt acti v ati o n. O nc o g e n e. 
2 0 1 0; 2 9: 3 3 5 – 3 4 4.
L y usti k m a n Y, M o m ot a H, P a o W, H oll a n d E. C o nstit uti v e Acti v ati o n of R af- 1 I n d uc es Gli o m a 
F or m ati o n i n Mic e. N e o pl asi a. 2 0 0 8 M a y; 1 0( 5): 5 0 1 – 5 1 0. d oi: 1 0. 1 5 9 3/ n e o. 0 8 2 0 6
A n dr e ws LJ, T h or nt o n Z A, S ai nc h er S S, Y a o I Y, D a ws o n S, Mc G ui n n ess L A, et al. Pr ev al e nc e of 
B R A F V 6 0 0 i n gli o m a a n d us e of B R A F I n hi bit ors i n p ati e nts wit h B R A F V 6 0 0 m ut ati o n- p ositi v e 
gli o m a: s yst e m atic r e vi e w. N e ur o O nc ol 2 0 2 2; 2 4: 5 2 8 – 4 0.
M a ki n o Y, Ar a k a w a Y, Y os hi o k a E, S h of u d a T, Mi n a mi g uc hi S, K a w a uc hi T, et al. I nfr e q u e nt R A S 
M ut ati o n Is N ot Ass oci at e d Wit h S p ecific Hist ol o gic al P h e n ot y p e i n Gli o m as. B M C C a nc er ( 2 0 2 1) 
2 1( 1): 1 0 2 5. d oi: 1 0. 1 1 8 6/s 1 2 8 8 5- 0 2 1- 0 8 7 3 3- 4
P hilli ps H S, K h ar b a n d a S, C h e n R, F orr est W F, S ori a n o R H, W u T D, et al. M ol ec ul ar s u bcl ass es of 
hi g h- gr a d e gli o m a pr e dict pr o g n osis, d eli n e at e a p att er n of dis e as e pr o gr essi o n, a n d r es e m bl e 
st a g es i n n e ur o g e n esis. C a nc er C ell 2 0 0 6; 9: 1 5 7 – 7 3.
V er h a a k R G, H o a dl e y K A, P ur d o m E, W a n g V, Qi Y, Wil k ers o n M D, et al. I nt e gr at e d g e n o mic 
a n al ysis i d e ntifi es cli nic all y r el e v a nt s u bt y p es of gli o bl ast o m a c h ar act eriz e d b y a b n or m aliti es i n 
P D G F R A, I D H 1, E G F R, a n d N F 1. C a nc er C ell 2 0 1 0; 1 7: 9 8 – 1 1 0.
Xi a oji n g W a n g, S h e n g pi n g Mi n, H o n gli Li u, N a n W u, Xi nc h e n g Li u, T a o W a n g, W ei Li, Y u a n bi n g 
S h e n, H o n gt a o W a n g, Z h o n g qi n g Qi a n, H u a n b ai X u, C h e n gli n g Z h a o, Y u qi n g C h e n. Nf 1 l oss 
pr o m ot es Kr as- dri v e n l u n g a d e n oc arci n o m a a n d r es ults i n Ps at 1- m e di at e d gl ut a m at e 
d e p e n d e nc e. E M B O M ol M e d. 2 0 1 9 J u n; 1 1( 6): e 9 8 5 6. P u blis h e d o nli n e 2 0 1 9 A pr 2 9. d oi: 
1 0. 1 5 2 5 2/ e m m m. 2 0 1 8 0 9 8 5 6
B e h n a n J, Fi n occ hi ar o G, H a n n a G. T h e L a n dsc a p e of t h e M es e nc h y m al Si g n at ur e i n Br ai n 
T u m o urs. Br ai n ( 2 0 1 9) 1 4 2( 4): 8 4 7 – 6 6. d oi: 1 0. 1 0 9 3/ br ai n/ a wz 0 4 4
Li pi ns ki C A, Tr a n NL, Vis o C, Kl oss J, Y a n g Z, B er e ns M E, L oft us J C. Ext e n d e d s urvi v al of P y k 2 or 
F A K d efici e nt ort h ot o pic gli o m a x e n o gr afts. J N e ur o o nc ol. 2 0 0 8 N o v; 9 0( 2): 1 8 1- 9.
L oft us J C, Y a n g Z, Tr a n NL, Kl oss J, Vis o C, B er e ns M E, Li pi ns ki C A. T h e P y k 2 F E R M d o m ai n as a 
t ar g et t o i n hi bit gli o m a mi gr ati o n. M ol C a nc er T h er. 2 0 0 9 J u n; 8( 6): 1 5 0 5- 1 4.

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 3 9 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
R ol ó n- R e y es K, K uc h er y a v y k h Y V, C u b a n o L A, I n y us hi n M, S k atc h k o v S N, E at o n MJ, H arris o n J K, 
K uc h er y a v y k h L Y. Micr o gli a Acti v at e Mi gr ati o n of Gli o m a C ells t hr o u g h a P y k 2 I ntr ac ell ul ar 
P at h w a y. PL o S O n e. 2 0 1 5 J u n 2 2; 1 0( 6). P MI D: 2 6 0 9 8 8 9 5 P M CI D: P M C 4 4 7 6 5 9 0
P a uli n o V M, Y a n g Z, Kl oss J, E n nis MJ, Ar mstr o n g B A, L oft us J C, Tr a n NL. T R O Y ( T N F R S F 1 9) is 
o v er ex pr ess e d i n a d v a nc e d gli al t u m ors a n d pr o m ot es gli o bl ast o m a c ell i n v asi o n vi a P y k 2- R ac 1 
si g n ali n g. M ol C a nc er R es. 2 0 1 0 N o v; 8( 1 1): 1 5 5 8- 6 7.
S at h or ns u m et e e S, Hj el m el a n d A B, K eir S T, McL e n d o n R E, B att D, R a ms e y T, Y us uff N, R as h e e d 
B K, Ki er a n M W, L af or m e A, Bi g n er D D, Fri e d m a n H S, Ric h J N. A AL 8 8 1, a n o v el s m all m ol ec ul e 
i n hi bit or of R A F a n d v asc ul ar e n d ot h eli al gr o wt h f act or r ec e pt or acti viti es, bl oc ks t h e gr o wt h of 
m ali g n a nt gli o m a. C a nc er R es. 2 0 0 6; 6 6: 8 7 2 2 – 8 7 3 0.
Cl o u g h es y T F, W e n P Y, R o bi ns HI, C h a n g S M, Gr o v es M D, Fi n k KL, J u nc k L, Sc hiff D, A br e y L, 
Gil b ert M R, Li e b er m a n F, K u h n J, D e A n g elis L M, M e ht a M, R aiz er JJ, Y u n g W K, Al d a p e K, Wri g ht 
J, L a m b or n K R, Pr a d os M D. P h as e II tri al of ti pif ar ni b i n p ati e nts wit h r ec urr e nt m ali g n a nt 
gli o m a eit h er r ec ei vi n g or n ot r ec ei vi n g e nz y m e-i n d uci n g a nti e pil e ptic dr u gs: a N ort h A m eric a n 
Br ai n T u m or C o ns orti u m St u d y. J. Cli n. O nc ol. 2 0 0 6; 2 4: 3 6 5 1 – 3 6 5 6.
Li u J, X u e L, X u X, L u o J, Z h a n g S. F A K-t ar g eti n g P R O T A C d e m o nstr at es e n h a nc e d a ntit u m or 
acti vit y a g ai nst K R A S m ut a nt n o n-s m all c ell l u n g c a nc er. Ex p C ell R es. 2 0 2 1 N o v 
1 5; 4 0 8( 2): 1 1 2 8 6 8. d oi: 1 0. 1 0 1 6/j. y excr. 2 0 2 1. 1 1 2 8 6 8.
Hir at a E, Gir otti M R, Vir os A, H o o p er S, S p e nc er- D e n e B, M ats u d a M, L ar ki n J, M ar ais R, S a h ai E. 
I ntr a vit al i m a gi n g r e v e als h o w B R A F i n hi biti o n g e n er at es dr u g-t ol er a nt micr o e n vir o n m e nts wit h 
hi g h i nt e gri n b et a 1/ F A K si g n ali n g. C a nc er C ell. 2 0 1 5 A pr 1 3; 2 7( 4): 5 7 4- 8 8. d oi: 
1 0. 1 0 1 6/j.cc ell. 2 0 1 5. 0 3. 0 0 8.
J o n es S, Si u L, B e n d ell J, Cl e ar y J, R az a k A, I nf a nt e J, P a n d y a S, B e d ar d P, Pi erc e K, H o u k B, 
R o b erts W, S hr e e v e S, S h a pir o G. A p h as e I st u d y of V S- 6 0 6 3, a s ec o n d- g e n er ati o n f oc al 
a d h esi o n ki n as e i n hi bit or, i n p ati e nts wit h a d v a nc e d s oli d t u m ors. I n v est N e w Dr u gs. 2 0 1 5 
Oct; 3 3( 5): 1 1 0 0- 7.
S hi miz u T, F u k u o k a K, T a k e d a M, I w as a T, Y os hi d a T, H or o bi n J, K e e g a n M, V aic k us L, C h a v a n A, 
P a d v al M, a n d N a k a g a w a K. A first-i n- Asi a n p h as e 1 st u d y t o ev al u at e s af et y, p h ar m ac o ki n etics 
a n d cli nic al acti vit y of V S- 6 0 6 3, a f oc al a d h esi o n ki n as e ( F A K) i n hi bit or i n J a p a n es e p ati e nts wit h 
a d v a nc e d s oli d t u m ors. C a nc er C h e m ot h er P h ar m ac ol. 2 0 1 6; 7 7: 9 9 7 – 1 0 0 3.
M arti n ez- G arci a M, et al. First-i n- h u m a n, p h as e I d os e- esc al ati o n st u d y of t h e s af et y, 
p h ar m ac o ki n etics, a n d p h ar m ac o d y n a mics of R O 5 1 2 6 7 6 6, a first-i n-cl ass d u al M E K/ R A F 
i n hi bit or i n p ati e nts wit h s oli d t u m ors. Cli n C a nc er R es. 2 0 1 2 S e p 1; 1 8( 1 7): 4 8 0 6- 1 9.
G u o C, C h é n ar d- P oiri er M, R o d a D, d e Mi g u el M, H arris SJ, C a n dil ej o I M, Sris k a n d ar aj a h P, X u 
W, Sc ar a nti M, C o nst a nti ni d o u A, Ki n g J, P ar m ar M, T ur n er AJ, C arr eir a S, Riis n a es R, Fi n n er a n L,

Pr ot oc ol Titl e : A si n gl e- d os e st u d y of or all y a d mi nistr at e d d ef acti ni b or a v ut o m eti ni b i n
p ati e nts wit h gli o bl ast o m a
P a g e 4 0 of 3 8
Pr ot oc ol V ersi o n: 6 8/ 3 0/ 2 0 2 4
H all E, Is hi k a w a Y, N a k ai K, T u n ari u N, B as u B, K ais er M, L o p ez J S, Mi nc h o m A, d e B o n o J S, 
B a n erji U. I nt er mitt e nt sc h e d ul es of t h e or al R A F- M E K i n hi bit or C H 5 1 2 6 7 6 6/ A V U T O M E TI NI B i n 
p ati e nts wit h R A S/ R A F- m ut a nt s oli d t u m o urs a n d m ulti pl e m y el o m a: a si n gl e-c e ntr e, o p e n- 
l a b el, p h as e 1 d os e- esc al ati o n a n d b as k et d os e- ex p a nsi o n st u d y. L a nc et O nc ol. 2 0 2 0 
N o v; 2 1( 1 1): 1 4 7 8- 1 4 8 8. d oi: 1 0. 1 0 1 6/ S 1 4 7 0- 2 0 4 5( 2 0) 3 0 4 6 4- 2.
Q uilli n J, P at el R, H erz b er g E, Alt o n D, Bi kz h a n o v a G, G eisl er L, Ols o n J: A p h as e 0 a n al ysis of 
ix az o mi b i n p ati e nts wit h gli o bl ast o m a. M ol ec ul ar a n d Cli nic al O nc ol o g y 1 4: 4 3, 2 0 2 0.