Conditions: Mydriasis Dilation
Keywords: Nyxol Pharmacologically Induced Mydriasis Dilation
Allocation: RANDOMIZED | Intervention model: PARALLEL
Enrollment: 185
Healthy volunteers: yes

Brief summary:
The objectives of this study are:

* To evaluate the efficacy of Nyxol to expedite the reversal of pharmacologically-induced mydriasis across multiple mydriatic agents with an emphasis on phenylephrine
* To evaluate the efficacy of Nyxol to return subjects to baseline accommodation after worsening (with cycloplegic agents tropicamide and Paremyd)
* To evaluate the safety of Nyxol
* To evaluate any additional benefits of the reversal of pharmacologically-induced mydriasis

Detailed description:
A randomized, parallel arm, double-masked, placebo-controlled Phase 3 study in at least 168 randomized subjects (160 completed), evaluating the safety and efficacy of Nyxol in subjects with pharmacologically-induced mydriasis.

Following the successful completion of screening, each subject will be stratified by eye color and then simultaneously be randomized to mydriatic agent (unmasked) and treatment (masked).

Treatment randomization will be 1:1, Nyxol or placebo (vehicle).

Stratification by iris color will be 1:1, light or dark irides.

The mydriatic agent randomization will be 3:1:1 (2.5% phenylephrine, 1% tropicamide, and Paremyd). That is, approximately 60% of the randomized subjects will receive one drop of 2.5% phenylephrine 1 hour before treatment (96 completed subjects), approximately 20% will receive one drop of 1% tropicamide 1 hour before treatment (32 completed subjects), and approximately 20% will receive Paremyd 1 hour before treatment (32 completed subjects).

At the treatment visit, subjects who have been randomized and stratified by iris color (1:1 \[light/dark\]) will receive one of three approved mydriatic agents approximately 1 hour prior to receiving study treatment. Measurements will be measured before (-1 hour /baseline) and 60 minutes after (maximum/0 minutes) the mydriatic agent instillation in each eye (i.e. right before the study treatment is administered), and at 30 minutes, 60 minutes, 90 minutes, 2 hours, 3 hours, 4 hours, and 6 hours after treatment dosing. Measurements will include pupil diameter (PD), distance and near visual acuity (VA), accommodation, and redness in each eye.

At the Follow-Up Visit, which is 1 day after Visit 1, measurements will again be recorded 24 hours after treatment dosing.

Arms:
arm 1: 2 drops in study eye and 1 drop in non-study eye, 1 hour post pharmacologically-induced mydriasis arm 2: 2 drops in study eye and 1 drop in non-study eye, 1 hour post pharmacologically-induced mydriasis

Interventions: intervention 1: Phentolamine Ophthalmic Solution 0.75% intervention 2: Phentolamine Ophthalmic Solution Vehicle (Placebo)
Intervention descriptions: intervention 1: 0.75% phentolamine ophthalmic solution (Nyxol), a non-selective alpha-1 and alpha-2 adrenergic antagonist intervention 2: Topical sterile ophthalmic solution

Protocol document:
Prot_000.pdf: 
 
 
 
C O N FI D E N TI A L  
 
Cli nic al St u d y P h ase 3  P r ot oc ol  
O PI -N Y X R M -3 0 1  
MI R A -2  
 
R a n d o mize d,  P ar allel  Ar m , D o u ble -M as ke d,  Pl ace b o -C o ntr olle d  St u dy  o f the  
S afety  a n d  Effic acy  o f Ny x ol  (0. 7 5 %  P he nt ol a mi ne  O p ht h al mic  S ol uti o n)  to  
Reverse  P h ar m ac ol o gic ally -I n d uce d My dri asis  in  He alt hy  S u bjects  
 
 
Oc u p hire P har m a, I nc.  
3 7 0 0 0 Gra n d Ri ver A ve n ue, S uite 1 2 0  
Far mi n gt o n Hills , MI 4 8 3 3 5  
 
 
 
 
 
Versi o n:  
0 2  
Ori gi n al:  
A u g ust  2 0 , 2 02 0  
A me n d me nt  1 : 
 
Fe br u ar y 5, 2 0 2 1  
 
 
C O N FI D E N TI A L  
T his d oc u me nt c o ntai ns Oc u p hire P har m a, I nc.  ( Oc u p hire) i nf or mati o n t hat is c o nfi de ntial, a 
tra de secret a n d/ or pr o pri eta y i n nat ure. It is l oa ne d t o y o u f or y o ur c o nfi d e ntial use o n be half of 
Oc u p hire a n d is n ot t o be p h ot oc o pie d, discl ose d or tra ns mitte d t o a n y ot her pers o n or part y w h o 
is n ot c o vere d b y a C o nfi de ntial Discl os ure A gree me nt wit h Oc u p hire. As t he Pri nci pal 
I n v esti gat or y o u are res p o nsi ble f or t he safe kee pi n g a n d ret ur n of t his d oc u me nt t o Oc u p hire 
u p o n re q uest. Y o u will b e se nt u p date d i nf or m ati o n a n d/ or a me n d me nts as t he y bec o me 
a vaila ble.
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
2  
0 2 /0 5 /2 1  
 
S U M M A R Y O F C H A N G E S  F O R A M E N D M E N T 1  
B ol d  u n derli ne d  te xt s h o ws a d diti o ns;  stri ket hr o u g h te xt s h o ws deleti o ns.  F or  c ha n ges t hat affect  
m ulti ple  secti o ns of  t he pr ot oc ol,  t he c ha n ge  is liste d o nce  at t he first i nsta nce bel o w,  a n d eac h 
s u bse q ue nt pr ot oc ol  secti o n i nc or p orati n g t hat c ha n ge  is als o liste d at t hat p oi nt.  A d mi nistrati ve  a n d 
mi n or  e diti n g  c ha n ges n ot  affect i n g t he c o nte nt  or  c o n d uct  of  t he pr ot oc ol  ha ve  bee n  ma de;  t hese are  
n ot  liste d. N o ne  of  t he c ha n ges ma de  affect  t he ris ks t o t he s u bjects i n t his st u d y. 
Secti o n : L oc ati o n  
Descri pti o n  of  C h a n ge  
R ati o n ale  f or 
C h a n ge  
S y n o psis:  Desi g n,  
p. 1 1  
A  ra n d o mize d, parallel  ar m,  d o u ble -mas ke d,  place b o -c o ntr olle d  
P hase  3  st u d y i n at least 1 6 8  ra n d o mize d s u bjects ( 1 6 0 c o m plete d  
e v al u a ble  f or effic ac y ), e val uati n g  t he safet y a n d  efficac y  of  
N y x ol  i n s u bjects wit h  p har mac ol o gicall y -i n d uce d m y driasis.  
Gl o bal c ha n ge t o 
clarif y  t hat 
e val ua ble s u bjects , 
w hic h are defi ne d 
wit hi n t he pr ot oc o l, 
will be use d f or 
efficac y a n al ysis 
S y n o psis:  Desi g n,  
p. 1 1  
T hat  is, a p pr o xi matel y  6 0 %  of  t he ra n d
mize d s u bjects will  
recei ve o ne  dr o p  of  2. 5 %  p he n yle p hri ne  1  o ur  bef ore  treat me nt 
( 9 6 c o m plete d  e v al u a ble  s u bjects), a p pr o xi ma el y  2 0 %  will  
recei ve o ne  dr o p  of  1 %  tr o pica mi de 1  h o r  bef ore  treat me nt ( 3 2 
c o m plete d  e v al u a ble  s u bjects), a n d  a p pr o xi matel y  2 0 %  will  
recei ve Pare m y d  1  h o ur  bef ore  treat me nt ( 3 2 c o m plete d  e v al u a ble  
s u bjects). 
S y n o psis:  Patie nt  
P o p ula ti o n, p. 1 2  
1 6 8 ra n d o mize d ( 1 6 0 c o m plete d  e al u a ble ) healt h y s u bjects. 
S y n o psis:  Visit  
Sc he d ule,  p. 1 4  
Ra n d o mizati o n  will  be  stratifie d 3: 1: 1  b y  m y driatic  a ge nt  ( 2. 5 % 
p he n yle p hri ne,  1 %  tr o pica i de, a n d  Pare m y d),  wit h  
ra n d o mizati o n i nt o ea h  m y driatic  a ge nt ca p pe d at a p pr o xi matel y  
9 6 : 3 2: 3 2 c o m plete d  e v al u a ble  s u bjects. 
S y n o psis:  Esti mate d  
T otal  Sa m ple  Size,  
p. 1 4  
A p pr o xi matel y  1 6 8  ra n d o mize d healt h y  s u bjects, wit h  
a p pr o xi matel y  1 6 0  c o m plete d  e v al u a ble  s u bjects. 
S y n o psis:  Sa m ple  
Size  J ustificati o n, 
p. 1 4  
9. 1  Sa m ple  Size,  p. 4 1  
A  sa m ple si e of  1 6 0  c o m plete d  e v al u a ble  s u bjects ( 8 0 per  
tr
t me nt gr o u p)  is nee de d  f or t he st u d y. T he  pri mar y  efficac y  
e n d p oi nt  will  be  met  if s u bjects s h o w a p ositi ve  effect  f or N y x ol  
( α =  0. 0 5  si g nifica nce, t w o-taile d). O ne  h u n dre d  a n d  si xt y 
c o m pl te d  e v al u a ble  s u bjects will  pr o vi de  
 
 bet wee n  t he N y x ol  a n d  place b o  ar ms i n 
perce nta ge  of  s u bjects ret ur ni n g t o ≤ 0. 2  m m  baseli ne  p u pil  
dia meter  at 9 0  mi n utes.  
S y n o psis:  D urati o n  of  
S u bject  Partici pati o n  
a n d  St u d y,  p. 1 7  
T he  e x ec uti o n  of  t he e ntire  st u d y (first s u bject scree n t hr o u g h last 
ra n d o mize d s u bject c o m plete d e v al u a ble ) is e x pecte d  t o be  
a p pr o xi matel y  3  t o 6  m o nt hs.  
4. 1 0  E val ua ble  
s u bjects, p. 3 0  
4. 1 0  E v al u a ble  s u bjects 
S u bjects  are  e v al u a ble  f or effic ac y  if t he y recei ve d  o ne  or  t w o 
dr o ps  of  st u d y tre at m e nt a n d  h a d  a  p u pil  di a m eter  
m e as ure m e nt  at  t he 9 0 -mi n ute  ti m e p oi nt  at  Visit  1 . 
A p pe n di x  2,  p. 4 9  
R A N D O MI Z A TI O N  S C H E M A  F O R  E V A L A U B L E  
S U B J E C T S  B Y  I N V E S TI G A TI O N A L T R E A T M E N T,  
M Y D RI A TI C  A G E N T,  A N D  I RI S C O L O R  
 
 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
3  
0 2 /0 5 /2 1  
 
S y n o psis:  Sec o n dar y  
Efficac y  E n d p oi nts,  
p. 1 5  
4. 1  Pri mar y  a n d  
sec o n dar y e n d p oi nts,  
p. 2 4  
7. 2  Assessi n g,  
rec or di n g, a n d  
a n al yzi n g  of  efficac y  
para meters,  p. 3 4  
S u bject  will  be  all o we d  t o accli mate  t o t hese li g hti n g c o n diti o ns  
( wit h t he e y es o pe n  n or mall y  f or a mi ni m u m  
) pri or  t o 
t he p u pil  dia meter,  B C D V A  a n d  D C N V A  (safet y meas ures) , a n d  
acc o m m o dati o n  meas ure me nts  at all sc he d ule d ti me p oi nts. S u bject  
will  sit i n t he e xa m  c hair  faci n g t he ill u mi nate d c hart d uri n g  t he 
accli mati o n  peri o d  a n d f or all assess me nts
 
 a n d  t he 
sc h e d ule d re mai ni n g  safet y assess me nts  (e. g. 
 
, a d verse  e ve nts,  s u bject q uesti o n naire,  etc.). 
T o clarif y t hat 
r o o m li g hts 
s h o ul d be o n 
d uri n g 
acc o m m o dati o n 
meas ure me nts  
9. 2  A nal ysis  
P o p ulati o ns,  p. 4 1  
Per  Pr ot oc ol  P o p ulati o n  ( P P):  T he  per  pr ot oc ol  ( P P) p o p ulati o n  
i ncl u ded s all s u bjects i n t he F A S mI T T  
 
, ha d  all sc he d ule d p u pil  dia meter  
meas ure me nts  d uri n g  Visit  1,  h a d  a n  i ncre ase of  >  0. 2  m m  i n P D  
i n t he st u d y e ye at  Ti m e  0  mi n utes  c o m p are d  t o B aseli ne  ( Ti m e 
– 1  h o ur),  a n d  ha d  n o  maj or  pr ot oc ol  d vi ati o ns.  T he  P P  p o p ulati o n  
will  be  use d  f or t he pri mar y  e n d p oi nt a na ysis a n d  t o a nal yze  
selecte d sec o n d ar y efficac y  e n d p oi nts. 
T o clarif y t he P P 
p o p ulati o n as 
ha vi n g a n i ncrease 
of > 0. 2 m m i n 
P D i n t he st u d y 
e ye at Ti me 0 
mi n utes c o m pare d 
t o Baseli ne  
S y n o psis  Sec o n dar y  
Efficac y  E n d p oi nts,  
p. 1 5  
4. 1  Pri mar y  a n d  
sec o n dar y e n d p oi nts,  
p. 2 3  
Sec o n dar y  efficac y  e n d p oi nts  (f or t he st u d y e ye; a n d  f or t he n o n -
st u d y e y e; a n d  f or t he best  of  eit her  e ye) will  i ncl u de: 
Gl o bal c h a n ge per 
F D A fee d bac k t o 
defi ne a n d use t he 
mI T T p o p ulati o n 
as well as 
i m p utati o n 
met h o ds f or 
missi n g data f or 
pri mar y efficac y 
a nal ysis.  
 
T o har m o nize t he 
pr ot oc ol wit h t he 
fi nal versi o n of 
t he Statistical 
A nal ysis P la n 
( V 1. 0) 
 
 
S y n o psis:  Sec o n dar y  
Efficac y  E n d p oi nts,  
p. 1 6  
4. 1. 1  Pri mar y  a n d  
sec o n dar y e n d p oi nts , 
p. 2 4  
7. 1  S pecificati o n  of  
t he efficac y  
para meters,  p. 3 3  
All  of  t he efficac y e n d p oi nts  will  be  a nal yze d  b y  F ull  A nal ysis  Set  
( F A S) a n d  Per  Pr ot oc ol  ( P P).T he  M o difie d  I nte nt-t o-Tre at  
( mI T T) will  be  use d  f r t he pri m ar y  e n d p oi nt  a n al ysis  a n d  t o 
a n al yze  selecte d sec n d ar y effic ac y  e n d p oi nts.  T he  Per  Pr ot oc ol  
( P P) p o p ul ati o n  will  be  use d  t o a n al yze  selecte d sec o n d ar y 
effic ac y  e n d p oi nts  S o m e  All of  t he efficac y  e n d p oi nts  will  be  
a n al yze d  o verall,  b y  m y driatic  a ge nt,  a n d  b y  li g ht/ dar k iri des. 
9  S T A TI S TI C S,  p. 4 1  
A det aile d prese nt ati o n of t he st atistic al a p pr o ac h is o utli ne d i n 
t he St atistic al A n al ysis Pl a n. 
9. 2  A nal ysis  
P o p ulati o ns,  p. 4 1  
F ull  A nal ysis  Set  M o difie d  I nte nt t o Tre at  (F A S mI T T ): T he  
F A S mI T T  will  i ncl u de all ra n d o mize d s u bjects w h o  recei ve d o ne  
or  t w o dr o ps  of  st u d y treat me nt a n d  t he n ha d  at least o ne  sc he d ule d 
p u pil  dia meter  meas ure me nt  d uri n g  Visit  1.  T he  F A S mI T T  will  be  
use d  f or t he pri m ar y  e n d p oi nt  a n al ysis  a n d  t o a nal yze  selecte d 
sec o n d ar y  efficac y  e n d p oi nts. 
9. 3. 2  De m o gra p hics  
a n d  baseli ne  
c h aracteristics,  p. 4 2  
De m o gra p hic  a n d  Baseli ne  c haracteristics  s uc h as a ge,  race a n d  
se x, will  be  s u m marize d b y  treat me nt gr o u p  usi n g  t he F A S mI T T , 
P P , S P,  a n d  t he A R P.   
9. 3. 4  Me dical  hist or y  
a n d  
pri or/c o nc o mita nt  
me dicati o ns,  p. 4 2  
Me dical  hist or y  will  be  c o de d  usi n g  t he latest versi o n  of  Me dical  
Dicti o nar y  f or Re g ulat or y  Acti vities  ( Me d D R A) a n d  will  be  
s u m marize d b y  treat me nt gr o u p  usi n g  t he F A S S P .  
Pri or  me dicati o ns  ( me dicati o ns wit h  a n  e n d  date  pri or  t o t he date  of  
ra n d o mizati o n) a n d  c o nc o mita nt  me dicati o ns  ( me dicati o ns wit h  a 
start or  e n d  date  after  t he date  of  ra n d o mizati o n) will  be  c o de d  
usi n g  W H O Dr u g  a n d  will  b ot h  be  s u m marize d b y  treat me nt gr o u p  
usi n g  t he F A S S P . 
9. 3. 5  A nal ysis  of  
efficac y , p. 4 2  
Efficac y  will  be  assesse d  usi n g  t he F A S mI T T  a n d  P P  wit h  s u bjects 
i ncl u de d i n t he treat me nt ar m  i n w hic h  t he y were  ra n d o mize d. 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
4  
0 2 /0 5 /2 1  
 
9. 3. 5  A nal ysis  of  
efficac y,  p. 4 2  
F or t he a nal ysis of t he pri mar y efficac y e n d p oi nt , o bser ve d case 
data will be use d ( n o i m p utati o n will be perf or me d f or missi n g 
data) f or t he pri mar y a nal ysis  i m p ut ati o n will be perf or m e d f or 
missi n g d at a as descri be d i n t he S t atistic al A n al ysis P l a n. 
9. 3. 5  A nal ysis  of  
efficac y,  p. 4 2  
If warra nte d, co nfir mat or y a nal yses usi n g t he A R P wit h i m p utati o n 
f or missi n g data will als o be perf or me d f or t he sec o n dar y efficac y 
e n d p oi nts.  
9. 3. 5  A nal ysis  of  
efficac y,  p. 4 3  
T he  a n al ysis will  be  perf or me d  usi n g  t he F A S mI T T  a n d  P P,  wit h  
s u bjects i ncl u de d i n t heir ra n d o mize d treat me nt re gar dless of  t he 
treat me nt t he y act uall y recei ve d. 
9. 3. 5  A nal ysis  of  
efficac y,  p. 4 3  
Eac h  A N C O V A  will  be  perf or me d  usi n g  t he F A S mI T T  a n d  P P  
wit h  s u bjects i ncl u de d i n t heir ra n d o mize d treat me nt re gar dless of  
t he treat me nt t he y act uall y  recei ve d. 
9. 3. 5  A nal ysis  of  
efficac y,  p. 4 3  
F or t hese e n d p oi nts, t he F A S mI T T  a n d P P will be use d wit h 
s u bjects i ncl u de d i n t heir ra n d o mize d treat me nt re gar dless of t he 
treat me nt t he y act uall y recei ve d. 
9. 3. 5  A nal ysis  of  
efficac y,  p. 4 3  
A c o m p aris o n of t he st u d y a n d n o n -st u d y e ye f or e ac h s u bject 
will be c o m plete d f or t he pri m ar y effic a y e n d p oi nt, as well as 
b y m y dri atic a ge nt.  
9. 3. 5  A nal ysis  of  
efficac y,  p. 4 3  
Ot her s u b gr o u ps, s uc h as a ge, se x, a n d ra e  ma y be a nal yze d as 
well. If t here is s ufficie nt sa m ple, a nal ysis of safet y a n dselecte d 
efficac y e n d p oi nts will be c o m p lete d f or t he s u b gr o u p of pe diatric 
s u bjects. 
9. 3. 5  A nal ysis  of  
efficac y,  p. 4 4  
O bser ve d  val ues  a n d  c ha n ge  fr o m baseli ne  (-1  h o ur)  i n c o nj u ncti val  
h y pere mia  at eac h  ti me p oi nt ( 0 mi n,  3 0  mi n utes,  6 0  mi n utes,  9 0  
mi n utes,  2  h o urs,  3,  h o urs,  4  h o urs,  6  h o urs,  a n d  2 4  h o urs),  will  be  
s u m marize d f or t he st u d y ye , a n d  t he n o n -st u d y e ye, a n d t he best  
of  eit her  e ye. Treat me nts  will  be  c o m pare d  usi n g  t he sa me 
A N C O V A  m o del  pr p ose d  f or t he c o nti n u o us  sec o n dar y efficac y  
e n d p oi nts.  
 
 
Vis ual  ac uit y  ass ss me nts  ( B C D V A a n d  D C N V A)  will  als o  be  
s u m marize d t eac h  time p oi nt  ( 0 mi n utes,  9 0  mi n utes,  6  h o urs,  a n d  
2 4  h urs)  usi n g  letters, a n d  l o g M A R u nits , a n d  t he n u m ber  of  li nes 
rea d. Letters  will  be  rec or de d, a n d  later c o n verte d t o l o g M A R a n d  
n u m ber  of  li nes rea d b y  pr o gra m mi n g  f or s uc h a nal yses.  O ne  letter 
is e q ui ale nt  t o 0. 0 2  l o g M A R. As  a  refere nce , fi ve letters is 
re prese nte d b y  1  li ne. Treat me nts  will  be  c o m pare d  usi n g  t he sa me 
A N C O V A  m o del  pr o p ose d  f or t he c o nti n u o us  sec o n dar y efficac y  
e n d p oi nts.  
 
 
. 
F or  I O P, Baseli ne  is defi ne d  as t he scree ni n g val ue.  O bser ve d  
val ues  a n d  c h a n ge  fr o m baseli ne  i n I O P at 6  h o urs  will  be  
s u m marize d f or t he st u d y e ye, a n d  t he n o n -st u d y e ye, a n d t he best  
of  eit her  e ye. 
9. 4  Pr o ce d ure  f or 
acc o u nti n g f or 
missi n g,  u n use d,  or  
s p uri o us data , p. 4 4  
F or  t he s u m marizati o n a n d  a nal ysis of  t he pri m ar y  efficac y data  
e n d p oi nt , t he f oc us will  be  o n  o bser ve d  case data  o nl y.  N o  
i m p utati o n will  be  perf or me d  f or missi n g  data.  As  a p pr o priate,  
c o nfir mat or y  efficac y  a nal yses  will  be  perf or me d  usi n g  i m p utati o n 
f or missi n g  data;  details  of  t he i m p utati o n, if perf or me d,  will  be  
i ncl u de d i n t he st u d y Statistical  A nal ysis  Pla n  i m p ut ati o n will  be  
perf or m e d  f or missi n g  d at a  as  descri be d  i n t he St atistic al  
A n al ysis  Pl a n . F or  t he s u m marizati o n a n d  a n al ysis  of  sec o n d ar y 
effic ac y  e n d p oi nts  a n d  safet y data,  o bser ve d  case data  o nl y  will  be  
use d.  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2




Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
6  
0 2 /0 5 /2 1  
 
 
I N V E S TI G A T O R’ S A G R E E M E N T  
 
O PI -N Y X R M -3 0 1  
MI R A -2  
 
R a n d o mize d,  P ar allel  Ar m,  D o u ble -M as ke d , Pl ace b o -C o ntr olle d  St u dy  o f the   
S afety  a n d  Effic acy  o f N y x ol  ( 0. 7 5 % P he nt ol a mi ne  O p ht h al mic  S ol uti o n)  to   
Reverse  P h ar m ac ol o gic ally -I n d uce d My dri asis  in  He alt hy  S u bjects  
 
Versi o n:  
 
 
0 2  
 
 
Ori gi n al:  
 
 
A u g ust  2 0 , 2 02 0  
A me n d me nt 1:  
 
Fe br u ar y 5, 2 0 2 1  
 
I n v esti gat or A gree me nt: 
I, t he u n d ersi g ne d, h a ve re vie we d t his pr ot oc ol a n d I a gree t o c o n d uct t his pr ot oc ol i n acc or da nce 
wit h G o o d C li nical Practice, t he et hical pri nci ples set f ort h i n t he Decl arati o n of Helsi n ki a n d 
wit h t he U. S. C o de  of Fe deral Re g ulati o ns g o ver ni n g t he pr otecti o n of h u m a n s u bjects 
(2 1  C F R  5 0), I nstit uti o nal Re vie w B o r ds ( 2 1 C F R 5 6) a n d t he o bli gati o ns of cli nical 
i n vesti gat ors (2 1  C F R 3 1 2).  
 
Si g nat ure: _ _ _ _ _ _ _ _ _ _
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Date : _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 
 
Pr i nte d Na me: _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 
 
 
 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
7  
0 2 /0 5 /2 1  
 
P R O C E D U R E S I N C A S E O F E M E R G E N C Y  
 
E M E R G E N C Y C O N T A C T I N F O R M A TI O N  
R ole i n St u d y  
N a me  
C o nt act I nf or m ati o n  
Cli nical St u d y Lea der  
 
  
 
 
E mail: 
 
Me dical M o nit or  
 
 
 
 
E mail: 
 
 
 
 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
8  
0 2 /0 5 /2 1  
 
A B B R E VI A TI O N S  A N D  T E R M S  
A b brevi ati o n  
F ull ter m  
A E  
A d verse E v e nt  
A N C O V A  
A nal ysis of C o varia nce  
A R P  
All Ra n d o mize d  P o p ulati o n  
B C D V A  
Best C orrecte d Dista n ce Vis ual Ac uit y  
B P  
Bl o o d Press ure  
° C 
De gree Ce nti gra de  
C C L R U  
C or nea a n d C o ntact L e ns Researc h U nit  
C F R  
C o de of Fe d eral Re g ulati o ns  
C H F  
C o n gesti ve Heart Fail ure  
CI  
C o nfi de nce I nter val  
C R O  
Cli nical Researc h Or ga ni zati o n  
C S R  
Cli nical St u d y Re p ort  
° F 
De gree Fa hre n h eit  
D B  
Data base  
D C N V A  
Dista nce C orrecte d Near Vis ual Ac uit y  
e C R F 
Electr o nic Case Re p ort F or m  
E T D R S  
Earl y Treat me nt  Dia betic Reti n o pat h y St u d y  
F D A  
F o o d a n d Dr u g A d mi nistrati o n  
G C P  
G o o d Cli nical Practice  
H R  
Heart Rate  
I B 
I n v esti gat ors’ Br oc h ure 
I C H 
I nter nati o nal C o u ncil f or Har m o nisati o n 
I N D 
I n v esti gati o nal Ne w Dr u g A p plicati o n  
I O P 
I ntr ao c ular Press ur e  
I R B 
I nstit uti o nal Re vie w B oar d 
I T T 
I nte nt-T o -Treat  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
9  
0 2 /0 5 /2 1  
 
I U D 
I ntr a-Uteri ne De vice  
L D P E  
L o w -De nsit y P ol yet h yle ne  
L S M  
Least S q uares M ea n  
M a x 
Ma xi m u m  
Me d D R A  
Me dical Dicti o nar y f or R e g ulat or y Acti vities  
mI T T  
M o difie d I nte nt -t o-Treat  
N y x ol  
0. 7 5 % P he nt ola mi ne O p ht hal mic S ol uti o n or 1 % P he nt ola mi ne Mes ylate 
O p ht hal mic S ol uti o n  ( N y x ol ®) 
O D  
Oc ul us De xter  
O R  
O d ds Rati o  
O S  
Oc ul us Si nister  
O T C  
O ver T he C o u nter  
O U  
Oc ul us Uter q ue ( b ot h e y es)  
Pare m y d ®  
1 % H y dr o x ya m p het a mi ne H y dr o br o mi de/ 0. 2 5 % T r o pica mi de 
P D  
P u pil Dia meter  
R A F  
R o yal Air F orc
 
S A E  
Seri o us A d verse E v e nt  
S E  
Sta n dar d Err or  
S O C  
S yste m Or ga n Class  
S P  
Safet y P o p ulati o n  
T E A E  
Treat me nt E mer ge nt A d v erse E ve nt  
U S  
U nite d States  
U S P  
U nite d States P har mac o p eia  
V A  
Vis ual Ac uit y  
 
 
 
 
 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
1 0  
0 2 /0 5 /2 1  
 
1.  S T U D Y  S U M M A R Y  
St u d y N u m ber  
O PI -N Y X R M -3 0 1  
Cli nic al P h ase  
P hase  3  
T y pe of St u d y  
Ra n d o mize d, parallel ar m, d o u ble -mas ke d, place b o -c o ntr olle d 
st u d y of t he safet y a n d efficac y of N y x ol ( 0. 7 5 % P he nt ola mi ne 
O p ht hal mic S ol uti o n) t o re verse p har m ac ol o gicall y -i n d uce d 
m y driasis i n healt h y s u bjects.  
N a me of I n vesti g ati o n al 
Pr o d uct  
N y x ol ® E ye D r o ps –  0. 7 5 % P he nt ola mi ne O p ht hal mic S ol uti o n  
D ur ati o n of St u d y  
2 da ys, i ncl u di n g scree ni n g /treat me nt a n d f oll o w-u p . 
R ati o n ale  
N y x ol is a o nce -d ail y preser vati ve -free e ye dr o p f or m ulati o n of 
p he nt ola mi ne, w hic h is a n o n -selecti ve al p ha-1 a n d al p ha-2 
a dre ner gic a nt a g o nist t hat acts o n t he a dre ner gic rece pt ors a n d 
i n hi bits c o ntracti n of t he s m o ot h m uscle. P he nt ola mi ne 
i n hi bits c o ntracti o n of t he iris dilat or m uscle, res ulti n g i n a 
s maller p pil size. 
P har mac ol o gicall y -i n d uce d m y driasis is ac hieve d eit her b y 
sti m ulati n g t he iris dilat or m uscle wit h t he use of direct or i n direct 
al p ha 1 a g nists (e. g. p h e n yle p hri ne , h y dr o x y a m p heta mi ne ), or b y 
bl oc ki n g t he iris s p hi ncter m uscle wit h t he use of m uscari nic 
a nta g o nists  or c ycl o ple gi c dr u gs  (e. g., tr o pi ca mi de), or 
c o m bi nati o n  (e. g., Pare m y d ® , w hic h is 1 %  h y dr o x ya m p heta mi ne 
h y dr o br o mi de  a n d 0. 2 5 %  tr o pica mi de).  
N y x ol,  eit her b y  directl y  a nta g o nizi n g  t he al p ha -1  rece pt or or  b y  
i n directl y a nta g o nizi n g  t he p u pil  dilati o n  effect of  m uscari nic  
bl oc ki n g,  ca n e x p e dite t he re versal of  m y driasis  pri or  t o nat ural  
re versal. 
I n a re v ersal of  m y driasis  p hase  2  cli nical  st u d y ( MI R A-1) , N y x ol  
all o we d p u pil  dilati o n  of  m ore  s u bjects t o ret ur n t o baseli ne  faster, 
it was  ge nerall y  well -t olerate d i n t he e y e ( m ost c o m m o n  c o m plai nt  
was  mil d  t o m o derate  c o nj u ncti val  h y pere mia)  a n d t here were  n o  
cli nicall y mea ni n gf ul  s yste mic effects  as meas ure d  b y  heart  rate 
( H R) a n d bl o o d  pre ss ure ( B P). 
St u d y O bjecti ves  
T he o bjecti ves of t his st u d y are:  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
1 1  
0 2 /0 5 /2 1  
 
•  T o e val uate t he effi cac y of N y x ol t o e x pe dite t he re versal 
of p har mac ol o gic all y-i n d uce d m y driasis  acr oss m ulti ple 
m y driatic a ge nts  wit h a n e m p hasis o n p he n yle p hri ne  
•  T o e val uate t he effi cac y of N y x ol t o ret ur n s u bjects t o 
baseli ne acc o m m o dati o n after w orse ni n g (wit h c y cl o ple gi c 
a ge nts  tr o pica mi de a n d Pare m y d ) 
•  T o e val uate t he s afet y of N y x ol  
•  T o e val uate a n y a d diti o nal be nefits of t he re versal of 
p har mac ol o gic all y-i n d uced  m y driasis  
T he S p o ns or i nte n ds t o use t his first P hase 3 re gistrati o n st u d y t o 
e val uate N y x ol  f or t he i n dicati o n “t he treat me nt of 
p har mac ol o gicall y -i n d uce d m y driasis pr o d uce d b y a dre ner gic 
( p he n yle p hri ne) or paras y m pat h ol ytic (tr o pica mi de) a ge nts, or a 
co m bi nati o n t here of.”  
Desi g n  
A ra n d o mize d, p arallel ar m, d o u ble -mas ke d, place b o -c o ntr olle d 
P hase 3 st u d y i n at least 1 6 8  ra n d o mize d s u bjects ( 1 6 0 e val ua ble 
f or efficacy ), e v al uati n g t he safet y a n d efficac y of N y x ol i n 
s u bjects wit h p har mac ol o gicall y-i n d uce d m y driasis. 
F oll o wi n g t he s uccessf ul c o m pleti o n of scree ni n g , eac h s u bject 
will be stratifie d b y e ye c ol or a n d t he n si m ulta ne o usl y  be  
ra n d o mized  to  m y driatic a ge nt (u n mas ke d ) a n d tr eat me nt 
(mas ke d )  
Treat me nt ra n d o mizati o n will be 1: 1 , N y x ol or place b o ( ve hi cle). 
S tratificati o n b y iris c ol or will be 1: 1, li g ht or dar k iri des, as s h o w n 
i n A p pe n di x  1 . 
T he m y driatic a ge nt ra n d o mizati o n will be 3: 1: 1  ( 2. 5 % 
p he n yle p hri ne, 1 % tr o pi ca mi de, a n d Pare m y d ). T h at is, 
a p pr o xi matel y 6 0 % of t h e ra n d o mize d s u bj ects will recei ve o ne 
dr o p of 2. 5 % p he n yl e p hri ne 1 h o ur bef ore treat me nt ( 9 6 e val ua ble 
s u bjects), a p pr o xi matel y 2 0 % will recei v e o ne dr o p of 1 % 
tr o pica mi de 1 h o ur bef ore treat me nt ( 3 2 e val ua ble  s u bjects), a n d 
a p pr o xi matel y 2 0 % will recei v e Pare m y d 1 h o ur bef ore tr eat me nt 
( 3 2 e val ua ble s u bj ects). 
A ra n d o mizati o n sc h e ma b y i n vesti gati o nal treat me nt, m y driatic 
a ge nt, a n d iris c ol or  is s h o w n i n A p pe n di x 2 . 
T R E A T M E N T  ( N Y X O L O R P L A C E B O) WI L L B E 
A D MI NI S T E R E D T O B O T H E Y E S ( O U) . A D U L T S U BJ E C T S 
(≥ 1 8 Y E A R S  O L D) WI L L H A V E T W O D R O P S O F  
T R E A T M E N T  A D MI NI S T E R E D FI V E MI N U T E S A P A R T I N 
T H E S T U D Y E Y E ( O D) A N D O N E D R O P O F T R E A T M E N T  
A D MI NI S T E R E D I N T H E N O N -S T U D Y E Y E ( O S) O N E H O U R 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
1 2  
0 2 /0 5 /2 1  
 
A F T E R M Y D RI A TI C D R U G I N S TI L L A TI O N.  P E DI A T RI C 
S U BJ E C T S ( < 1 8 Y E A R S O L D) WI L L H A V E O N E  D R O P O F 
T R E A T M E N T  A D MI NI S T E R E D I N E A C H E Y E  O N E H O U R 
A F T E R M Y D RI A TI C D R U G I N S TI L L A TI O N.  
 
T he st u d y a n d n o n -st u d y e ye will 
b ot h be e val uate d at all assess me nts.  
P atie nt P o p ul at i o n  
1 6 8 ra n d o mize d ( 1 6 0 e val ua ble) healt h y s u bjects.  
I ncl usi o n Criteri a 
1.  Males or fe m ales ≥ 1 2  years of a g e.  
2.  Ot her wise healt h y a n d well c o ntr olle d s u bjects.  
3.  A bilit y t o c o m pl y wit h all pr ot oc ol -ma n date d pr o ce d ures 
i n de pe n de ntl y a n d t o tte n d all sc he d ule d offi ce visits.  
4.  A d ults  ( ≥ 1 8 years f a ge) w illi n g t o gi ve writte n i nf or me d 
c o nse nt t o partici pate i n t his st u d y.  C hil dre n a ge d 1 2 -1 7 
years t o pr o vi de i g ne d asse nt f or m, as well as a s e parat e 
pare ntal/ Le g al G uar di a n c o nse nt.  
E xcl usi o n Criteri a  
O p ht h al mic  (i n eit her e ye):  
1.  
Cli nicall y  si g nifica nt oc ular  disease  as dee me d  b y  t he 
I n v esti gat or (e. g., cat aract,  gla uc o ma,  c or neal  e de ma,  
u veitis,  se vere kerat o c o nj u ncti vitis  sicca) t hat mi g ht  
i nterfere wit h  t he st u d y. 
2  
U n willi n g  or  u na ble  t o disc o nti n ue  use  of  c o ntact  le nses at  
scree ni n g u ntil  st u d y c o m pleti o n.  
3  
U n willi n g  or  u na ble  t o s us pe n d use  of  t o pical me dicati o n  at 
scree ni n g u ntil  st u d y c o m pleti o n. 
4.  
Oc ular  tr a u ma, oc ular  s ur ger y or  n o n -refracti ve laser 
treat me nt wit h i n t he 6  m o nt hs  pri or  t o scree ni n g. 
5.  
Use  of  a n y t o pical prescri pti o n  or  o ver -t h e-c o u nt er ( O T C) 
o p ht hal mic  me dicati o ns  of  a n y ki n d  wit hi n  7  da ys  of  
scree ni n g, wit h  t he e x ce pti o n  of  a) li d scr u bs wit h  O T C  
pr o d ucts  (e. g., O C u S O F T ®  li d scr u b, Steri Li d ® , ba b y  
s ha m p o o, etc.) or  b)  O T C  l u bricati n g dr o ps  f or dr y  e ye  
(preser ve d  a n d u n preser ve d  artifici al  tears), w hi c h  ma y  
ha ve  bee n  use d  pri or  t o, b ut  n ot  at scree ni n g u ntil  st u d y 
c o m pleti o n . 
6.  
Rece nt  or  c urre nt  e vi de nce  of  o c ular  i nfecti o n or  
i nfla m mati o n i n eit her e ye (s uc h as c urre nt  e vi d e nce of  
cli nicall y  si g nifica nt ble p haritis,  c o nj u ncti vitis,  or  a hist or y  
of  her pes  si m ple x or  h er pes  z oster keratitis  at Scree ni n g  i n 
eit her e ye).  
7.  
Hist or y  of  dia betic  reti n o pat h y or  dia betic  mac ular  e de ma. 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
1 3  
0 2 /0 5 /2 1  
 
8.  
Cl ose d  or  ver y  n arr o w  a n gles t hat i n t he I n vesti gat or’s 
o pi ni o n  are p ote ntiall y  o ccl u da ble  if t he s u bject’s p u pil  is 
dilate d.  
9.  
Hist or y  of  a n y tr a u matic (s ur gical or  n o ns ur gi cal)  or  n o n -
tra u matic c o n diti o n  affecti n g  t he p u pil  or  iris (e. g., 
irre g ularl y s ha pe d p u pil,  ne ur o ge nic  p u pil  dis or der,  iris 
atr o p h y, iri d ot o m y, iri dect o m y, etc.). 
1 0.  K n o w n  aller g y or  c o ntr ai n dicati o n  t o a n y  c o m p o ne nt  of  t h e 
m y driatic  a ge nts  or  t he v e hicle  f or m ulati o n. 
1 1.  Hist or y  of  ca uterizati o n  of  t he p u n ct u m  or  p u n ctal  pl u g  
(silic o ne or  c olla ge n) i nserti o n or  re m o val. 
 
 
 
S yste mic:  
1.  
K n o w n  h y perse nsiti vit y  or  c o ntr ai n dicati o n  t o α - a n d/ or β  
a dre n oce pt or  a nta g o nists  (e. g., c hr o ni c o bstr ucti ve  
p ul m o nar y  disease  or  br o nc hial  ast h ma;  a b n or m all y  l o w 
bl o o d  press ure  ( B P) or  heart  rate ( H R); sec o n d- or  t hir d-
de gree  h eart  bl oc ka ge  or  C o n gesti ve  Heart  Fail ure  ( C H F); 
se vere di a betes).  
2.  
Cli nicall y  s g nifica nt s yste mic disease  (e. g., u nc o ntr olle d  
dia bete
 m yast he nia  gra vis,  ca ncer, he patic,  re nal, 
e n d ocri ne  or  car di o vasc ul ar dis or ders)  t hat mi g ht  i nterfere 
wit h  t he st u d y. 
3  
I nitiati o n of  treat me nt wit h  or  a n y c ha n ges  t o t he c urre nt  
d osa ge,  dr u g  or  re gi me n of  a n y s yste mic a dre ner gic or  
c h oli ner gic dr u gs  wit hi n  7  da ys  pri or  t o scree ni n g, or  
d uri n g  t he st u d y (see A p pe n di x  3 ). 
4.  
Partici pati o n  i n a n y i n vesti gati o nal st u d y wit hi n  3 0  da ys  
pri or  t o scree ni n g. 
5.  
Fe males  of  c hil d beari n g  p ote ntial  w h o  are pre g na nt,  
n ursi n g,  pla n ni n g  a pre g na nc y,  or  n ot  usi n g  a m e dicall y  
acce pta ble f or m of  birt h  c o ntrol.  Acce pta bl e  met h o ds  
i ncl u de t he use  of  at least o ne  of  t he f oll o wi n g: i ntra uteri ne 
de vice  (I U D), h or m o nal  ( oral, i njecti o n, pat c h,  i m pla nt, 
ri n g), barrier  wit h  s per mici de (c o n d o m, dia p hra g m),  or  
a bsti ne nce.  A  fe male is c o nsi dere d  t o b e  of  c hil d beari n g  
p ote ntial  
u nless  
s he 
is 
pre -me nstr ual,  
1  
year  
p ost me n o pa usal , or  3  m o nt hs  p ost -s ur gical sterilizati o n. 
All  fe males of  c hil d beari n g  p ote ntial  i ncl u di n g t h ose wit h  
p ost -t u bal li gati o n m ust  ha ve  a ne gati ve  uri ne  pre g na nc y  
test res ult at Visit  1/ Scree ni n g  a n d m ust  i nte n d t o n ot  
bec o me  pre g na nt  d uri n g  t he st u d y. 
6.  
Resti n g  H R  o utsi de  t he n or mal  ra n ge ( 5 0-1 1 0  b eats  per  
mi n ute)  at t he Scree ni n g  Visit.  H R  ma y  be  re peate d o nl y  
o nce  if o utsi de  t he n or mal  ra n ge f oll o wi n g at least a 5 -
mi n ute  rest peri o d  in  t he sitti n g p ositi o n.  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
1 4  
0 2 /0 5 /2 1  
 
7.  
H y perte nsi o n  wit h  resti n g diast olic  B P  >  1 0 5  m m H g  or  
s yst olic B P  >  1 6 0  m m H g  at t he Scree ni n g  Visit.  B P  ma y  
be  re p eate d o nl y  o nce  if o utsi de  t he s pecifie d ra n ge 
f oll o wi n g at least a 5 -mi n ute  rest peri o d  i n t he sitti n g 
p ositi o n.  
Visit Sc he d ule  
 
 
  
 
 
 
 
 
 
 
 
  
 
 
 
 
 
  
 
 
  
N u m ber of 
I n vesti g ati o n al Sites 
A p pr o xi matel y  1 2  sites 
Esti m ate d T ot al S a m ple 
Size  
A p pr o xi matel y 1 6 8  ra n d o mize d healt h y s u bjects, wit h 
a p pr o xi matel y 1 6 0 e val u a ble s u bjects. 
S a m ple Size J ustific ati o n  A  sa m ple size of  1 6 0  e val ua ble s u bjects ( 8 0 per  tr eat me nt gr o u p)  is 
nee de d  f or t he st u d y. T he  pri mar y  efficac y  e n d p oi nt  will  be  met  if 
s u bjects s h o w a p ositi ve  effect  f or N y x ol  ( α =  0. 0 5  si g nifica nce, 
t w o-taile d). O ne  h u n dre d  a n d six t y e val ua ble s u bj ects will  pr o vi de  
at least 
 
 
 
.  
All  s u bjects will  be
 
 
 F urt her m ore,  s u bjects will  be  ra n d o mize d at a 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A
o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
1 5  
0 2 /0 5 /2 1  
 
rati o of  3: 1: 1  t o o ne  of  t hree m y driatic  a ge nts  (2. 5 %  p he n yle p hri ne,  
1 %  tr o pica mi de, a n d Pare m y d ).  
It is ass u me d  t hat t here will  be  a 5 %  dr o p -o ut  d uri n g  Visit  1 . T o  
acc o u nt  f or t his dr o p -o ut,  a t otal of  1 6 8  s u bjects will  be  
ra n d o mize d i nt o t he st u d y. 
Pri m ar y Effic ac y 
E n d p oi nt  
Perce nta ge  of  s u bjects’ st u d y e yes ret ur ni n g t o ≤ 0. 2  m m  fr o m 
baseli ne  p u pil  dia meter  at 9 0  mi n utes.  
Sec o n d ar y Effic ac y 
E n d p oi nts  
Sec o n dar y efficac y e n d p oi nts (f or t he st u d y e ye  a n d t he n o n-st u d y 
e ye) will i ncl u de: 
•  Perce nta ge of s u bjects ret ur ni n g t o ≤ 0. 2 m m  fr o m baseli ne 
p u pil di a meter  at eac h  re mai ni n g ti me p oi nt (3 0 mi n utes , 
6 0 mi n utes , 2 h o urs, 3 h o urs, 4 h o urs, 6 h o urs , 2 4 h o urs) 
•  C ha n ge (i n m m) fr o m m a x p u pil di a m eter  ( 0 mi n utes) at 
eac h ti me p oi nt ( 3 0 mi n utes , 6 0 mi n utes , 9 0 mi n utes , 2 
h o urs, 3 h o urs  4 h o urs, 6 h o urs , 2 4 h o urs) 
•  Perce nt a g e of s u bjects wit h u nc ha n ge d acc o m m o d ati o n  
fr o m bas eli ne (-1 h o ur) at t he f oll o wi n g ti me p oi nts (0 mi n, 
6 0 mi n utes , 9 0 mi n utes , 2 h o urs, 3 h o urs, 4 h o urs , 6 h o urs )  
•  Ti me  ( h o urs) t o ret ur n t o ≤ 0. 2  m m  fr o m baseli ne  p u pil  
di
meter  (ti me s a vi n gs a n al ysis )  
 
Meas ure me nts:  
 P u pil dia meter will be meas ure d wit h a Ne ur O ptics VI P -
3 0 0  p u pill o meter ( m m).  
 Acc o m m o dati o n will be meas ure d b y t he 
 
 
 
 
 
 
T he p h ot o pic li g hti n g c o n diti o ns i n t he r o o m will be defi ne d as 
 
 
 
 
 
 
 
 
 
 s h o ul d be 
 f or acc o m m o d ati o n meas ure me nts a n d t he re mai ni n g 
safet y assess me nts (e. g. c o nj u ncti val h y pere mia, a d verse e v e nts , 
s u bject q uesti o n naire, et c.). T he s u bject will be i n t he sa me r o o m 
f or all assess m e nts, a n d ever y eff ort will be ma de t o ha ve t he sa me 
pers o n perf or m t he meas ure me nts at all ti me p oi nts. 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
1 6  
0 2 /0 5 /2 1  
 
S o me  of t he efficac y e n d p oi nts will be a nal yze d o verall, b y 
m y driatic a ge nt, a n d b y li g ht/ dar k iri des at all ti me p oi nts . Eac h 
m y driatic a ge nt will be a nal yze d i n di vi d uall y, a n d a n a d diti o nal 
a nal ysis c o m bi ni n g 1 % tr o pica mi de a n d t he Pare m y d s u bjects i nt o 
a “tr o pica mi de” gr o u p will be perf or me d.  E x pl orat or y a nal yses 
ma y  be perf or me d t o c o m pare 
 
t. 
T he M o difie d I nte nt -t o-Treat ( mI T T ) will be use d f or t he pri mar y 
e n d p oi nt a nal ysis a n d t o a nal yze s electe d s ec o n dar y efficac y 
e n d p oi nts . T he Per Pr ot oc ol (P P ) p o p ulati o n will be use d t o 
a nal yze s electe d s ec o n dar y efficac y e n d p oi nts.  
S afet y E n d p oi nts  
T he pri mar y safet y meas ures are c o nj u ncti v al h y pere mi a, 
s u bjecti ve oc ul ar t oler a bilit y, a n d a d verse e ve nts  ( A Es).  
A nal ysis f or c o nj u ncti val h y pere mia  i ncl u des cha n ge fr o m baseli ne 
(-1 h o ur) i n t he c o nj u ncti val h y pere mia gra di n g  scale ( C C L R U 
i ma ges) at each ti me p oi nt ( 0 mi n, 3 0 mi n utes , 6 0 mi n utes , 9 0 
mi n utes , 2 h o urs, 3, h o urs, 4 h o urs, 6 h o urs , 2 4 h o urs), f or st u d y 
e ye a n d n o n -st u d y e ye. A n y i ncreas e of t w o u nits i n c o nj u ncti val 
h y pere mia bet w ee n c o ns ec uti ve ti me p oi nts is c o nsi dere d a n A E.  
Ot her safet y meas ures i ncl u de i m pair me nt  i n V A (B C D V A a n d 
D C N V A )  i ntr a oc ul ar press ure (I O P), a n d s yste mic s afet y as 
me as ure d b y H R a n d B P . Uri ne pre g na nc y tests f or fe mal es of 
c hil d bear n g p ote ntial will be perf or me d at scree ni n g . 
S u bject  Q uesti o n n aire  val ues will be s u m marize d b y treat me nt 
gr o u p a n d ti me p oi nt ( -1 h o ur, 0 mi n utes, 6 0 mi n utes, 9 0 mi n utes, 3 
h o urs, 6 h o urs, a n d 2 4 h o urs) . 
Mea s ure me nts: 
•  C o nj u ncti val h y pere mia  will be assesse d vis uall y wit h  a 
gra di n g scale  ( 0-3)  usi n g i ma ges fr o m t he C C L R U  
•  S u bjecti ve oc ular t olera bilit y meas ure d o n a 4 -p oi nt scale 
( 0-3)  
•  B C D V A Dista nce will b e meas ure d bi n oc ularl y i n 
p h ot o pic c o n diti o ns b y Sta n dar d E T D R S ill u mi nate d c hart 
( o n wall or sta n d) at 4 meters  (letters rec or de d, lat er 
c o n verte d t o l o g M A R a n d n u m ber of li nes )  
•  D C N V A Rea di n g/ Near will be meas ure d bi n oc ul arl y i n 
p h ot o pic c o n diti o ns b y Near Vis ual A c uit y C hart i n t he 
Precisi o n Visi o n S mall 9 1 4 Ill u mi nat or Ca bi net (li g ht b o x) 
at 1 6 i nc hes ( ~4 0  c m) (letters rec or de d, later c o n v erte d t o 
l o g M A R a n d n u m ber of li nes) 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
1 7  
0 2 /0 5 /2 1  
 
•  I O P is meas ure d wit h t he T o n o-Pe n  
•  S u bject  Q uesti o n naire  will be a  brief  s y m pt o m s ur ve y 
(A p pe n di x 4 ) 
St u d y Me dic ati o ns, D ose 
a n d M o de of 
A d mi nistr ati o n  
N y x ol ® E ye Dr o ps  (P he nt ola mi ne O p ht hal mic S ol uti o n) : 
T w o  dr o p s ( d ose d 
 a part) of  N y x ol  i n t he st u d y e ye ( O D) 
( 1-h o ur  p ost  m y driatic  dr u g  i nstillati o n) a n d o ne  dr o p  of  N y x ol  i n 
t he n o n -st u d y e ye ( O S). N ote:  Pe diatric  s u bjects will  recei ve o nl y  
o ne  dr o p  i n eac h e ye.  
 
Place b o  (N y x ol  ve hicl e):  
T w o  dr o p s ( d ose d 
 a part) of  P lace b o i n t he st u d y e ye 
( O D) ( 1-h o ur  p ost  m y dri atic  dr u g  i nstillati o n) a n d o ne  dr o p  of  
P lace b o i n t he n o n -st u d y e ye ( O S). N ot e:  Pe diatric  s u bjects will  
recei v e o nl y  o ne  dr o p  i n eac h e ye.  
D ur ati o n of S u bject 
P artici p ati o n  
a n d St u d y  
T he t otal le n gt h of s u bject partici pati o n is 
 
 
 
 
 
 
T he e xec uti o n of t he e ntire st u d y (first s u bject scree n t hr o u g h last 
ra n d o mize d s u bject e val u a ble) is e x pecte d t o be a p pr o xi matel y 3 t o 
6 m o nt hs.  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
1 8  
0 2 /0 5 /2 1  
 
T A B L E O F C O N T E N T S  
S U M M A R Y  O F  C H A N G E S  F O R  A M E N D M E N T  1  ........................................................................................ 2  
S P O N S O R  SI G N A T U R E  &  C O N T A C T S  .......................................................................................................... 5  
I N V E S TI G A T O R’ S A G R E E M E N T  ................................................................................................................... 6  
A B B R E VI A TI O N S  A N D  T E R M S  ....................................................................................................................... 8  
1.  
S T U D Y  S U M M A R Y  .................................................................................................................................. 1 0  
2.  
I N T R O D U C TI O N...................................................................................................................................... 2 0  
2. 1.  
I n vesti g ati o n al pr o d ucts  ................................................................................................................... 2 0  
2. 2.  
Fi n di n gs  fr o m n o ncli nic al  a n d  cli nic al st u dies ............................................................................... 2 0  
2. 3.  
Desi g n  j ustific ati o n ........................................................................................................................... 2 1  
2. 4.  
R o ute  of  a d mi nistr ati o n,  d os a ge  re gi m e n,  a n d  tre at m e nt peri o d  ................................................. 2 2  
2. 5.  
C o m pli a nce  ....................................................................................................................................... 2 3  
2. 6.  
St u d y  p o p ul ati o n ............................................................................................................................... 2 3  
3.  
O B J E C TI V E S  A N D  P U R P O S E  .......................................... ................................................................. 2 3  
4.  
S T U D Y  D E SI G N  ................................................................. ......... .......................................................... 2 3  
4. 1.  
Pri m ar y  a n d  sec o n d ar y e n d p oi nts  ..................... 
...
............................................................. 2 3  
4. 2.  
Descri pti o n  a n d  sc he d ule of  visits  a n d  pr oce d ures  .... ................................................................... 2 5  
4. 3.  
Me as ures  t a k e n t o mi ni mize/ a v oi d  bi as  ...... .... 
.................................................................... 2 7  
4. 4.  
St u d y  m e dic ati o ns  ..................................... ....... .............................................................................. 2 7  
4. 4. 1.  Pac k a gi n g  a n d  la beli n g ....................................... ............................................................................ 2 7  
4. 4. 2.  St ora g e  of  st u d y me dicati o n  a n d  dis pe nsi n g  ..................................................................................... 2 8  
4. 4. 3.  St u d y  me dicati o n  a d mi nistrati o n  ........ .......... ................................................................................... 2 8  
4. 4. 4.  St u d y  me dicati o n  acc o u nta bilit y  .. . ........ ....................................................................................... 2 8  
4. 4. 4. 1.  Recei pt  a n d dis p ositi o n  of  st u d y me dicati o n  .............................................................................. 2 8  
4. 4. 4. 2.  Ret ur n  of  st u d y me dicati o n  .. .... ............................................................................................. 2 8  
4. 5.  
E x pecte d  d ur ati o n  of  s u bje
 p artici p ati o n  .................................................................................... 2 8  
4. 6.  
R a n d o miz ati o n  a n d  pr oce d ure  f or bre a ki n g  t he c o de ................................................................... 2 8  
4. 7.  
C ollecti o n  of  d at a  ....... ......... ........ ................................................................................................ 2 9  
4. 8.  
C o m plete d  s u bject ......... ... ........................................................................................................... 2 9  
4. 9.  
N o n -c o m pleti n g  s u bject...... .. ....................................................................................................... 2 9  
4. 9. 1.  St u d y  me dicati o n  disc o n n uati o n  ...................................................................................................... 2 9  
4. 9. 2.  Reas o ns  f or wit h dra wal  fr o m st u d y ................................................................................................... 3 0  
4. 9. 3.  E ntire  st u d y ter mi nate d ...................................................................................................................... 3 0  
4. 9. 4.  Acti o ns  after  disc o nti n uati o n  ............................................................................................................. 3 0  
4. 1 0.  
E v al u a ble  s u bjects ............................................................................................................................ 3 0  
4. 1 1.  
C o m plete d  st u d y ............................................................................................................................... 3 0  
4. 1 2.  
Pr o ce d ure  after  t he c o m pleti o n  of  t he st u d y .................................................................................. 3 0  
5.  
S U B J E C T  I N C L U SI O N A N D  E X C L U SI O N  C RI T E RI A  ..................................................................... 3 0  
5. 1  
S u bject  i ncl usi o n criteri a  ................................................................................................................. 3 0  
5. 2  
S u bject  e x cl usi o n  criteri a  ................................................................................................................. 3 1  
6.  
T R E A T M E N T  O F  S U B J E C T S  ................................................................................................................ 3 2  
6. 1.  
Tre at m e nt  a d here nce  ....................................................................................................................... 3 2  
6. 2.  
C o nc o mit a nt  m e dic ati o ns  ................................................................................................................ 3 2  
7.  
A S S E S S M E N T  O F  E F FI C A C Y  ............................................................................................................... 3 3  
7. 1.  
S pecific ati o n  of  t he effic acy  p ar a m eters  ......................................................................................... 3 3  
7. 2.  
Assessi n g,  rec o r di n g,  a n d  a n al yzi n g  of  effic ac y  p ar a m eters  ......................................................... 3 3  
7. 2. 1.  Scree ni n g/ Da y  1  ................................................................................................................................. 3 4  
7. 2. 2.  Treat me nt  Visit  1/ Da y  1  ..................................................................................................................... 3 4  
7. 2. 3.  F oll o w -U p  Visit  / Da y  2  ..................................................................................................................... 3 5  
7. 2. 4.  U nsc h e d ule d  Visits  ............................................................................................................................ 3 5  
7. 2. 5.  Visit  variati o n ..................................................................................................................................... 3 6  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
1 9  
0 2 /0 5 /2 1  
 
8.  
A S S E S S M E N T  O F  S A F E T Y  .................................................................................................................... 3 6  
8. 1.  
S pecific ati o n  of  s afet y p ar a m eters  .................................................................................................. 3 6  
8. 2.  
Assessi n g,  rec o r di n g,  a n d  a n al yzi n g  s afet y p ar a m eters  ................................................................ 3 6  
8. 3.  
A d verse  e v e nts a n d  seri o us a d verse  e ve nts .................................................................................... 3 7  
8. 3. 1.  A d verse  e v e nt defi niti o ns  ................................................................................................................... 3 7  
8. 3. 2.  F oll o w -u p  of  s u bjects after  a d verse  e ve nts ........................................................................................ 4 1  
9.  
S T A TI S TI C S  .............................................................................................................................................. 4 1  
9. 1.  
S a m ple  Size  ....................................................................................................................................... 4 1  
9. 2.  
A n al ysis  P o p ul ati o ns  ........................................................................................................................ 4 1  
9. 3.  
St atistic al  Met h o ds  ........................................................................................................................... 4 2  
9. 3. 1.  Ge n eral  c o nsi derati o ns ....................................................................................................................... 4 2  
9. 3. 2.  De m o gra p hic  a n d  baseli ne  c haracteristics  ......................................................................................... 4 2  
9. 3. 3.  S u bject  dis p ositi o n  ............................................................................................................................. 4 2  
9. 3. 4.  Me dical  hist or y  a n d  pri or/c o nc o mita nt  me dicati o ns  .......................................................................... 4 2  
9. 3. 5.  A nal ysis  of  efficac y ........................................................................................................................... 4 2  
9. 3. 6.  A nal ysis  of  safet y ............................................................................................................................... 4 3  
9. 4.  
Pr o ce d ure  f or acc o u nti n g  f or missi n g,  u n use d,  or  s p uri o us d at a  ................................................. 4 4  
9. 5.  
Pr o ce d ure  f or re p orti n g  de vi ati o ns  fr o m t he st atistic al pl a n  ........................................................ 4 4  
1 0.  
DI R E C T  A C C E S S  T O  S O U R C E  D A T A  A N D  D O C U M E N T S  ............................................................ 4 4  
1 1.  
Q U A LI T Y  C O N T R O L  A N D  Q U A LI T Y  A S S U R A N C E  ... ................................................................... 4 5  
1 2.  
E T HI C A L  C O N SI D E R A TI O N S  A N D  G C P  C O M P LI A N C E  .............................................................. 4 5  
1 2. 1.  
G C P  c o m pli a nce  ........................................... 
............................................................................. 4 5  
1 2. 2.  
I nstit uti o n al Re vie w  B o ar d  (I R B) ..... 
...................................................................................... 4 5  
1 2. 3.  
Pr ot oc ol  De vi ati o ns/ Vi ol ati o ns  ........ ......... ................................................................................... 4 5  
1 2. 4.  
I nf o r m e d c o nse nt  a n d  asse nt  re q uire m e nts  ................................................................................... 4 6  
1 3.  
D A T A  H A N D LI N G  A N D  R E C O R D  K E E PI N G  .................................................................................... 4 6  
1 3. 1.  
D at a  e ntr y ............................. ....... .................................................................................................. 4 6  
1 3. 2.  
D at a  q u alit y  c o ntr ol  a n d  re p rti n g  ................................................................................................. 4 6  
1 3. 3.  
Arc hi vi n g  of  d at a  ......... 
..... ................................................................................................... 4 7  
1 3. 4.  
Rec or ds  rete nti o n  ...... ......... ........ ................................................................................................. 4 7  
1 3. 5.  
A m e n d m e nts  t o t he pr ot oc ol  ............................................................................................................ 4 7  
A P P E N DI X  1:  I RI S C O L O R  C H
R T  ... .......................................................................................................... 4 8  
A P P E N DI X  2:  R A N D O MI Z A TI O N  S C H E M A  F O R  E V A L U A B L E  S U B J E C T S  B Y  I N V E S TI G A TI O N A L 
T R E A T M E N T,  M Y D RI A TI C  A G E N T,  A N D  I RI S C O L O R  ......................................................................... 4 9  
A P P E N DI X  3:  A D R E N E R GI C  A N D  C H O LI N E R GI C  D R U G S  ................................................................... 5 0  
A P P E N DI X  4:  S U B J E C T  Q U E S TI O N N AI R E  ................................................................................................ 5 1  
A P P E N DI X  5:  C O N J U N C TI V A L  H Y P E R E MI A  G R A DI N G  S C A L E  U SI N G  I M A G E S F R O M  C C L R U 5 2  
A P P E N DI X  6:  VI S U A L  A C UI T Y  C H A R T S  .................................................................................................... 5 3  
 
 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
2 0  
0 2 /0 5 /2 1  
 
2.   I N T R O D U C TI O N 
2. 1.  I nvesti g ati o n al pr o d ucts  
T he test pr o d uct is N y x ol ®  E ye Dr o ps –  0. 7 5 % P h e nt ola mi ne O p ht hal mic S ol uti o n ( N y x ol), a 
n o n -selecti ve al p ha-1 a n d al p ha -2 a dre ner gic a nta g o nist. N ote t hat t he c o nce ntrati o n of 0. 7 5 % 
refers t o p he nt ola mi ne free base, a n d is t he sa me a n d use d i n place of 1 % p he nt ola mi ne mes ylate 
dr u g s u bsta nce, w hi c h w as h o w N y x ol ha d bee n d escri be d i n  all pri or st u di es. Place b o c o ntr ol is 
N y x ol ve hicle al o ne.  
2. 2.  Fi n di n gs  fr o m n o ncli nic al  a n d  cli nic al st u dies 
Detaile d fi n di n gs fr o m n o ncli nical a n d cli nical st u dies a n d p ote ntial ris k are pr o vi de d i n t he 
I n v esti gat ors’ Br oc h ure (I B) (2 0 2 0 ).  
N y x ol has bee n assesse d i n se ve n i n vesti gat or -i nitiate d a n d s p o ns or e d P hase 1 a n d P hase 2 
cli nical trials. Acr oss all trials, 1 6 8 of 2 3 2 a d ult pat ie nts were e x p ose d t o at least o ne d ose of 
p he nt ola mi ne mes ylate o p ht hal mic s ol uti o n . 
I n pri or cli nical trials, N y x ol  has de m o nstrate d a c o nsiste nt a bilit y t o decrease p u pil b y 
a p pr o xi matel y 2 0 % ( ~ 1 –  1. 5 m m) i n b ot h mes o pic a n d p h ot o pic c o n diti o ns . Ke y p upil dia meter 
data are s u m marize d b el o w ( Ta ble 1 ). 
T a ble  1 :  Effic ac y of 1 % N y x ol i n Re d uci n g P u pil Di a meter i n Mes o pic C o n diti o ns i n P h ase 
2 Tri als  
 
St u d y  
Gr o u p  
Mes o pic C o n diti o ns  
Pre -Tre at me nt 
( B aseli ne) P u pil 
Di a meter  
P ost -
Tre at me nt 
P u pil 
Di a meter  
C h a n ge ( %)  
p -v al ue 
c o m p are d 
t o b aseli ne 
p -v al ue 
c o m p are d 
t o pl ace b o 
N Y X -S N V 1 
Place b o ( N = 1 6)  
6. 6 m m  
6. 4 m m  
-0. 2 m m ( -3 %)  
p = 0. 0 8  
p < 0. 0 0 0 1  
 
1. 0 % N y x ol ( N = 3 2)  
6. 5 m m  
5. 2 m m  
-1. 3 m m ( -2 0 %)  
p < 0. 0 0 0 1  
N Y X -0 1a 2 2 
Place b o ( N = 3 8)  
6. 2 5 m m  
6. 3 1 m m  
0. 0 7 m m ( + 1 %)  
p = 0. 6  
p < 0. 0 0 0 1  
1. 0 % N y x ol ( N = 4 0)  
6. 1 7 m m  
5. 3 1 m m  
-0. 8 6 m m ( -1 4 %)  
p < 0. 0 0 0 1  
N Y X G -2 0 1  
Place b o ( N = 2 0)  
4. 5 7 m m  
4. 5 2 m m  
-0. 0 5 m m ( -1 %)  
p = 0. 6 1 7 8  
p < 0. 0 0 0 1  
1. 0 % N y x ol ( N = 1 9)  
4. 6 9 m m  
3. 7 0 m m  
-1. 0 0 m m ( -2 1 %)  
p < 0. 0 0 0 1  
N y x ol was o bser v e d t o be well -t olerate d at si n gle d oses u p t o a n d i ncl u di n g 1. 0 % dail y i n eac h 
e ye. T his i ncl u des 5 9 patie nts w h o recei ve d m ulti ple d oses of u p t o 1 % N y x ol f or at least 1 4 
da ys. Safet y of t he patie nts i n t hese trials was e val uate d b y A E m o n it ori n g, p h ysical 
e xa mi nati o ns, a n d vital si g n assess me nts. Acr oss all trials, n o healt h y v ol u nteers or patie nts 
re p orte d a treat me nt-e mer ge nt S A E. N o deat hs o cc urre d i n a n y of t he trials. N o cli nicall y 
mea ni n gf ul c ha n g es were o bser ve d i n p h ysical e xa mi nat i o ns or vital si g ns, i ncl u di n g bl o o d 
press ure a n d heart rate. A Es re p orte d were mil d t o m o derate i n i nte nsit y wit h t he m ost c o m m o n 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
2 1  
0 2 /0 5 /2 1  
 
bei n g tra nsie nt c o nj u ncti val h y pere mia a n d oc ular irritati o n; h o we ver, N y x ol d osi n g at or near 
be dti me was o bser v e d t o miti ga te or mi ni mize t hese si de effects d uri n g t he da yti me. 
P h ase 2  Revers al of P h ar m ac ol o gic ally I n d uce d My dri asis St u d y ( MI R A-1) : 
I n O PI -N Y X R M -2 0 1  ( MI R A-1) , 3 2  ot her wise healt h y s u bjects ra n d o mize d at Visit 1 i n a 1: 1 
rati o t o recei ve 1 dr o p O U of 1 % N y x ol  ( 1 % P he nt ola mi ne Mes ylate O p ht hal mic S ol uti o n)  or 1 
dr o p of place b o ( ve hicl e) at Visit 1 a n d 1 dr o p of t he alter nati ve st u d y me di cati o n at Visit 2. O ne 
h o ur bef ore a d mi nistrati o n of st u d y me dicati o n, s u bjects recei ve d 1 dr o p O U of a m y driatic 
a ge nt (2. 5 % p h e n yle p hri ne  or  1 %  tr o pica mi de). E ac h s u bject r ecei ve d t he s a me m y driatic a g e nt 
t hr o u g h o ut t he st u d y. P u pil dia meter was me as ure d 3 0 mi n utes, 1, 2, 4, a n d 6 h o urs after d osi n g 
at Visit  1 a n d 2 a n d c o m pare d t o t he bas eli ne.  
I n t his st u d y, N y x ol de m o nstrate d a statisticall y si g nifica nt re d u cti o n i n P D  fr o m ma xi m u m 
ti me p oi nt c o m pare d t o place b o  at 1 h o ur, 2 h o urs, 4 h o urs, a n d 6 h o urs.  T hese statisticall y 
si g nifica nt a n d cli nical m ea ni n gf ul (t y picall y great er t ha n 1 m m re d u cti o n) P D re d ucti o ns were 
o bser ve d at eac h  ti me p oi nt, i ncl u di n g w he n stratifie d b y b ot h par s y m pat h etic (tr o pica mi de) a n d 
a dre ner gic ( p he n yle p hri n e) a ge nts.  
A d diti o nall y, N y x ol was effecti ve at ret ur ni n g a st atisticall y si g nifica nt perce nt of s u bjects’ e yes 
t o baseli ne P D c o m pare d wit h place b o treat me nt at e ver y ti me p oi nt wit h  esti ma ble res ults, 
re gar dl ess of w het h er t he P D t hres h ol d use d was ≤ 0. 5 m m a b o ve baseli ne ( 2-h o ur ti me p oi nt) or 
t he m ore stri n ge nt, ≤ 0 m m a b o ve baseli ne ( 4 -h o ur a n d 6 -h o ur  ti me p oi nts). T he re d u cti o n i n ti me 
nee de d t o a c hie ve re vers al of m y driasis i n eit her e ye w s statisticall y  si g nifica nt wit h N y x ol 
treat me nt c o m pare d wit h place b o treat me nt, re g ar dless of t he t hres h ol d e xa mi ne d or t h e 
m y driatic a ge nt use d. I n t his st u d y, N y x ol le d t o a n a vera g e ti me -sa vi n gs of 2. 2 h o urs t o ret ur n 
P D t o ≤ 0 m m a b o ve baseli ne. A d diti o nall y, 4 0 % of s u bjects gi ve n  p he n yle p hri ne a n d 3 0 % of 
s u bjects gi ve n tr o pica mi de ac hie v e d a ti me sa vi n gs of m ore t ha n 4 h o urs wit h N y x ol , base d o n a 
p ost -h oc a nal ysis . 
I n a p ost-h oc a nal ysis of t he pla n ne d P h se 3 pri m ar y e n d p oi nt (t he  p erce nt of s u bjects t hat 
ret ur ne d t o wit hi n ≤ 0. 2 m m of baseli ne p u pil dia meter), a statisticall y si g nifica nt perce nta ge of 
s u bjects i n t he N y x ol  gr o u p  c o m pare d t o place b o after b ei n g dilate d  wit h eit her p he n yle p hri ne or 
tr o pica mi de ret ur n e d t o b aseli ne at 2 h o urs (2 9 %  vs 1 3 %, res pecti vel y; p  = . 0 3 ). T his differe nce 
wi de ne d ( 6 8 % f or N y x ol  a n d 2 3 % f or pl ace b o) at 4 h o urs ( p  < . 0 0 0 1).   
I n t his st u d y, there were n o se vere a d vers e e v e nts, wit h o nl y mil d t o m o derate c o nj u ncti val 
h y pere mia t hat res ol ve d i n m ost patie nts b y 6 h o urs.  
2. 3.  Desi g n  j ustific ati o n 
P u p il size is u n der t he c o ntr ol of t w o o p p osi n g sets of m uscles –  t he circ ul ar c o nstrict or m uscles 
c o ntr olle d b y t he c h oli ner gic ner v o us s yste m  a n d t he ra dial dilat or m uscles, c o ntr olle d b y t he 
a dre ner gic ner v o us s yste m. 3, 4 T he ra dial dilat or m uscles c o ntai n pre d o mi na ntl y α -1 a dre n er gic 
rece pt ors t hat ca n b e i n hi bite d b y α -1 a nta g o nists 5; t heref ore it is p ossi ble t o i n hi bit dilati o n of 
t he p u pil t hr o u g h bl oc ka de of t he ra dial dilat or m uscles.  
P har mac ol o gicall y i n d uc e d m y driasis is ac hie ve d eit her b y sti m ulati n g t he iris dilat or m uscle 
wit h t he use of direct or i n direct al p ha -1  a g o nists ( e. g., p he n yle p hri ne , h y dr o x ya m p heta mi ne ), b y 
bl oc ki n g t he iris s p hi ncter m uscle wit h t he use of m uscari nic a n ta g o nists or c ycl o ple gic dr u gs  
(e. g., tr o pica mi de), or c o m bi nati o n (e. g., Pare m y d ® , w hic h is 1 % h y dr o x ya m p heta mi ne 
h y dr o br o mi de  a n d 0. 2 5 %  tr o pica mi de). A t y pical  i n d uce d m y driasis dilates t he p u pil t o 6 -8 m m, 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
2 2  
0 2 /0 5 /2 1  
 
a size s uita ble f or o p ht hal mic e xa mi nati o n of t he reti na a n d ot her str u ct ures of t he i nteri or of t he 
e ye.  
S uc h p har mac ol o gicall y i n d uce d m y driasis ca n l ast fr o m a fe w h o urs (t y picall y 6 h o urs) t o da ys, 
de pe n di n g o n t he pi g me ntati o n of t he iris, t he a ge of  t he s u bject, a n d ot her u n k n o w n-t o-date 
fact ors. T h e si de effects of s uc h dilati o n are s e nsiti vit y t o li g ht a n d bl urre d visi o n. Als o, ma n y 
dilati n g a ge nts  ca use c ycl o ple gia  (l oss of acc o m m o dati o n), t he te m p orar y paral ysis of t he m uscle 
w hic h all o ws t he e ye  t o f oc us o n near o bj ects. Accelerati n g m y driatic re v ersal after a n e y e e x a m 
ma y be  be neficial f or m a n y s u bjects. 
P he nt ola mi ne is a n o n -selecti ve al p ha-1 a n d al p ha -2 a dre n er gic a nta g o nist acti n g o n  a dre n er gic 
rece pt ors a n d is k n o w n t o i n hi bit c o ntracti o n of t he iris dilat or m uscle, res ulti n g i n a s maller 
p u pil size. T o c o u nteract t he dilat or y effects of m y driatic a ge nt , N y x ol is pr o p ose d t o be i nstille d 
i n t he e yes p ost  e ye e xa m all o wi n g a ra pi d re v ersal  of m y driasis , t here b y mi ni mizi n g t he si de 
effects a n d disc o mf ort p ost e xa m.  
Alt h o u g h acc o m m o dati o n will be st u die d acr oss v ari o us m y driatic a ge nts, a decreas e i n t he 
a m plit u de of acc o m m o dati o n wit h m y driasis w o ul d o nl y be e x pect e d wit h t he use of m y driatic 
a ge nts t hat i ncl u de tr o pic a mi de. Tr o pica mi de is a m uscari nic a nta g o nist t hat elicits c ycl o ple gia, 
or paral ysis of t he ciliar y m uscle of t he e y e, res ulti n g i n a l oss of acc o m m o dati o n. P he n yle p hri ne 
is a s y m pat hetic a g o nist t hat s h o ws m y driatic effect t hr o u g h direct acti o n o n s y m pat hetic ner ve 
rece pt ors l ocate d o n t he p u pillar y dilat or m uscle of t he iris, a n d as s uc h w o ul d n ot be e x pecte d t o 
ha ve materi al c ha n g es i n acc o m m o dati o n.  
I n a pre vi o us P hase 2  re v ersal of p har mac ol o gicall y i n d uce d m y driasis cli nical st u d y  ( MI RA -1) , 
N y x ol was effecti ve at i n d uci n g re versal of m y dri asis  wit h b ot h a dre ner gi c  ( p he n yle p hri ne) a n d 
paras y m pat hetic (tr o pica mi de) m y driatic a ge nts  as descri be d a b o ve. T he pl ace b o o utc o m es i n 
t his st u d y de m o nstrate d  t hat t he nat ural r e versal of m y driasis ta kes l o n ger wit h tr o pica mi de t ha n 
wit h  p he n yle p hri ne. Des pite t his differe nce  N y x ol was  a ble t o re v erse m y driasis faster i n t he 
vast maj orit y of e yes r e g ar dless of t he m y driatic a ge nt use d , b ut faster f or p he n yle p hri ne as 
e x pecte d gi v e n t he p ha r mac ol o g y of N y x ol   
Al p ha -1 a dre ner gi c a nta g o nists ha ve bee n s h o w n t o be safe a n d effecti ve f or t he p har mac ol o gical 
re vers al of m y driasis. I n 1 9 9 0, t he F D A a p pr o ve d Da pi praz ol e H y dr o c hl ori de O p ht hal mic 
S ol uti o n 0. 5 % ( Re v -E yes) f or t his i n dicati o n, h o we ver , t he pr o d uct was wit h dra w n a n d 
disc o nti n ue d b y t he ma n ufact ur er f or reas o ns n ot relate d t o safet y or efficac y. Ma n y pe o ple w h o 
u n der g o p u pil dilati o n f or a n a n n ual o p ht hal mic e xa mi nati o n or ot her o p ht hal mic pr oce d ure 
re q uiri n g p u pil dilati o n c o nti n ue t o re q uest a n o pti o n f or ra pi d r e versal of t he m y driasis.  
2. 4.  R o ute  of  a d mi nistr ati o n,  d os a ge  re gi me n, a n d  tre at me nt peri o d  
As t he i nte n de d r o ute of a d mi nistrati o n f or N y x ol  is t o pical oc ular, t his is t he r o ute t o be use d i n 
t his st u d y.  
T he d ose f or o n e dr o p of N y x ol selecte d f or t his st u d y, 0. 7 5 %, was bas e d u p o n : 1) precli nical 
safet y st u dies, 2) t he res ults of t he pre vi o us o p ht hal mic cli nical st u dies descri be d a b o ve  a n d i n 
t he I B, a n d 3) cli nical st u dies c o n d ucte d wit h var yi n g d oses of dr u gs i n t he sa me class .6  
S U BJ E C T S ≥ 1 8 Y E A R S O L D WI L L R E C EI V E T W O D R O P S ( D O S E D 5  MI N U T E S  
A P A R T)  O F T R E A T M E N T ( N Y X O L O R P L A C E B O) I N T H EI R S T U D Y E Y E A N D O N E 
D R O P O F T R E A T M E N T I N T H EI R N O N -S T U D Y E Y E. S U BJ E C T < 1 8 Y E A R S O L D WI L L 
R E C EI V E O N E D R O P O F T R E A T M E N T I N T H EI R S T U D Y E Y E A N D O N E D R O P O F 
T R E A T M E N T I N T H EI R N O N -S T U D Y E Y E . 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
2 3  
0 2 /0 5 /2 1  
 
N ote t hat 0. 7 5 % P he nt ola mi ne O p ht hal mic S ol uti o n, w hic h e x presses t he p he nt ola mi ne mes ylate 
c o nce ntr ati o n i n free b as e, is t he ne w n o me ncl at ure bei n g use d i n place of 1 % P he nt ola mi ne 
Mes ylate O p ht hal mic S ol uti o n , w hic h was h o w N y x ol ha d bee n des cri be d i n pri or st u dies, 
i ncl u di n g MI R A-1 . 
2. 5.  C o m pli a nce  
T his st u d y will be c o n d ucte d i n c o m plia nce wit h t he pr ot oc ol a n d i n acc or da nce wit h G o o d 
Cli nical Practice, t he et hi cal pri nci ples set f ort h i n t he Declarati o n of Helsi n ki a n d wit h t he U. S. 
C o de of Fe d eral Re g ulati o ns.  
2. 6.  St u dy  p o p ul ati o n  
A sa m ple size of a p pr o xi matel y 1 6 8 healt h y s u bjects ≥ 1 2  years of a ge will be ra n d o mize d i n a 
1: 1 rati o t o o ne of t w o mas ke d treat me nt ar ms ( N y x ol or Place b o), wit h t h e e x pectati o n t hat 
a p pr o xi matel y 1 6 0  s u bjects will c o m plete t he st u d y . Ra n d o mizati o n will be stratifie d 1: 1 b y 
li g ht/ dar k c ol or iri des a n d f urt her ra n d o mize d 3: 1: 1 t o u n mas ke d m y driatic a ge nt  (2. 5 % 
p he n yle p hri ne , 1 % tr o pi ca mi de, a n d Pare m y d ). T h e s u bjects will be recr uite d fr o m 
a p pr o xi matel y 1 2 i n vesti gati o nal sites.  
3.  O B J E C TI V E S  A N D  P U R P O S E  
T he MI R A -2 st u d y is a r a n d o mize d, parallel ar m, d o u ble -mas ke d, place b o -c o ntr olle d st u d y of 
t he safet y a n d effi cac y of N y x ol ( 0. 7 5% P he nt ola mi ne O p ht hal mic S ol uti o n) t o re verse 
p har mac ol o gicall y -i n d uc e d m y driasis i n healt h y s u bjects. 
T he o bjecti ves of t his st u d y are:  
•  T o e val uate t he effi cac y of N y x ol t o e x pe dite t he re versal of p har m ac ol o gi c all y-i n d uce d 
m y driasis  acr oss m ulti ple m y driatic a g nts wit h a n e m p hasis o n p he n yle p hri ne  
•  T o e val uate t he effi cac y of N y x ol o ret ur n s u bjects t o baseli ne acc o m m o d ati o n after 
w orse n i n g ( wit h c y cl o ple gic a ge nts tr o pica mi de a n d Pare m y d) 
•  T o e val uate t he s afet y of N y x ol  
•  T o e val uate a n y a d diti o nal be nefits of t he re versal of p har mac ol o gicall y -i n d uce d m y driasis 
T he S p o ns or i nte n ds t o use t his first P hase 3 re gistrati o n st u d y t o e val uate N y x ol f or t he 
i n dicati o n “t he treat me nt of p har mac ol o gicall y-i n d uce d m y driasis pr o d uce d b y a dre ner gic 
( p he n yle p hri ne) or paras y m pat h ol ytic (tr o pica mi de) a ge nts, or a c o m bi nati o n t here of.” 
4.  S T U D Y  D E SI G N  
4. 1.  Pri m ary  a n d  sec o n d ary e n d p oi nts 
Effic ac y:  
T he pri mar y eff ic ac y e n d p oi nt is t he perce nta ge of s u bjects’ st u d y e yes ret ur ni n g t o ≤ 0. 2 m m 
baseli ne p u pil dia meter at 9 0 mi n utes.  
T he  st u d y e ye is defi n e d  as t he ri g ht e ye ( O D). T h e  n o n -st u d y e ye is defi ne d  as t he left e y e ( O S). 
T he  st u d y a n d n o n -st u d y e ye will  b ot h  be  e val u ate d  at all assess me nts.  Sec o n dar y efficac y 
e n d p oi nts (f or t he st u d y e ye  a n d f or t h e n o n -st u d y e ye) will i ncl u de: 
•  Perce nt of s u bje cts ret ur ni n g t o ≤ 0. 2 m m  fr o m baseli ne p u pil di a m eter  at eac h 
re mai ni n g ti me p oi nt (3 0 mi n utes , 6 0 mi n utes , 2 h o urs, 3 h o urs, 4 h o urs, 6 h o urs, 2 4 
h o urs)  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
2 4  
0 2 /0 5 /2 1  
 
•  C ha n ge (i n m m) fr o m ma x p u pil di a meter  ( 0 mi n utes) at eac h ti me p oi nt ( 3 0 mi n utes , 6 0 
mi n utes , 9 0 mi n utes , 2 h o urs, 3 h o urs, 4 h o urs, 6 h o urs, 2 4 h o urs) 
•  Perce nt a g e of s u bjects wit h u nc ha n ge d acc o m m o d ati o n  fr o m baseli ne (-1 h o ur) at eac h 
ti me p oi nt ( 0 mi n, 6 0 mi n utes , 9 0 mi n utes , 2 h o urs, 3 h o urs, 4 h o urs , 6 h o urs)  
•  Ti me  (h o urs ) t o ret ur n t o ≤ 0. 2  m m  fr o m baseli ne  p u pil  dia meter  (ti me s a vi n gs a n al ysis )  
Meas ure me nts:  
•  P u pil  dia meter  will  be  meas ure d  wit h  a Ne ur O ptics  VI P -3 0 0  p u pill o meter  ( m m). 
•  Acc o m m o dati o n  will  be  meas ure d  b y  t he R A F  Near  P oi nt  R ule  (meas ure d  i n ce nti meters 
a n d t he n c o n v erte d  t o Di o pters ). U nc h a n ge d  acc o m m o dati o n  fr o m Baseli ne  (-1  h o ur)  is 
defi ne d  as a c ha n g e fr o m baseli ne  val ue  ≥ -1,  as m eas ure d  i n di o pters.  
 
 
 
 
 pri or t o t he p u pil dia meter, 
B C D V A  a n d D C N V A  (s afet y meas ures) at all sc h e d ule d ti me p oi nts. S u bject will sit i n t he e xa m 
c hair faci n g t he ill u mi nate d c hart d uri n g t he accli mati o n peri o d a n d f or all assess me nts.  
 
 f or acc o m m o dati o n meas ure me nts a n d  t he re mai ni n g safet y 
assess me nts (e. g. c o nj u n cti val h y pere mia, a d verse e ve nts, brief s u bject q u esti o n naire, etc.). T he 
s u bject will be i n t he sa me r o o m f or all ass ess me nts, a n d e ver y eff ort will be ma de t o ha ve t he 
sa me pers o n p erf or m t he meas ure me nts at all ti me p oi nts. 
S o me  of t he effic ac y e n d p oi nts will be a nal yze d o verall, b y m y driatic a g e nt, a n d b y li g ht/ dar k 
iri des at all ti me p oi nts. Eac h m y driatic a g e nt will be a nal yze d i n di vi d uall y, a n d a n a d diti o nal 
a nal ysis c o m bi ni n g 1 % tr o pica mi de a n d t he Pare m y d s u bjects i nt o a “tr o pica mi de” gr o u p will be 
perf or me d. E x pl orat or y a nal yses ma y  be perf or me d t o c o m pare efficac y e n d p oi nts bet wee n t he 
st u d y e ye a n d n o n-st u d y e ye wit hi n t he sa me s u bj ect. 
S afet y:  
T he pri mar y safet y meas ures are c o nj u ncti val h y p ere mia, s u bjecti ve oc ular t olera bilit y , a n d A Es. 
O t her safet y meas ur es i ncl u de i m pair me nt i n V A (B C D V A  a n d D C N V A ), i ntra oc ular press ure 
(I O P), s u bject q uesti o n naire, a n d s yste mic safet y as meas ure d b y H R a n d B P. Uri ne pre g na n c y 
tests f or fe males of c hil d beari n g p ote ntial will be perf or me d at scree ni n g pri or t o d osi n g of st u d y 
me dicati o n . 
Meas ure me nts:  
•  C o nj u ncti val h y pere mia  will be assesse d vis uall y wit h a 4-p oi nt c o nj u ncti val h y pere mia 
gra di n g scale ( 0 -3) wit h C C L R U i ma ges ( A p pe n di x 5 )  
•  S u bjecti ve oc ular t olera bilit y meas ure d o n a 4 -p oi nt scale  ( 0-3)  
•  B C D V A Dista nce will b e meas ur e d bi n oc ularl y i n p h ot o pic c o n diti o ns b y Sta n dar d 
E T D R S ill u mi nate d c hart ( o n wall or sta n d) at  
 (letters rec or de d, lat er c o n verte d 
t o l o g M A R a n d n u m ber of li nes) 
•  D C N V A Rea di n g/ Near will be meas ure d bi n oc ul arl y i n p h ot o pic c o n diti o ns b y Near 
Vis ual Ac uit y C hart i n t he 
 at 
 (letters rec or de d, later c o n vert e d t o l o g M A R a n d n u m ber of li nes) 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
2 5  
0 2 /0 5 /2 1  
 
•  I O P is meas ure d wit h t he T o n o-Pe n  
•  S u bject Q uesti o n naire will be a brief s y m pt o m s ur ve y  
4. 2.  Descri pti o n  a n d  sc he d ule of  visits  a n d  pr oce d ures  
A sa m ple size of a p pr o xi matel y 1 6 8  healt h y s u bjects ≥1 2  years of a ge will be ra n d o mize d i n a 
1: 1 rati o t o o ne of t w o treat me nt ar ms ( N y x ol or  Pl ace b o) , wit h t he e x pectati o n t hat 
a p pr o xi matel y 1 6 0  s u bjects will c o m plete t he st u d y. 
 
 a n d 3 : 1: 1 b y m y driatic a ge nt (2. 5 %  p h e n yle p hri ne , 1 % tr o pica mi de, 
a n d Pare m y d ). S u bjects will recei ve t heir m y driatic a ge nt 1 h o ur bef ore treat me nt.  St u d y 
pr oce d ures are s h o w n i n detail i n  Ta ble 2 . 
 
 
 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2




Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
2 7  
0 2 /0 5 /2 1  
 
4. 3.  Me as ures  t a ke n t o mi ni mize/ av oi d  bi as  
T his is a place b o -c o ntr olle d, d o u ble -mas ke d,  1: 1 r a n d o mize d, 2 -ar m P hase 3  st u d y.  
 
4. 4.  St u dy  me dic ati o ns  
St u d y Me dic ati o n I de ntific ati o n 
Est a blis he d n a me  
P he nt ola mi ne mes ylate –  pare nt p he nt ola mi ne  
C A S re gistr y n u m ber  
6 5 -2 8 -1  –  pare nt 5 0 -6 0 -2  
C he mic al cl ass  
A n al p ha-a dre n er gic a nt a g o nist,  it is a me m ber 
of t he f oll o wi n g classes: i mi daz oles, of 
p he n ols, is a tertiar y a mi n o c o m p o u n d a n d a 
s u bstit ute d a nili ne. 
C he mic al n a me  
3 -[ N-( 4, 5-di h y dr o -1 H -i mi daz ol-2 -yl met h yl) -4 -
met h yla nili n o] p he n ol ; met ha nes ulf o nic aci d 
M olec ul ar f or m ul a  
C 1 8H 2 3N 3O 4S –  pare nt C 1 7H 1 9N 3O  
M olec ul ar wei g ht  
3 7 7. 1 4 0 –  pare nt 2 8 1 .3 5 2  
Dr u g n a me/f or m ul ati o n  
N y x ol / a q ue o us is ot o nic s ol uti o n  
C o nce ntr ati o n acti v e  
1 % –  pare nt 0. 7 5 %  
M a n uf act urer dr u g s u bst a nce  
M a n uf act urer dr u g pr o d uct, pl ace b o  Bi oc o nce pt La b orat ories I n c.  
S t or a ge re q uire me nts 
 
 
  
St ore d i n a sec ure d l ocati o n (l oc ke d) wit h n o 
access f or u na ut h orize d p ers o n nel.  
 
F or m ul ati o n  
N y x ol  (0. 7 5 %  P he nt ola mi ne O p ht hal mic S o l uti o n) is a clear, c ol orless t o sli g htl y br o w n,  
sterile, n o n-pres er ve d, is ot o nic, b uffere d a q ue o us s ol uti o n c o ntai ni n g 1 % p he nt ola mi ne 
mes ylate (  e q ui val e nt t o 0. 7 5 %  p he nt ola mi ne free base ), ma n nit ol a n d s o di u m acetate. 
Place b o f or N y x ol  is a clear, c ol orless, sterile, n o n -preser ve d, is ot o nic, b uffere d a q u e o us 
s ol uti o n c o ntai ni n g ma n nit ol a n d s o di u m acetate. T he p H of t he st u d y me dicati o ns ma y be 
a dj uste d wit h  
 ( U S P) a n d/or  
l ( U S P) t o  
.   
4. 4. 1.  P ac k a gi n g  a n d  l a beli n g 
T he i n vesti gati o nal pr o d ucts, acti ve a n d place b o, are b ot h pac ka ge d i n 1-ml L D P E dr o p per 
b ottles c o ntai ni n g 0. 6 ml s ol uti o n f or si n gle -d ose use. Eac h b ottle  is wra p pe d wit h a n 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
2 8  
0 2 /0 5 /2 1  
 
al u mi n u m f oil o ver wra p i m per mea ble t o wat er a n d o x y ge n,  a n d will be la bele d wit h a n 
i n vesti gati o nal la bel s h o wi n g t he st u d y pr ot oc ol n u m ber a n d ot her rele va nt i nf or mati o n, 
i ncl u di n g a state me nt “ Ca uti o n –  Ne w Dr u g –  Li mite d b y Fe deral ( U S) La w t o 
I n v esti gati o nal Use ”.  
4. 4. 2.  St or a ge  of  st u d y me dic ati o n  a n d  dis pe nsi n g  
Pri or t o dis pe nsi n g , all i n vesti gati o nal material m ust be st ore d i n a sec ure l o cati o n wit h 
strictl y li mite d access d oc u me nte d b y si g nat ure of a ut h orize d pers o ns w h o ma y dis pe nse 
i n vesti gati o nal materials. 
 
 St u d y 
me dicati o n m ust  n ot be fr oze n a n d m ust be pr otecte d  fr o m li g ht. All i n vesti gati o nal st u d y 
me dicati o n m ust be st ore d i n a sec ure facilit y, wit h access li mi te d t o t he I n vesti gat or a n d 
a ut h orize d staff.  
4. 4. 3.  St u d y  m e dic ati o n  a d mi nistr ati o n  
St u d y me dicati o n  will be a d mi nistere d b y t he I n vesti gat or or desi g nee  at t he site o n 
Treat me nt Visit 1 .  
4. 4. 4.  St u d y  me dic ati o n  acc o u nt a bilit y  
4. 4. 4. 1.  Recei pt  a n d  dis p ositi o n  of  st u dy me dic ati o n  
T he I n vesti gat or or d esi g nee ( e. g., st u d y c o or di nat or or p har macist) will mai ntai n a f ull 
acc o u nta bilit y rec or d f or t he st u d y me dicati o n a n d will be res p o nsi ble f or rec or di n g t he 
recei pt, dis pe nsi n g, a n d ret ur n of all s u p plies of t h e st u d y me dic ati o n usi n g t he i n ve nt ories 
s u p plie d b y Oc u p hire . T h e I n vesti gat or or d esi g ne e will acc o u nt f or all st u d y me dicati o n. 
T he m o nit or will re vie w dis pe nsi n g a n d st u d y me dicati o n acc o u nta bilit y rec or ds d uri n g 
site visits a n d at t he c o m pleti o n of t h  st u d y a n d note a n y discre p a ncies.   
4. 4. 4. 2.  Ret ur n  of  st u dy me dic ati o n  
W he n t he st u d y is c o m pl te d or is t r mi nate d b y Oc u p hire, all st u d y material i ncl u di n g 
use d a n d u n use d st u d y m e dicati o n b ottles will be ret ur ne d t o Oc u p hir e ( or its desi g nee) or 
destr o ye d u n der t he direct i o n of sa me. All st u d y me dicati o n acc o u nti n g pr o ce d ures m ust 
be c o m plete d bef ore t h e st u d y is c o nsi dere d c o m plete d . A fi nal st u d y me di cati o n 
dis p ositi o n will be c o m plete d b y t he st u d y c o or di n at or.  
4. 5.  E x pecte d  d ur ati o n  of  s u bject p artici p ati o n  
T he t otal le n gt h of s u bject partici pati o n is 
, as s u m marize d b el o w: 
•  
 
 
 
 
 
T he  e x ec uti o n  of  t h e e ntire st u d y (first s u bj ect s cree ne d t hr o u g h last ra n d o mize d s u bject 
c o m plete d) is e x pecte d  t o be  a p pr o xi matel y  3  t o 6  m o nt h s. 
4. 6.  R a n d o miz ati o n  a n d  pr oce d ure  f or bre a ki n g  t he c o de 
A  ra n d o mizati o n c o d e f or all ocati n g  s u bjects t o tr eat me nt will  be  pre p are d  b y  a mas ke d  
bi ostatisticia n  n ot  c o n necte d wit h  t he st u d y. S u bjects  will  be  ra n d o mize d b y  m y driatic  
a ge nt ( u n mas ke d) a n d str atifie d b y  li g ht or  dar k  iri des. 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
2 9  
0 2 /0 5 /2 1  
 
At  t he i nitiati o n of  st u d y relate d pr oce d ures,  e v er y  p ote ntial  s u bject is assi g ne d  a 
S cree ni n g  n u m ber  i n n u merical  or der.  O nce  a s u bject is q ualifie d  f or t he st u d y, t he s u bject 
is assi g ne d  a r a n d o miz ati o n n u m ber  i n t he or der  pr o vi de d  b y  t he bi ostatisticia n.  
T he  st u d y me dicati o ns  will  be  mas ke d  t o b ot h  I n v esti gat or a n d st u d y s u bjects, as well  as 
Oc u p hire . O nl y  i n case of  me dical  e mer ge nc y  or  o cc urre nce  of  S A E s will  t he 
ra n d o mizati o n c o de b e  u n mas ke d  b y  t he Me dical  M o nit or  a n d ma de  a v aila ble t o t he 
I n v esti gat or, Oc u p hir e,  a n d/ or ot her  pers o n nel  i n v ol ve d i n t he m o nit ori n g  or  c o n d uct  of  
t his st u d y. 
4. 7.  C ollecti o n  of  d at a  
St u d y s pecific data t hat h as bee n o utli ne d i n t he pr ot oc ol will be e ntere d i nt o t he cli nical 
data base b y i n di vi d ual(s) desi g nate d b y t he I n v esti gat or. Data is verifie d el ectr o nicall y 
usi n g a series of o nli ne pr o gra m me d e dit c h ec ks t hat ha ve bee n create d b y t he Cli nical 
Data Ma na g er a n d pr o gra m me d b y t h e Cli nical Data Pr o gra m mer or Desi g nee. Dat a 
discre pa n cies will be br o u g ht t o t he atte nti o n of t he cli nic l tea m a n d i n v esti gate d b y t he 
st u d y m o nit or a n d site staff. St u d y m o nit ors  will re vie w a n d v erif y all dat a c ollecte d i n t he 
electr o nic Cas e Re p ort F o r m (eC R F ) a gai nst  a n y a p plica ble  s o urce d oc u m e ntati o n d uri n g 
re m ote re vi e w or sc h e d ul e d m o nit ori n g visits. T he st u d y m o nit or will w or k cl osel y wit h 
t he site staff t o a d dress a n y discre pa n cies w hic h ha ve bee n f o u n d s o t hat pr o per res ol uti o ns 
ca n be ma d e a n d  d o c u me nte d i n t he cli nical data b ase. A n a u dit trail wit hi n t he s yste m will 
trac k all c h a n ges ma de t o t he data. 
4. 8.  C o m plete d  s u bject 
A c o m plete d s u bject is d efi ne d as o n e w h o c o m pl etes all pla n ne d d osi n g a n d pr oce d ures 
t hr o u g h t he e n d of Visit 2 .   
4. 9.  N o n -c o m pleti n g  s u bject 
A n o n -c o m pleti n g s u bject is defi ne d as o ne w h o e xits t he st u d y b y t heir o w n v oliti o n or at 
t he discreti o n of t he I n v esti gat or a n d/ or t he Me dical M o nit or  pri or t o c o m pleti n g all of t he 
st u d y pr oce d ures re q uir e d i n t his pr ot oc ol. A n y su bject  ma y deci de t o v ol u ntaril y 
wit h dra w fr o m t he st u d y at a n y ti me wit h o ut prej u dice.  
4. 9. 1.  St u d y  me dic ati o n  disc o nti n u ati o n  
T he st u d y me dicati o n ma y be disc o nti n ue d f or t he f oll o wi n g reas o ns:  
•  A d verse E ve nts: A E s i n cl u de cli nicall y si g nifica nt la b orat or y a b n or m alities a n d 
i nterc urre nt diseases r e p orte d b y t he s u bject or o bs er ve d b y t he In vesti gat or wit h 
d oc u me ntati o n o n t he eC R F  
•  De at h:  If a s u bject dies, t he A E  t hat ca us e d t he deat h s h o ul d be d oc u me nte d o n t he 
eC R F a n d be  n ote d as seri o us a n d fatal . 
•  Dis all o we d c o nc urre nt me dic ati o n: A n y me dicati o n n ot all o we d b y t he pr ot oc ol 
w o ul d be a pr ot oc ol vi olati o n . 
•  L ac k of effic a c y:  A s u bject ma y elect t o disc o nti n ue partici pati o n i n t he st u d y f or 
a percei ve d lac k of efficac y . 
•  I n vesti g at or decisi o n:  A s u bject ma y be disc o nti n ue d f or reas o ns ot her t h a n t h ose 
b ullete d pre vi o usl y if t he In v esti gat or t hi n ks it is n ot i n t he best i nterest of t he 
s u bject t o c o nti n ue. 
•  Ot her:  If t h ere is a n y ot h er  reas o n f or s u bject disc o nti n uati o n  t his s h o ul d be  n ote d  
o n  t he eC R F . 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
3 0  
0 2 /0 5 /2 1  
 
T he  reas o n f or pre mat ure  st u d y me dicati o n  disc o nti n uati o n  s h o ul d be  e ntere d o nt o  t he 
a p pr o priate eC R F.   
4. 9. 2.  Re as o ns  f or w it h dr a w al fr o m st u d y 
•  S u bject  wit h dra ws c o nse nt . 
•  S u bject  is l ost t o f oll o w-u p . 
•  S u bject  wit h dra ws f or ot her reas o n . 
4. 9. 3.  E ntire  st u d y ter mi n ate d 
T he e ntire st u d y ma y be t er mi nate d b y I n v esti gat ors or Oc u p hir e . Pr o m pt, writte n n otice of 
reas o na bl e ca us e t o t he ot her part y (Oc u p hire  or I n ves ti gat ors, res pecti vel y) is re q uire d. 
Pr o m pt n otice t o t he I R B a n d t o re g ulat or y a ut h orities is als o re q uire d.  
4. 9. 4.  Acti o ns  after  disc o nti n u ati o n  
All s u bjects w h o disc o nti n ue st u d y me dicati o n d ue t o a re p ort of a n A E m ust  be f oll o we d -
u p a n d pr o vi de d a p pr o pri ate me dical care u ntil t heir si g ns a n d s y m pt o ms ha ve re mitte d or 
sta bilize d or u ntil me dical assess me nts  ha ve ret ur n e d t o acce pta ble or pre -st u d y li mits. 
F or a n y s u bject w h o c h o oses t o wit h dra w c o nse nt or w h o is n o n -c o m plia nt, e ver y p o ssi ble 
eff ort s h o ul d be ma d e b y t he I n v esti gat or t o ass ur e t h e 6 h o ur meas ure me nts are assess e d 
pri or t o disc o nti n uati o n , i n a d diti o n t o a f oll o w-u p  tele p h o ne call t hat i ncl u des assess me nts 
f or A E s, c o n c o mita nt me dicati o ns a n d s u bject e v al u ate d c o nj u ncti val h y pere mia .  
4. 1 0.  Ev al u a ble  su bjects  
S u bject s are e v al ua bl e f or effic ac y if t h e y  recei ve d o ne or t w o dr o ps of st u d y treat me nt 
a n d ha d a p u pil dia meter meas ure me nt at t he 9 0 -mi n ute ti me p oi nt at Visit 1.   
4. 1 1.  C o m plete d  st u dy 
T he  st u d y is c o m plete d  w he n  all ra n d o mize d s u bjects ha ve  c o m plet e d  t he st u d y, all eC R Fs  
ha ve  bee n  c o m plete d,  a n d all eC R F  data  e nt ere d  i nt o t he data base.  Fi nal  D B  l oc k will  
occ ur  after t he l ast ra n do mize d  s u bject c o m pletes  last visit , all data  has  bee n  e ntere d a n d 
all q ueries  res ol v e d.  
4. 1 2.  Pr oce d ure  after  t he c o m pleti o n  of  t he st u dy 
W he n t he st u d y is c o m plete d, t he C R O will pr o vi de Oc u p hire a n d t he I n v esti gat or w it h a 
brief (i.e., o n e t o t hree p a ges) re p ort , c o ntai ni n g a descri pti o n of t he st u d y, t he n u m ber of 
s u bjects e nr olle d, t he n u m ber of s u bjects c o m plet e d  t he n u m ber of s u bjects w h o dr o p pe d 
o ut a n d w h y, effi cac y fi n di n gs a n d A E s. 
5.  S U B J E C T  I N C L U SI O N A N D  E X C L U SI O N  C RI T E RI A  
5. 1  
S u bject  i ncl usi o n criteri a 
1.  Males  or  fe m ales ≥ 1 2  years  of  a g e. 
2.  Ot her wise  healt h y  a n d w ell  c o ntr olle d  s u bjects.  
3.  A bilit y  t o c o m pl y wit h  all pr ot oc ol -ma n date d  pr oce d ures  i n de pe n de ntl y a n d t o 
atte n d all sc he d ule d offic e  visits .  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
3 1  
0 2 /0 5 /2 1  
 
4.  A d ults  (≥ 1 8  years  of  a ge) willi n g  t o gi ve  writte n  i nf or me d c o nse nt  t o partici pate  i n 
t his st u d y. C hil dre n  a ge d 1 2 -1 7  years  t o pr o vi de  si g ne d asse nt  f or m, as well  as a 
se parat e pare ntal/ Le gal  G uar dia n  c o ns e nt.  
5. 2  
S u bject  excl usi o n  criteri a 
E xcl u de d fr o m t he st u d y will be i n di vi d uals wit h t he f oll o wi n g c haracter istics: 
O p ht h al mic  (i n eit her e ye):  
1.  
Cli nicall y si g nifica nt oc ular disease as dee m e d b y t he I n vesti gat or (e. g., cat aract, 
gla uc o ma, c or neal e de ma, u veitis, se vere k erat oc o nj u ncti vitis sicca) t hat mi g ht 
i nterfere wit h t he st u d y. 
2.  
U n willi n g or u na ble t o disc o nti n ue use of c o nt act l e nses at scree ni n g u ntil st u d y 
c o m pleti o n.  
3.  
U n willi n g or u na ble t o s us pe n d use of t o pical me di cati o n at scree ni n g u ntil st u d y 
c o m pleti o n.  
4.  
Oc ular tr a u ma, oc ular s ur ger y or n o n -refracti ve l as er treat me nt wit hi n t he 6 
m o nt hs pri or t o scree ni n g.  
5.  
Use of a n y t o pical prescri pti o n or o ver -t he-c o u nter ( O T C) o p ht hal mic 
me dicati o ns of a n y ki n d wit hi n 7 da ys of scree ni n g, wit h t he e xce pti o n of a) li d 
scr u bs wit h O T C pr o d ucts (e. g., O C u S O F T ®  li d scr u b, Steri Li d® , ba b y s ha m p o o, 
etc.) or b) O T C l u bricati n g dr o ps f or dr y e ye ( preser ve d a n d u n preser ve d 
artificial tears), w hic h m a y ha ve b ee n use d pri or t o, b ut n ot at scree ni n g u ntil 
st u d y c o m pleti o n. 
6.  
Rece nt or c urre nt e vi de n ce of oc ular i nf ecti o n or i nfla m mati o n i n eit her e y e (s uc h 
as c urre nt e vi de n ce of cli nicall y si g nifica nt ble p haritis, c o nj u ncti vitis, or a hist or y 
of her pes si m ple x or her pes z oster k eratitis at Scree ni n g i n eit her e y e).  
7.  
Hist or y of dia betic reti n o pat h y or dia betic mac ular e de ma.  
8.  
Cl ose d or ver y narr o w a n gles t hat i n t he I n v esti gat o r’s o pi ni o n are p ote ntiall y 
occl u da ble if t h e s u bject’s p u pil is dilate d.  
9.  
Hist or y of a n y tra u m atic (s ur gical or n o ns ur gical) or n o n -tr a u matic c o n diti o n 
affecti n g t he p u pil or iris (e. g., irre g ularl y s h a pe d p u pil, ne ur o ge nic p u pil dis or der, 
iris atr o p h y, iri d ot o m y, iri dect o m y, etc.).  
1 0.  K n o w n aller g y or c o ntr ai n dicati o n t o a n y c o m p o n e nt of t he m y driatic a ge nts or t he 
ve hicle f or m ulati o n.  
1 1.  
Hist or y of ca uterizati o n of t he p u nct u m or p u nctal pl u g (silic o ne or c olla ge n) 
i nserti o n or re m o val. 
S yste mic:  
1.  
K n o w n  h y perse nsiti vit y  or  c o ntrai n dicati o n  t o α - a n d/ or β  a dre n oce pt or  
a nta g o nists  (e. g., c hr o ni c  o bstr ucti ve  p ul m o nar y  disease  or  br o nc hial  ast h m a;  
a b n or mall y  l o w bl o o d  press ure  ( B P) or  h eart  rate ( H R); sec o n d- or  t hir d-de gree  
heart  bl oc k a ge  or  C o n gesti ve  Heart  Fail ure  ( C H F); se vere di a betes).  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
3 2  
0 2 /0 5 /2 1  
 
2.  
Cli nicall y  si g nifica nt s yste mic disease  (e. g., u nc o ntr olle d  dia betes,  m yast he nia  
gra vis,  ca n cer, h e patic,  re nal, e n d ocri n e or  car di o v asc ular  dis or d ers)  t hat mi g ht  
i nterfere wit h  t he st u d y. 
3.  
I nitiati o n of  tr eat me nt wit h  or  an y  c ha n g es  t o t he c urre nt  d osa g e,  dr u g  or  
re gi me n of  a n y s yste mic a dre n er gic or  c h oli ner gic  dr u gs  wit hi n  7  da ys  pri or  t o 
scree ni n g, or  d uri n g  t he st u d y (see A p pe n di x  3 ). 
4.  
Partici pati o n  i n a n y i n vesti gati o nal st u d y wit hi n  3 0  da ys  pri or  t o scree ni n g. 
5.  
Fe males  of  c hil d beari n g  p ote ntial  w h o  are pre g na nt,  n ursi n g,  pla n ni n g  a 
pre g na n c y,  or  n ot  usi n g  a me dicall y  acce pta ble f or m of  birt h  c o ntr ol. 
Acce pt a ble  met h o ds  i ncl u de t he use  of  at least o n e  of  t he f oll o wi n g: 
i ntra uteri ne de vi ce  (I U D), h or m o nal  ( oral, i njecti o n, patc h,  i m pla nt, ri n g), 
barrier  wit h  s per mici de (c o n d o m, dia p hra g m),  or  a bsti ne nce.  A  fe male is 
c o nsi dere d  t o be  of  c hil d beari n g  p ote ntial  u nless  s he is pre -me nstr ual,  1  year  
p ost me n o pa usal  or  3  m o nt hs  p ost -s ur gical sterilizati o n. All  fe males of  
c hil d beari n g  p ote ntial  i ncl u di n g t h ose wit h  p ost -t u bal li gati o n m ust  ha ve  a 
ne gati ve  uri ne  pre g na n c y  test res ult at Visit  1/ Scree ni n g  a n d m ust  i nte n d t o n ot  
bec o me  pre g na nt  d uri n g  t he st u d y. 
6.  
Resti n g  H R  o utsi de  t he n or mal  ra n ge ( 5 0-1 1 0  beats  per  mi n ute)  at t he 
Scree ni n g  Visit.  H R  ma y  be  re p eate d o nl y  o nce  if o utsi de  t he n or mal  ra n g e 
f oll o wi n g at least a 5 -mi n ute  rest peri o d  i n t he sitti n g p ositi o n.  
7.  
H y perte nsi o n  wit h  resti n g diast olic  B P  >  1 0 5  m m H g  or  s yst olic B P  >  1 6 0  
m m H g  at t he Scree ni n g  Visit.  B P  ma y  be  re peate d o nl y  o nce  if o utsi de  t he 
s pecifie d ra n ge f oll o wi n g at least a 5 -mi n ute  rest peri o d  i n t he sitti n g p ositi o n.  
6.  T R E A T M E N T  O F  S U B J E C T S  
A p pr o xi matel y 1 6 8 healt h y s u bj cts ≥ 1 2  years of a ge will be ra n d o mize d i n a 1: 1 rati o t o 
o ne o f t w o treat me nt ar ms ( N y x ol or Place b o), wit h t he e x pect ati o n t hat a p pr o xi matel y 
1 6 0  s u bjects will c o m plete t he st u d y. Ra n d o mizati o n will be stratifie d 1: 1 b y li g ht/ dar k 
c ol or iri des a n d f urt her ra n d o mize d 3: 1: 1 t o m y driatic a ge nt ( 2. 5 % p he n yle p hri ne, 1 % 
tr o pica mi de, a n d Pare m y d) . S u bjects will recei ve t heir m y driatic a ge nt 1 h o ur bef ore 
treat me nt o n  Treat me nt Visit 1 o n Da y 1. T here will be n o treat me nt a d mi nistere d o n 
F oll o w -U p Visit / Da y 2.  
6. 1.  Tre at me nt  a d here nce  
All s u bjects will be treate d b y t he I n vesti gat or or d esi g nee at t he st u d y cli ni c o n Visit 1 .  
6. 2.  C o nc o mit a nt  me dic ati o ns  
As n ote d i n t he e xcl usi o n criteria ( Secti o n 5. 2 ), u s e of a n y t o pical prescri pti o n or o ver-t he-
c o u nter ( O T C) o p ht hal mic me dicati o ns of a n y ki n d wit hi n 7 da ys of scree ni n g are 
pr o hi bite d , wit h t he e xce pti o n of a) li d scr u bs wit h O T C pr o d ucts (e. g., O C u S O F T ® li d 
scr u b, Steri Li d ®, ba b y s ha m p o o, etc.) or b) O T C l u bricati n g dr o ps f or dr y e ye ( preser ve d 
a n d u n preser ve d artificial tears), w hic h m a y ha ve bee n use d pri or t o, b ut n ot at scree ni n g 
u ntil st u d y c o m pleti o n.  A lar ge n u m ber of dr u gs, b ot h prescri pti o n a n d o ver -t he-c o u nter 
( O T C), c o ntai n acti ve i n gre die nts t hat ca n affect P D. T his w o ul d i ncl u de ma n y e ye dr ops, 
s uc h as Visi ne, t hat w o ul d be use d t o re d u ce re d ness, m ost c o u g h or c ol d pr e parati o ns, 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
3 3  
0 2 /0 5 /2 1  
 
a nti hista mi nes a n d br o nc h o dilat ors, m ost n ose -dr o ps, m ost bl o o d -press ure me dicati o ns, 
ma n y dr u gs use d f or mi grai nes, a n d ma n y ot h er pr o d ucts .  
A d diti o nall y, initiati o n of treat me nt wit h or a n y c ha n ges t o t he c urre nt d os a ge, dr u g or 
re gi me n of a n y s yste mic a dre ner gic or c h oli ner gic dr u gs (see A p pe n di x 3 ) wit hi n 7 da ys 
pri or t o scree ni n g, or d uri n g t he st u d y , is pr o hibite d . H o we ver , a s u bject ca n be treate d 
wit h a s yste mic a dre n o ce pt or a nta g o nist , f or e x a m ple, as l o n g as t he partic ular a ge nt a n d 
its d ose a n d re gi me n ha d bee n c o nsiste nt f or t h e 7 da ys pri or t o Scree ni n g, a n d t here was 
n o reas o n t o belie v e t hat alterati o n w o ul d be necessar y at s o me p oi nt later d uri n g t he 
st u d y. 
If t here is a ny q uesti o n a b o ut w het her a me dic ati o n is acce pt a ble, t he Me dic al M o nit or 
s h o ul d be c o ns ulte d bef ore pr ocee di n g 
Use of all me dicati o ns s h o ul d be d oc u me nte d o n t he a p pr o pri ate eC R F. I n v esti gat ors are 
e nc o ura g e d t o c o ntact t he Me dical M o nit or f or a n y q uesti o ns re gar di n g all o we d 
me dicati o ns. J u d g me nt of c o nti n ue d st u d y partici pati o n  b y t he s u bject, a n d i ncl usi o n of 
t his s u bject’s s u bse q ue nt visits i n t he safet y a n d efficac y a nal ysis will be ma de b y 
Oc u p hire.  
All me dicati o ns w hic h t he s u bject has ta k e n wit hi n 3 0 da ys  pri or t o t he Scree ni n g Visit 
a n d d uri n g t he st u d y will be rec or de d i n t he eC R F. T he na me of t h e dr u g, d ose, r o ute of 
a d mi nistrati o n, d urati o n of treat me nt a n d i n dicati o n  will be rec or de d f or eac h me dicati o n. 
F or c o m bi nati o n pr o d u cts (e. g., C o ntac ® , C os o pt® ), t he bra n d n a me is re q uire d. F or 
n o n -c o m bi nati o n pr o d ucts, t he ge neric n a m  is desire d. T he use of r o uti ne o p ht hal mic 
dia g n ostic p har mace utical a ge nts (e. g., fl u orescei n a n d l ocal a nest hetic) will be all o we d, 
a n d s h o ul d be d oc u me nte d. A n y c ha n ge i n d osi n g para meters s h o ul d als o be rec or de d i n 
t he eC R F.  
7.  A S S E S S M E N T  O F  E F FI C A C Y  
7. 1.  S pecific ati o n  of  t he effic acy p ar a meters  
T he pri mar y effic ac y e n d p oi nt is t he p erce nta ge of s u bjects’ st u d y e yes wit h t w o dr o ps of 
treat me nt ret ur ni n g t o ≤ 0. 2 m m baseli ne p u pil dia meter at 9 0 mi n utes. 
T he st u d y e ye is defi n e d as t he ri g ht e ye ( O D). T h e n o n -st u d y e ye is defi ne d as t he left e y e 
( O S). T he st u d y a n d n o n-st u d y e ye will b ot h be e v al uate d at all assess m e nts. N ote t hat 
s u bjects ≥ 1 8 years ol d will recei ve t w o dr o ps ( d ose d
) of tr eat me nt i n t heir 
st u d y e ye a n d o ne dr o p of treat me nt i n t heir n o n-st u d y e ye. S u bjects < 1 8 years ol d will 
recei v e o ne dr o p of treat me nt i n t heir st u d y e ye a n d o ne dr o p of treat me nt i n t heir n o n-
st u d y e ye. Sec o n dar y efficac y e n d p oi nts ca n be f o u n d i n Secti o n 4. 1 .  
T he M o difie d I nte nt -t o-Treat ( mI T T ) will be use d f or t he pri mar y e n d p oi nt a nal ysis a n d t o 
a nal yze s electe d s ec o n dar y effic ac y e n d p oi nts . T he Per Pr ot oc ol (P P ) p o p ulati o n will be 
use d t o a nal yze s electe d sec o n dar y effi cac y e n d p oi nts.  S o me  of  t he efficac y  e n d p oi nts  will  
be  a nal yz e d  o verall,  b y  m y driatic  a ge nt, a n d b y  li g ht/dar k  iri des.  
7. 2.  Assessi n g,  rec or di n g, a n d  a n alyzi n g  of  effic acy p ar a meters  
P u pil dia meter will be meas ure d at Treat me nt Visit 1  a n d F oll o w -U p  Visit 2 . 
Acc o m m o dati o n will be meas ure d at Treat me nt Visit 1.  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
3 4  
0 2 /0 5 /2 1  
 
•  P u pil dia meter will be meas ure d wit h t he 
 ( m m) 
•  Acc o m m o dati o n will be meas ure d b y t he 
 
 U nc h a n ge d acc o m m o dati o n fr o m 
Baseli ne ( -1 h o ur) is defi ne d as a c h a n ge fr o m bas eli ne val ue ≥ -1, as meas ure d i n 
di o pters.  
T he p h ot o pic li g hti n g c o n diti o ns i n t he r o o m will be defi ne d as 
 
 
 
. S u bject will sit i n t he e xa m c hair at a dista n ce of  
 fr o m t he 
dista nce E T D R S 
 f or all ass ess me nts. S u bject will be all o we d t o 
accli mate t o t hese li g hti n g c o n diti o ns (
  
 t he p u pil dia meter, B C D V A 
a n d D C N V A  (s afet y meas ures) at all sc h e d ule d ti me p oi nts. 
 
 f or acc o m m o dati o n meas ure m e nts a n d  t he re mai ni n g safet y assess me nts (e. g. 
c o nj u ncti val h y pere mia, a d verse e ve nts, s u bject q uesti o n naire, etc.). T he s u bject will be i n 
t he sa me r o o m f or all ass ess me nts, a n d e ver y eff ort will be ma de t o ha ve t h e same pers o n 
perf or m t he meas ure m e nts at all ti me p oi nts.  
7. 2. 1.  Scree ni n g / D a y 1  
I n di vi d uals w h o are p ote ntial s u bjects are i de ntifi e d b y t he st u d y ce nter t o sc he d ule t he 
S cree ni n g V isit. The  Scree ni n g V isit s h o ul d occ ur t he sa me  da y as Treat me nt Visit 1, 
w here t he d os e of st u d y treat me nt is gi ve n.  
O nce s u bjects arri ve at t he st u d y ce nter, a me m ber of t he st u d y ce nter st aff will i nter vie w 
t he i n di vi d ual as t o t heir q ualificati o ns f or partici pati o n i n t he st u d y, a n d if t he s u bject 
wis hes t o c o nti n ue, t he I nf or me d C o se nt f or m is si g ne d, a n d a Scree ni n g n u m ber is 
assi g ne d . C hil dre n a ge d 1 2 -1 7 years t o pr o vi d e asse nt. Scree ni n g i ncl u des a n e x pla nati o n 
of t he st u d y, a me dical a n d o p ht hal mi  hist or y, de m o gra p hics, a re vie w of 
pri or/c o nc o mita nt me dicati o ns  a uri ne pre g n a nc y test (f or fe m ales of c hil d beari n g 
p ote ntial ), a n d H R / B P  T he sec o n d ste p i n scree ni n g i ncl u des pr oc e d ures s uc h as I O P 
assess me nt a n d an  o p ht h al mic e xa mi nati o n  t hat i ncl u des bi o micr osc o p y a n d direct or 
i n direct o p ht hal m osc o p y  wit h o ut dilati o n .  
I n v esti gat ors are ca uti o n e d t o a p pr o priat el y n ote all o bser vati o ns of c o nj u n cti val 
h y pere mia (als o calle d c o nj u ncti val er yt he ma) o n t he bi o micr osc o p y eC R F at scree ni n g.  
7. 2. 2.  Tre at me nt  Visit  1/ D a y  1  
Treat me nt Visit 1 s h o ul d be t he sa me da y as Scree ni n g. O nce t he s u bject h as c o m plete d 
t he Scree ni n g ass ess me nts part of t he visit a n d it is c o nfir me d t hat he/s he m eets all of t he 
i ncl usi o n criteria b ut n o n e of t he e x cl usi o n criteria, t he visit will t he n tra nsiti o n t o t he 
Treat me nt Visit 1 assess me nts. As part of t he Treat me nt Visit 1 , t he s u bject:   
• 
 
 
• 
 
 
 
 
 
  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
3 5  
0 2 /0 5 /2 1  
 
• 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
. 
 
. A d ult  
s u bjects (≥ 1 8  years  ol d)  will  ha ve  t w o dr o ps  ( d ose d 5  mi n utes  a part) of  treat me nt 
a d mi nistere d  i n t he St u d y  E ye  ( O D) a n d o n  dr o p  of  treat me nt a d mi nistere d  i n t he N o n -
St u d y  E ye  ( O S) o ne  h o ur  after m y driatic  dr u g  i nstillati o n. Pe diatric  s u bjects (< 1 8  years  
ol d ) will  ha ve  o ne  dr o p  o f treat me nt a d mi nistere d  i n eac h e y e o ne  h o ur  after m y driatic  
dr u g  i nstillati o n. 
 
7. 2. 3.  F oll o w -U p  Visit  / D a y  2   
O n Da y  2 , t he s u bject will ret ur n t o t he cli nic f or  a F oll o w -U p  Visit .  
As part of t he F oll o w-U p Visit , t he s u bject: 
• Will  be  ass esse d  at 2 4  h o urs  +  6  h o urs  rel ati ve t o st u d y treat me nt a d mi nistrati o n  d uri n g  
Treat me nt  Visit  1  f or t h e f oll o wi n g: c o nc o mita nt  me dicati o ns,  H R/ B R,  p u pil  dia meter , 
B C D V A , D C N V A , c o nj u ncti val  h y pere mia , A Es , a n d s u bject q uesti o n nair e . 
7. 2. 4.  U nsc he d ule d  Visits  
A n U nsc he d ule d V isit ma y be a n y visit t o t he I n v esti gat or ot her t ha n t he s pecific visits 
re q ueste d i n t he pr ot o c ol as p ossi bl y re q uire d f or t he s u bject’s o p ht hal mic c o n diti o n. T he 
I n v esti gat or will perf or m all pr oce d ures necessar y t o e val uate t he st u d y partici pa nt at t hese 
visits a n d rec or d a n y A Es  i n t he eC R F . 
As n ote d  i n Secti o n 4. 9. 4 , e ver y p ossi ble eff ort s h o ul d be ma de b y In vesti g at ors t o ass ure 
t hat s u bjects w h o disc o nti n ue earl y fr o m t he st u d y ha ve a tel e p h o ne f oll o w-u p t hat 
i ncl u des assess me nts of A Es , c o nc o mita nt me dicati o ns a n d s u bject-e v al uate d c o nj u ncti val 
h y pere mia . 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
3 6  
0 2 /0 5 /2 1  
 
7. 2. 5.  Visit  v ari ati o n  
Visits o n Da y 2  ma y occ ur bet wee n 2 4 t o 3 0 h o urs after t he b aseli ne ( 0 mi n ute ) ti me p oi nt 
o n Treat me nt D a y 1 . 
8.  A S S E S S M E N T  O F  S A F E T Y  
8. 1.  S pecific ati o n  of  s afety p ar a meters  
T he assess me nt of s afet y a n d t olera bilit y is t he sec o n dar y o bjecti ve of t his st u d y. T he 
assess me nt of safet y will be e val uat e d b y:  
•  C o nj u ncti val h y pere mia  meas ure d wit h C C L R U i ma ges usi n g a 4 -p oi nt scale  ( 0-3)  
–  A p pe n di x 5 . 
o  N o ne ( 0) = N or mal A p pears w hite wit h a s mall n u m ber of c o n ju ncti val 
bl o o d vessel  easil y o bser ve d  
o  Mil d ( + 1) = Pr o mi ne nt , pi n kis h-re d c ol or of b ot h t he b ul bar a n d p al pe bral 
c o nj u ncti va  
o  M o derate ( + 2) = Bri g ht, scarlet r e d c ol or of t h e b ul bar a n d pal pe bral 
c o nj u ncti va  
o  Se vere ( + 3) = Beef y re d wit h petec hiae, d ar k re d b ul bar a n d pal pe bral 
c o nj u ncti va wit h e vi de nce of s u bc o nj u ncti val he m orr ha ge   
•  S u bjecti ve oc ular t olera bilit y  meas ure d  o n a 4 -p oi nt scale  
o  0 –  N o disc o mf ort  
o  1 –  Mil d disc o mf ort  
o  2 –  M o derate disc o mf ort  
o  3 –  Se vere disc o mf ort  
•  B C D V A Dista nce will be meas ure d bi n oc ularl y i n p h ot o pic c o n diti o ns b y Sta n dar d 
E T D R S  
 at 
 –  A p pe n di x  6  
•  D C N V A Rea di n g/ Near will be meas ure d bi n oc ul arl y i n p h ot o pic c o n diti o ns b y Near 
Vis ual Ac uit y C hart i n t he 
 
 –  A p pe n di x  6  
•  I O P is meas ure d wit h t he T o n o-Pe n  
•  H R  a n d B P  (As per t he site’s n or mal e q ui p me nt a n d pr oce d ure s) 
•  A Es  
•  Brief S u bject  Q uesti o n naire  –  A p pe n di x 4  
8. 2.  Assessi n g,  rec or di n g, a n d  a n alyzi n g  s afety p ar a meters  
T he ti mi n g f or rec or di n g safet y para meters ma y be f o u n d i n Secti o n 4. 2 , Ta ble 2 : 
Scree ni n g a n d M y driatic/ Treat me nt Sc he d ul e . 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
3 7  
0 2 /0 5 /2 1  
 
8. 3.  A dverse  eve nts  a n d  seri o us  a dverse  eve nts  
All A Es a n d S A Es t hat occ ur f oll o wi n g c o ns e nt a n d u ntil t he fi nal st u d y visit s h o ul d be 
c ollecte d a n d rec or de d o n t he A E or S A E e C R F p a ge . O nl y treat me nt-e mer ge nt a d verse 
e ve nts/a d vers e reacti o ns ( T E A Es) will be s u m marize d (see Secti o n 9. 3. 5 ). 
All A Es /a d verse reacti o ns occ urri n g d uri n g t he st u d y (i.e. o nce t he s u bj ect has si g ne d t he 
i nf or me d c o nse nt/asse nt) m ust be d oc u me nte d, re gar dless of t he ass u m pti o n of ca usal 
relati o ns hi p, o n t he res pecti ve eC R F. All  treat me nt-e mer ge nt  A Es / a d verse reacti o ns m ust 
be d oc u me nte d fr o m t he ti me t he s u bject recei ves t he st u d y me dicati o n u ntil t he s u bject’s 
partici pati o n i n t he st u d y has bee n c o m plet e d. If a s u bject has o n g oi n g A Es /a d verse 
reacti o ns at t he ti me of st u d y c o m pleti o n  or disc o nti n uati o n fr o m t he st u d y , t he o n g oi n g 
A Es /a d verse reacti o ns m ust  be f oll o we d -u p a n d pr o vi de d a p pr o priate me di cal care u ntil 
t he si g ns a n d s y m pt o ms ha ve re mitte d or sta bilize d or u ntil me dical assess me nts  ha ve 
ret ur ne d t o a cce pta ble or pre-st u d y li mits. 
D oc u me ntati o n of A Es /a d verse reacti o ns i ncl u des start date a n d e n d date, s e verit y, 
relati o ns hi p t o st u d y me dicati o ns, acti o n(s) ta ke n, seri o us ness a n d o utc o me.  
8. 3. 1.  A d verse  e v e nt defi niti o ns  
T he f oll o wi n g defi niti o ns of ter ms a p pl y t o t his secti o n:  
A dverse eve nt . A n A E is a n y u nt o war d me dical occ urre n ce ass ociate d wit h t he use of a 
st u d y me dicati o n i n h u ma ns, w het her or n ot c o nsi dere d dr u g relate d. A n A E ca n t heref ore 
be a n y u nfa v ora ble a n d u ni nte n de d si g n (i ncl u di n g a b n or mal la b orat or y fi n di n gs), 
s y m pt o m, or disease te m p oraril y ass oci t e d wit h t he use of t he st u d y me dicati o n, w het her 
or n ot relate d t o t he st u d y me dicati o n. St u d y me dicati o n i ncl u de s t he i n vesti gati o nal dr u g 
u n der e val u ati o n a n d t he c o m parat or pr o d uct or v e hicle place b o t hat is gi ve n or 
a d mi nistere d d uri n g a n y p hase of t he st u d y.  
Me dical c o n diti o ns/ diseases prese nt bef ore st arti n g t he i n vesti gati o nal treat me nt are o nl y 
c o nsi dere d A E s if t he y w o se n after starti n g t he i n vesti gati o nal treat me nt. A b n or mal test 
res ults c o nstit ute A Es o nl y if t he y i n d uce cli nical si g ns or s y m pt o ms, are c o nsi dere d 
cli nicall y si g nifica nt, or re q uire t hera p y. 
T he occ urre n ce of A Es s h o ul d be s o u g ht b y o pe n -e nde d q uesti o ni n g of t he s u bject at eac h 
visit d uri n g t he st u d y. At eac h cli nic assess me nt /visit, st u d y pers o n nel s h o ul d as k t he 
f oll o wi n g q uesti o n: “ Ha v e y o u ha d a n y pr o ble ms si nce y o ur last assess me nt /visit ?”. A Es 
als o ma y be dete cte d w h e n t he y are v ol u nte ere d b y t he s u bject d uri n g or bet wee n visits or 
t hr o u g h st u d y assess me nts. 
Life -t hre ate ni n g a dverse eve nt or life-t hre ate ni n g s us pecte d a dverse r e acti o n. A n A E  or 
s us pecte d a d v erse reacti o n is c o nsi dere d “life-t hreate ni n g” if, i n t he vie w of eit her t he 
In v esti gat or or O c u p hire , its occ urre n ce places t he s u bject or s u bject at i m me diate ris k of 
deat h. It d oes n ot i ncl u de a n A E  or s us pecte d a d v erse reacti o n t hat, h a d it o cc urre d i n a 
m ore se vere f or m, mi g ht ha ve ca use d deat h.  
Seri o us a dverse eve nt or seri o us s us pecte d a dverse re acti o n . A n A E  or s us pecte d a d v erse 
reacti o n is c o nsi dere d ‘‘seri o us’’ if, i n t he vie w of eit her t he In v esti gat or or Oc u p hire,  it 
res ults i n a n y of t he f oll o wi n g o utc o mes:  
•  Deat h  
•  L ife t hreat e ni n g a d vers e e ve nt  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
3 8  
0 2 /0 5 /2 1  
 
•  In p atie nt  h os pitalizati o n  or  pr ol o n gati o n  of  e xisti n g  h os pitalizati o n  
•  P ersiste nt  or  si g nifica nt i nca pacit y or  s u bsta ntial disr u pti o n  of  t he a bilit y t o c o n d uct  
n or mal  life f u ncti o ns  
•  C o n g e nital  a n o mal y/ birt h  defect   
•  Ot her  me dicall y  i m p orta nt seri o us e ve nt  
I m p ort a nt me dical e v e nts t hat ma y n ot res ult i n deat h, be life-t hr eate ni n g, or re q uire 
h os pitalizati o n ma y be c o nsi dere d seri o us w he n, base d u p o n a p pr o priate me dical 
j u d g me nt, t he y ma y je o par dize t he patie nt or s u bj ect a n d ma y re q uire m e dical or s ur gical 
inter ve nti o n t o pre ve nt o ne of t he o utc o mes liste d i n t his defi niti o n.  
Treat me nt o n a n o ut p atie nt e mer ge nc y b asis t hat d oes n ot res ult i n h os pital a d missi o n, or a 
h os pitalizati o n t hat is electi ve or is a pre pl a n ne d tr eat me nt f or a pre -e xisti n g c o n diti o n  t hat 
is u nrelate d t o t he me dicati o n u n der st u d y a n d has n ot w orse ne d si nce t he start of t he 
st u d y, is n ot c o nsi dere d a n S A E. 
S us pecte d a dverse re a cti o n  mea ns a n y A E  f or w hic h t here is a reas o n a ble p ossi bilit y t hat 
t he dr u g ca us e d t he A E . F or t he p ur p os es of I N D safet y re p orti n g, “reas o n a ble p ossi bilit y” 
mea ns t here is e vi d e nce t o s u g gest a ca us al relati o ns hi p bet wee n t he dr u g a n d t he A E . 
S us pecte d a d verse reacti o n i m plies a lesser de gree of certai nt y a b o ut ca usalit y t ha n 
a d verse reacti o n, w hic h mea ns a n y A E  ca use d b y a dr u g.  
U ne x pecte d a d verse eve nt or u ne x pecte d s us pecte d a dverse r e acti o n . A n A E  or s us pecte d 
a d verse reacti o n is c o nsi dere d  “u n e x pecte d ” if i  is n ot liste d i n t he In vesti gat or s’ B r oc h ure 
or is n ot liste d at t he s pecificit y or se verit y t hat has bee n o bser ve d; or, if a n In v esti gat or s’ 
B r oc h ure is n ot re q uir e d or a vaila ble  is n ot c o nsiste nt wit h t he ris k i nf or mati o n descri be d 
i n t he ge neral i n vesti gati o nal pla n or else w h ere i n t he c urre nt a p plicati o n, as a me n de d. 
“ U ne x pecte d,” as use d i n t his defi niti o n, als o refers t o A Es  or s us pecte d a d verse reacti o ns 
t hat are me nti o ne d i n t he In v esti gat or s’ B r oc h ure as occ urri n g wit h a class of dr u gs or as 
a ntici pate d fr o m t he p h ar mac ol o gical pr o perties of t he dr u g b ut are n ot s pecificall y 
me nti o ne d as occ urri n g wit h t he partic ular dr u g u n der i n vesti gati o n.  
T he st u d y me dicati o n rel ati o ns hi p f or eac h A E /a d verse reacti o n s h o ul d be deter mi ne d b y 
t he I n v esti gat or usi n g t hese e x pla nati o ns: 
•  N ot rel at e d  
•  U nli kel y rel ate d  
•  P ossi bl y rel ate d  
•  Pr o b a bl y rel ate d  
•  Defi nitel y rel at e d  
•  U n k n o w n  
U nless t he relati o ns hi p is c o nsi dere d t o be “ N ot relate d” or “ U nli kel y relate d” a n d t here is 
a n y vali d reas o n, e ve n if u n deter mi ne d , f or s us p ecti n g a p ossi ble ca use-a n d -effect 
relati o ns hi p bet wee n t h e st u d y me dicati o n a n d t he occ urre nce of t he A E, t h e n t he A E 
s h o ul d be c o nsi dere d “ rel ate d”.  
If t he rel ati o ns hi p bet wee n t he A E/ S A E a n d t he i n vesti gati o nal pr o d uct is deter mi ne d b y 
Oc u p hire  t o be a n yt hi n g ot her t ha n “N ot relate d” or “ U nli kel y rel ate d” t he e ve nt will be 
c o nsi dere d t o be relate d t o t he i n vesti gati o nal pr o d uct f or t he p ur p os es of e x pe dite d 
re g ulat or y re p orti n g. 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
3 9  
0 2 /0 5 /2 1  
 
Severity  of a n A E  is defi ne d as a q ualitati ve assess me nt of t he le vel of disc o mf ort of a n 
A E  as is deter mi ne d b y t he I n vesti gat or or re p ort e d t o hi m/ her b y t he s u bject. T he 
assess me nt of se verit y is ma de irres p ecti ve of st u d y me dicati o n relati o ns hi p or seri o us ness 
of t he e ve nt a n d s h o ul d b e e val uat e d acc or di n g t o t he f oll o wi n g scale:  
•  1 = Mil d: prese nt, b ut n ot distressi n g, a n d n o disr u pti o n of n or mal dail y acti vit y  
•  2 = M o derat e: disc o mf ort s ufficie nt t o re d uce or affect n or m al dail y acti vit y  
•  3 = Se vere: i nca pacitati n g, wit h  i na bilit y t o w or k or perf or m n or mal dail y acti vit y 
A  c ha n ge i n s e verit y f or a re p ort e d A E  will  re q uir e an  e n d d ate  f or t h e pre vi o us  se verit y 
a n d a ne w  start a n d e n d d ate  f or t he n e w  se v erit y. F or  e x a m ple,  a c h a n ge i n se verit y ma y  
g o  fr o m mil d  t o se vere or  fr o m se vere t o m o derat e.  I n eit her case t he start or  e n d 
ti mes/dates  s h o ul d be  rec or de d.  
T he  ter m “se v ere”  is use d  t o descri be  t he i nte nsit y of  a n e ve nt/reacti o n;  t he e ve nt/reacti o n  
itself ma y  be  of  relati vel y mi n or  me dical  si g nifica nce (s uc h as a s e vere hea dac h e) . T his  is 
n ot  t he sa me as a “ Seri o us”  A d vers e  E ve nt,  w hic h  is base d  o n  a s u bject/e v e nt o utc o me  or  
acti o n criteria us u all y  ass ociate d  wit h  e ve nts  t hat p ose  a t hreat t o t he s u bject’s life or  vital  
f u ncti o ns. “ Seri o us ness ” ( N O T se v erit y) ser ves as a g ui de  fo r d efi ni n g  re g ul at or y re p orti n g 
o bli gati o ns.  
Acti o n  ta ke n i n res p o nse t o a n A E  is c o de d as:  
•  D ose  i ncrease d: A n  i n di cati o n t hat a me dicati o n  sc he d ule was  m o difie d  b y  a d diti o n; 
eit her b y  c ha n gi n g  t he fre q ue nc y, str e n gt h or  a m o u nt.  
•  D ose  n ot  c ha n g e d:  A n  i n dicati o n t hat a me dicati o n  sc he d ule was  m ai ntai ne d.  
•  D os e re d u ce d: A n  i n dicati o n t hat a me dicati o n  sc he d ule was  m o difie d  b y  s u btracti o n, 
eit her b y  c ha n gi n g  t he fre q ue nc y, str e n gt h or  a m o u nt.  
•  D ose  i nterr u pt e d: A
 i n di cati o n t hat a me dicati o n  s c he d ule was  m o difie d  b y  te m p oraril y 
ter mi nati n g a prescri be d  re gi me n o f me dicati o n.  
•  Dr u g  wit h dra w n:  A n  i n dicati o n t hat a me dicati o n  sc he d ule was  m o difie d  t hr o u g h 
ter mi nati o n of  a pres cri b e d  re gi me n of  m e dicati o n.  
•  N ot  a p plica ble:  Det er mi nati o n  of  a val ue  is n ot  rel e va nt i n t he c urre nt  c o nte xt  
•  U n k n o w n:  N ot  k n o w n,  n ot  o bser ve d,  n ot  rec or de d, or  ref use d 
A d diti o nal Ot her Acti o n Ta ke n:  
•  C o nc o mita nt  Me dicati o n  
•  H os pitalizati o n  
O utc o me  of  a n A E  is c o d e d as: 
•  Fatal : T he  ter mi nati o n of  life as a res ult of  a n a d v erse e v e nt.  
•  N ot  rec o v ere d/ n ot res ol v e d: O ne  of  t he p ossi ble  res ults of  a n a d v erse e ve nt o utc o me  
t hat i n dicates t hat t he e ve nt has  n ot  i m pr o ve d or  rec u perat e d. 
•  Rec o vere d/res ol ve d : O ne  of  t he p ossi ble  res ults of  a n a d verse e v e nt o utc o me  t hat 
i n dicates t hat t he e ve nt h as  i m pr o ve d or  rec u perat e d. 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
4 0  
0 2 /0 5 /2 1  
 
•  Rec o vere d/ Res ol ve d  wit h  se q uelae: O n e of  t h e p ossi ble  res ults of  a n a d verse e v e nt 
o utc o me  w here  t he s u bject rec u p erate d b ut  retai ne d pat h ol o gical  c o n diti o ns  res ulti n g 
fr o m t he pri or  diseas e  or  i nj ur y. 
•  Rec o veri n g/res ol vi n g:  O ne  of  t he p ossi ble  res ults of  a n a d verse e ve nt o utc o me  t hat 
i n dicates t hat t he e ve nt is i m pr o vi n g 
•  U n k n o w n:  N ot  k n o w n,  n ot  o bser ve d,  n ot  rec or de d, or  ref use d 
I n pre vi o us cli nical st u di es of N y x ol  t he m ost fre q ue ntl y re p orte d A E  was c o nj u ncti val 
h y pere mia.  
I n t he prese nt st u d y, a n y i ncre ase of t w o u nits i n c o nj u ncti v al h y pere mi a bet wee n 
c o nsec uti ve ti me p oi nts is c o nsi dere d a n A E.  
In vesti g at ors are c a uti o ne d t o use t he a p pr o pri ate ver b ati m ter m o n t he A E  f or m t o 
descri be t his o bser v ati o n:  
•  Re d ness rel ate d t o i nstill ati o n t h at is tr a nsie nt (i.
 is n o l o n ger prese nt 2 
h o urs after i nstill ati o n) = “c o nj u ncti v al er yt h
m a u p o n i nstill ati o n ”  
•  Re d ness t h at is N O T tr a nsie nt (i.e., is prese nt 2 h o urs after i nstill ati o n) = 
“c o nj u ncti v al h y pere mi a ”  
E x pe dite d  re p orti n g  of  Seri o us  a n d  U ne x pecte d  A d verse  E ve nts : A ll S A Es  (relat e d a n d 
u nrelate d)  will  be  rec or de d f oll o wi n g s u bject si g nat ure of  t he i nf or me d c o nse nt /asse nt a n d 
u ntil  t he F oll o w -U p  Visit  (Da y  2 ). A n y  S A Es  “s us pecte d” t o b e  relat e d t o t he st u d y 
me dicati o n  a n d disc o vere d  b y  t he I n vesti gat or at a n y ti me after  t he st u d y s h o ul d be  
re p orte d. 
A n y  S A E  t hat occ urs  m ust  be  re p orte d t o t he C R O  wit hi n  2 4  h o urs  of  its occ urre nce  or  
wit hi n  2 4  h o urs  of  lear ni n g of  its occ urre nce.  Rec urre nt  e pis o des,  c o m plicati o ns,  or  
pr o gressi o n  of  t he i nitial S A E  m ust  be  re p orte d t o t he C R O  as f oll o w-u p  t o t he ori gi nal  
e pis o de wit hi n  2 4  h o urs  of  t h  I n vesti gat or recei vi n g t he i nf or mati o n. I nf or mati o n a b o ut 
all S A Es  will  be  c ollecte d  a n d rec or de d o n  t he S A E  f or m. All  perti ne nt  me dical  rec or ds 
a n d i nf or mati o n c ollecte d  d uri n g  t he treat me nt a n d f oll o w-u p  of  t he s u bject s h o ul d be  
mai ntai ne d  at t he site wit h  a c o p y e maile d t o
 T he  
I n v esti gat or m ust  assess t he S A E  relati o ns hi p a n d c o m plete t he S A E  f or m. T he  C R O  ma y  
re q uest a d diti o nal  i nf or m ati o n. F oll o w -u p  i nf or mati o n (e. g., disc har ge  s u m mar y) will  be  
retai ne d i n t he s u bject’s c hart a n d a c o p y will  be  e maile d t o 
 I n a d diti o n,  all S A Es  s h o ul d be  rec or de d o n  t he A E  eC R F  
pa ge  wit h  t he seri o us q uesti o n  mar ke d  “ Yes ”.  
It is t he In v esti gat or’s  res p o nsi bilit y t o n otif y  t he a p pr o vi n g  I R B of  a n y S A Es  o n  a ti mel y 
basis  as i nstr ucte d b y  Oc u p hire  f oll o wi n g Oc u p hir e ’s deter mi nati o n  of  ca us alit y.  All  
s u bjects w h o  e x perie nce  a n S A E  s h o ul d be  f oll o w e d cli nicall y a n d u n der g o  t he a p pr o priat e 
d ia g n ostic e val uati o ns  u ntil  sta bilizati o n or  res ol uti o n of  t he e ve nt. Oc u p hire  will  re p ort all 
S A Es  t o t he U S  F o o d  a n d Dr u g  A d mi nistrati o n  ( F D A) o n  t h e a p pr o priate  sc he d ule 
de pe n di n g  if t he e v e nt  is dr u g  relate d or  n ot  dr u g  relate d, e x p ecte d, u ne x pecte d  ( bas e d o n  
t he a vaila ble i nf or m ati o n i n t he I n v esti gat ors’ Br oc h ure).  
A n y  deat h  occ urri n g  d uri n g  t he st u d y a n d f oll o w-u p  peri o d  m ust  be  re p ort e d as a n S A E.  
F or  a n y d eat h  occ urri n g  t hr o u g h t he e n d of  t he st u d y, re gar dl ess of  t he de gree  of  
relati o ns hi p t o st u d y me dicati o n , t he S A E  res ulti n g i n t he deat h  m ust  be  re p orte d t o the  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
4 1  
0 2 /0 5 /2 1  
 
C R O . A  deat h  occ urri n g  after c o m pleti o n  of  t he st u d y i ncl u di n g t he Safet y  F oll o w -u p  
Visit s, t hat is n ot  reas o na bl y ass ociate d wit h  st u d y me dicati o n  a d mi nistrati o n,  d oes  n ot  
re q uire c o m plet i o n of  t he S A E  f or m. 
8. 3. 2.  F oll o w -u p  of  s u bjects after  a d verse  e v e nts  
If a n A E /a d v erse reacti o n occ urs, t he I n vesti gat or will i nstit ute s u p p ort a n d/ or treat me nt as 
dee me d a p pr o priate. All  S A Es  o n g oi n g  at t he ti me of t he last visit, or disc o nti n uati o n fr o m 
t he st u d y, will be f oll o we d u p  u ntil t he A E /a d verse reacti o n is res ol v e d or sta bilize d, t he 
s u bject is l ost t o f oll o w-u p, or t here is ot her res ol uti o n t o t he e ve nt.  
9.  S T A TI S TI C S  
A detaile d pres e ntati o n of t he statistical a p pr oac h  is o utli ne d i n t he Statistical A nal ysis 
Pla n.  
9. 1.  S a m ple  S ize 
A  sa m ple size of  1 6 0  e val ua ble s u bjects ( 8 0 per  tr eat me nt gr o u p)  is nee de d  f or t he st u d y.  
T he pri mar y effic ac y e n d p oi nt will be met if s u bjects  s h o w a p ositi ve effect f or N y x ol ( α = 
0. 0 5  si g nifica nce, t w o-taile d). O ne h u n dre d a n d si xt y  e val ua ble s u bjects will pr o vi de at 
least 
 
 
 
 T he 
ass u m pti o ns f or t his p o wer calc ulati o n are esti mat e d fr o m t he MI R A -1 P hase 2 res ults a n d 
a dj uste d f or t he 3: 1: 1 m y driatic a ge nt gr o u ps.  
All  s u bjects will  be  ra n d o mize d i nt o t he st u d y i n a 1: 1  rati o t o o ne  of  t he t w o treat me nt 
ar ms ( N y x ol or  pla ce b o) , wit h  a 1: 1  stratificati o n b y  li g ht/ dar k iri des. F urt her m ore,  
su bjects  will  be  ra n d o mi ze d i nt o t he st u d y at a rati o of  3: 1: 1  t o o ne  of  t he t hree m y driatic  
a ge nts,  2. 5 %  p he n yle p hri ne,  1 %  tr o pica mi de, a n d Pare m y d.  T heref ore,  if 9 6  s u bjects are 
ra n d o mize d t o 2. 5 %  p h e n yle p hri n
 t he n 3 2  s u bjects will  be  assi g ne d  t o 1 %  tr o pica mi de 
a n d 3 2  s u bjects t o Pare m y d  res ulti n g i n 1 6 0  t otal s u bjects.  
It is ass u me d t hat t here will be a 5 % dr o p-o ut bef ore t he 9 0 mi n ute efficac y assess me nt. 
T o acc o u nt f or t his dr o p -o ut, a t otal of 1 6 8 s u bjects will be ra n d o mize d i nt o t he st u d y.  
9. 2.  A n alysis  P o p ul ati o ns  
M o difie d I nte nt -t o-Tre at  (mI T T ): T he mI T T  will  i ncl u de all ra n d o mize d s u bjects w h o 
recei v e d o ne or t w o dr o ps of st u d y treat me nt a n d t he n ha d at least o ne sc he d ule d p u pil 
dia meter meas ure me nt d uri n g  Visit 1. T h e mI T T  will be use d f or t he pri m ar y e n d p oi nt 
a nal ysis a n d t o a nal yz e s electe d sec o n dar y efficac y e n d p oi nts.  
Per Pr ot oc ol P o p ul ati o n ( P P):   T he p er pr ot oc ol ( P P) p o p ulati o n i ncl u de s all s u bjects i n 
t he mI T T w h o ha d  t w o dr o ps of st u d y treat me nt i n t heir St u d y E ye, ha d all sc he d ule d p u pil 
dia meter meas ure me nts d uri n g Vi sit 1, ha d a n i ncrease of > 0. 2 m m i n  P D i n t he st u d y e ye 
at Ti me 0 mi n utes  c o m pare d t o Baseli ne ( Ti me – 1 h o u r), a n d ha d n o m aj or pr ot oc ol 
de viati o ns. T he P P p o p ulati o n will be use d t o a nal yze selecte d sec o n dar y efficac y 
e n d p oi nts .  
All R a n d o mize d P o p ul ati o n ( A R P ):  T h e A R P  will i ncl u de all ra n d o mize d s u bjects. T his 
p o p ulati o n is als o k n o w n as t he i nte nt -t o-treat (I T T) p o p ulati o n . T he A R P  will be use d i n 
c o nfir mat or y effic ac y a n al yses . 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr
d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
4 2  
0 2 /0 5 /2 1  
 
S afet y P o p ul ati o n  ( S P): T he S P  will i ncl u de all ra n d o mize d s u bjects w h o ha ve recei ve d 
at least o ne dr o p  of st u d y treat me nt. T he S P  will  be use d t o s u m marize s afet y varia bles.  
9. 3.  St atistic al  Met h o ds   
9. 3. 1.  Ge ner al  co nsi der ati o ns  
All c o nti n u o us varia bles will  be s u m marize d b y tr eat me nt a n d ti me p oi nt (as a p plica ble) 
usi n g descri pti ve statistics ( n, mea n, me dia n, sta n d ar d de viati o n, mi ni m u m, a n d 
ma xi m u m). All cate g oric al varia bles will  be s u m marize d b y tr eat me nt a n d ti me p oi nt (as 
a p plica ble) usi n g fre q ue n c y c o u n ts a n d perce nt a ges.  
All st u d y data will be liste d b y treat me nt, s u bject a n d ti me p oi nt (as a p plica ble).  
All statistical tests will be perf or me d usi n g a  si g nifica nce le vel of 5 % (t w o-taile d). T he p -
val ues f or t he a nal ysis of sec o n dar y effi cac y e n d p oi nts wil l be c o nsi dere d descri pti ve. 
9. 3. 2.  De m o gr a p hic  a n d  b aseli ne  ch ar acteristi cs  
De m o gra p hic a n d B aseli ne c haracteristics s uc h as a ge, race n d se x, will b e s u m marize d 
b y treat me nt gr o u p usi n g t he mI T T , P P, S P , a n d t he A RP  T h ese data will als o be pr o vi de d 
i n b y-s u bject listi n gs. 
9. 3. 3.  S u bject  d is p ositi o n 
S u bject  dis p ositi o n, i ncl u di n g ra n d o mizati o n, a n d c o m pleti o n  a n d wit h dra wal fr o m t he 
st u d y will be s u m marize d usi n g t he A R P . T hese data will als o be pr o vi de d i n b y -s u bject 
listi n gs. 
9. 3. 4.  Me dic al  h ist or y a n d  p ri or/co c o mit a nt  m e dic ati o ns  
Me dical hist or y will be c o de d usi n g t he latest versi o n of Me dical Dicti o nar y f or 
Re g ulat or y Acti vities ( Me d D R A)  a n d will be s u m marize d b y tr eat me nt gr o u p usi n g t he 
S P .  
Pri or me dicati o ns ( me dic ati o s wit h a n e n d date pri or t o t he date of ra n d o mizati o n) a n d 
c o nc o mita nt me dicati o ns ( me dicati o ns wit h a start or e n d date after t h e date of 
ra n d o mizati o n) will be c o de d usi n g W H O Dr u g a n d will b ot h be s u m marize d b y treat me nt 
gr o u p usi n g t he S P . 
Me dical hist or y  a n d pri or a n d c o nc o mita nt me dicati o ns wil l als o be pr o vi de d i n b y -s u bject 
listi n gs.  
9. 3. 5.  A n al ysis  of  effic ac y 
Efficac y will be assesse d usi n g t he mI T T  a n d P P wit h s u bjects i ncl u de d i n t he treat me nt 
ar m i n w hic h t he y were ra n d o mize d . F or  t he a n al ysis of t he pri mar y efficac y e n d p oi nt, 
i m p utati o n will be perf or me d f or missi n g data as descri be d i n t he S tatistical A nal ysis P la n. 
If t he a n al ysis usi n g t he P P s h o ws a p ositi ve effect f or N y x ol at t he 0. 0 5 le vel of 
si g nifica nce, t he pri mar y e n d p oi nt will be c o nsi dere d met. C o nfir mat or y a n al yses ma y  be 
perf or me d usi n g t he A R P , wit h i m p utati o n perf or me d f or missi n g data. F or t he a nal ysis of 
t he sec o n dar y efficac y e n d p oi nts, o nl y o bser ve d case data will be use d.  
F or all efficac y e n d p oi nts, B aseli ne is defi ne d  as -1  h o ur  pri or t o treat me nt o n Visit 1 . Ma x 
ti me p oi nt is defi ne d as ti me 0 mi n utes, d uri n g w hic h ma xi m u m p u pil dia meter is e x pecte d.  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
4 3  
0 2 /0 5 /2 1  
 
All efficac y data will be s u m marize d b y treat me nt gr o u p, st u d y da y a n d ti me p oi nt ( -1  h o ur  
[ baseli ne], 0 mi n utes, 3 0 mi n utes, 6 0 mi n utes , 9 0 mi n utes , 2 h o ur s, 3 h o urs,  4 h o ur s, 6 
h o ur s, a n d 2 4 h o urs ), as a p pr o priate. 
T he pri mar y effic ac y e n d p oi nt is t he p erce nta ge of s u bjects  ret ur ni n g t o ≤ 0. 2 m m baseli ne 
p u pil dia meter at 9 0 mi n utes  p ost -treat me nt i n t he st u d y e ye. T he pri mar y efficac y 
e n d p oi nt will be a nal yze d usi n g a l o gistic re gressi o n m o del wit h treat me nt, m y driatic 
a ge nt, a n d li g ht/ dar k iri d es as fact ors a n d t he baseli ne  p u pil dia meter as a c o variate. T h e 
perce nt a ge of s u bjects i n eac h tr eat me nt gr o u p meeti n g t he criteria, t he o d ds rati o ( O R) 
wit h 9 5 % c o nfi de nce i nter val  (CI ) a n d p-v al ue will be pr o vi de d.  T he a n al ysis  will be 
perf or me d usi n g t he mI T T a n d P P , wit h s u bjects i ncl u de d i n t heir ra n d o mi ze d treat me nt 
re gar dl ess of t he treat me nt t he y act uall y recei ve d.  
Sec o n dar y  effic ac y  e n d p oi nts  are i n dicate d i n Secti o n 4. 1 .  
E ac h of t he c o nti n u o us sec o n dar y effi cac y e n d p oi nts  wi ll be a nal yze d usi n g a nal ysis of 
c o varia n ce ( A N C O V A)  wit h c ha n ge fr o m baseli n e as t he de p e n de nt varia ble, treat me nt , 
m y driatic a ge nt , a n d li g ht/ dar k iri des as fact ors , a n d t he res pecti ve baseli n e (-1 h o ur) val ue 
i ncl u de d as t he c o v ariate. N ote t hat m ost  sec o n dar y efficac y e n d p oi nts are i n relati o n t o 
baseli ne ( -1 h o ur), w h ereas s o me p u pil dia meter e n d p oi nt s are i n relati o n t o ma x ( 0 
mi n ute). Eac h A N C O V A will be perf or me d us i n g t he mI T T  a n d P P wit h s u bjects i ncl u de d 
i n t heir ra n d o mize d tr eat me nt re gar dl ess of t he trea me nt t he y act uall y r ecei ve d. T he 
o ut p ut fr o m eac h A N C O V A will i ncl u de t he L S M a n d S E f or b ot h treat me nt gr o u ps, al o n g 
wit h t he place b o -c orrecte d L S M, its 9 5 % CI a n d ass ociate d p -val ue.  
A c o m paris o n of t he st u d y a n d n o n -st u d y e ye f or eac h s u bject will be c o m pl ete d f or t he 
pri mar y efficac y e n d p oi nt, as well as b y m y driatic a ge nt.  
F or eac h of t h e sec o n d ar y e n d p oi nts  r late d t o perce nt of s u bjects a c hie vi n g certai n 
criteria, t he a nal ysis will be perf or me d usi n g a l o gistic re gressi o n m o del wit h treat me nt, 
m y driatic a ge nt , li g ht/ dar k iri des, a n d t he res p ecti ve baseli ne  as a c o v ariate . F or eac h 
a nal ysis, t he perce nta ge of s bjects i n eac h treat m e nt gr o u p meeti n g t he criteria, t he O R  
wit h 9 5 % CI a n d p -val ue will be pr o vi de d. F or t h ese e n d p oi nts, t he mI T T a n d P P  will be 
use d wit h s u bjects i ncl u de d i n t heir ra n d o mize d tr eat me nt re g ar d less of t he treat me nt t he y 
act uall y recei ve d.  
I n a d diti o n, eac h s ec o n dar y effic ac y e n d p oi nt will be a nal yze d b y li g ht/ dar k iri des a n d b y 
m y driatic a ge nt usi n g t he sa me m o del i n dicate d a b o ve  b ut wit h o ut iri des or m y driatic 
a ge nt as a fact or , as a p pr o priate. Eac h m y driatic a ge nt will be a nal yze d i n di vi d uall y, a n d 
a n a d diti o nal a nal ysis c o m bi ni n g 1 % tr o pica mi de a n d t he Pare m y d s u bjects i nt o a 
“tr o pica mi de” gr o u p will be perf or me d.  Ot her s u b gr o u ps, s uc h as a g e, se x, a n d race, ma y 
be a nal yz e d as w ell . If t h ere is s uffici e nt sa m ple, a nal ysis  of selecte d  efficac y  e n d p oi nts 
will be c o m plete d f or t he s u b gr o u p of  pe diatric s u bjects.   
9. 3. 6.  A n al ysis  of  safet y  
N o statistical a nal ysis of safet y data will be perf or me d i n t his st u d y. Safet y will be 
assesse d usi n g t he S P wit h s u bjects i ncl u de d i n t he treat me nt gr o u p t he y act uall y recei v e d, 
re gar dl ess of t heir ra n d o mize d treat me nt. O bs er ve d case d ata will be use d; n o i m p utati o n 
will be perf or me d f or missi n g safet y data.  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
4 4  
0 2 /0 5 /2 1  
 
F or H R a n d B P, Baseli ne is defi ne d as t he scree ni n g val ue. H R a n d B P val ues  a n d c ha n g e 
fr o m B aseli ne i n t he val u es will be s u m marize d b y treat me nt gr o u p a n d ti m e p oi nt 
(scree ni n g, 6 h o urs, a n d 2 4  h o u rs).  
O bser ve d v al ues a n d c ha n ge fr o m baseli ne ( -1 h o ur) i n c o nj u ncti val h y pere mia at eac h 
ti me p oi nt ( 0 mi n, 3 0 mi n utes , 6 0 mi n utes , 9 0 mi n utes , 2 h o urs, 3, h o urs, 4 h o urs, 6 h o urs, 
a n d 2 4 h o urs), will be s u m marize d f or t h e st u d y e ye  a n d t he n o n-st u d y e y e. Treat me nts 
will be c o m pare d usi n g t he sa me A N C O V A m o del pr o p ose d f or t h e c o nti n u o us sec o n dar y 
effic ac y e n d p oi nts . C o nj u ncti val h y pere mia will als o be s u m marize d cate g ori call y. 
Vis ual ac uit y assess me nts ( B C D V A  a n d D C N V A ) will als o be s u m marize d at eac h 
ti me p oi nt (0 mi n utes, 9 0 mi n utes , 6 h o urs, a n d 2 4 h o urs), usi n g letters a n d l o g M A R u nits. 
Letters will be rec or de d, a n d later c o n v erte d t o l o g M AR  b y pr o gra m m i n g f or s uc h 
a nal yses. O ne l etter is e q ui vale nt t o 0. 0 2 l o g M A R. As a refere nce f i ve letters is 
re pres e nte d b y 1  li ne. Treat me nts will be c o m pare d usi n g t he sa me A N C O V A m o del 
pr o p ose d f or t he c o nti n u o us sec o n dar y effi cac y e n d p oi nts.  D C N V A will als o be a nal yze d 
b y m y driatic a ge nt usi n g a m o del t hat d oes n ot i ncl u de  t he m y driatic a ge nt as a fact or.  
F or I O P, Baseli ne is d efi n e d as t he scree ni n g val u e  O bser ve d val ues a n d c ha n ge fr o m 
baseli ne i n I O P at 6 h o urs will be s u m marize d f or t he st u d y e ye  a n d t he n o n-st u d y e ye. 
Oc ular t olera bilit y val ues will be s u m marize d b y treat me nt gr o u p at ti me p oi nt 0 m i n utes.  
S u bject  Q uesti o n naire val ues will be s u m marize d b y treat me nt gr o u p ( o verall a n d b y 
m y driatic a ge nt ) a n d ti me p oi nt  (-1 h o ur, 0 mi n utes, 6 0 mi n utes , 9 0 mi n utes , 3 h o urs, 6 
h o urs, a n d 2 4 h o urs) . 
Ver bati m descri pti o ns of A Es will be c o de d usi n g  Me d D R A.  O nl y t reat me nt-e mer ge nt 
A Es  (T E A Es;  t h ose t hat occ ur after t he first d ose of st u d y me dicati o n or i ncre asi n g i n 
severity after i niti ati o n of st u dy me dic ati o n) will be s u m marize d . T E A Es a n d seri o us A Es 
( S A Es) will be s u m mariz e d b y treat me nt gr o u p, b y s yste m or ga n class ( S O C), se verit y, 
a n d relati o ns hi p t o st u d y me dicati o n. Deat hs, wit h dra wal fr o m st u d y me dicati o n d ue t o 
A Es, a n d wit h dra w al fr o m t he st u d y d ue t o A Es will eac h b y s u m marize d b y treat me nt 
g r o u p. N ote t hat i n Me d D R A, oc ular e v e nts are c o de d t o t he S O C of “s pecial se nses” . 
T h us , usi n g S O C i n t he s u m maries will pr o vi de a se parati o n of o c ular a n d n o n -oc ular A Es . 
All safet y data will be pr o vi de d i n b y -s u bject listi n gs. Safet y listi n gs f or pe diatric s u bjects 
ma y be  pr o vi de d.  
9. 4.  Pr oce d ure  f or acc o u nti n g  f or missi n g,  u n use d,  or  s p uri o us d at a  
F or t he s u m mariz ati o n a n d a nal ysis of t he pri mar y efficac y e n d p oi nt , i m p utati o n will be 
perf or me d f or missi n g data as descri b e d i n t he Statistical A nal ysis Pla n . F or t he 
s u m marizati o n a n d a n al ysis of sec o n dar y effi cac y e n dp oi nts a n d safet y dat a, o bser ve d case 
data o nl y will be use d . 
9. 5.  Pr oce d ure  f or re p orti n g devi ati o ns  fr o m t he st atistic al pl a n  
A n y de viati o ns fr o m t he S tatistical A nal ysis P la n will be descri be d a n d a j ustificati o n 
gi ve n i n t he fi nal Cli nical St u d y Re p ort ( C S R).  
1 0.  DI R E C T  A C C E S S  T O  S O U R C E  D A T A  A N D  D O C U M E N T S  
T he I n vesti ga t or will per mit st u d y-relate d m o nit ori n g visits, a u dits, I R B re vie w, a n d 
re g ulat or y i ns pecti o n(s) b y pr o vi di n g direct access t o s o urce data a n d d oc u me nts. 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
4 5  
0 2 /0 5 /2 1  
 
1 1.   Q U A LI T Y  C O N T R O L  A N D  Q U A LI T Y  A S S U R A N C E  
T he pr o gress of t he st u d y will be m o nit ore d b y o n -site, writte n, a n d tele p h o ne 
c o m m u nicati o ns bet wee n pers o n nel at t he I n v esti gat or’s site a n d t he Me di cal M o nit or . 
S h o ul d t he C O VI D -1 9 p a n de mic restri ct m o nit ors fr o m tra veli n g t o a site, re m ote re vie w 
will be c o n d ucte d t o t he e xte nt p ossi ble, w hile still e ns uri n g t he st u d y is m o nit ore d 
a p pr o priatel y p er a p plica ble re g ulati o ns a n d g ui deli nes . T he I n vesti gat or will all o w 
Oc u p hire, t he St u d y M o nit or, a n d t he Me dical M o nit or t o i ns pect all eC R Fs, s u bject 
rec or ds (s o urce d oc u me nts), si g ne d c o nse nt /asse nt f or ms, rec or ds of st u d y me dicati o n 
recei pt, st ora g e, pre parati o n, a n d dis p ositi o n, a n d re g ulat or y files relate d t o t his st u d y.  
1 2.   E T HI C A L  C O N SI D E R A TI O N S  A N D  G C P  C O M P LI A N C E   
1 2. 1.  G C P  c o m pli a nce  
T he pr o p ose d st u d y is s u bject t o all a p plica ble g o ver n me ntal r ules a n d re g ulati o ns 
c o ncer ni n g t he c o n d uct of cli nical trials o n h u ma n s u bjects. T his i ncl u des, b ut is n ot 
necessaril y li mite d t o , t he a p pr o val of I nstit uti o nal Re vie w B oar ds, t he Helsi n ki 
Decl arati o n, U. S. F D A L a w, I C H G C P g ui deli nes, o btai ni n g pr os pecti ve i nf or me d 
c o nse nt, m o nit ori n g of t he c o n d uct of t he st u d y a n d t he c o m plete ness of t he eC R Fs  b y 
Oc u p hire or its desi g nee(s), a n d a p pr o priate rec or d rete nti o n b y t he I n vesti gat or.  
1 2. 2.  I nstit uti o n al Revie w  B o ar d  (I R B) 
T his pr ot oc ol, materials use d t o recr uit s u bjects, a n d materials use d t o d oc u me nt 
c o nse nt /asse nt m ust be a p pr o ve d b y t he I R B pri or t o i nitiati o n of t he st u d y. Writte n I R B 
a p pr o val m ust a de q u atel y i de ntif y t he pr ot oc ol a n d i nf or me d c o nse nt /asse nt. I n a d diti o n t o 
a p pr o vi n g t he pr ot oc ol, t he I R B m ust als o a p pr o ve t he S u bject  I nf or m ati o n a n d 
C o nse nt /Asse nt /Pare ntal C o nse nt  F or m, as well as a n y a d vertisi n g t o ols t hat will be use d 
f or t he st u d y. C o pies of all a p pr o v d m teri als, all c orres p o n d e nce wit h t he I R B a n d 
writte n a p pr o val fr o m t he I R B m ust be ma de a vaila ble t o Oc u p hire, pri or  t o t he start of 
s u bject e nr oll me nt i nt o t he st u d y.  
1 2. 3.  Pr ot oc ol  Devi ati o ns/ Vi ol ati o ns  
T he I n vesti gat or s h o ul d n ot de viate fr o m t he re q uire me nts of t his pr ot oc ol wit h o ut pri or 
writte n a p pr o val of t h e Me dical M o nit or or S p o ns or e xce pt i n t he e ve nt of a me dical 
e mer ge n c y.  
A re p orta bl e  pr ot oc ol de viati o n is defi ne d as n o na d here nce t o t he pr ot oc ol t hat i n v ol ves 
i ncl usi o n/e xcl usi o n criteria, affects s u bject safet y, ri g hts or welfare, or has t he p ote ntial t o 
affect t he i nte grit y of t he data. E xa m ples of maj or pr ot oc ol de viati o ns  i ncl u de stu d y 
e nr oll me nt  b y i neli gi ble s u bject , loss of ke y data s uc h as e q ui p me nt  malf u n cti o n  (e. g., 
p u pill o meter), a n d/ or use  of a pr o hi bite d me dicati o n d uri n g t he st u d y .  
All pr ot oc ol de viati o ns will be re p orte d b y e nt eri n g t he e ve nt i n t he a p pr o priate e C R F 
pa ge. Pr ot oc ol de viati o ns s h o ul d be re p orte d t o t he I R B i n acc or da n ce wit h I R B -s pecific 
g ui deli nes. If t here is a n y q uesti o n as t o w het her t he de viati o n is re p orta bl e, Oc u p hire or 
desi g nee a n d t he I R B s h o ul d be c o ntacte d.  
All c ha n ges t o t he pr ot oc ol will b e ma de b y t he S p o ns or or desi g nee as a n a p pr o ve d 
a me n d me nt t o t he pr ot oc ol, s u b mitte d t o t he F D A, a n d a p pr o ve d b y t he I R B pri or t o 
i m ple me ntati o n.  
C ha n ges i m ple me nte d wit h o ut pri or a p pr o val will be c o nsi dere d Pr ot oc ol Vi olati o ns.  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
4 6  
0 2 /0 5 /2 1  
 
1 2. 4.   I nf or me d c o nse nt  a n d  ass e nt re q uire me nts 
Writte n i nf or me d c o ns e nt will be o btai ne d fr o m eac h a d ult s u bject. A si g n e d asse nt f or m 
will be o btai ne d f or all mi n ors a ges 1 2 -1 7, as well as a se parate pare nt al/ Le gal G uar dia n 
c o nse nt. A c o p y of t h e si g ne d a n d date d c o nse nt /asse nt d oc u me nt will be gi ve n t o eac h 
s u bject or pare nt g uar dia n . T he ori gi nal si g ne d a n d date d i nf or me d c o nse nt/asse nt 
d oc u me nt m ust be mai ntai ne d i n t he st u d y files at t he I n v esti gat or’s site.   
T he In vesti gat or is r es p o nsi ble f or e ns uri n g t hat n o s u bject is s u bject to a n y st u d y -relate d 
e xa mi nati o n or acti vit y b ef ore t h at s u bject has gi v e n i nf or me d c o ns e nt /asse nt. T he s u bject 
m ust gi ve writte n c o nse nt /asse nt after t he recei pt of detaile d i nf or mati o n. T he ver bal 
e x pla nati o n will c o ver all t he ele me nts s pecifi e d i n t he w ritte n i nf or mati o n pr o vi de d f or 
t he s u bject. 
It s h o ul d be e m p hasize d t hat t he s u bject is at li bert y t o wit h dra w c o nse nt/asse nt t o 
partici pate at a n y ti me, wit h o ut pe nalt y or l oss of be nefits t o w hic h t he s u bject is ot her wise 
e ntitle d. S u bject s w h o ref use t o gi ve, or wit h dra w, writte n i nf or me d c o ns e nt/asse nt ma y 
n ot be i ncl u de d or c o nti n ue d i n t his st u d y, b ut t his will n ot i m pact o n t heir s u bse q ue nt 
care.  
T he In vesti gat or will i nf or m t he s u bject of t he ai ms, met h o ds, a ntici pate d be nefits a n d 
p ote ntial hazar ds of t he st u d y, i ncl u di n g a n y disc o mf ort it ma y e ntail.  T he s u bject m ust be 
gi ve n e ver y o p p ort u nit y t o clarif y a n y p oi nts he/ s h e d oes n ot u n dersta n d a n d if necessar y, 
as k f or m ore i nf or mati o n. At t he e n d of t he i nte vi e w, t he s u bject ma y be gi ve n ti me t o 
reflect if t his is re q uire d, or if t he s u bject re q uests m ore ti me. S u bject s a n d/ or le gal 
g uar dia n will be re q uire d t o si g n a n d  dat  t he i nf or me d c o nse nt f or m.  
A c o p y of t he si g ne d a n d date d c o nse nt /asse nt d o c u me nt will be gi ve n t o eac h s u bject. T he 
ori gi nal si g ne d a n d date d i nf or me d c o nse nt /asse nt d oc u me nt m ust be mai ntai ne d i n t he 
st u d y files at t he I n vesti gat or’s site. Si g ne d i nf or m e d c o nse nt/asse nt m ust b e o b tai ne d pri or 
t o t he c o n d ucta nce of a n y st u d y pr oce d ures. 
1 3.   D A T A  H A N D LI N G  A N D  R E C O R D  K E E PI N G  
All pr oce d ures f or t he h a n dli n g a n d a nal ysis of data will be c o n d ucte d usi n g g o o d 
c o m p uti n g practices me eti n g I C H a n d U. S. F D A g ui d eli nes f or t he ha n dli n g a n d a nal ysis 
of data f or cli nical trials.  
1 3. 1.   D at a  entry  
St u d y s pecific data t hat h as bee n o utli ne d i n t he pr ot oc ol will be e ntere d i nt o t he cli nical 
data base b y i n di vi d ual(s) desi g nate d b y t he I n v esti gat or.  
1 3. 2.  D at a  q u ality  c o ntr ol  a n d  re p orti n g 
Data is verifie d electr o nicall y usi n g a series of pr o gra m me d e dit c hec ks t h at ha ve bee n 
creat e d b y t he Cli nical D ata Ma na g er a n d pr o gra m me d b y t he Cli nical Data Pr o gra m mer 
or Desi g nee. Data discre pa ncies will be br o u g ht t o t he atte nti o n of t he cli nical tea m a n d 
i n vesti gate d b y t he C R A a n d site staff. C R As will re vie w a n d v erif y all data c ollecte d i n 
t he eC R F a gai nst s o urce d oc u me ntati o n d uri n g sc he d ule d m o nit ori n g visits. T he C R A will 
w or k cl osel y wit h t he site staff t o a d dress a n y dis cre pa n cies w hic h ha ve b ee n f o u n d s o t hat 
pr o per res ol uti o ns ca n be ma de a n d d oc u m e nte d i n t he cli nical data bas e. A n a u dit trail 
wit hi n t he s yste m will trac k all c ha n g es ma de t o t h e data.  
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
4 7  
0 2 /0 5 /2 1  
 
1 3. 3.   Arc hivi n g  of  d at a  
Arc hi ve d versi o ns of t he data base will be s a ve d b y  Oc u p hire c o nsiste nt wit h I C H G o o d 
Cli nical Practices ( G C P) G ui deli nes, c o m pl yi n g wit h w hic he ver of t h e re q uire me nts is 
l o n ger. Oc u p hir e will n otif y t he In vesti gat or w h e n d oc u me nts s h o ul d be ret ur ne d.  
1 3. 4.   Rec or ds  rete nti o n 
T he I n vesti gat or’s site a n d cli nical la b orat or y will retai n all rec or ds relate d t o t he st u d y i n 
c o m plia nce wit h I C H G C P  G ui deli nes.  
1 3. 5.   A me n d me nts  t o t he pr ot oc ol  
M o dificati o ns of t he si g ne d pr ot oc ol are o nl y p ossi ble b y a p pr o ve d pr ot o c ol a me n d me nts 
a n d wit h t he a gree me nt of all res p o nsi ble pers o ns. T he pr oce d ure f or a p pr o val of a 
pr ot oc ol a me n d me nt is i de ntical t o t hat f or a p pr o val of t he pr ot oc ol. T he I R B m ust be 
i nf or me d of all pr ot oc ol a me n d me nts a n d s h o ul d be as ke d f or its o pi ni o n as t o w het her a 
f ull re-e v al uati o n of t he et hical as pects of t he st u d y is ne essar y b y t h e c o m mittee. T his 
s h o ul d be f ull y d oc u me nte d. 
T he In vesti gat or m ust n ot i m ple me nt a n y de viati o n fr o m or c ha n ge t o t he pr ot oc ol, wit h o ut 
disc ussi o n wit h, a n d a gree me nt b y Oc u p hire a n d pri or re vie w a n d d oc u me nte d 
a p pr o val/fa v ora ble o pi ni o n of t he a me n d me nt fr o m t he rele va nt et hics c o m mittee, e xce pt 
w here it is necessar y t o eli mi nate a n i m me diate hazar d t o st u d y s u bjects, or w here t he 
c ha n ge(s) i n v ol ves o nl y l o gistical or a d mi nistrati ve as pects o f t h e st u d y (e. g., c ha n ge i n 
m o nit or, c ha n ge of tel e p h o ne n u m ber ). 
Pr ot oc ol a me n d me nts will be s u b mitte d t o t he a p pr o priate a ut h orit y(ies) as re q uire d b y t he 
a p plica ble re g ulat or y re q uire me nt(s).  
 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
4 8  
0 2 /0 5 /2 1  
 
A P P E N DI X  1 : I RI S C O L O R  C H A R T  
 
St u d y e nr ollme nt i ncl u des b ot h li g ht a n d dar k -c ol ore d e yes . E x a m ples  of li g ht iri des ( 
 
) a n d dar k  iri des (
 f or t he p ur p oses of t his st u d y are det aile d i n t he 
c hart bel o w .  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
p
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
4 9  
0 2 /0 5 /2 1  
 
A P P E N DI X  2 : R A N D O MI Z A TI O N  S C H E M A  F O R  E V A L U A B L E  S U B J E C T S  B Y  
I N V E S TI G A TI O N A L T R E A T M E N T,  M Y D RI A TI C  A G E N T , A N D  I RI S C O L O R   
 
 
 
 
 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2


Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
5 0  
0 2 /0 5 /2 1  
 
A P P E N DI X  3 : A D R E N E R GI C  A N D  C H O LI N E R GI C  D R U G S  
T he f oll o wi n g dr u gs are e xa m ples of dr u gs w hic h ca n n ot be use d wit hi n 7  da ys pri or t o Scree ni n g, or 
d uri n g t he st u d y u nless  t he dr u g, d ose a n d re gi me n has bee n c o nsiste nt f or t he 7  da ys pri or t o 
Scree ni n g. T his list is n ot i ncl usive of all dr u gs i n t hese cl asses. If t here is a ny d o u bt, ple ase 
c o ns ult wit h t he Me dic al M o nit or. 
Al p h a -1 -a g o nists  
Met h yl 
n ore pi ne p hri ne  
Na p haz oli ne  
O x y metaz oli ne  
Tetra h y dr oz oli ne  
P he n yle p hri ne  
X yl o metaz oli ne  
 
Al p h a -2 -a g o nists  
Bri m o ni di ne  
Cl o ni di ne  
G ua nfaci ne  
G ua na be n z  
G ua n o xa be n z  
G ua net hi di ne  
X ylazi ne  
Tiza ni di ne  
Met h yl d o pa  
N o n -selective al p h a-
a nt a g o nists  
P he n o x y be nza mi ne  
T olaz oli ne  
La betal ol  
Car ve dil ol  
 
Al p h a -1 -a nt a g o nists   
Alf uz osi n  
Praz osi n  
D o xaz osi n  
Ta ms ul osi n  
Teraz osi n  
 
Al p h a -2 -a nt a g o nists  
Ati pa mez ole  
I d az o xa n 
Y o hi m bi ne   
 
 
Acetylc h oli ne 
rece pt or a g o nists  
Pil ocar pi ne  ( M3 
rece pt ors) 
 
Acetylc h oli ne 
rece pt or 
a nt a g o nists  
Sc o p ola mi ne  
Dic ycl o veri ne  
T olter o di ne  
O x y b ut y ni n  
I pratr o pi u m 
Ma m ba T o xi n 
( M T7) 
Pire nze pi ne  
Tele nze pi n e  
 
A ntiverti g o  
meclizi ne  
Sc o p ola mi ne  
G astr oi ntesti n al  
atr o pi ne  
bella d o n na  
 
P ar ki ns o nis m  
a ma nta di ne  
be nztr o pi ne  
bi peri de n  
tri he x y p he ni d yl 
 
 
 
 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2




Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
5 2  
0 2 /0 5 /2 1  
 
A P P E N DI X  5 : C O N J U N C TI V A L  H Y P E R E MI A  G R A DI N G  S C A L E  U SI N G  I M A G E S 
F R O M  C C L R U  
 
 
 
 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2




Oc u p hire  P har ma,  I nc. 
 
 
 
 
   Cli nical  Pr ot oc ol  
O PI -N Y X R M -3 0 1  
 
 
 
 
 
  MI R A -2  
C O N FI D E N TI A L  
5 4  
0 2 /0 5 /2 1  
 
Bi bli o gr a p h y  
1.  Mc D o nal d, M. P he nt ola mi ne Mes ylate Treat me nt of Se vere Ni g ht Visi o n C o m plai nts,.  
2.  H olla da y, J., Me yer, A. & Pitlic k, B. P he nt ola mi ne Mes ylate O p ht hal mic S ol uti o n 1 % O nce 
Dail y Re d uces P u pil Dia meter a n d I m pr o v es Ni g ht Visi o n Dist ur ba nces.  
3.  Stei n ha uer, S. R., Sie gle, G. J., C o n dra y, R. & Pless, M. S y m pat hetic a n d paras y m pat hetic 
i n ner vati o n of p u pillar y dilati o n d uri n g s ustai ne d pr ocessi n g. I nter n ati o n al j o ur n al of 
psyc h o p hysi ol o gy : offici al j o ur n al of t he I nter n ati o n al Or g a niz ati o n of Psyc h o p hysi ol o gy  5 2 , 
7 7 – 8 6 ( 2 0 0 4).  
4.  Y os hit o mi, T., It o, Y. & I n o mat a, H. A dre ner gic e xcitat or y a n d c h oli ner gic i n hi bit or y 
i n ner vati o ns i n t he h u ma n iris dilat or. Ex peri me nt al eye res e arc h  4 0 , 4 5 3– 9 ( 1 9 8 5).  
5.  Y u, Y. & K oss, M. C. al p ha( 1 A) -a dre n oce pt ors me diate s y m pat heticall y e v o ke d p u pillar y 
dilati o n i n rats. T he J o ur n al of p h ar m ac ol o gy a n d ex peri me nt al t her a p e utics  3 0 0 , 5 2 1– 5 
( 2 0 0 2). 
6.  H o ga n, T. S., Mc Da niel, D. D  Bartlett, J. D., Hart, K. K. & Pa g giari n o, D. A. D ose -Res p o nse 
St u d y of Da pi praz ole H Cl i n t he Re versal of M y driasis I n d u ce d b y 2. 5 % P h e n yle p hri ne. 
J o ur n al of Oc ul ar P h ar m ac ol o gy a n d T her a pe utics  1 3 , 2 9 7– 3 0 2 ( 1 9 9 7).  
 
O PI- N Y X R M- 3 0 1 _ Pr ot oc ol A m d 1 _ 0 5 Fe b 2 0 2 1 | V V- T M F- 1 3 1 2 8 | 1. 0
A p pr o ve d
D o w nl oa de d b y Es meral da Car d osa o n 1/ 3 1/ 2 0 2 2

SAP_001.pdf: 
  
 
CONFIDENTIAL 
 
 
 
 
Ocuphire Pharma, Inc. 
 
STATISTICAL ANALYSIS PLAN 
 
 
 
Protocol Title: 
Randomized, Parallel Arm, Double-Masked, Placebo-Controlled 
Study of the Safety and Efficacy of Nyxol (0.75% Phentolamine 
Ophthalmic Solution) to Reverse Pharmacologically-Induced 
Mydriasis in Healthy Subjects 
Study Number: 
OPI-NYXRM-301 (MIRA-2) 
Phase: 
Phase 3 
Sponsor: 
Ocuphire Pharma, Inc. 
37000 Grand River Avenue, Suite 120 
Farmington Hills, MI 48335 USA 
 
Author: 
Summit Analytical, LLC 
8354 Northfield Blvd. 
Bldg. G Suite 3700 
Denver, CO 80238 
SAP Date: 
2021-02-09 
Status: 
FINAL V1.0 
 
 
 


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1. 
TABLE OF CONTENTS 
1. 
TABLE OF CONTENTS ............................................................................................................................... 2 
2. 
LIST OF ABBREVIATIONS AND DEFINITION OF TERMS................................................................... 5 
3. 
INTRODUCTION ......................................................................................................................................... 7 
3.1. 
Preface .............................................................................................................................................. 7 
3.2. 
Purpose of Analyses .......................................................................................................................... 7 
3.3. 
Summary of Statistical Analysis Changes to the Protocol ................................................................ 7 
4. 
STUDY OBJECTIVES AND ENDPOINTS ................................................................................................. 8 
4.1. 
Study Objectives ............................................................................................................................... 8 
4.2. 
Study Endpoints ................................................................................................................................ 8 
4.2.1. 
Primary Endpoints ........................................................................................................... 8 
4.2.2. 
Secondary Endpoints ....................................................................................................... 8 
5. 
STUDY METHODS .................................................................................................................................... 10 
5.1. 
General Study Design and Plan....................................................................................................... 10 
5.2. 
Inclusion – Exclusion Criteria and General Study Population ........................................................ 13 
5.3. 
Randomization and Blinding........................................................................................................... 13 
5.4. 
Analysis Variables .......................................................................................................................... 14 
6. 
SAMPLE SIZE ............................................................................................................................................ 16 
7. 
GENERAL CONSIDERATIONS ............................................................................................................... 17 
7.1. 
Analysis Populations ....................................................................................................................... 17 
7.1.1. 
Modified Intention-to-Treat (mITT) ............................................................................. 17 
7.1.2. 
Per Protocol Population (PP) ......................................................................................... 17 
7.1.3. 
All Randomized Population (ARP) ............................................................................... 17 
7.1.4. 
Safety Population (SP) .................................................................................................. 17 
7.2. 
Covariates and Subgroups ............................................................................................................... 17 
7.2.1. 
Planned Covariates ........................................................................................................ 17 
7.2.2. 
Planned Subgroups ........................................................................................................ 17 
7.3. 
Management of Analysis Data ........................................................................................................ 18 
7.3.1. 
Data Handling ............................................................................................................... 18 
7.3.2. 
Missing Data ................................................................................................................. 18 
7.3.2.1. 
Handling of Missing Date Values ............................................................ 18 
7.3.2.2. 
Missing Baseline Data ............................................................................. 19 
7.3.2.3. 
Imputation Methods ................................................................................. 19 
7.3.3. 
Handling of Early Termination Visit Information ......................................................... 20 
7.3.4. 
Pooling of Investigational Sites ..................................................................................... 21 
7.3.5. 
Coding Conventions for Events and Medications ......................................................... 21 
7.3.6. 
Analysis Software ......................................................................................................... 21 
7.3.7. 
Study Data ..................................................................................................................... 21 
7.4. 
Planned Study Analyses .................................................................................................................. 22 
7.4.1. 
Statistical Summaries: Descriptive and Inferential ....................................................... 22 
7.4.2. 
Interim Analyses and Data Monitoring ......................................................................... 23 
7.4.3. 
Final Analysis and Publication of Study Results .......................................................... 23 
7.5. 
Multiple Testing Procedures ........................................................................................................... 23 
7.6. 
Baseline Values ............................................................................................................................... 23 
8. 
SUMMARY OF STUDY DATA ................................................................................................................ 24 
8.1. 
Subject Disposition ......................................................................................................................... 24 


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8.2. 
Protocol Deviations ......................................................................................................................... 24 
8.3. 
Demographics and Baseline Characteristics ................................................................................... 24 
8.4. 
Medical History .............................................................................................................................. 25 
8.5. 
Prior and Concurrent Medications .................................................................................................. 25 
8.6. 
Treatment Administration ............................................................................................................... 26 
9. 
EFFICACY ANALYSES ............................................................................................................................ 27 
9.1. 
Clinical Efficacy ............................................................................................................................. 27 
10. SAFETY ANALYSES................................................................................................................................. 30 
10.1. 
Adverse Events ............................................................................................................................... 30 
10.2. 
Deaths, Serious Adverse Events and Other Significant Adverse Events ........................................ 31 
10.2.1. 
Deaths............................................................................................................................ 31 
10.2.2. 
Serious Adverse Events ................................................................................................. 31 
10.2.3. 
Adverse Events Leading to Withdrawal from the Study ............................................... 31 
10.2.4. 
Adverse Events Leading to Discontinuation of Study Medication ............................... 31 
10.3. 
Conjunctival Hyperemia ................................................................................................................. 31 
10.4. 
Subjective Ocular Tolerability ........................................................................................................ 31 
10.5. 
Visual Acuity .................................................................................................................................. 32 
10.6. 
Vital Signs ....................................................................................................................................... 32 
10.7. 
IOP .................................................................................................................................................. 32 
10.8. 
Subject Questionnaire ..................................................................................................................... 32 
10.9. 
Other Safety Measures .................................................................................................................... 32 
11. REFERENCES ........................................................................................................................................... 33 
12. APPENDICES ............................................................................................................................................ 34 
12.1. 
List of Planned Tables ..................................................................................................................... 34 
12.2. 
List of Planned Listings .................................................................................................................. 37 
12.3. 
List of Planned Figures ................................................................................................................... 38 
 
 


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LIST OF TABLES 
 
Table 1 
Screening and Mydriatic/Treatment Schedule .............................................................. 11 
 
 
LIST OF FIGURES 
 
Figure 1 
Randomization schema for evaluable subjects by investigational 
treatment, mydriatic agent, and irides type ................................................................... 14 
Figure 2 
SDTM, ADaM, and TFL Development and Validation ................................................ 22 
 
 
 


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2. 
LIST OF ABBREVIATIONS AND DEFINITION OF TERMS 
Only abbreviations and terms relevant to the SAP are repeated herein. The reader is referred 
to the protocol for the complete and comprehensive list of abbreviations and definitions of 
terms for the study. 
Abbreviation/Term 
Definition 
ADaM
Analysis Data Model
AE
adverse event
ANCOVA
analysis of covariance
ARP
All Randomized Population
ATC
Anatomical Therapeutic Chemical
BCDVA
best-corrected distance visual acuity 
BP
blood pressure
CCLRU
Cornea and Contact Lens Research Unit
CDISC
Clinical Data Interchange Standards Consortium
CI
confidence interval
CRF
Case Report Form
CSR
Clinical Study Report
DD
Drug Dictionary
DCNVA
distance-corrected near visual acuity
EDC
electronic data capture
HR
heart rate
IOP
Intraocular pressure
ITT
Intention-To-Treat
LOCF
last observation carried forward
logMAR
logarithm of the minimum angle of resolution
LSM
least squares mean
MAR
missing at random
max
maximum
MCMC
Markov Chain Monte Carlo
MedDRA
Medical Dictionary for Regulatory Activities
mITT
Modified Intention-To-Treat
Nyxol
Phentolamine Mesylate Ophthalmic Solution 1% (Nyxol®)
OD
right eye
OR
odds ratio
OS
left eye


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Abbreviation/Term 
Definition 
PD
pupil diameter
PP
Per Protocol
PT
preferred term
SAE
serious averse event
SAP
Statistical Analysis Plan
SDTM
Study Data Tabulation Model
SE
standard error
SOC
system organ class
SP
Safety Population
TEAE
treatment-emergent adverse event
TFL
tables, figures, and listings
VA
visual acuity
WHO
World Health Organization


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3. 
INTRODUCTION 
3.1. 
Preface 
This document presents a statistical analysis plan (SAP) for Ocuphire Pharma, Inc. Protocol 
OPI-NYXRM-301 (MIRA-2) (Randomized, parallel arm, double-masked, placebo-
controlled study of the safety and efficacy of Nyxol (0.75% Phentolamine Ophthalmic 
Solution) to reverse pharmacologically-induced mydriasis in healthy subjects). 
Reference materials for this statistical plan include the protocol OPI-NYXRM-
301(20AUG2020) and as amended version 1 05FEB2021 and Case Report Forms (CRFs; 
Version 07NOV2020).  
The SAP described hereafter is an a priori plan. The SAP will be finalized and approved 
prior to unmasking of any study data.     
For the reasons stated here, the conduct of the study in the field is considered to be 
independent of any study outcome that might materialize upon enactment of the currently 
proposed statistical plan.  
3.2. 
Purpose of Analyses 
The MIRA-2 study is a randomized, parallel arm, double-masked, placebo-controlled study 
of the safety and efficacy of Nyxol (0.75% Phentolamine Ophthalmic Solution) to reverse 
pharmacologically-induced mydriasis in healthy subjects. 
The Sponsor intends to use this first Phase 3 registration study to evaluate Nyxol for the 
indication “the treatment of pharmacologically-induced mydriasis produced by adrenergic 
(phenylephrine) or parasympatholytic (tropicamide) agents, or a combination thereof.” 
Post-hoc exploratory analyses not identified in this SAP may be performed to further 
examine the study data. These analyses will be clearly identified, where appropriate, in the 
final CSR. Additional analyses not prospectively identified in this SAP may also be 
completed for publications, or regulatory or funding inquiries. These analyses, if performed, 
may not be reported in the CSR but will be fully detailed in the document containing the 
additional analyses. 
3.3. 
Summary of Statistical Analysis Changes to the Protocol 
The analyses described in this analysis plan are consistent with the analyses described in the 
study protocol. 


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4. 
STUDY OBJECTIVES AND ENDPOINTS 
Study objectives and endpoints defined in the protocol include safety and efficacy endpoints. 
Objectives and pre-specified endpoints are as follows: 
4.1. 
Study Objectives 
The objectives of this study are as follows: 
 To evaluate the efficacy of Nyxol to expedite the reversal of pharmacologically-
induced mydriasis across multiple mydriatic agents with an emphasis on 
phenylephrine 
 To evaluate the efficacy of Nyxol to return subjects to baseline accommodation after 
worsening (with cycloplegic agents tropicamide and Paremyd) 
 To evaluate the safety of Nyxol 
 To evaluate any additional benefits of the reversal of pharmacologically-induced 
mydriasis  
4.2. 
Study Endpoints 
4.2.1. 
Primary Endpoints 
The primary efficacy endpoint is the percentage of subjects’ study eyes returning to ≤ 0.2 
mm from baseline pupil diameter at 90 minutes. 
The primary safety measures are: 
 Conjunctival hyperemia 
 Subjective ocular tolerability 
 Adverse events (AEs) 
4.2.2. 
Secondary Endpoints 
Secondary endpoints for efficacy and safety assessments include the following: 
Efficacy: 
Secondary efficacy endpoints will be analyzed by study eye and non-study eye unless 
otherwise indicated, and will include: 
 Percentage of subjects returning to ≤ 0.2 mm from baseline pupil diameter at each 
remaining timepoint (30 minutes, 60 minutes, 2 hours, 3 hours, 4 hours, 6 hours, 24 
hours) 


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 Change (in mm) from max pupil diameter (0 minutes) at each timepoint (30 minutes, 
60 minutes, 90 minutes, 2 hours, 3 hours, 4 hours, 6 hours, 24 hours) 
 Percentage of subjects with unchanged accommodation from baseline (-1 hour) at 
the following timepoints (0 min, 60 minutes, 90 minutes, 2 hours, 3 hours, 4 hours, 6 
hours)  
 Change (in diopters) from baseline accommodation (-1 hour) at each time point (0 
minutes, 60 minutes, 90 minutes, 2 hours, 3 hours, 4 hours, 6 hours) 
 Time (hours) to return to ≤ 0.2 mm from baseline pupil diameter (time savings 
analysis)  
Safety and Tolerability: 
 Best-corrected distance visual acuity (BCDVA) 
 Distance corrected near visual acuity (DCNVA) 
 Vital signs (heart rate [HR] and blood pressure [BP]) 
 Intraocular pressure (IOP) 
 Subject questionnaire of symptoms 
 Urine pregnancy tests for females of childbearing potential 
 
 


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5. 
STUDY METHODS 
5.1. 
General Study Design and Plan 
A randomized, parallel arm, double-masked, placebo-controlled Phase 3 study in at least 168 
randomized subjects (160 evaluable for efficacy, where evaluable is defined as receiving one 
or two drops of study medication and have a pupil diameter measurement at the 90 minute 
time point at Visit 1), evaluating the safety and efficacy of Nyxol in subjects with 
pharmacologically-induced mydriasis. 
Following the successful completion of screening, each subject will be stratified by eye color 
and simultaneously be randomized to mydriatic agent (unmasked) and treatment (masked). 
At Visit 1 subjects will be screened for study eligibility. After screening, eligible subjects 
will be randomized 1:1 to one of the two treatment arms (Nyxol or Placebo). 
At the same visit, subjects who have been randomized and stratified by irides type (1:1 
[light/dark]) will receive one of three approved mydriatic agents approximately 1 hour prior 
to receiving study treatment. Randomization will be stratified 3:1:1 by mydriatic agent (2.5% 
phenylephrine, 1% tropicamide, and Paremyd), with randomization into each mydriatic agent 
capped at approximately 96:32:32 evaluable subjects. 
Each subject will receive one drop of mydriatic agent in each eye. If the drop is missed, the 
Investigator should give the drop again.  
At Visit 1, measurements will be completed before (-1 hour /baseline) and 60 minutes after 
(maximum/0 minutes) the mydriatic agent instillation in each eye (i.e., right before the study 
treatment is administered), and at 30 minutes, 60 minutes, 90 minutes, 2 hours, 3 hours, 4 
hours, and 6 hours after treatment dosing.  
At the Follow-Up Visit, which is one day after Visit 1, measurements will be measured 24 
hours after treatment dosing. 
The study eye is defined as the right eye (OD). The non-study eye is defined as the left eye 
(OS). The study and non-study eye will both be evaluated at all assessments. Adult subjects 
(≥ 18 years old) will have two drops (dosed 5 minutes apart) of treatment administered in the 
study eye (OD) and one drop of treatment administered in the non-study eye (OS) one hour 
after mydriatic drug instillation. Pediatric subjects (<18 years old) will have one drop of 
treatment administered in each eye one hour after mydriatic drug instillation.  
The schedule for assessments and timing of events is presented in Table 1. 


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Table 1 
Screening and Mydriatic/Treatment Schedule 
  
Day (D)
Visit 
Hour 
Informed Consent/ Assent 
Screening # assigned 
Med/Ophth History 
Demographics 
Prior/Concomitant Medications* 
Urine Pregnancy Test** 
HR/BP 
 
 
 
 
 
Adverse Events 
Ocular Tolerability^ 
Mydriatic Agent 
IOP^ 
Biomicroscopy 
Ophthalmoscopy^ 
Questionnaire^ 
Randomization # Assigned 
Treatment: Nyxol/ Placebo 
Screening Visit if subject qualifies becomes the first Treatment Visit. The mydriatic drug(s) will be given at -1 hour. 
*Investigators to note changes to concomitant medications at any time throughout the visit. 
**UPT is for females of childbearing potential. 
^Measurements will be performed as follows: 
    
 
    
 
    
 
    
 


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    IOP – Tono-Pen 
    Ophthalmoscopy – Direct or indirect ophthalmoscopy without dilation. Use of 90D lens (indirect) is allowed 
    Questionnaire – Subject Questionnaire will be a brief symptom survey.


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5.2. 
Inclusion – Exclusion Criteria and General Study Population 
The study population will be approximately 168 normal healthy subjects at least 12 years of 
age, with approximately 160 evaluable subjects. Written informed consent will be obtained 
from each adult subject. A signed assent form will be obtained for all minors ages 12-17, as 
well as a separate parental/Legal Guardian consent.  
The inclusion and exclusion criteria defined in the protocol apply to all subjects and are not 
repeated herein the SAP. Reference is made to the final protocol for the specific inclusion 
and exclusion criteria for study subjects. 
5.3. 
Randomization and Blinding 
A randomization code for allocating subjects to treatment will be prepared by a masked 
biostatistician not connected with the study. At the initiation of study related procedures, 
every potential subject is assigned a Screening number in numerical order per strata. Once a 
subject is qualified for the study, the subject is assigned a randomization number in the order 
provided by the biostatistician. 
 
. The mydriatic agent randomization will be approximately 3:1:1 
(2.5% phenylephrine, 1% tropicamide, and Paremyd). That is, approximately 60% of the 
randomized subjects will receive one drop of 2.5% phenylephrine 1 hour before treatment 
(96 evaluable subjects), approximately 20% will receive one drop of 1% tropicamide 1 hour 
before treatment (32 evaluable subjects), and approximately 20% will receive Paremyd 1 
hour before treatment (32 evaluable subjects). 
A randomization schema for evaluable subjects by investigational treatment, mydriatic agent, 
and irides type is are described in Figure 1. 


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Figure 1 
Randomization schema for evaluable subjects by investigational 
treatment, mydriatic agent, and irides type 
 
The study medications will be masked to both Investigator and study subjects, as well as 
Ocuphire. Only in case of medical emergency or occurrence of serious adverse events 
(SAEs) will the randomization code be unmasked by the study pharmacist and made 
available to the Investigator, Ocuphire, and/or other personnel involved in the monitoring or 
conduct of this study. Rules for unmasking a subject for safety reasons are fully described in 
the protocol and not repeated herein this SAP. 
5.4. 
Analysis Variables 
Variables to be summarized include demographics and baseline characteristics, medical 
(non-ocular) and ocular history, concomitant medications, and study drug accountability.  
Efficacy variables include: 
 
 
 
 
Safety variables include: 
 Conjunctival hyperemia (eye redness) measured with a Cornea and Contact Lens 
Research Unit (CCLRU) card 4-point scale: 


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o None (0) = Normal Appears white with a small number of conjunctival blood 
vessel easily observed. 
o Mild (+1) = Prominent, pinkish-red color of both the bulbar and palpebral 
conjunctiva. 
o Moderate (+2) = Bright, scarlet red color of the bulbar and palpebral 
conjunctiva. 
o Severe (+3) = Beefy red with petechiae, dark red bulbar and palpebral 
conjunctiva with evidence of subconjunctival hemorrhage.   
 Subjective ocular tolerability measured on a 4-point scale: 
o 0 – No discomfort 
o 1 – Mild discomfort 
o 2 – Moderate discomfort 
o 3 – Severe discomfort 
 DCNVA (i.e., Near VA) 
 BCDVA (i.e., Distance VA) 
 AEs 
 Vital signs (HR and BP) 
 IOP 
 Subject questionnaire 
 Urine pregnancy tests for females of childbearing potential 


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6. 
SAMPLE SIZE 
A sample size of 160 evaluable subjects (80 per treatment group) is needed for the study.  
The primary efficacy endpoint will be met if subjects show a positive effect for Nyxol (α = 
0.05 significance, two-tailed). 
 
 
 
 The assumptions for this 
power calculation are estimated from the MIRA-1 (OPI-NYXRM-201) Phase 2 results. Data 
from this study on the effects of treatment to reverse pharmacologically induced mydriasis 
were adjusted to take into account the increased dose of Nyxol used in this MIRA-2 Phase 3 
study (2 drops vs 1 drop) and adjusted for the 3:1:1 mydriatic agent groups, among other 
factors.  
 
 Furthermore, subjects will 
be randomized into the study at a ratio of approximately 3:1:1 to one of the three mydriatic 
agents, 2.5% phenylephrine, 1% tropicamide, and Paremyd. Therefore, if 96 evaluable 
subjects are randomized to 2.5% phenylephrine, then 32 subjects will be assigned to 1% 
tropicamide and 32 subjects to Paremyd, resulting in 160 evaluable subjects.  
It is assumed that there will be a 5% drop-out before the 90-minute efficacy assessment; 
therefore 168 subjects will be randomized to obtain 160 evaluable subjects for the primary 
analysis. 


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7. 
GENERAL CONSIDERATIONS 
7.1. 
Analysis Populations 
The following analysis populations will be defined for this study.  
7.1.1. 
Modified Intention-to-Treat (mITT) 
The mITT will include all randomized subjects who received one or two drops of study 
treatment and then had at least one scheduled pupil diameter measurement during Visit 1. 
The mITT will be used for the primary endpoint analysis and to analyze selected secondary 
efficacy endpoints, with subjects included in their randomized treatment regardless of the 
treatment they actually received. 
7.1.2. 
Per Protocol Population (PP) 
The PP population will include all subjects in the mITT who have two drops of study 
treatment in their Study Eye, have all scheduled pupil diameter measurements during Visit 1, 
had an increase of >0.2 mm in pupil diameter in the study eye at time 0 minutes compared to 
baseline (-1 hour), and have no major protocol deviations. The PP population will be used to 
analyze selected secondary efficacy endpoints, with subjects included in their randomized 
treatment regardless of the treatment they actually received. 
7.1.3. 
All Randomized Population (ARP) 
The ARP will include all randomized subjects. This population is also known as the 
Intention-To-Treat (ITT) population. The ARP will be used in confirmatory efficacy 
analyses, with subjects included in their randomized treatment regardless of the treatment 
they actually received. 
7.1.4. 
Safety Population (SP) 
The SP will include all randomized subjects who have received at least one drop of study 
treatment. The SP will be used to summarize safety variables, using the actual treatment a 
subject received. 
7.2. 
Covariates and Subgroups 
7.2.1. 
Planned Covariates 
Planned covariates include baseline values for the given assessment.  
7.2.2. 
Planned Subgroups 
Subgroup analyses by irides type (light, dark), and by mydriatic agent (phenylephrine, 
tropicamide, Paremyd) will be completed for efficacy endpoints. An additional mydriatic 


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agent subgroup, combining 1% tropicamide and the Paremyd subjects into a “tropicamide” 
group, will be used for efficacy endpoints.  
Other possible subgroups include age, sex, and race. If there is sufficient sample, analysis of 
safety and efficacy endpoints will be completed for the subgroup of pediatric subjects. 
7.3. 
Management of Analysis Data 
7.3.1. 
Data Handling 
Data from unscheduled visits will not be included in the analysis of efficacy or safety but will 
be listed.   
7.3.2. 
Missing Data 
The primary efficacy endpoint is the percentage of subjects’ study eyes returning to  
≤ 0.2 mm baseline pupil diameter at 90 minutes in the study eye. For the analysis of the 
primary efficacy endpoint, imputation will be performed for missing efficacy data as 
specified in Section 7.3.2.3 for the analysis using the mITT. Confirmatory analyses will be 
performed using the ARP, also using imputation for missing data.   
Otherwise there will be no substitutions made to accommodate missing data points for 
efficacy data. All data recorded on the CRF will be included in data listings that will 
accompany the CSR. 
Safety data will be imputed in limited situations. If the severity of an AE is missing, then the 
severity will remain missing. If relationship of the AE to study drug is missing, the 
relationship will remain missing. Missing or partial dates for AEs or concomitant 
medications will be imputed as described in Section 7.3.2.1. Otherwise, all summaries of 
safety endpoints will be completed using observed cases in the SP; no imputation will be 
completed. 
7.3.2.1. 
Handling of Missing Date Values 
Partial or Missing Dates 
The following conventions will be used to impute missing portions of dates for AEs and 
concomitant medications, if warranted. Note that the imputed values outlined here may not 
always provide the most conservative date. In those circumstances, the imputed value may be 
replaced by a date that will lead to a more conservative analysis. 
A. 
Start Dates 
1) 
If the year is unknown, then the date will not be imputed and will 
be assigned a missing value.  
2) 
If the month is unknown, then: 


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i) If the year matches the first dose date year, then impute 
the month and day of the first dose date. 
ii) Otherwise, assign ‘January.’  
3) 
 If the day is unknown, then: 
i) If the month and year match the first dose date month 
and year, then impute the day of the first dose date. 
ii) Otherwise, assign the first day of the month. 
B. 
Stop Dates 
1) 
If the year is unknown, then the date will not be imputed and will 
be assigned a missing value. 
2) 
If the month is unknown, then assign ‘December.’ 
3) 
If the day is unknown, then assign the last day of the month. 
 
7.3.2.2. 
Missing Baseline Data 
Every effort will be made to ensure that accurate baseline information on the subjects is 
collected. In the event that a subject is missing baseline information, the subject will be 
included in the SP for assessment of safety and excluded from the primary analyses. Each 
case of missing baseline data will be evaluated for potential inclusion in the exploratory 
endpoints. All baseline data will be observed cases, without imputation. 
7.3.2.3. 
Imputation Methods 
Imputation for efficacy data will only be performed for the primary efficacy endpoint using 
the mITT and ARP. If 5% or fewer data are missing at the 90-minute time point in all 
treatment groups, an analysis with last observation carried forward (LOCF) for missing data 
will be applied within a treatment group. Neither the baseline (-1 hour) nor max pupil 
diameter (0 minutes) time points will be used for imputation. If more than 5% of data are 
missing at the 90-minute time point in any treatment group, multiple imputation will be 
employed to analyze incomplete data sets under the assumption that the mechanism 
responsible for the missing data is at worst characterized as missing at random (MAR).    
Multiple imputation is a simulation-based approach where missing values are replaced by 
multiple Bayesian draws from the conditional distribution of missing data given the observed 
data and covariates, creating multiple completed data sets. These completed datasets can then 
be analyzed using standard analysis methods. Rubin (1987) presented rules for how to 
combine the multiple sets of estimates to produce overall estimates, confidence intervals 
(CIs), and tests that adequately incorporate missing data uncertainty. 
Missing values for pupil diameter will be imputed simultaneously based on an underlying 
joint normal distribution using a Markov Chain Monte Carlo (MCMC) method.   
The imputations will be done separately for each treatment group and will include the 
following variables in the imputation model: pupil diameter at Day 1 (30 minutes, 60 


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minutes, 90 minutes, 2 hours, 3 hours, 4 hours, 6 hours, 24 hours). No imputation will be 
applied to the max pupil diameter (0 minutes) time point. 
The number of imputations will be set to 500. The outcomes of interest (change from 
baseline) will be calculated from these imputed datasets. The treatment difference for each 
imputed dataset will be evaluated using mixed models. See Section 9.1 for details on these 
models. The estimates and standard errors (SEs) of the differences in LSM (least squares 
means) based on the 500 imputed datasets are then combined by applying Rubin’s rules for 
multiple imputed datasets. T-tests are also provided. SAS Proc MI, Proc MIXED, and Proc 
MIANALYZE will be utilized for these analyses. The averaged difference in LSM with the 
corresponding 95% CI will also be presented.  
Example SAS code is provided below: 
 
 
 
 
 
      
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
          
 
          
 
          
 
          
 
          
 
          
  
         
 
 
 
 
 
 
 
    
 
 
 
      
 
 
      
 
 select ParameterEstimates; 
 
   run; 
   
7.3.3. 
Handling of Early Termination Visit Information 
In the event that a subject is terminated early from this study on Day 1, the early termination 
data for safety variables will be assigned to the closest scheduled time point on Day 1. If the 


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closest time point has valid data, the early termination data will be assigned to the next 
available time point. 
7.3.4. 
Pooling of Investigational Sites 
The data from all study centers will be pooled together for all planned analyses. 
7.3.5. 
Coding Conventions for Events and Medications 
All AEs and medical history will be mapped to the Medical Dictionary for Regulatory 
Activities (MedDRA Version 22.0) system for reporting (preferred term and body system). 
Prior and concomitant medications will be coded using WHO-DD (World Health 
Organization Drug Dictionary) (Version March 2019, C Format). 
7.3.6. 
Analysis Software 
Data manipulation, tabulation of descriptive statistics, calculation of inferential statistics, and 
graphical representations will be performed primarily using SAS (release 9.4 or higher) for 
Windows. If the use of other software is warranted, the final CSR will detail what software 
was used and for what purposes. 
7.3.7. 
Study Data 
Study data identified in the schedule for time and events (Table 1) are collected, and source 
verified, on the electronic data capture (EDC) Datatrak One version 14.5.5.  
All study data will be formulated into regulatory-compliant data sets to provide transparency, 
traceability, and integrity of trial analysis results from the collection source. Observed study 
data will be mapped to the Clinical Data Interchange Standards Consortium (CDISC) Study 
Data Tabulation Model (SDTM) and serve as the source data from the trial. All study 
analyses will be completed using analysis data sets that are derived from the SDTM and 
follow the CDISC Analysis Data Model (ADaM) architecture.  
The methods for programming the CDISC SDTM and ADaM data sets are described in 
Figure 2. 


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Figure 2 
SDTM, ADaM, and TFL Development and Validation 
 
7.4. 
Planned Study Analyses 
7.4.1. 
Statistical Summaries: Descriptive and Inferential 
Categories for data presentation and analysis will consist of each treatment group (Nyxol or 
Placebo).  
All statistical tests will be two-sided and a difference resulting in a p-value of less than or 
equal to 0.05 will be considered statistically significant. All p-values will be rounded to and 
displayed in four decimals. If a p-value less than 0.0001 occurs, it will be shown in tables as 
<0.0001. 
Descriptive summaries of variables will be provided where appropriate. For continuous 
variables, the number of non-missing values (n), mean, standard deviation, median, 
minimum, and maximum will be tabulated by treatment group. For categorical variables, the 
counts and proportions of each value will be tabulated by treatment group. Expansion of 


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descriptive table categories within each treatment may occur if such elaborations are thought 
to be useful. 
All study-related data collected will be presented in listings. Study-related data not subject to 
analysis according to this plan will not appear in any tables or graphs but will be included in 
the data listings. 
7.4.2. 
Interim Analyses and Data Monitoring 
No formal interim analysis or safety monitoring committee is planned for this study.  
7.4.3. 
Final Analysis and Publication of Study Results 
The final analysis will be completed after all subjects have completed the study.   
7.5. 
Multiple Testing Procedures 
There will be no adjustments for multiplicity and no formal multiple testing procedures are to 
be implemented with this analysis plan. 
7.6. 
Baseline Values 
Baseline values are the values obtained prior to any drug administration on Day 1 (study drug 
or mydriatic agent), usually the -1 hour time point. If the Day 1 value is missing or the 
assessment is not completed at the -1 hour time point, the value at Screening will be treated 
as the baseline. 
 


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8. 
SUMMARY OF STUDY DATA 
8.1. 
Subject Disposition 
A summary of the analysis sets includes the number and percentage of subjects by treatment 
group and overall for the following categories: subjects in the ARP, subjects in the SP, 
subjects in the mITT Population, and subjects in the PP Population. All percentages will be 
based on the number of subjects in the ARP.  
End of trial information will also be summarized in this table, including the number of 
subjects completing the study, the number of subjects who prematurely discontinued the 
study with reasons for withdrawal, the number of subjects completing the study medication 
dosing, and the number of subjects who prematurely discontinued the study medication with 
reasons for study medication discontinuation.   
A by-subject data listing of study completion information including the reason for premature 
study withdrawal, if applicable, will be presented. 
8.2. 
Protocol Deviations 
Major protocol deviations, as determined by a Sponsor blinded review of the data prior to 
database lock and unblinding of the study, may result in the removal of a subject’s data from 
the PP Population. The Sponsor or designee will be responsible for producing the final 
deviation file; this file will include a description of the protocol deviation and clearly identify 
whether this violation warrants exclusion from the PP Population. This file will be finalized 
prior to database lock. 
All protocol deviations will be presented in a by-subject data listing, with a flag to indicate if 
a deviation was considered major.  
8.3. 
Demographics and Baseline Characteristics 
Subject demographic data and baseline characteristics will be tabulated and summarized 
descriptively by treatment group and overall. The demographic data and baseline 
characteristics will be summarized for the mITT Population, PP Population, SP, and ARP. 
The demographics consist of age (year), sex, race, ethnicity, and study eye (OD), iris color 
(light blue, dark blue, blue with peripupillary brown, uniform green, green with brown iris 
ring, central brown and peripheral green, brown with some peripheral green, or brown), 
irides type (light or dark), eyeglasses-wearing status (yes or no) (distance vision or near 
vision), and mydriatic agent (phenylephrine, tropicamide, or Paremyd). A subject’s age in 
years is calculated using the date of the informed consent and date of birth. Age will be 
summarized using descriptive statistics. The number and percentage of subjects by sex, race, 
ethnicity, study eye, iris color, irides type, distance vision/near vision correction status, and 
mydriatic agent will also be reported. Percentages will be based on the total number of 
subjects in the study population presentation. 


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The following baseline characteristics will be summarized for study eye and non-study eye, 
using descriptive statistics: 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
 
  
 
 
All demographic and baseline information will be presented in by-subject listings. 
8.4. 
Medical History  
The number and percent of subjects with individual medical histories will be summarized for 
all subjects by treatment group and overall. Non-ocular and ocular medical history will be 
summarized separately.  
Medical history will be coded using the MedDRA Version 22.0. The number and percentage 
of subjects with any medical history will be summarized overall and for each system organ 
class (SOC) and preferred term (PT). Percentages will be calculated based on number of 
subjects in the SP.   
Subject medical history data including specific details will be presented in by-subject 
listings. 
8.5. 
Prior and Concurrent Medications 
The number and percentages of all concomitant medications will be summarized by 
treatment group, Anatomical Therapeutic Chemical (ATC) level 4, and PT. The total number 
of concomitant medications and the number and percentages of subjects with at least 1 
concomitant medication will be summarized by treatment group. All summaries will be 
performed using the SP.   
A concomitant medication is defined as any medication taken on or after the day of first 
exposure to study drug. 
Prior medications are defined as any medication that has a start and stop date prior to the day 
of first exposure to any study drug, collected from up to 30 days prior to Screening. The total 


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number of prior medications and the number and percentages of subjects with at least 1 prior 
medication will be summarized by treatment group. 
8.6. 
Treatment Administration 
Treatment administration data for both the study drug and the mydriatic agent on Day 1 will 
be presented in by-subject listings.  


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9. 
EFFICACY ANALYSES 
Unless otherwise noted, efficacy will be assessed using the mITT and PP populations, with 
subjects included in their randomized treatment regardless of the treatment they actually 
received. For the analysis of the primary efficacy endpoint, imputation will be performed for 
missing data as described in Section 7.3.2.3. If the analysis using the mITT Population shows 
a positive effect for Nyxol at the 0.05 level of significance, the primary endpoint will be 
considered met.  
Confirmatory analysis of the primary efficacy endpoint will be performed using the ARP, 
also using imputation for missing data. For the analysis of the secondary efficacy endpoints, 
only observed case data will be used.  
All efficacy assessment data, regardless of whether they are included in the analysis, will be 
presented in by-subject listings. If there is sufficient sample, analysis of efficacy endpoints 
will be completed for the subgroup of pediatric subjects. 
9.1. 
Clinical Efficacy   
For all efficacy endpoints, Baseline is defined as -1 hour prior to treatment on Day 1. This is 
the time when the mydriatic agent is administered, and the pupil diameter measurement is 
considered normal. Max timepoint is defined as time 0 minutes, during which maximum 
pupil diameter is expected; this is also the timepoint at which the treatment is administered 
(Nyxol or Placebo).  
All efficacy data will be summarized by treatment group and timepoint (-1 hour [baseline], 0 
minutes, 30 minutes, 60 minutes, 90 minutes, 2 hours, 3 hours, 4 hours, 6 hours, and 24 
hours), as appropriate. 
Primary Efficacy Endpoint: 
The primary efficacy endpoint is the percentage of subjects returning to ≤ 0.2 mm baseline 
pupil diameter at 90 minutes post-treatment in the study eye. 
 
 
 The primary efficacy endpoint will 
be analyzed using a logistic regression model with 
 
. The percentage of subjects in 
each treatment group meeting the criteria, the odds ratio (OR) with 95% CI, and p-value will 
be provided. Example SAS code is as follows: 
  
  
 
 
 


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By including the mydriatic agent and irides type as factors in the primary efficacy analysis 
model, the model efficiency as well as a change in the treatment effect will be increased. 
Including these factors in the model will also make the results more generalizable to other 
studies in which the sample characteristics may differ from the current study [2]. 
Additionally, a sensitivity analysis will be applied to the primary efficacy endpoint for the 
mITT and PP populations, which uses a 
 
 
.  
In addition, the primary efficacy endpoint will be analyzed by mydriatic agent and by 
light/dark irides using the same model indicated above but without mydriatic agent or irides 
as a factor, as appropriate. For these subgroup analyses, observed case data only will be used; 
that is, missing values will not be imputed. Each mydriatic agent will be analyzed 
individually, and an additional mydriatic agent subgroup, combining 1% tropicamide and the 
Paremyd subjects into a “tropicamide” group, will be analyzed.   
In addition, the primary efficacy endpoint will be analyzed for the pediatric population, as 
well as by mydriatic agent. 
A comparison of the study and non-study eye for each subject will be completed for the 
primary efficacy endpoint, as well as by mydriatic agent. This analysis will be by treatment, 
and will be analyzed using a logistic regression model with eye type (study eye or non-study 
eye), mydriatic agent, and light/dark irides as fixed effects, subject as a random effect, and 
the average baseline pupil diameter across eye type as a covariate. The percentage of eyes 
meeting the criteria, the OR with 95% CI, and p-value will be provided. Example SAS code 
is as follows: 
  
  
 
 
 
 
 
Secondary Efficacy Endpoints: 
Secondary efficacy endpoints are indicated in Section 4.2.2. Secondary efficacy endpoints 
will be analyzed by study eye and non-study eye, unless otherwise indicated. Binocular 
accommodation will be analyzed separately.  
Each of the continuous secondary efficacy endpoints will be analyzed using analysis of 
covariance (ANCOVA), with change from baseline as the dependent variable, treatment, 
mydriatic agent, and light/dark irides as factors, and the respective baseline value included as 
the covariate. Note that most secondary efficacy endpoints are in relation to baseline (-1 
hour), whereas some pupil diameter endpoints are in relation to max (0 minute).  


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Each ANCOVA will be performed using the mITT and PP populations. The output from 
each ANCOVA will include the LSM and SE for both treatment groups, along with the 
placebo-corrected LSM, its 95% CI and associated p-value. Line graphs displaying the mean 
and SE of the change from maximum (0 minute) pupil diameter value will be presented.  
For each of the secondary endpoints related to percent of subjects achieving certain criteria, 
the analysis will be performed using a logistic regression model with 
 
 For each 
analysis, the percentage of subjects in each treatment group meeting the criteria, the OR with 
95% CI and p-value will be provided. The analysis of the time (hours) to return to ≤ 0.2 mm 
from baseline pupil diameter (time savings analysis) endpoint will be performed using a Cox 
proportional hazards regression model with 
 
. Subjects who do not return to ≤ 0.2 mm from 
baseline pupil diameter by the 6 hour time point will have their time to return censored at 8 
hours. Example SAS code is as follows: 
 
 
 
 
 
 
 
 
  
 
 
The output from the model will include the hazard ratio comparing treatment groups, its 95% 
CI and associated p-value. Survival plots will also be generated. Time to return to baseline (-
1 hour) pupil diameter will be measured beginning at the Max (0 hour) time point. 
In addition, each secondary efficacy endpoint will be analyzed by 
 
 
, as appropriate. Each mydriatic agent will be analyzed individually, and an 
additional mydriatic agent subgroup, combining 1% tropicamide and the Paremyd subjects 
into a “tropicamide” group, will be analyzed. Analyses of endpoints related to 
accommodation will only be completed by mydriatic agent. 
 
 
Exploratory analyses may be performed to compare efficacy endpoints between the study eye 
and non-study eye within the same subject. Categorical variables will be analyzed as 
described above for the primary efficacy endpoint. Continuous variables will be analyzed 
using a mixed model with eye type (study eye or non-study eye), mydriatic agent, and 
light/dark irides as fixed effects, subject as a random effect, and the average baseline pupil 
diameter across eye type as a covariate. 


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10. 
SAFETY ANALYSES 
All safety analyses will be conducted using the SP. All safety analyses will be completed 
using the actual treatment a subject received. Observed case data will be used; no imputation 
will be performed for missing safety data except for the limited situations described in 
Section 7.3.2. 
All safety data will be presented in by-subject listings. Unscheduled assessments will not be 
summarized but will be included in the listings.  
 
10.1. 
Adverse Events 
AEs will be coded using MedDRA, Version 22.0.   
Treatment-emergent adverse events (TEAEs) are defined as any AE that begins or worsens 
after initiation of the investigational product and through the subject’s last study visit (study 
completion or early termination). 
If the onset of an AE is on or after the date of first dose of study medication or is increasing 
in severity after first dose of study medication, then the AE will be considered treatment 
emergent. 
Only TEAEs will be summarized; all AEs (TEAE, non-TEAE) will be included in a by-
subject listing. A separate listing of AEs for pediatric subjects will be provided. 
The number and percent of subjects with any TEAEs will be summarized by SOC and PT by 
treatment group and overall. At each level of tabulation (e.g., at the PT level), subjects will 
be counted only once if they had more than one such event reported during the AE collection 
period. A separate summary by SOC, PT, and treatment group will be completed by 
mydriatic agent. 
 
Note that in MedDRA, ocular events are coded to the SOC of “Eye Disorders”. Thus, using 
SOC in the summaries will provide a separation of ocular and non-ocular adverse events. 
The following summary tables will be presented for TEAE data:  
 
Overall summary of TEAEs  
 
Summary table of TEAEs by SOC and PT  
 
Summary table of TEAEs by SOC, PT, and by greatest relationship level to study 
drug (not related, unlikely related, possibly related, probably related, definitely 
related, or unknown) 
 
Summary table of TEAEs by SOC, PT, and maximum severity (mild, moderate, 
severe) 


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 
Summary table of serious TEAEs by SOC and PT  
 
Summary table of TEAEs leading to withdrawal from the study by SOC and PT 
 
Summary table of TEAEs leading to study medication discontinuation by SOC and 
PT 
 
10.2. 
Deaths, Serious Adverse Events and Other Significant Adverse Events 
10.2.1. 
Deaths 
The AE listing will include all AEs, including deaths, regardless of causality; one of the 
columns in the listing will specify whether the AE was fatal.     
10.2.2. 
Serious Adverse Events 
The AE listing will include all AEs, including SAEs; one of the columns in the listing will 
specify whether the AE was an SAE.     
10.2.3. 
Adverse Events Leading to Withdrawal from the Study 
The AE listing will include all AEs, including AEs leading to withdrawal from the study; one 
of the columns in the listing will specify whether the AE led to withdrawal from the study.   
10.2.4. 
Adverse Events Leading to Discontinuation of Study Medication 
The AE listing will include all AEs, including AEs leading to discontinuation of study 
medication; one of the columns in the listing will specify whether the AE led to 
discontinuation of study medication.   
10.3. 
Conjunctival Hyperemia 
Results from the conjunctival hyperemia assessment, measured with a CCLRU card 4-point 
scale, 
 
 
e (-1 
hour). Separate summaries will be created for the study eye and the non-study eye. 
Additionally, 
 
 
 
10.4. 
Subjective Ocular Tolerability 
Results from the subjective ocular tolerability assessment, measured on a 4-point scale, will 
be summarized descriptively using counts and percentages for each treatment group at the 0 
minute time point. Additionally, the categories “No Discomfort” and “Mild Discomfort” will 
be pooled into a single category and summarized descriptively, as will the categories 


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“Moderate Discomfort” and “Severe Discomfort”. Treatments will be compared for the two 
pooled categories using a Fisher’s exact test. Separate summaries will be created for the 
study eye and the non-study eye. 
10.5. 
Visual Acuity 
Visual acuity assessments (BCDVA and DCNVA) will be summarized at the following 
timepoints (0 minutes, 90 minutes, 6 hours, and 24 hours), using letters and logMAR units. 
Only letters will be recorded in the CRF and will be converted to logMAR programmatically 
as follows: 
 
 
 
 
 
 Separate summaries will be created for the study eye, the non-study eye, and both eyes. 
Treatments will be compared using the same ANCOVA model proposed for the continuous 
secondary efficacy endpoints. In addition, DCNVA will be analyzed by mydriatic agent 
using the same model as for the efficacy variables. 
10.6. 
Vital Signs 
Descriptive statistics of observed values will be presented for vital sign data at each time 
point (Screening, 6 hours, and 24 hours), including systolic BP (mmHg), diastolic BP 
(mmHg), and HR (bpm) by treatment group and overall. Changes from baseline to each 
scheduled post-baseline time point will be presented.  
10.7. 
IOP 
Observed values and change from baseline in IOP at 6 hours will be summarized for the 
study eye and the non-study eye. Treatments will be compared using the same ANCOVA 
model proposed for the continuous secondary efficacy endpoints. 
10.8. 
Subject Questionnaire 
Subject questionnaire values will be summarized for each timepoint (-1 hour, 0 minutes, 60 
minutes, 90 minutes, 3 hours, 6 hours, and 24 hours) by treatment group. A separate 
summary will be created for each mydriatic agent. 
10.9. 
Other Safety Measures 
Urine pregnancy tests for females of childbearing potential will be presented in by-subject 
listings. Results from biomicroscopic and ophthalmoscopic examinations, which are 
completed only at Screening, will also be presented in by-subject listings. 


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11. 
REFERENCES 
[1] ICH E9 Expert Working Group. Statistical Principles for Clinical Trials: ICH 
Harmonized Tripartite Guideline, September 1998 
[2] Hauck WM, Anderson S, and Marcus SM, Should We Adjust for Covariates in Nonlinear 
Regression Analyses of Randomized Trials? Controlled Clin Trials 1998;19:249–256 
 
 


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12. 
APPENDICES 
12.1. 
List of Planned Tables 
The list of planned tables includes all of the main tables to be presented for the study. 


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12.2. 
List of Planned Listings 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Listing 
Description of Listing 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
12.3. 
List of Planned Figures