Conditions: Mydriasis Dilation
Keywords: Nyxol Pharmacologically Induced Mydriasis Dilation
Allocation: RANDOMIZED | Intervention model: PARALLEL
Enrollment: 368
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
* To evaluate the systemic exposure of Nyxol on pharmacokinetic (PK) sampling

Detailed description:
A randomized, parallel arm, double-masked, placebo-controlled Phase 3 study in at least 330 randomized subjects, 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 2:1, Nyxol or placebo (vehicle). Stratification by iris color will be 1:1, light or dark rides. The mydriatic agent randomization will be 3:1:1 (2.5% phenylephrine, 1% tropicamide, and Paremyd).

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.

Blood sampling for Nyxol PK measurements will be conducted in a subset of approximately 30 adult subjects at approximately two select study sites.

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

Protocol document:
SAP_002.pdf: 




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1.  
T A B L E O F C O N T E N T S  
1.  T A B L E O F C O N T E N T S  ................................................................................................... 2 
2.  LI S T O F A B B R E VI A TI O N S A N D D E FI NI TI O N O F T E R M S  ....................................... 5 
3.  I N T R O D U C TI O N .............................................................................................................. 7 
3. 1.  
Preface ...................................................................................................................... 7 
3. 2.  
P ur p ose of A nal yses ................................................................................................. 7  
3. 3.  
S u m mar y of Statistical A nal ysis C ha n ges t o t he Pr ot oc ol....................................... 7  
4.  S T U D Y O BJ E C TI V E S A N D E N D P OI N T S  ...................................................................... 8 
4. 1.  
St u d y O bjecti ves ...................................................................................................... 8  
4. 2.  
St u d y E n d p oi nts ....................................................................................................... 8  
4. 2. 1.  
Pri mar y E n d p oi nts .................................................................................... 8  
4. 2. 2.  
Sec o n dar y E n d p oi nts ................................................................................ 8  
5.  S T U D Y M E T H O D S  ......................................................................................................... 1 0 
5. 1.  
Ge ner al St u d y Desi g n a n d Pla n ............................................................................. 1 0  
5. 2.  
I n cl usi o n – E xcl usi o n Criteria a n d Ge n eral St u d y P o p ulati o n .............................. 1 3 
5. 3.  
Ra n d o mizati o n a n d Bli n di n g ................................................................................. 1 3  
5. 4.  
A nal ysis Varia bles  ................................................................................................. 1 3 
6.  S A M P L E SI Z E  ................................................................................................................. 1 5 
7.  G E N E R A L C O N SI D E R A TI O N S  .................................................................................... 1 6 
7. 1.  
A na l ysis P o p ulati o ns .............................................................................................. 1 6 
7. 1. 1.  
M o difie d I nte nti o n -t o-Treat ( mI T T)  ....................................................... 1 6 
7. 1. 2.  
Per Pr ot oc ol P o p ulati o n ( P P) .................................................................. 1 6 
7. 1. 3.  
All Ra n d o mize d P o p ulati o n ( A R P) ........................................................ 1 6 
7. 1. 4.  
Safet y P o p ulati o n ( S P) ............................................................................ 1 6  
7. 1. 5.  
P K P o p ulati o n ......................................................................................... 1 6  
7. 2.  
C o variates a n d S u b gr o u ps...................................................................................... 1 7  
7. 2. 1.  
Pla n ne d C o variates .................................................................................. 1 7 
7. 2. 2.  
Pla n ne d S u b gr o u ps.................................................................................. 1 7  
7. 3.  
Ma na ge me nt of A nal ysis Data  ............................................................................... 1 7 
7. 3. 1.  
Data Ha n dli n g  ......................................................................................... 1 7 
7. 3. 2.  
Missi n g Data  ........................................................................................... 1 7 
7. 3. 2. 1.  
Ha n dli n g of Missi n g Date Val ues ......................................... 1 8  
7. 3. 2. 2.  
Missi n g Baseli ne Data  .......................................................... 1 8 
7. 3. 2. 3.  
I m p utati o n Met h o ds .............................................................. 1 8 
7. 3. 3.  
Ha n dli n g of Earl y T er mi nati o n Visit I nf or m ati o n .................................. 2 0  
7. 3. 4.  
P o oli n g of I n vesti gati o nal Sites .............................................................. 2 0  
7. 3. 5.  
C o di n g C o n ve nti o ns f or E ve nts a n d Me dicati o ns .................................. 2 0  
7. 3. 6.  
A nal ysis S oft ware ................................................................................... 2 0  
7. 3. 7.  
St u d y Data ............................................................................................... 2 0  
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7. 4.  
Pla n ne d St u d y A nal yses ......................................................................................... 2 1  
7. 4. 1.  
Statistical S u m maries: Descri pti ve a n d I nfere ntial  ................................. 2 1 
7. 4. 2.  
I nteri m A nal yses a n d Data M o nit ori n g................................................... 2 2 
7. 4. 3.  
Fi nal A nal ysis a n d P u blicati o n of St u d y Res ults .................................... 2 2  
7. 5.  
M ulti ple Testi n g Pr oce d ures .................................................................................. 2 2  
7. 6.  
Baseli ne Val ues  ...................................................................................................... 2 2 
8.  S U M M A R Y O F S T U D Y D A T A  ..................................................................................... 2 3 
8. 1.  
S u bject Dis p ositi o n  ................................................................................................ 2 3 
8. 2.  
Pr o t oc ol De viati o ns ................................................................................................ 2 3 
8. 3.  
De m o gra p hics a n d Baseli ne C haract eristics  .......................................................... 2 3 
8. 4.  
Me dical Hist or y  ..................................................................................................... 2 4 
8. 5.  
Pri or a n d C o nc urr e nt Me dicati o ns ......................................................................... 2 4  
8. 6.  
Treat me nt A d mi nistrati o n  ...................................................................................... 2 5 
9.  E F FI C A C Y A N A L Y S E S  ................................................................................................. 2 6 
9. 1.  
Cli nical Efficac y  .................................................................................................... 2 6 
1 0.  S A F E T Y  A N A L Y S E S  ..................................................................................................... 3 0 
1 0. 1.  A d verse E v e nts  ...................................................................................................... 3 0 
1 0. 2.  Deat hs, Seri o us A d verse E ve nts a n d Ot her Si g nifi ca nt A d vers e E ve nts............... 3 1  
1 0. 2. 1.  Deat hs  ..................................................................................................... 3 1 
1 0. 2. 2.  Seri o us A d verse E v e nts .......................................................................... 3 1  
1 0. 2. 3.  A d verse E v e nts Lea di n g t o Wit h dra wal fr o m t he St u d y ........................ 3 1  
1 0. 2. 4.  A d verse E v e nts Lea di n g t o Disc o nti n uati o n of St u d y 
Me dicati o n  .............................................................................................. 3 1 
1 0. 3.  C o nj u ncti val H y pere mia ........................................................................................ 3 1  
1 0. 4.  S u bjecti ve Oc ular T ol era bilit y  ............................................................................... 3 1 
1 0. 5.  Vis ual Ac uit y  ......................................................................................................... 3 2 
1 0. 6.  Vital Si g ns  .............................................................................................................. 3 2 
1 0. 7.  I O P ......................................................................................................................... 3 2 
1 0. 8.  S u bject Q uesti o n naire ............................................................................................ 3 2  
1 0. 9.  Ot her Safet y M eas ur es  ........................................................................................... 3 3 
1 1.  P K  A N A L Y S E S  ............................................................................................................... 3 4 
1 2.  R E F E R E N C E S  ................................................................................................................. 3 6 
1 3.  A P P E N DI C E S  .................................................................................................................. 3 7 
1 3. 1.  List of Pla n ne d Ta bles ........................................................................................... 3 7  
1 3. 2.  List of Pla n ne d Listi n gs ......................................................................................... 4 4  
1 3. 3.  Hierar c h y of Se q u e ntial Testi n g  ............................................................................ 4 6 
 
 
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LI S T O F T A B L E S  
 
Ta ble 1  
Scree ni n g a n d M y driatic/ Treat me nt Sc he d ul e  ................................................. 1 1 
 
 
LI S T O F FI G U R E S  
 
Fi g ure 1  
S D T M, A Da M, a n d T F L De vel o p me nt a n d V ali dati o n .................................. 2 1  
 
 
 
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5.  
S T U D Y M E T H O D S  
5. 1.  
Ge ner al St u d y Desi g n a n d Pl a n  
T his is a  ra n d o mize d, par allel ar m, d o u ble-mas k e d, place b o -c o ntr olle d P has e 3 st u d y i n 
a p pr o xi matel y 3 3 0 ra n d o mize d s u bjects e val u ati n g t he safet y a n d effi cac 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 ra n d o mize d t o m y driatic a ge nt ( u n mas k e d) a n d treat me nt 
( mas ke d). 
At Visit 1 s u bjects will be scree ne d f or st u d y eli gi bilit y. After scree ni n g, eli gi ble s u bjects   
will be ra n d o mize d 2: 1 t o o ne of t he t w o treat me nt ar ms ( N y x ol or Place b o). S u bjects w h o 
ha ve bee n ra n d o mize d a n d stratifie d b y iri des t y pe ( 1: 1 [li g ht/ dar k]) will recei ve o ne of t hree 
a p pr o ve d m y driatic a ge nts a p pr o xi matel y 1 h o ur pri or t o recei vi n g st u d y treat me nt. 
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 mi de, a n d Par e m y d). 
S u bjects will ha ve 2 dr o ps of st u d y treat me nt ( N y x ol or place b o) a d mi nistere d i n t he st u d y 
e ye (ri g ht e y e [ O D]). E ac h dr o p will be i nstille d 5 mi n utes a part. S u bjects will ha ve o nl y 1 
dr o p of st u d y treat me nt a d mi nistere d i n t he fell o w e ye (left e ye [ O S]).   
T he st u d y e ye a n d fell o w e ye will b ot h be e val u ate d at all assess me nts u nless ot her wise 
s pecifie d.   
Bl o o d sa m pli n g f or N y x ol P K meas ure me nts will be c o n d ucte d i n a s u bset of a p pr o xi matel y 
3 0 a d ult s u bjects at a p pr o xi matel y 2 select st u d y sites.  
At Visit 1, meas ure me nts will be ma de bef ore ( -1 h o ur / baseli ne) a n d 6 0 mi n utes after ( ma x 
p u pil dilati o n/ 0 mi n utes) t he m y driatic a ge nt i nstillati o n i n eac h e ye (i .e., ri g ht bef ore t he 
st u d y treat me nt is a d mi nistere d). A d diti o nall y, meas ure me nts will be ta ke n at 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, a n d 6 h o urs aft er st u d y treat me nt d osi n g.   
At Visit 2 ( F oll o w - U p Visit), w hic h is 1 da y after Visit 1, meas ure me nts will be ma de   
2 4 h o urs after st u d y tr eat me nt d osi n g. 
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 fell o w e y e is defi ne d as t he left e ye ( O S). 
T he st u d y a n d fell o w e y e will b ot h be e val uate d at all assess me nts. All  s u bjects will ha ve t w o 
dr o ps ( d ose d 5 mi n utes a part) of tr eat me nt a d mi nistere d i n t he st u d y e ye ( O D) a n d o n e dr o p 
of treat me nt a d mi nistere d i n t he fell o w  e ye ( O S) o ne h o ur aft er m y driatic dr u g i nstillati o n.  
T he sc he d ule f or ass ess me nts a n d ti mi n g of e ve nts is prese nte d i n Ta ble 1 . 
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5. 2.  
I ncl usi o n – E xcl usi o n Criteri a a n d Ge ner al St u d y P o p ul ati o n  
T he st u d y p o p ulati o n will be a p pr o xi matel y 3 3 0 n or mal healt h y s u bjects at least 1 2 years of 
a ge. Writte n i nf or me d c o nse nt will be o btai ne d fr o m eac h a d ult s u bject. A si g ne 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 w ell as a se parate p are nt al/ Le gal G uar dia n 
c o nse nt.  
T he i ncl usi o n a n d e xcl usi o n criteria defi ne d i n t he pr ot oc ol a p pl y t o all s u bjects a n d are n ot 
re peat e d her ei n t he S A P. Refere n ce is ma d e t o t he fi nal pr ot oc ol f or t he s p ecific i ncl usi o n 
a n d e xcl usi o n criteria f or st u d y s u bjects. 
5. 3.  
R a n d o miz ati o n a n d Bli n di n g  
A ra n d o mizati o n c o d e f or all ocati n g s u bjects t o trea t 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. At t he i nitiati o n of st u d y relate d pr oce d ures, 
e ver y p ote ntial s u bject is assi g ne d a Scree ni n g n u m ber i n n u merical or d er per strata. O n ce a 
s u bject is q ualifie d f or t h e st u d y, t he s u bject is assi g ne d a ra n d o mizati o n n u m ber i n t he or der 
pr o vi de d b y t he bi ostatisticia n. 
Treat me nt r a n d o mizati o n will be 2: 1, N y x ol or P lace b o ( ve hi cle). Stratificati o n b y iri des t y pe 
will be 1: 1, li g ht or dar k. T he m y driatic a ge nt ra n d o mizati o n will be a p pr o xi matel y  3: 1: 1 
( 2. 5 % p he n yle p hri ne, 1 % tr o pica mi de, a n d Pare m y d). T hat is, a p pr o xi matel y 6 0 % of t he 
ra n d o mize d s u bjects will recei v e o ne dr o p of 2. 5 % p he n yle p hri ne 1 h o ur bef ore treat me nt 
( 1 9 8 s u bjects), a p pr o xi matel y 2 0 % will recei ve o n e dr o p of 1 % tr o pi ca mi de 1 h o ur bef ore 
treat me nt (6 6 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 (6 6 s u bjects). 
T he st u d y treat me nt ( N y x ol or Place b o) will be m as ke d t o b ot h I n vesti gat or a n d st u d y 
s u bjects, as well as O c u p hire. O nl y i n case of me dical e mer g e nc y or o cc urre nce of s eri o us 
a d verse e ve nts ( S A Es ) will t he ra n d o mizati o n c o d e be u n mas ke d b y t he st u d y p har macist a n d 
ma de a vaila ble t o t he I n v esti gat or, Oc u p hire, a n d/ or ot her pers o n n el 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. R ules f or u n mas ki n g a s u bject f or safet y reas o ns are 
f ully descri be d i n t he pr ot oc ol a n d n ot re peate d herei n t his S A P. 
5. 4.  
A n al ysis V ari a bles 
Varia bles t o be s u m marize d i ncl u de d e m o gra p hics a n d baseli ne c h aracteristics, me dical 
( n o n-oc ular)  a n d o c ular  hist or y, c o nc o mita nt me dicati o ns, a n d st u d y dr u g acc o u nta bilit y.  
Efficac y vari a bles i ncl u d e:  
•  P u pil dia meter  
•  Acc o m m o dati o n 
•  P u pillar y li g ht refle x  
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•  B C D V A u n der glar e c o n diti o ns ( B A T) 
•  G lare disc o mf ort meas ur e d o n a 4 -p oi nt scal e:  
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  
S afet y varia bl es  i ncl u de:  
•  C o nj u ncti val h y pere mia ( e ye re d n ess) meas ure d wit h a C or nea a n d C o ntact Le ns 
Researc h U nit (C C L R U ) car d 4- p oi nt scale: 
o  N o ne ( 0) = N or mal A p p ears w hite wit h a s mall n u m ber of c o nj u ncti val bl o o d 
vessel easil y o bser v e 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 re 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 oler a 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  
•  D C N V A (i.e., N ear V A)  
•  B C D V A (i.e., Dista nce V A)  
•  A Es  
•  Vital si g ns ( H R a n d B P) 
•  I O P 
•  S u bject q uesti o n naire 
P K  varia bles i ncl u de:  
•  N y x ol plas ma  c o nce ntrati o n  
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7.  
G E N E R A L C O N SI D E R A TI O N S  
7. 1.  
A n al ysis P o p ul ati o ns 
T he  f oll o wi n g a nal ysis p o p ulati o ns will be defi ne d f or t his st u d y.  
7. 1. 1.  
M o difie d I nte nti o n -t o-Tre at ( mI T T)  
T he  mI T T  P o p ulati o n will  i ncl u de all ra n d o mize d s u bjects w h o  recei ve d 2 dr o ps of st u d y 
treat me nt i n t he st u d y e y e a n d ha d  at least 1 sc he d ule d p ost-treat me nt P D  meas ure me nt  
d uri n g Visit  1. T he  mI T T  P o p ulati o n 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 
anal yze  s electe d s ec o n dar y efficac y e n d p oi nts, wit h  s u bjects i ncl u de d i n t heir ra n d o mize d 
treat me nt re g ar dless of t h e treat me nt t he y act uall y recei v e d. 
7. 1. 2.  
Per Pr ot oc ol  P o p ul ati o n (P P ) 
T he  P P  P o p ulati o n i ncl u des all s u bjects i n t he mI T T  P o p ulati o n w h o  ha d  2 dr o ps of st u d y 
treat me nt i n t he st u d y e y e, ha d  all sc he d ul e d P D  meas ure me nts  d uri n g Visit  1, ha d  a n 
i ncreas e of > 0. 2 m m  i n P D  i n t he st u d y e ye at 0 mi n utes  c o m pare d  t o bas eli ne  (- 1 h o ur), a n d 
ha d  n o maj or  pr ot oc ol de viati o ns c o nsi dere d  t o ha ve si g nifica nt i mpact  o n treat me nt 
o utc o me. 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, 
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 v e d. 
7. 1. 3.  
All R a n d o mize d P o p ul ati o n (A R P) 
T he  A R P  wil l 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  ma y  be use d  i n c o nfir mat or y  efficac y a nal yses, 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 v e d. 
7. 1. 4.  
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  recei ve d at least 1 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 safet y varia bl es,  usi n g t he treat me nt t he y act uall y 
recei v e d. 
7. 1. 5.  
P K  P o p ul ati o n 
T he P K P o p ulati o n will i ncl u de all N y x ol s u bjects w h o ha d at least o ne P K sa m ple ta ke n at 
a n y p ost-treat me nt ti me p oi nt. T he  P K  p o p ulati o n will  be use d  t o s u m marize P K  varia bles,  
usi n g t he treat me nt t he y act uall y recei ve d. 
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7. 3. 3.  
H a n dli n g of E arl y Ter mi n ati o n Visit I nf or m ati o n 
I n t he e ve nt t hat a s u bj ect is ter mi nate d earl y fr o m t his st u d y o n Da y 1, t he earl y ter mi nati o n 
data f or safet y varia bles will be assi g ne d t o t he cl osest sc he d ule d ti me p oi nt o n Da y 1. If t he 
cl osest ti me p oi nt has vali d data, t he earl y ter mi nati o n data will be assi g ne d t o t he ne xt 
a vaila ble ti me p oi nt. 
7. 3. 4.  
P o oli n g of I n vesti g ati o n al Sites 
T he data fr o m all st u d y ce nters  will be p o ole d t o g et her f or all pla n ne d a n al yses . 
7. 3. 5.  
C o di n g C o n ve nti o ns f or E ve nts a n d Me dic ati o ns  
All A E s a n d me dical hist or y  will be ma p pe d t o t he Me dical Dicti o nar y f or Re g ulat or y 
Acti vities ( Me d D R A Versi o n 2 4. 1) s yste m f or re p orti n g ( preferre d ter m a n d b o d y s yste m). 
Pri or a n d c o nc o mita nt m e dicati o ns will  be c o de d usi n g W H O -D D ( W orl d Healt h 
Or ga nizati o n Dr u g Dicti o nar y) ( Gl o bal Versi o n 2 0 2 0- 0 9- 0 1). 
7. 3. 6.  
A n al ysis S oft w are 
Data ma ni p ulati o n, ta b ulati o n of descri pti ve statistics, calc ulati o n of i nfere ntial statistics, a n d 
gra p hical r e pres e ntati o ns will be perf or me d pri maril y usi n g S A S ( release 9. 4 or hi g her ) f or 
Wi n d o ws. If t he use of ot her s oft ware is warra nt e d, t he fi nal C S R  will detail w hat s oft ware 
was use d a n d f or w hat p ur p oses. 
7. 3. 7.  
St u d y  D at a  
St u d y data i de ntifie d i n t he sc he d ule f or ti me a n d e ve nts (Ta ble 1 ) are c ollecte d, a n d s o urce 
verifie d, o n t he electr o ni c data ca pt ure ( E D C) Dat atra k O ne v ersi o n 1 4. 5. 5. A nal ysis of 
plas ma sa m ples f or N y x ol c o nce ntrati o n det er mi nati o ns will be perf or me d b y a ce ntral P K 
la b orat or y ( Micr o C o nsta nts n o w Bi o Agil y ti x). 
All st u d y data will be f or m ulate d i nt o re g ulat or y-c o m plia nt data sets t o pr o vi de tra ns pare nc y, 
tracea bilit y, a n d i nte grit y of trial a nal ysis res ults fr o m t he c ollecti o n s o urce. O bser ve d st u d y 
data will be ma p pe d t o t h e Cli nical Data I nterc ha n ge Sta n dar ds C o ns orti u m ( C DI S C ) St u d y 
Data Ta b ulati o n M o del ( S D T M) a n d ser v e as t he s o urce dat a fr o m t he trial. All st u d y 
a nal yses will be c o m plet e d usi n g a nal ys is data sets t hat are deri ve d fr o m t h e S D T M a n d 
f oll o w t he C DI S C A nal ysis Data M o del ( A D a M) arc hitect ure.   
T he met h o ds f or pr o gra m mi n g t he C DI S C S D T M a n d A Da M dat a sets are descri be d i n 
Fi g ure 1.  
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descri pti ve ta ble cate g ori es wit hi n eac h treat me nt ma y occ ur if s u c h ela b orati o ns are t h o u g ht 
t o be usef ul. 
All st u d y-relate d data c ollecte d will be pres e nte d i n listi n gs. St u d y-relate d data n ot s u bject t o 
a nal ysis acc or di n g t o t his pla n will n ot a p pear i n a n y ta bles or gra p hs b ut will be i ncl u de d i n 
t he data listi n gs. 
7. 4. 2.  
I nteri m A n al yses a n d D at a M o nit ori n g 
N o f or mal i nteri m a nal ysis or safet y m o nit ori n g c o m mittee is pla n ne d f or t his st u d y.  
7. 4. 3.  
Fi n al A n al ysis a n d P u blic ati o n of St u d y Res ults  
T he f i nal a n al ysis will be c o m plete d after all s u bjects ha ve c o m plete d t h e st u d y.   
7. 5.  
M ulti ple Testi n g Pr oce d ures  
T here will be n o a dj ust me nts f or m ulti plicit y a n d n o f or mal m ulti ple testi n g pr oce d ures are t o 
be i m ple me nte d wit h t his a nal ysis pla n.  
7. 6.  
B aseli ne V al ues  
Baseli ne val ues are t he v al ues o btai ne d pri or t o a n y dr u g a d mi nistrati o n o n Da y 1 (st u d y dr u g 
or m y driatic a ge nt), us u all y at Scree ni n g or t he - 1 h o ur ti me p oi nt. 
 
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8.  
S U M M A R Y O F S T U D Y D A T A  
8. 1.  
S u bject Dis p ositi o n  
A s u m mar y of t he a n al ysis sets i nc l u des t he n u m ber a n d perce nta ge of s u bj ects b y treat me nt 
gr o u p a n d o ver all f or t he f oll o wi n g cate g ories: s u bjects i n t he A R P, s u bjects i n t he S P, 
s u bjects i n t he mI T T  P o p ulati o n, s u bjects i n t he P P P o p ulati o n, a n d s u bjects i n t he P K 
P o p ulati o n. All perce nta ges will be base d o n t he n u m ber of s u bjects i n t he A R P.  
E n d of trial i nf or mati o n will als o be s u m marize d i n t his ta ble, i ncl u di n g t he n u m ber of 
s u bjects c o m pleti n g t he st u d y, t he n u m ber of s u bjects w h o pre m at urel y dis c o nti n ue d t he 
st u d y wit h reas o ns f or wit h dra wal, t he n u m ber of s u bjects c o m pleti n g t he st u d y me dicati o n 
d osi n g, a n d t he n u m ber of s u bjects w h o pre m at urel y disc o nti n ue d t he st u d y me dicati o n wit h 
reas o ns f or st u d y me dicati o n disc o nti n uati o n.   
A b y -s u bject data listi n g of st u d y c o m pleti o n i nf or mati o n i ncl u di n g t he reas o n f or pre mat ure 
st u d y wit h dra wal, if a p plica ble, will be pres e nte d. 
8. 2.  
Pr ot oc ol D e vi ati o ns 
Maj or pr ot oc ol de viati o ns, as deter mi ne d b y a S p o ns or bli n de d re vie w of t he data pri or t o 
data base l oc k a n d u n bli n di n g of t he st u d y, ma y res ult i n t he re m o val of a s u bject’s data fr o m 
t he P P P o p ulati o n. T he S p o ns or or desi g nee will be res p o nsi ble f or pr o d uci n g t he fi nal 
de viati o n file; t his file will i ncl u de a descri pti o n of t he pr ot oc ol de viati o n a n d clearl y i de ntif y 
w het her t his vi olati o n warra nts e xcl usi o n fr o m t he P P P o p ulati o n. T his file will be fi nalize d 
pri or t o data base l o c k. 
All pr ot oc ol de viati o ns will be prese nte d i n a b y- s u bject data listi n g, wit h a fla g t o i n dicate if 
a de viati o n was c o nsi d er e d maj or .  
8. 3.  
De m o gr a p hics a n d B aseli ne C h ar acteristics  
S u bject de m o gra p hic dat a a n d baseli ne c haracteristics will be ta b ulate d a n d s u m marize d 
descri pti vel y b y treat me nt gr o u p a n d o verall. T he de m o gra p hic dat a a n d baseli ne 
c haracteristics will be s u m marize d f or t h e mI T T  P o p ulati o n, P P  P o p ulati o n , S P, P K 
p o p ulati o n, a n d A R P. If t he mI T T p o p ulati o n is e q ui vale nt t o a n y of t he ot h er p o p ulati o ns, 
t he n o nl y t he mI T T v ersi o n will be ge nerat e d rat h er t ha n re peati n g e q ui v ale nt s u m maries 
T he de m o gra p hi cs c o nsist of a ge ( y ear), se x, race, et h nicit y, a n d st u d y e ye ( O D), iris c ol or 
(li g ht bl ue, dar k bl ue, bl u e wit h peri p u pillar y br o w n, unif or m gree n , gree n wit h br o w n iris 
ri n g, ce ntral br o w n a n d p eri p her al gree n , br o w n wit h s o me peri p heral gree n, or br o w n), 
iri des t y pe (li g ht or dar k), e ye glasses- weari n g stat us ( yes or n o) ( dista n ce visi o n or n ear 
visi o n), a n d m y driatic a g e nt ( p he n yle p hri n e, tr o pica mi de, or Pare m y d). A s u bject’s a ge i n 
year s is cal c ulate d usi n g t he date of t he i nf or me d c o nse nt a n d date of birt h. A ge will be 
s u m marize d usi n g descri pti ve statistics. T he n u m ber a n d perce nta ge of s u bjects b y se x, race, 
et h nicit y, st u d y e ye, iris c ol or, iri des t y pe, dista nce visi o n/ near visi o n c orrecti o n stat us, a n d 
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m y driatic a ge nt  will als o be re p ort e d. Perce nta ges will  be base d o n t he t otal n u m ber of 
s u bjects i n t he st u d y p o p ulati o n prese ntati o n. 
T he f oll o wi n g baseli ne c haract eristics will be s u m marize d  f or st u d y e ye a n d fell o w e ye, 
usi n g descri pti ve statistics : 
•  P u pil dia meter ( - 1 H o ur) 
•  M a x p u pil dia meter ( 0 Mi n utes)  
•  Acc o m m o dati o n (- 1 H o ur)  
•  Acc o m m o dati o n ( 0 Mi n utes) 
•  B C D V A ( - 1 H o ur) 
•  B C D V A ( 0  Mi n utes) 
•  D C N V A ( - 1 H o ur) 
•  D C N V A ( 0 Mi n utes)   
•  I O P (Scree ni n g ) 
Al l de m o gra p hic a n d b as eli ne i nf or mati o n will be prese nte d i n b y- s u bject listi n gs. 
8. 4.  
Me dic al Hist or y  
T he n u m ber a n d p erce nt of s u bjects wit h i n di vi d ual me dical hist ories will be s u m marize d f or 
all s u bjects  b y treat me nt gr o u p a n d o verall. N o n -o c ular a n d o c ular me dical hist or y will be 
s u m marize d se paratel y.  
Me dical hist or y will be c o de d usi n g t he Me d D R A Versi o n 2 4. 1. T he n u m b er a n d perce nta ge 
of s u bjects wit h a n y me dical hist or y will be s u m marize d o v erall a n d f or eac h s yste m or ga n 
class ( S O C) a n d pr eferre d ter m ( P T). Perce nta ges will be calc ulate d b ase d o n n u m ber of 
s u bjects i n t he S P.   
S u bject me dical hist or y data i ncl u di n g s pecific d etails will be prese nte d i n b y- s u bject 
listi n gs. 
8. 5.  
Pri or a n d C o nc urre nt Me dic ati o ns  
T he n u m ber a n d p erce nta ges of all c o nc o mita nt m e dicati o ns will be s u m marize d b y 
treat me nt gr o u p, A n at o mical T hera pe utic C he mical ( A T C) le vel 4, a n d P T. T he t otal n u m ber 
of c o nc o mita nt me dicati o ns a n d t he n u m ber a n d p erce nt a ges of s u bjects wit h at least 1 
c o nc o mita nt me dicati o n will be s u m marize d b y treat me nt gr o u p. All s u m maries will be 
perf or me d usi n g t he S P.   
A c o nc o mita nt me dicati o n is defi ne d as a n y me di cati o n ta ke n o n or after t h e da y of first 
e x p os ure t o st u d y dr u g. 
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Eac 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 will 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, 
, a n d t he res pecti ve baseli n e val ue i ncl u de d as 
t he c o variat e. Note t hat m ost sec o n dar y efficac y e n d p oi nts are i n rel ati o n t o baseli ne  
(- 1 h o ur), w h er eas s o me p u pil dia meter  e n d p oi nts are i n relati o n t o ma x ( 0 mi n utes).  
Eac h A N C O V A will be perf or me d usi n g t he mI T T  a n d P P  p o p ulati o ns. 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 orr ecte d L S M, its 9 5 % CI a n d ass ociate d p - val ue. Li ne gra p hs dis pla yi n g t he mea n 
a n d S E of t he c ha n g e fr o m ma xi m u m ( 0 mi n utes) p u pil dia meter val ue will be prese nt e d.  
F or eac h of t h e sec o n d ar y e n d p oi nts relate d t o perce nt of s u bjects ac 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,  
 
, a n d t he res p ecti ve baseli ne  as a c o variate. F or eac h 
a nal ysis, t he perce nta ge of s u 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 u e will be pr o vi de d.  
T he a nal ysis of t he 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 
sa vi n gs a nal ysis) e n d p oi nt will be perf or me d f or t he P P P o p ulati o n usi n g a C o x pr o p orti o nal 
hazar ds re gr essi 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 baseli ne 
p u pil dia meter  as a c o variate.  S u bjects w h o d o n ot ret ur n t o ≤ 0. 2 m m fr o m base li ne p u pil 
dia meter b y t he 6 h o ur ti me p oi nt will ha ve t heir ti me t o ret ur n ce ns ore d at 8 h o urs. E xa m ple 
S A S c o de is as f oll o ws: 
pr oc p hr e g d at a = effic ac y d at a pl ots ( o v erl a y)=s ur vi v al;  
 
cl ass trt n a m e a g e nt iri d es ; 
 
m o d el ti m et or et ur n *c nsr( 1 ) = trt n a m e a g e nt iri d es b as e;  
 
h az ar dr ati o trt n a m e; 
 
b as eli n e c o v ari at es = effic ac y d at a o ut = Pr e d 1 s ur vi v al = _ all _/ dir a dj gr o u p =trt n a m e;  
 
o ds o ut p ut  h az ar dr ati os = hr;  
r u n; 
T he o ut p ut fr o m t he m o del will i ncl u de t he hazar d rati o  c o m pari n g treat me nt gr o u ps, its 9 5 % 
CI a n d ass ociate d p -val u e . S ur vi val pl ots, ge nerat e d fr o m t he C o x m o del d escri be d a b o ve, 
will als o be ge ner ate d.  Ti me t o ret ur n  t o baseli ne (- 1 h o ur) p u pil dia meter will be meas ure d 
be gi n ni n g at t he Ma x ( 0 h o ur) ti me p oi nt. 
I n a d diti o n, eac h s ec o n dar y efficacy e n d p oi nt will be a nal yze d b y 
 
 usi n g t he sa me m o del i n dicate d a b o ve b ut wit h o ut m y driatic a ge nt or iri des 
as a fact or, as a p pr o priat e. Eac h m y dri atic a ge nt will be a nal yze d i n di vi d u all y, a n d a n 
a d diti o nal m y driatic a ge nt s u b gr o u p, c o m bi ni n g 1 % tr o pica mi de a n d t h e Pare m y d s u bjects 
i nt o a “tr o pica mi de” gr o u p, will be  a nal yze d. A nal yses of e n d p oi nts relate d t o 
acc o m m o dati o n will o nl y be c o m plete d b y m y driatic a ge nt. Acc o m m o dati o n will be 
prese nte d usi n g di o pters, w hic h will be c o n vert e d fr o m c m usi n g di o pters = 1 0 0/c m. 
E x pl orat or y a nal ys es 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 y e 
a n d fell o w e y e wit hi n t he sa me s u bject.  Cate g ori cal varia bles will be a n al yze d as descri be d 
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a b o ve f or t h e pri mar y efficac y e n d p oi nt. C o nti n u o us varia bles will be a n al yze d usi n g a 
mi xe d m o del wit h e ye t y pe (st u d y e ye or fell o w e ye), my driatic a g e nt , a n d li g ht/ dar k iri des 
as fi xe d effects, s u bject as a ra n d o m effect,  a n d t h e a vera ge b aseli ne  p u pil dia meter acr oss 
e ye t y pe as a c o variate.  
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1 0.  
S A F E T Y A N A L Y S E S  
All safet y a nal yses will be c o n d ucte d usi n g t he S P. All safet y a nal yses will be c o m plete d 
usi n g t he act ual treat me nt a s u bject recei ve d . 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 e x ce pt f or t he li mite d sit uati o ns descri be d i n 
Secti o n 7. 3. 2 . 
All safet y data will be pr ese nte d i n b y -s u bject listi n gs. U nsc he d ule d ass ess me nts will n ot be 
s u m marize d b ut will be i ncl u de d i n t he listi n gs.  
1 0. 1.  
A d verse E ve nts  
A Es will be c o de d usi n g Me d D R A, Versi o n 2 4. 1.    
Treat me nt -e mer ge nt a d v erse e v e nts ( T E A Es) are defi ne d as a n y A E  t hat b e gi ns or w orse ns 
after i nitiati o n of t he i n vesti gati o nal pr o d uct a n d t hr o u g h t he s u bject’s last st u d y visit (st u d y 
c o m pleti o n or earl y ter mi nati o n ). 
If t he o nset of a n A E is o n or after t h e date of first d ose of st u d y me dicati o n  or is i ncreasi n g 
i n se verit y after first d ose of st u d y me dicati o n, t he n t he A E will be c o nsi dere d  treat me nt 
e mer ge nt.  
O nl y T E A Es will be s u m marize d; all A Es ( T E A E, n o n - T E A E) will be i ncl u de d i n a b y-
s u bject listi n g. A se parat e listi n g of A Es f or pe diatric s u bjects will be pr o vi de d. 
T he n u m ber a n d p e rce nt of s u bjects wit h a n y T E A Es will be s u m marize d b y S O C  a n d P T  b y 
treat me nt gr o u p a n d o v er all. At eac h le v el of ta b ul ati o n ( e. g., at t he P T  le vel), su bjects  will 
be c o u nte d o nl y o nce if t he y ha d m ore t ha n o n e s uc h e ve nt r e p orte d d uri n g t he A E c ollecti o n 
peri o d. A se parate s u m m ar y b y S O C, P T , a n d treat me nt gr o u p will be c o m plete d b y 
m y driatic a ge nt.  
N ote t hat i n Me d D R A, o c ular e v e nts are c o de d t o t he S O C of “ E ye Dis or d ers ”. 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 oc ular a n d n o n- oc ular a d verse e v e nts . 
T he f oll o wi n g s u m mar y ta bles will be prese nt e d f or T E A E  data:  
•  
O verall s u m mar y of T E A Es  
•  
S u m mar y of T E A Es b y S O C a n d P T  
•  
S u m mar y of T E A Es b y S O C, P T , a n d b y greatest  relati o ns hi p le vel t o st u d y dr u g 
( n ot relate d, u nli kel y r elate d, p ossi bl y relate d, pr o ba bl y relate d, d efi nitel y relate d, or 
u n k n o w n) 
•  
S u m mar y of T E A Es b y S O C, P T , a n d ma xi m u m se verit y ( mil d, m o derate, se vere) 
•  
S u m mar y of seri o us T E A Es b y S O C a n d P T  
•  
S u m mar y of T E A Es lea di n g t o wit h dra wal fr o m t he st u d y b y S O C a n d P T  
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 2 0 2 2- 0 3- 1 4 
 
C O N FI D E N TI A L  
 
P a g e  | 3 1  
•  
S u m mar y of T E A Es lea di n g t o st u d y me dicati o n  disc o nti n uati o n b y S O C a n d P T 
 
1 0. 2.  
De at hs, Seri o us A d vers e E ve nts a n d Ot her Si g nific a nt A d verse E ve nts  
1 0. 2. 1.  
De at hs  
T he A E listi n g will i ncl u de all A Es, i ncl u di n g deat hs, re gar dless of ca us alit y; o ne of t he 
c ol u m ns i n t he listi n g will s pecif y w het her t he A E was fatal.     
1 0. 2. 2.  
Seri o us A d verse E ve nts  
T he A E listi n g will i ncl u de all A Es,  i ncl u di n g S A Es; o ne of t he c ol u m ns i n t he listi n g will 
s pecif y w het h er t he A E was a n S A E.     
1 0. 2. 3.  
A d verse E ve nts Le a di n g t o Wit h dr a w al fr o m t he St u d y  
T he A E listi n g will i ncl u de all A Es, i ncl u di n g A Es lea di n g t o wit h dra wal fr o m t he st u d y; o ne 
of t he c ol u m ns i n t he listi n g will s pecif y w het her t he A E le d t o wit h dra w al fr o m t he st u d y.   
1 0. 2. 4.  
A d verse E ve nts Le a di n g t o Disc o nti n u ati o n of St u d y Me dic ati o n  
T he A E listi n g will i ncl u de all A Es, i ncl u di n g A Es lea di n g t o disc o nti n uati o n of st u d y 
me dicati o n; o ne of t he c ol u m n s i n t he listi n g will s pecif y w het h er t he A E l e d t o 
disc o nti n uati o n of st u d y me dicati o n.   
1 0. 3.  
C o nj u ncti v al H y pere mi a  
R es ults fr o m t he co nj u ncti val h y pere mia assess me nt, meas ure d wit h a C C L R U car d 4 - p oi nt 
scale, will be s u m marize d descri pti vel y usi n g c o u nts a n d perce nta ges f or eac h treat me nt 
gr o u p at eac h ti me p oi nt (- 1 h o ur, 0 mi n utes, 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) f or t he o bser ve d val u e a n d c h a n ge fr o m baseli ne  
(- 1 h o ur). Se p arat e s u m ma ries will be create d f or t he st u d y e ye a n d the fell o w  e ye. T he 
cate g orical s u m mar y will be re peate d f or pe diatric s u bjects.  
A d diti o nall y , c o nj u ncti val h y pere mia will be s u m marize d as a c o nti n u o us varia ble. 
Treat me nts will be c o m p are 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. 
1 0. 4.  
S u bjecti ve O c ul ar T oler a bilit y  
Res ults fr o m t he s u bjecti ve oc ular t olera bilit y  assess me nt, meas ure d o n a 4- p oi nt scale , will 
be s u m marize d d escri pti vel y usi n g c o u nts a n d perce nta ges f or eac h treat me nt gr o u p at t he  
0 mi n ute ti me p oi nt . A d diti o nall y, t he cate g ories “ N o Disc o mf ort” a n d “ Mil d Disc o mf ort” 
will be p o ole d i nt o a si n gle cate g or y a n d s u m mari ze d descri pti vel y, as will t he cate g ori es 
“ M o derat e Disc o mf ort” a n d “ Se vere Disc o mf ort”. Treat me nts will be c o m p are d f or t he t w o 
Pa ge 3 2


Oc u p hire P har m a, I nc. 
 
Fi nal  Versi o n 1. 0 
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P a g e  | 3 2  
p o ole d cate g ori es usi n g a Fis her’s e xact test. Se parate s u m maries will be create d f or t he 
st u d y e ye a n d t he f ell o w e ye.  
1 0. 5.  
Vis u al Ac uit y  
Vis ual ac uit y assess me nts will be s u m marize d at the ti me p oi nts - 1 h o ur, 0 mi n utes,  
6 0 mi n utes, 2 h o urs, 6 h o urs, a n d 2 4 h o urs f or B C D V A a n d - 1 h o ur, 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 f or D C N V A, usi n g letters a n d l o g M A R u nits. O nl y letters will 
be rec or de d  i n t he C R F a n d will be  c o n verte d t o l o g M A R pr o gra m maticall y  as f oll o ws: 
l o g M A R = 0. 0 2 *( S - # Letters), w here S = 5 5 f or B C D V A a n d S = 7 0 f or D C N V A. T he 
val ues of S ar e t he n u m ber of letters rea d e q ui vale nt t o a S nelle n Ac uit y of 2 0/ 2 0. F or 
e xa m ple, f or B C D V A, if t he # letters = 5 5, t he n L o g M A R = 0. 0 2 *( 5 5 - 5 5) = 0. 0 0; if # letters 
> 5 5, t he n L o g M A R is ne gati ve. As a refere n ce  5 letters is e q ui vale nt t o 1 li ne. Se parate 
s u m maries will be create d f or t he st u d y e ye, t he fell o w e ye, a n d b ot h e y es. Treat me nts will 
be c o m par e d  f or t he Safet y P o p ulati o n usi n g t he sa me A N C O V A m o d el 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 b ot h c ha n ge fr o m baseli ne (- 1 h o ur) a n d c h a n ge 
fr o m ma xi m u m ( 0 mi n utes). 
I n a d diti o n, D C N V A  will be a nal yze d b y m y driati c a ge nt, a n d B C D V A a n d D C N V A will be 
a nal yze d b y a ge gr o u p ( < 1 8, 1 8 – 3 0, 3 0 – 4 4, 4 5 – 6 4, a n d ≥ 6 5 ), usi n g t he sa me m o dels as f or 
t he efficac y v aria bles. All D C N V A ta bles will be re peat e d f or t he P P P o p ulati o n. 
1 0. 6.  
Vit al Si g ns  
Descri pti ve statistics of o bser ve d val u es will be prese nte d f or vital si g n dat a at eac h ti me 
p oi nt ( Scree ni n g, 6 h o urs, a n d 2 4 h o urs), i ncl u di n g s yst olic B P ( m m H g), diast olic B P 
( m m H g), a n d H R ( b p m) b y treat me nt gr o u p a n d o verall . C ha n ges fr o m bas eli ne t o eac h 
sc he d ule d p ost-bas eli ne ti me p oi nt will be prese nte d. T he s u m mar y will be re peat e d f or 
pe diatric s u bjects.  
1 0. 7.  
I O P 
O bser ve d v al 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 f ell o w e ye. Tr eat me nts will be c o m pa re 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. T he s u m mar y will be re peat e d f or 
pe diatric s u bjects.  
1 0. 8.  
S u bject Q uesti o n n aire  
S u bject q uesti o n naire val ues will be s u m marize d f or eac h ti me p oi nt (- 1 h o ur, 0 mi n utes,  
6 0 mi n utes, 2 h o urs, 4 h o urs, a n d 2 4 h o urs) b y treat me nt gr o u p. A se parate s u m mar y will be 
creat e d f or eac h m y driatic a ge nt.  
Pa ge 3 3


Oc u p hire P har m a, I nc. 
 
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P a g e  | 3 3  
1 0. 9.  
Ot her S afet y Me as ures  
Uri ne pre g n a nc y tests f or fe males of c hil d beari n g p ote ntial will be prese nte d i n b y- s u bject 
listi n gs. Res ults fr o m bi o micr osc o pic a n d o p ht hal m osc o pic e xa mi nati o n s, w hic h are 
c o m plete d o nl y at Scree ni n g, will als o be prese nte d i n b y- s u bject listi n gs. 
Pa ge 3 4


Oc u p hire P har m a, I nc. 
 
Fi nal  Versi o n 1. 0 
St u d y O PI -N Y X R M - 3 0 2 Statistical A nal ysis Pla n  
 2 0 2 2- 0 3- 1 4 
 
C O N FI D E N TI A L  
 
P a g e  | 3 4  
1 1.  
P K A N A L Y S E S  
All P K  a nal yses will be c o n d ucte d usi n g t he P K p o p ulati o n. All P K a nal yses will be 
c o m plete d us i n g t he act u al treat me nt a s u bject recei ve d. O bser v e d case dat a will be use d; n o 
i m p utati o n will be perf or me d f or missi n g P K  data . 
All P K data will be prese nte d i n b y -s u bject listi n gs. U nsc he d ule d ass ess me nts will n ot be 
s u m marize d b ut will be i ncl u de d i n t he listi n gs.  
Bl o o d sa m pli n g f or N y x ol P K meas ure me nts will be c o n d ucte d i n a s u bset of a p pr o xi matel y 
3 0 a d ult s u bjects at a p pr o xi matel y 2 select st u d y sites. A nal ysis of plas ma s a m ples f or N y x ol 
c o nce ntr ati o n deter mi nati o ns will be perf or me d b y a ce ntral P K la b orat or y. N y x ol plas ma 
c o nce ntr ati o ns will be s u m marize d b y ti me p oi nt (1 5 mi n utes, 6 0 mi n utes, a n d 3 h o urs) usi n g 
descri pti ve statistics. F or c o nce ntr ati o ns t hat are b el o w t he l o wer li mit of q ua ntificati o n 
( L L O Q), t he s u m mar y will use o ne-half t he L L O Q as t he a nal ysis val ue.  
T he f oll o wi n g P K para m eters will be calc ulate d f or p he nt ola mi ne usi n g a m o del-i n de pe n de nt 
a p pr oac h (i.e., n o nc o m p art me ntal a nal ysis) b y a v ali date d p har mac o ki netic s oft ware 
( P h oe ni x Wi n N o nli n Pr ofessi o nal v 8. 3 or hi g her); Tma x , Cma x , A U C( 0-T) , a n d A U C( 0-3). T he 
a p pare nt ter mi nal eli mi nati o n rate c o nsta nt λ (t he rate f or t h e l o g li near p orti o n of t he plas ma 
c o nce ntr ati o n pr ofile ter mi nal p hase) will n ot be calc ulate d si nce a mi ni m u m of t hree 
c o nce ntr ati o ns i n t he ter mi nal p hase ar e re q uire d f or cal c ulati o n of λ. C o nse q ue ntl y, t he T 1/ 2, 
A U C ( 0-i nf), C L/ F, a n d V d/ F will n ot be calc ul ate d.   
A n y ta bles a n d listi n gs of P K para meters will be c o m plete d b y a P K ve n d or a n d will n ot be 
i ncl u de d i n t he S A P T L Fs. 
Defi niti o ns f or Pl as m a P K P ar a meter s: 
A U C ( 0-3) is t he area u n d er t he plas ma c o n ce ntrati o n-ti me c ur ve fr o m ti me zer o t o 3 h o urs as 
calc ulate d b y t he tra pez oi dal r ule; li near - u p l o g- d o w n.   
A U C ( 0-T)  is t he area u n d er t he plas ma c o n ce ntrati o n-ti me c ur ve fr o m ti me zer o t o t he last 
meas ura ble pl as ma c o nce ntrati o n ( C T) as calc ulate d b y t he tra pez oi d al r ule; li near - u p l o g-
d o w n.   
A U C ( 0-i nf) is t he area u n d er t he plas ma c o nce ntrati o n-ti me c ur ve fr o m ti me zer o t o i nfi nit y.  
It is calc ulate d as t h e s u m of t he area fr o m ti me zer o t o t he ti me of t he last q ua ntifia ble 
plas ma c o nce ntrati o n ( C T ) a n d t he area fr o m T t o i nfi nit y, calc ulate d as t he last q ua ntifia ble 
plas ma c o nce ntrati o n di vi de d b y λ , w here λ  is t he ter mi nal eli mi nati o n rate c o nsta nt as 
f oll o ws: 
λ
T
T
C
A U C
A U C
+
=
−
−
)
0
(
i nf)
0(
 
C L/ F  is cleara n ce di vi de d b y bi oa vaila bilit y. It is deter mi ne d b y di vi di n g t he t otal d ose b y 
A U C ( 0-i nf).  
Pa ge 3 5


Oc u p hire P har m a, I nc. 
 
Fi nal  Versi o n 1. 0 
St u d y O PI -N Y X R M - 3 0 2 Statistical A nal ysis Pla n  
 2 0 2 2- 0 3- 1 4 
 
C O N FI D E N TI A L  
 
P a g e  | 3 5  
C m a x  is t he ma xi m u m o bser ve d c o n ce ntrati o n. 
T 1/ 2  is ter mi nal half-life calc ulate d b y l n( 2)/ λ  w here λ  is t he rate c o nsta nt f or t he 
l o g-li near p orti o n of t he t er mi nal p hase. A mi ni m u m of t hree val u es i n t he 
p ost- distri b uti o n p hase of t he plas ma c o nce ntrati o n- ti me c ur ve are re q uire d f or 
calc ulati o n of λ . 
T m a x  is t he ti me t o reac h Cma x . 
V d/ F  is t he v ol u me of distri b uti o n base d o n t he ter mi nal p hase di vi de d b y bi oa vaila bilit y.  
It is deter mi ne d b y di vi di n g C L/ F b y λ .  
λ  is t he a p pare nt ter mi nal eli mi nati o n rate c o nsta nt. 
 
 
 
 
Pa ge 3 6


Oc u p hire P har m a, I nc. 
 
Fi nal  Versi o n 1. 0 
St u d y O PI -N Y X R M - 3 0 2 Statistical A nal ysis Pla n  
 2 0 2 2- 0 3- 1 4 
 
C O N FI D E N TI A L  
 
P a g e  | 3 6  
1 2.  
R E F E R E N C E S  
[ 1] IC H E 9 E x pert W or ki n g Gr o u p. Statistical Pri nci ples f or Cli nical Trials: I C H 
Har m o nize d  Tri partite G ui deli ne, Se pte m ber 1 9 9 8 
[ 2] Ha u c k W M, A n d ers o n S, a n d Marc us S M, S h o ul d We A dj ust f or C o variates i n N o nli near 
Re gressi o n A nal yses of Ra n d o mize d Trials? C o nt r olle d Cli n Tri als 1 9 9 8; 1 9: 2 4 9 – 2 5 6 
 
 
Pa ge 3 7





























Prot_000.pdf: 
 
 
 
CONFIDENTIAL  
 
 
Clinical Study Phase 3 Protocol 
OPI-NYXRM-302 
MIRA-3 
 
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 
 
 
Ocuphire Pharma, Inc. 
37000 Grand River Avenue, Suite 120 
Farmington Hills, MI 48335 
 
 
 
 
 
Version: 
01 
Original: 
23 September 2021 
 
 
CONFIDENTIAL 
This document contains Ocuphire Pharma, Inc. (Ocuphire) information that is confidential, a 
trade secret and/or proprietary in nature. It is loaned to you for your confidential use on behalf 
of Ocuphire and is not to be photocopied, disclosed or transmitted to any other person or party 
who is not covered by a Confidential Disclosure Agreement with Ocuphire. As the Principal 
Investigator you are responsible for the safekeeping and return of this document to Ocuphire 
upon request. You will be sent updated information and/or amendments as they become 
available.


Ocuphire Pharma, Inc. 
 
 
 
 
 
OPI-NYXRM-302
 
 
 
 
 
 
               MIRA-3 
CONFIDENTIAL 
2  
09/23/21 
 
SPONSOR SIGNATURE & CONTACTS 
Study 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-302 
Original Protocol: 
Version 01; 23 September 2021 
Person authorized to sign the protocol and protocol amendment(s) for the Sponsor, Ocuphire 
Pharma, Inc. 
Mina Sooch, MBA 
President & CEO  
Ocuphire Pharma, Inc. 
Telephone: 248-681-9815 (Office) 
 
Telephone: 248-980-6538 (Mobile) 
Email: msooch@ocuphire.com 
 
____________________________________ 
Signature  
 
____________________________________ 
Date 
 
Medical Monitor 
Charles Slonim, MD 
Oculos Development Services, LLC 
Telephone: 813-690-0255 (Mobile) 
Email: chuck.slonim@oculoscr.com 
 
____________________________________ 
Signature  
 
____________________________________ 
Date 
 


Ocuphire Pharma, Inc. 
 
 
 
 
 
OPI-NYXRM-302
 
 
 
 
 
 
               MIRA-3 
CONFIDENTIAL 
3  
09/23/21 
 
INVESTIGATOR’S AGREEMENT 
 
OPI-NYXRM-302 
MIRA-3 
 
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 
 
Version: 
 
 
01 
 
 
Original: 
 
 
23 September 2021 
 
Investigator Agreement: 
I, the undersigned, have reviewed this protocol and I agree to conduct this protocol in accordance 
with Good Clinical Practice, the ethical principles set forth in the Declaration of Helsinki and 
with the U.S. Code of Federal Regulations governing the protection of human subjects 
(21 CFR 50), Institutional Review Boards (21 CFR 56) and the obligations of clinical 
investigators (21 CFR 312). 
 
Signature: ____________________________ Date: _________________________ 
 
Printed Name: _________________________ 
 
 
 


Ocuphire Pharma, Inc. 
 
 
 
 
 
OPI-NYXRM-302
 
 
 
 
 
 
               MIRA-3 
CONFIDENTIAL 
4  
09/23/21 
PROCEDURES IN CASE OF EMERGENCY 
 
EMERGENCY CONTACT INFORMATION 
Role in Study 
Name 
Contact Information 
Clinical Study 
Leader 
Roselyn Judd 
Director of Clinical Operations  
Oculos Development Services, LLC 
586-212-0237 (Mobile) 
Email: roselyn.judd@oculoscr.com 
Medical 
Monitor 
Charles Slonim, MD 
Chief Medical Officer 
Oculos Development Services, LLC 
813-690-0255 (Mobile) 
Email: 
chuck.slonim@oculoscr.com 
 
 
 


Ocuphire Pharma, Inc. 
 
 
 
 
 
OPI-NYXRM-302
 
 
 
 
 
 
               MIRA-3 
CONFIDENTIAL 
5  
09/23/21 
ABBREVIATIONS AND TERMS 
Abbreviation 
Full term 
AE 
adverse event 
ANCOVA 
analysis of covariance 
ARP 
All Randomized Population 
BAT 
Brightness Acuity Tester  
BCDVA 
best-corrected distance visual acuity 
BP 
blood pressure 
°C 
degree Centigrade 
CCLRU 
Cornea and Contact Lens Research Unit 
CI 
confidence interval 
CRA 
clinical research associate 
CRO 
clinical research organization 
°F 
degree Fahrenheit 
DCNVA 
distance-corrected near visual acuity 
eCRF 
electronic Case Report Form 
ETDRS 
Early Treatment Diabetic Retinopathy Study 
FDA 
Food and Drug Administration 
GCP 
Good Clinical Practice 
HR 
heart rate 
IB 
Investigators’ Brochure 
ICH 
International Council for Harmonisation 
IOP 
intraocular pressure 
IRB 
Institutional Review Board 
ITT 
Intent-to-Treat 
IUD 
intrauterine device 
LC-MS/MS 
liquid chromatography-tandem mass spectrometry 


Ocuphire Pharma, Inc. 
 
 
 
 
 
OPI-NYXRM-302
 
 
 
 
 
 
               MIRA-3 
CONFIDENTIAL 
6  
09/23/21 
LSM 
least squares mean 
MedDRA 
Medical Dictionary for Regulatory Activities 
mITT 
Modified Intent-to-Treat 
Nyxol 
0.75% Phentolamine Ophthalmic Solution or 1% Phentolamine 
Mesylate Ophthalmic Solution (Nyxol®) 
OD 
oculus dexter (right eye) 
OR 
odds ratio 
OS 
oculus sinister (left eye) 
OTC 
over-the-counter 
OU 
oculus uterque (both eyes) 
Paremyd® 
1% hydroxyamphetamine hydrobromide/0.25% tropicamide 
PD 
pupil diameter 
PK 
pharmacokinetic 
PP 
Per Protocol 
RAF 
Royal Air Force 
SAE 
serious adverse event 
SOC 
system organ class 
SP 
Safety Population 
TEAE 
treatment-emergent adverse event 
US 
United States 
USP 
United States Pharmacopeia 
 
 


Ocuphire Pharma, Inc. 
 
 
 
 
 
OPI-NYXRM-302
 
 
 
 
 
 
               MIRA-3 
CONFIDENTIAL 
7  
09/23/21 
1. STUDY SUMMARY 
 
Study Number  
OPI-NYXRM-302  
Clinical Phase  
Phase 3  
Type of Study  
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  
Name of Investigational 
Product  
Nyxol® Eye Drops – 0.75% Phentolamine Ophthalmic Solution   
Duration of Study   
2 days, including screening/treatment and follow-up  
Rationale  
Nyxol is a once-daily preservative-free eye drop formulation of 
phentolamine, which is a non-selective alpha-1 and alpha-2 adrenergic 
antagonist that acts on the adrenergic receptors and inhibits contraction 
of the smooth muscle. Phentolamine inhibits contraction of the iris 
dilator muscle, resulting in a smaller pupil size. Pharmacologically 
induced mydriasis is achieved either by stimulating the iris dilator 
muscle with the use of direct or indirect alpha-1 agonists (eg, 
phenylephrine, hydroxyamphetamine), by blocking the iris sphincter 
muscle with the use of muscarinic antagonists or cycloplegic drugs (eg, 
tropicamide) or with a combination (eg, Paremyd®, which is 
1% hydroxyamphetamine hydrobromide and 0.25% tropicamide). 
Nyxol, either by directly antagonizing the alpha-1 receptor or by 
indirectly antagonizing the pupil dilation effect of muscarinic blocking, 
can expedite the reversal of pharmacologically induced mydriasis prior 
to natural reversal.  
Study Objectives  
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 systemic exposure of Nyxol based on 
pharmacokinetic (PK) sampling 
• 
To evaluate the safety of Nyxol  
• 
To evaluate any additional benefits of the reversal of 
pharmacologically induced mydriasis  
The Sponsor intends to use this 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.”  


Ocuphire Pharma, Inc. 
 
 
 
 
 
OPI-NYXRM-302
 
 
 
 
 
 
               MIRA-3 
CONFIDENTIAL 
8  
09/23/21 
Study Design  
This is a randomized, parallel-arm, double-masked, placebo-controlled 
Phase 3 study in approximately 330 randomized subjects evaluating 
the safety and efficacy of Nyxol in subjects with pharmacologically 
induced mydriasis. Following the successful completion of screening, each 
subject will be randomized to mydriatic agent (unmasked) and treatment 
(masked). Treatment randomization will be 2:1 (Nyxol or placebo 
[vehicle], respectively). Stratification by iris color will be 1:1 (light or 
dark irides) as shown in Appendix 1.  
The mydriatic agent randomization will be 3:1:1 (2.5% phenylephrine, 
1% tropicamide, or Paremyd, respectively). Approximately 60% of the 
randomized subjects will receive 1 drop of 2.5% phenylephrine in both 
eyes (OU) 1 hour before treatment (198 subjects), approximately 20% will 
receive 1 drop of 1% tropicamide OU 1 hour before treatment 
(66 subjects), and approximately 20% will receive 1 drop of Paremyd OU 
1 hour before treatment (66 subjects). Subjects will have 2 drops of study 
treatment (Nyxol or placebo) administered in the study eye (right eye 
[OD]). Each drop will be instilled 5 minutes apart. Subjects will have only 
1 drop of study treatment administered in the fellow eye (left eye [OS]).   
The study eye and fellow eye will both be evaluated at all assessments 
unless otherwise specified.   
Blood sampling for Nyxol PK measurements will be conducted in a subset 
of approximately 30 adult subjects at approximately 2 select study sites.   
Patient Population   
Approximately 330 healthy subjects  
Inclusion Criteria  
1. Males or females ≥ 12 years of age  
2. Ability to comply with all protocol-mandated procedures 
independently and to attend all scheduled office visits  
3. Adults (≥ 18 years of age) willing to give written informed consent 
to participate in this study. Children aged 12-17 years to 
provide signed assent form, as well as a separate parental/Legal 
Guardian consent 
Exclusion Criteria  
Ophthalmic (in either eye):  
1. Clinically significant ocular disease as deemed by the Investigator 
(eg, glaucoma, corneal edema, uveitis, severe keratoconjunctivitis 
sicca) that might interfere with the study  
2. Unwilling or unable to discontinue use of contact lenses at 
screening until study completion 
3. Unwilling or unable to suspend use of topical medication at 
screening until study completion  
4. Ocular trauma, ocular surgery, or non-refractive laser treatment 
within the 6 months prior to screening  
5. Use of any topical prescription or over-the-counter (OTC) 
ophthalmic medications of any kind within 7 days of screening, 


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with the exception of lid scrubs with OTC products (eg, 
OCuSOFT® lid scrub, SteriLid®, baby shampoo, etc.)   
6. Recent or current evidence of ocular infection or inflammation in 
either eye (such as current evidence of clinically significant 
blepharitis, conjunctivitis, or keratitis). Subjects must be symptom 
free for at least 7 days prior to screening 
7. History of herpes simplex or herpes zoster keratitis 
8. Closed or very narrow angle that in the Investigator’s opinion is 
potentially occludable if the subject’s pupil is dilated  
9. History of any traumatic (surgical or nonsurgical) or nontraumatic 
condition affecting the pupil or iris  
10. History of cauterization of the punctum or punctal plug (silicone or 
collagen) insertion or removal  
11. Known allergy, hypersensitivity, or contraindication to any 
component of the phentolamine ophthalmic solution or to any 
component of the mydriatic agents or vehicle formulation  
12. Prior participation in a study involving the use of Nyxol for the 
reversal of mydriasis 
Systemic:  
13. Known hypersensitivity or contraindication to α- and/or β-
adrenoceptor antagonists  
14. Clinically significant systemic disease (eg, uncontrolled diabetes, 
myasthenia gravis, cancer, hepatic, renal, endocrine, or 
cardiovascular disorders) that might interfere with the study  
15. Initiation of treatment with or any changes to the current dosage, 
drug, or regimen of any systemic adrenergic or cholinergic 
drugs within 7 days prior to screening or during the study 
(Appendix 5)  
16. Participation in any investigational study within 30 days prior to 
screening  
17. Females of childbearing potential who are pregnant, nursing, 
planning a pregnancy, or not using a medically acceptable form of 
birth control. Acceptable methods include the use of at least one of 
the following: intrauterine device (IUD), hormonal (oral, injection, 
patch, implant, ring), barrier with spermicide (condom, diaphragm), 
or abstinence. A female is considered to be of childbearing potential 
unless she is premenstrual, 1 year postmenopausal, or 3 months 
post-surgical sterilization. All females of childbearing 
potential, including those with post-tubal ligation, must have a 
negative urine pregnancy test result at Visit 1/Screening and must 
intend to not become pregnant during the study  
18. Resting heart rate (HR) outside the normal range (50-110 beats per 
minute) at the Screening Visit. Heart rate may be repeated only 


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once if outside the normal range following at least a 5-minute rest 
period in the sitting position  
19. Hypertension with resting diastolic blood pressure (BP) 
> 105 mmHg or systolic BP > 160 mmHg at the Screening Visit. 
Blood pressure may be repeated only once if outside the specified 
range following at least a 5-minute rest period in the sitting position 
Visit Schedule  
At Visit 1, subjects will be screened for study eligibility. After screening, 
eligible subjects will be randomized 2:1 to one of the 2 treatment arms 
(Nyxol or placebo, respectively) and will receive 1 of 3 approved mydriatic 
agents approximately 1 hour prior to receiving study treatment. 
Randomization will be 3:1:1 to mydriatic agent (2.5% phenylephrine, 1% 
tropicamide, or Paremyd, respectively). Each subject will receive 1 drop of 
mydriatic agent in each eye. At Visit 1, measurements as defined in Table 
1 will be made before (-1 hour /baseline) and 60 minutes after (max pupil 
dilation/0 minutes) the mydriatic agent instillation in each eye (ie, right 
before the study treatment is administered). Additionally, measurements 
will be taken at 30 minutes, 60 minutes, 90 minutes, 2 hours, 3 hours, 
4 hours, and 6 hours after study treatment dosing.   
At Visit 2 (Follow-Up Visit), which is 1 day after Visit 1, measurements as 
defined in Table 1 will be measured 24 hours after study treatment dosing.   
Number of  
Investigational Sites  
Approximately 16 sites  
Sample Size 
Justification  
A sample size of approximately 330 subjects (approximately 220 treated 
with Nyxol) in this study will result in a total of > 300 subjects treated with 
Nyxol in the reversal of mydriasis program (including prior studies); this 
number of subjects is needed to meet the minimum number of subjects 
exposed to Nyxol to assess safety in this population. The primary efficacy 
endpoint will be met if subjects show a positive effect for Nyxol (α = 0.05, 
two-tailed). A total of 330 subjects (Nyxol, n=220; placebo, n=110) will 
provide > 90% power to detect a difference of 25% between the Nyxol and 
placebo arms in percentage of subjects returning to ≤ 0.2 mm from baseline 
pupil diameter at 90 minutes. This calculation is based on a 20% placebo 
effect and a 45% Nyxol treatment effect.  
Primary Efficacy 
Endpoint  
Percentage of subjects’ study eyes returning to ≤ 0.2 mm from baseline 
(-1 hour) photopic pupil diameter at 90 minutes  
Secondary Efficacy 
Endpoints  
Secondary efficacy endpoints (for the study eye and the fellow eye, unless 
otherwise specified; and binocular for selected visual acuity 
measurements) will include:  
• 
Percentage of subjects returning to ≤ 0.2 mm from baseline 
(-1 hour) photopic pupil diameter at each remaining time point 
(0 minutes, 30 minutes, 60 minutes, 90 minutes [fellow eye], 
2 hours, 3 hours, 4 hours, 6 hours, and 24 hours)  


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• 
Change (in mm) in photopic pupil diameter from max pupil 
dilation (0 minutes) at each time point (30 minutes, 60 minutes, 
90 minutes, 2 hours, 3 hours, 4 hours, 6 hours, and 24 hours)  
• 
Time (hours) to return to ≤ 0.2 mm from baseline (-1 hour) 
photopic pupil diameter (time-savings analysis)   
• 
Percentage of subjects with unchanged accommodation from 
baseline (-1 hour) at 0 minutes, 90 minutes, 2 hours, 3 hours, and 
6 hours   
• 
Change (in diopters) in accommodation from max pupil dilation 
(0 minutes) at 90 minutes, 2 hours, 3 hours, and 6 hours  
• 
Change (in mm) in the pupillary light reflex from max pupil 
dilation (0 minutes) at 90 minutes, 3 hours, and 6 hours (study eye) 
• 
Change (in letters) in best-corrected distance visual acuity 
(BCDVA) under normal photopic lighting versus glare 
conditions (using Brightness Acuity Tester [BAT]) from baseline 
(-1 hour) at 0 minutes, 60 minutes, 2 hours, and 6 hours (study eye) 
• 
Percentage of subjects with unchanged (within ± 4 letters) BCDVA 
under glare conditions (BAT) from baseline (-1 hour) at 
0 minutes, 60 minutes, 2 hours, and 6 hours (study eye) 
• 
Change in glare discomfort from max pupil dilation (0 minutes) at 
60 minutes, 2 hours, and 6 hours (study eye) 
Measurements:  
• Pupil diameter and pupillary light reflex will be measured with 
a NeurOptics VIP-300 pupillometer (mm) 
• 
Accommodation will be measured by the Royal Air Force 
(RAF) Near Point Rule (measured in cm and then converted 
to diopters). Unchanged accommodation from baseline (-1 hour) is 
defined as a change from baseline value ≥ -1, as measured in 
diopters 
• 
Glare testing will be performed using the Marco BAT 2000 
• 
Glare discomfort will be measured on a 4-point scale from 0 (none) 
to 3 (severe) 
The Modified Intent-to-Treat (mITT) Population will be used for the 
primary endpoint analysis and to analyze selected secondary efficacy 
endpoints. The Per Protocol (PP) Population will be used to analyze 
selected secondary efficacy endpoints. Some of the efficacy endpoints will 
be analyzed overall, by mydriatic agent, and by light/dark irides at all time 
points. 
Safety Endpoints  
The primary safety measures are conjunctival hyperemia, impairment in 
visual acuity (BCDVA and distance-corrected near visual acuity 
[DCNVA]), subjective ocular tolerability, and adverse events (AEs). 
Other safety measures include intraocular pressure (IOP), subject 
questionnaire, and systemic safety as measured by HR and BP.  


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Safety analyses will include: 
• 
Change from baseline (-1 hour) in the conjunctival hyperemia 
grading (Cornea and Contact Lens Research Unit [CCLRU] 
images) at each time point (0 minutes, 30 minutes, 60 
minutes, 90 minutes, 2 hours, 3 hours, 4 hours, 6 hours, and 
24 hours) for the study eye and fellow eye   
• 
Change from baseline (-1 hour) in BCDVA at 0 minutes, 
60 minutes, 2 hours, 6 hours, and 24 hours for the study eye, fellow 
eye, and binocular 
• 
Change from baseline (-1 hour) in DCNVA at 0 minutes, 
90 minutes, 3 hours, 6 hours, and 24 hours for the study eye, fellow 
eye, and binocular 
• 
Subjective ocular tolerability at 0 minutes after administration of 
study medication for the study eye and fellow eye 
• 
Change from screening in IOP at 6 hours for the study eye and 
fellow eye 
• 
Change from baseline (-1 hour) in subject questionnaire responses 
at 0 minutes, 60 minutes, 2 hours, 4 hours, and 24 hours  
• 
Change from screening in vital signs (HR and BP) at 6 hours and 
24 hours  
Measurements:  
• 
Conjunctival hyperemia will be assessed visually with a 4-point 
grading scale (0-3) using CCLRU images (Appendix 2)  
• 
BCDVA will be measured in photopic conditions by a 
standard Early Treatment Diabetic Retinopathy Study (ETDRS) 
illuminated chart (on wall or stand) at 4 m (letters recorded)   
• 
DCNVA will be measured in photopic conditions by Near Visual 
Acuity Chart in the Precision Vision Small 914 Illuminator Cabinet 
(light box) at 16 inches (~40 cm) (letters recorded) 
• 
Subjective ocular tolerability will be measured on a 4-point scale 
(0-3)  
• 
IOP will be measured with the Tono-Pen or Goldmann applanation 
tonometer 
• 
Subject questionnaire will be a brief symptom survey (Appendix 3)  
Study Medications, Dose 
and Mode of 
Administration  
Nyxol® Eye Drops (Phentolamine Ophthalmic Solution):  
2 drops (dosed 5 minutes apart) of Nyxol in the study eye (OD) (1 hour 
post mydriatic drug instillation) and 1 drop of Nyxol in the fellow eye 
(OS). 
Placebo (Nyxol vehicle):  
2 drops (dosed 5 minutes apart) of placebo in the study eye (OD) (1 hour 
post mydriatic drug instillation) and 1 drop of placebo in the fellow eye 
(OS).   


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Pharmacokinetics 
At approximately 2 study sites, blood will be drawn from approximately 
30 adult subjects) for Nyxol PK measurement on Visit 1/Day 1 at 
15 minutes, 60 minutes, and 3 hours following administration of study 
medication. For adult subjects participating in PK sampling, a total of 
approximately 18 mL blood will be drawn (3 × 6 mL). 
Duration of Subject 
Participation and Study  
The total length of subject participation is 2 days, with 2 clinic visits, as 
summarized below:  
• 
Screening/Treatment Visit 1/Day 1   
• 
Follow-Up Visit 2/Day 2  
The execution of the entire study (first subject screen through last 
randomized subject) is expected to be approximately 4 to 6 months.  
 
 


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TABLE OF CONTENTS 
SPONSOR SIGNATURE & CONTACTS .......................................................................................................... 2 
INVESTIGATOR’S AGREEMENT.................................................................................................................... 3 
ABBREVIATIONS AND TERMS ....................................................................................................................... 5 
1. 
STUDY SUMMARY .................................................................................................................................... 7 
2. 
INTRODUCTION ...................................................................................................................................... 16 
2.1. 
Investigational products ................................................................................................................... 16 
2.2. 
Findings from nonclinical and clinical studies ............................................................................... 16 
2.3. 
Design justification ........................................................................................................................... 18 
2.4. 
Route of administration, dosage regimen, and treatment period ................................................. 19 
2.5. 
Compliance ........................................................................................................................................ 19 
2.6. 
Study population ............................................................................................................................... 19 
3. 
OBJECTIVES AND PURPOSE ............................................................................................................... 20 
4. 
STUDY DESIGN ........................................................................................................................................ 20 
4.1. 
Primary and secondary endpoints .................................................................................................. 20 
4.2. 
Description and schedule of visits and procedures ........................................................................ 22 
4.3. 
Measures taken to minimize/avoid bias .......................................................................................... 26 
4.4. 
Study medications ............................................................................................................................. 26 
4.4.1. Packaging and labeling ...................................................................................................................... 27 
4.4.2. Storage of study medication and dispensing ...................................................................................... 27 
4.4.3. Study medication administration ........................................................................................................ 27 
4.4.4. Study medication accountability ........................................................................................................ 27 
4.4.4.1. Receipt and disposition of study medication .............................................................................. 27 
4.4.4.2. Return of study medication ........................................................................................................ 27 
4.5. 
Expected duration of subject participation .................................................................................... 27 
4.6. 
Randomization and procedure for breaking the code ................................................................... 28 
4.7. 
Collection of data .............................................................................................................................. 28 
4.8. 
Study medication discontinuation ................................................................................................... 28 
4.8.1. Reasons for withdrawal from study .................................................................................................... 29 
4.8.2. Entire study terminated ...................................................................................................................... 29 
4.8.3. Actions after discontinuation ............................................................................................................. 29 
4.9. 
Completed study ............................................................................................................................... 29 
5. 
SUBJECT INCLUSION AND EXCLUSION CRITERIA ..................................................................... 29 
5.1. 
Subject inclusion criteria ................................................................................................................. 29 
5.2. 
Subject exclusion criteria ................................................................................................................. 29 
6. 
TREATMENT OF SUBJECTS ................................................................................................................ 31 
6.1. 
Treatment adherence ....................................................................................................................... 31 
6.2. 
Concomitant medications ................................................................................................................. 31 
7. 
ASSESSMENT OF EFFICACY ............................................................................................................... 32 
7.1. 
Specification of the efficacy parameters ......................................................................................... 32 
7.2. 
Assessing, recording, and analyzing efficacy parameters ............................................................. 32 
7.2.1. Screening/Day 1 ................................................................................................................................. 33 
7.2.2. Treatment Visit 1/Day 1 ..................................................................................................................... 33 
7.2.3. Follow-Up Visit/Day 2 ....................................................................................................................... 34 
7.2.4. Unscheduled Visits ............................................................................................................................. 34 
7.2.5. Visit Variation .................................................................................................................................... 34 
8. 
PHARMACOKINETIC ASSESSMENTS ............................................................................................... 34 
9. 
ASSESSMENT OF SAFETY .................................................................................................................... 35 
9.1. 
Specification of safety parameters .................................................................................................. 35 


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9.2. 
Assessing, recording, and analyzing safety parameters ................................................................ 35 
9.3. 
Adverse events and serious adverse events ..................................................................................... 35 
9.3.1. Adverse event definitions .................................................................................................................... 36 
9.3.2. Follow-up of subjects after adverse events ........................................................................................ 40 
10. 
STATISTICS .............................................................................................................................................. 40 
10.1. 
Sample size ........................................................................................................................................ 40 
10.2. 
Analysis Populations......................................................................................................................... 40 
10.3. 
Statistical methods ............................................................................................................................ 41 
10.3.1. General considerations .................................................................................................................... 41 
10.3.2. Demographic and baseline characteristics ...................................................................................... 41 
10.3.3. Subject disposition ........................................................................................................................... 41 
10.3.4. Medical history and prior/concomitant medications........................................................................ 41 
10.3.5. Analysis of efficacy ........................................................................................................................... 41 
10.3.6. Analysis of safety .............................................................................................................................. 43 
10.4. 
Procedure for accounting for missing, unused, or spurious data ................................................. 44 
10.5. 
Procedure for reporting deviations from the statistical plan ........................................................ 44 
11. 
DIRECT ACCESS TO SOURCE DATA AND DOCUMENTS ............................................................ 44 
12. 
QUALITY CONTROL AND QUALITY ASSURANCE........................................................................ 44 
13. 
ETHICAL CONSIDERATIONS AND GOOD CLINICAL PRACTICE COMPLIANCE ................ 44 
13.1. 
Good Clinical Practice compliance ................................................................................................. 44 
13.2. 
Institutional Review Board .............................................................................................................. 44 
13.3. 
Protocol deviations/violations .......................................................................................................... 45 
13.4. 
Informed consent and assent requirements .................................................................................... 45 
14. 
DATA HANDLING AND RECORD KEEPING .................................................................................... 46 
14.1. 
Data entry .......................................................................................................................................... 46 
14.2. 
Data quality control and reporting ................................................................................................. 46 
14.3. 
Archiving of data .............................................................................................................................. 46 
14.4. 
Records retention .............................................................................................................................. 46 
14.5. 
Amendments to the protocol ............................................................................................................ 46 
15. 
BIBLIOGRAPHY ...................................................................................................................................... 47 
APPENDIX 1: IRIS COLOR CHART .............................................................................................................. 48 
APPENDIX 2: CONJUNCTIVAL HYPEREMIA GRADING SCALE USING IMAGES FROM 
CCLRU……………………………………………………………………………………………………………49 
APPENDIX 3: SUBJECT QUESTIONNAIRE ................................................................................................. 50 
APPENDIX 4: RANDOMIZATION SCHEMA BY INVESTIGATIONAL TREATMENT, MYDRIATIC 
AGENT, AND IRIS COLOR .............................................................................................................................. 51 
APPENDIX 5: ADRENERGIC AND CHOLINERGIC DRUGS ................................................................... 52 
 
 


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2.  INTRODUCTION 
2.1. Investigational products 
The test product is Nyxol® Eye Drops – 0.75% Phentolamine Ophthalmic Solution (0.75% POS) 
(herein referred to as Nyxol), a non-selective alpha-1 and alpha-2 adrenergic antagonist. Note 
that the concentration of 0.75% refers to phentolamine free base and is the same as and used in 
place of “1% phentolamine mesylate ophthalmic solution”, which was how Nyxol had been 
described in all studies prior to the End of Phase 2 meeting in mid-2020. Placebo control is 
Nyxol vehicle alone. 
2.2. Findings from nonclinical and clinical studies 
Detailed findings from nonclinical and clinical studies and potential risk are provided in the 
Investigators’ Brochure (IB) (2021). 
Nyxol has been assessed in 8 investigator-initiated and sponsored Phase 1, 2, and 3 clinical trials. 
Across all trials, 335 of 567 subjects were exposed to at least 1 dose of Nyxol. 
In prior clinical trials, Nyxol has demonstrated a consistent ability to decrease pupil diameter 
(PD) by approximately 20% (~1 to 2 mm) in both mesopic and photopic conditions with rapid 
onset and durable effects to 24 to 36 hours (OP-NYX-01a2 CSR; OP-NYX-SNV CSR; OPI-
NYXG-201 CSR; OPI-NYXRM-201 CSR; Karpecki 2021; Pepose MIRA-2 2021; Pepose 
VEGA-1 2021; Pepose ORION 2021).    
Nyxol was observed to be well tolerated in all studies at single-doses and multiple daily doses for 
up to 14 days. Safety of the patients in these trials was evaluated by adverse event (AE) 
monitoring, ophthalmic examinations, and vital sign assessments. Across all trials, there were no 
treatment-related serious adverse events (SAEs). No deaths occurred in any of the trials. No 
clinically meaningful changes were observed in physical examinations or vital signs, including 
blood pressure (BP) and heart rate (HR). Adverse events reported were mild to moderate in 
severity, with the most common being transient conjunctival hyperemia and ocular irritation; 
however, Nyxol dosing at or near bedtime was observed to mitigate or minimize these side 
effects during the daytime. 
Phase 2 Reversal of Pharmacologically Induced Mydriasis Study (MIRA-1) 
In OPI-NYXRM-201 (MIRA-1), 32 healthy subjects were randomized at Visit 1 in a 1:1 ratio to 
receive 1 drop of 1% Nyxol (1% Phentolamine Mesylate Ophthalmic Solution) or 1 drop of 
placebo (vehicle) in both eyes (OU) at Visit 1 and 1 drop of the alternative study medication OU 
at Visit 2. One hour before administration of study medication, subjects received 1 drop OU of a 
mydriatic agent (2.5% phenylephrine or 1% tropicamide) to dilate their pupils. Each subject 
received the same mydriatic agent throughout the study. Pupil diameter (PD) was measured 
30 minutes and 1 hour, 2 hours, 4 hours, and 6 hours after dosing at Visit 1 and Visit 2 and 
compared to baseline. 
In this study, Nyxol demonstrated a statistically significant reduction in PD from max pupil 
dilation (0 minutes [60 minutes after mydriatic agent]) compared to placebo at 1 hour, 2 hours, 
4 hours, and 6 hours after treatment. These statistically significant (p<0.0001) and clinical 
meaningful (typically greater than 1 mm reduction) PD reductions were observed at each time 
point, including when stratified by both parasympathetic (tropicamide) and adrenergic 
(phenylephrine) agents.  


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Additionally, Nyxol was effective at returning a statistically significant percent of subjects’ eyes 
to baseline PD compared with placebo treatment at every time point with estimable results, 
regardless of whether the PD threshold used was ≤ 0.5 mm from baseline (2-hour time point) or 
the more stringent, ≤ 0 mm from baseline (4-hour and 6-hour time points). The reduction in time 
needed to achieve reversal of mydriasis in either eye was statistically significant with Nyxol 
treatment compared with placebo treatment, regardless of the threshold examined or the 
mydriatic agent used. In this study, Nyxol led to an average time-savings of 2.2 hours to return 
PD to ≤ 0 mm from baseline compared to placebo. Additionally, 40% of subjects given 
phenylephrine and 30% of subjects given tropicamide achieved a time-savings of more than 
4 hours with Nyxol, based on a post-hoc analysis. 
In a post-hoc analysis, a statistically significant difference was observed in the percentage of 
subjects returning to baseline in the Nyxol group compared to the placebo group (after dilation 
with either phenylephrine or tropicamide) at 2 hours (29% vs 13%, respectively; p=0.03). This 
difference favoring Nyxol widened (68% for Nyxol and 23% for placebo) at 4 hours (p<0.0001).  
In this study, there were no severe AEs, with only mild-to-moderate conjunctival hyperemia that 
resolved in most subjects within 6 hours.  
Phase 3 Reversal of Pharmacologically Induced Mydriasis Study (MIRA-2) 
The OPI-NYXRM-301 study (MIRA-2) was a phase 3, multicenter, randomized, placebo-
controlled, double-masked clinical trial. In MIRA-2, 185 healthy subjects aged ≥ 12 years from 
12 sites were randomized 1:1 to receive Nyxol or placebo (2 drops in the study eye and 1 drop in 
the fellow eye), 1 hour after pupil dilation with 1 of 3 mydriatic agents (2.5% phenylephrine, 1% 
tropicamide, or Paremyd). The primary endpoint of an increase in the percent of study eyes 
returning to baseline PD at 90 minutes after Nyxol compared to placebo was met (Figure 1A 
[green box]). For subjects in the Modified Intent-to-Treat (mITT) Population treated with Nyxol, 
49% had PD return to ≤ 0.2 mm from baseline at 90 minutes compared to only 7% of subjects 
treated with placebo (p<0.0001). At 60 minutes, 28% of study eyes in subjects who received 
Nyxol had returned to baseline PD, compared to only 2% of participants who received placebo. 
The differences in percent of patients returning to baseline were statistically significant across all 
time points from 60 minutes through 6 hours (p<0.0001; Figure 1A). Additionally, multiple 
secondary endpoints met statistical significance, including mean PD, which was significantly 
smaller with Nyxol compared with placebo at all timepoints after 30 minutes (p<0.0001; Figure 
1B). Finally, the mean time to return to baseline PD for subjects in the Per Protocol (PP) 
Population was ~2 hours in Nyxol-treated study eyes compared to ~6 hours in placebo-treated 
study eyes. Efficacy was demonstrated across the mydriatic agents and for subjects with either 
light or dark irides. 
Nyxol treatment was well tolerated in this study. Adverse effects were limited to predominantly 
mild, transient hyperemia and predominantly mild discomfort on drug instillation. Reduction in 
PD with Nyxol did not result in a decrease in best-corrected distance visual acuity (BCDVA). 
There were no discontinuations due to AEs or SAEs. Vital signs were not affected by Nyxol 
treatment. Nyxol was also well tolerated in the pediatric population, with AEs in a single subject, 
who reported transient mild pain upon Nyxol instillation OU.  
 
 


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Figure 1: MIRA-2 Pupil Diameter Results. A. Percent of Subjects Returning to ≤ 0.2 mm 
From Baseline. B. Mean Pupil Diameter Across Time Points. 
A.  
 
 
 
 
 
         B.  
 
2.3. Design justification 
Pupil size is under the control of 2 opposing sets of muscles – the circular constrictor muscles 
controlled by the cholinergic nervous system and the radial dilator muscles, controlled by the 
adrenergic nervous system (Steinhauer 2004; Yoshitomi 1985). The radial dilator muscles 
contain predominantly α-1 adrenergic receptors that can be inhibited by α-1 antagonists (Yu 
2002); therefore, it is possible to inhibit dilation of the pupil through blockade of the radial 
dilator muscles.  
Pharmacologically induced mydriasis is achieved either by stimulating the iris dilator muscle 
with the use of direct or indirect alpha-1 agonists (eg, phenylephrine, hydroxyamphetamine) or 
by blocking the iris sphincter muscle with the use of muscarinic antagonists or cycloplegic drugs 
(eg, tropicamide) or with a combination (eg, Paremyd®, which is 1% hydroxyamphetamine 
hydrobromide and 0.25% tropicamide). Pharmacologically induced mydriasis is commonly used 
to facilitate clinical examination of the retina and other intraocular structures. Typically, 
pharmacologically induced mydriasis dilates the pupil to 6 to 8 mm, with effects lasting 6 to 
24 hours. However, factors such as an individual’s iris pigmentation and age cause variations in 
effect duration, which ranges from hours to days. During this time, individuals may experience 
sensitivity to light, blurred vision, or cycloplegia (loss of accommodation via the temporary 
paralysis of the muscle that allows the eye to focus on near objects). Accelerating mydriatic 
reversal after an eye exam may reduce the duration of these side effects and be beneficial for 
many patients. 
Phentolamine is a non-selective alpha-1 and alpha-2 adrenergic antagonist acting on adrenergic 
receptors and is known to inhibit contraction of the iris dilator muscle, resulting in a smaller 
pupil size. To counteract the dilatory effects of mydriatic agent, Nyxol is proposed to be instilled 
in the eyes post eye exam, allowing a rapid reversal of mydriasis, thereby minimizing the 
duration of side effects and discomfort post exam.  
Although accommodation will be studied across various mydriatic agents, a decrease in the 
amplitude of accommodation with mydriasis would only be expected with the use of mydriatic 
agents that inhibit muscarinic receptors, including tropicamide and Paremyd. These drugs elicit 


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cycloplegia, or paralysis of the ciliary muscle of the eye, resulting in a loss of accommodation. 
Phenylephrine is a sympathetic agonist that shows mydriatic effect through direct action on 
sympathetic nerve receptors located on the pupillary dilator muscle of the iris, and as such would 
not be expected to cause material changes in accommodation. 
In previous reversal of pharmacologically induced mydriasis clinical studies (MIRA-1 and 
MIRA-2), Nyxol was effective at inducing reversal of mydriasis with both adrenergic 
(phenylephrine) and parasympathetic (tropicamide) mydriatic agents, as described previously. 
The placebo outcomes in these studies demonstrated that the natural reversal of mydriasis takes 
longer with tropicamide than with phenylephrine. Despite this difference, Nyxol was able to 
reverse mydriasis faster in the vast majority of eyes regardless of the mydriatic agent used, but it 
worked faster for phenylephrine (an alpha-1 agonist), as expected given the pharmacology of 
Nyxol (an alpha-1 and alpha-2 antagonist).  
Alpha-1 adrenergic antagonists have been shown to be safe and effective for the pharmacological 
reversal of mydriasis. In 1990, the United States (US) Food and Drug Administration (FDA) 
approved Dapiprazole Hydrochloride Ophthalmic Solution 0.5% (Rev-Eyes) for this indication; 
however, the product was withdrawn and discontinued by the manufacturer for reasons not 
related to safety or efficacy. Many people who undergo pupil dilation for an annual ophthalmic 
examination or other ophthalmic procedure continue to request an option for rapid reversal of the 
mydriasis. If Nyxol is a safe and effective treatment for the reversal of mydriasis, this may 
provide a new option for patients who experience side effects of pupil dilation. 
2.4. Route of administration, dosage regimen, and treatment period 
As the intended route of administration for Nyxol is topical ocular, this is the route to be used in 
this study.  
The dosing of Nyxol selected for this study, 0.75%, was based upon 1) preclinical safety studies, 
2) previous ophthalmic clinical studies of Nyxol described above and in the IB, and 3) clinical 
studies conducted with varying doses of drugs in the same class.  
Note that 0.75% Phentolamine Ophthalmic Solution, which expresses the phentolamine mesylate 
concentration in free base, is the new nomenclature being used in place of “1% Phentolamine 
Mesylate Ophthalmic Solution”, which was how Nyxol had been described in MIRA-1. 
2.5. Compliance 
This study will be conducted in compliance with the protocol and in accordance with 
International Council for Harmonisation (ICH) Guidelines for Good Clinical Practice (GCP), the 
ethical principles set forth in the Declaration of Helsinki, and with the US Title 21 Code of 
Federal Regulations. 
2.6. Study population 
A sample size of approximately 330 healthy subjects ≥ 12 years of age will be randomized in a 
2:1 ratio to 1 of 2 masked treatment arms (Nyxol or placebo, respectively). Randomization will 
be stratified 1:1 by light/dark color irides (Appendix 1) and further randomized 3:1:1 to 
unmasked mydriatic agent (2.5% phenylephrine, 1% tropicamide, or Paremyd, respectively). 
Subjects will be recruited from approximately 16 investigational sites. 


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3. OBJECTIVES AND PURPOSE 
The MIRA-3 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 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 systemic exposure of Nyxol based on pharmacokinetic (PK) sampling  
• To evaluate the safety of Nyxol 
• To evaluate any additional benefits of the reversal of pharmacologically induced mydriasis 
The Sponsor intends to use this 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.” 
4. STUDY DESIGN 
This is a randomized, parallel-arm, double-masked, placebo-controlled Phase 3 study in 
approximately 330 randomized subjects evaluating the safety and efficacy of Nyxol in subjects 
with pharmacologically induced mydriasis. Following the successful completion of screening, 
each subject will be randomized to mydriatic agent (unmasked) and treatment (masked). 
Treatment randomization will be 2:1 (Nyxol or placebo [vehicle], respectively). 
Stratification by iris color will be 1:1 (light or dark irides) as shown in Appendix 1.  
The mydriatic agent randomization will be 3:1:1 (2.5% phenylephrine, 
1% tropicamide, or Paremyd, respectively). That is, approximately 60% of the randomized 
subjects will receive 1 drop of 2.5% phenylephrine OU 1 hour before treatment (198 subjects), 
approximately 20% will receive 1 drop of 1% tropicamide OU 1 hour before treatment 
(66 subjects), and approximately 20% will receive 1 drop of Paremyd OU 1 hour before 
treatment (66 subjects). Study treatment (Nyxol or placebo) will be administered OU 1 hour after 
mydriatic drug instillation. Subjects will have 2 drops of study treatment (Nyxol or placebo) 
administered in the study eye (right eye [OD]). Each drop will be instilled 5 minutes apart.  
Subjects will have only 1 drop of study treatment administered in the fellow eye (left eye [OS]).   
The study eye and fellow eye will both be evaluated at all assessments unless otherwise 
specified.   
Blood sampling for Nyxol PK measurements will be conducted in a subset of approximately 
30 adult subjects at approximately 2 select study sites.   
4.1. Primary and secondary endpoints 
Efficacy: 
The primary efficacy endpoint is the percentage of subjects’ study eyes returning to ≤ 0.2 mm 
from baseline (-1 hour) photopic pupil diameter at 90 minutes. 


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The study eye is defined as the right eye (OD). The fellow eye (also referred to as the non-study 
eye in MIRA-2) is defined as the left eye (OS). Unless specified, the study eye and fellow eye 
will both be evaluated at all assessments. In addition, binocular will be evaluated for selected 
visual acuity measurements. 
Secondary efficacy endpoints will include: 
• Percentage of subjects returning to ≤ 0.2 mm from baseline (-1 hour) photopic pupil 
diameter at each remaining time point (0 minutes, 30 minutes, 60 minutes, 90 minutes 
[fellow eye], 2 hours, 3 hours, 4 hours, 6 hours, and 24 hours) 
• Change (in mm) in photopic pupil diameter from max pupil dilation (0 minutes) at each 
time point (30 minutes, 60 minutes, 90 minutes, 2 hours, 3 hours, 4 hours, 6 hours, and 
24 hours) 
• Time (hours) to return to ≤ 0.2 mm from baseline (-1 hour) photopic pupil diameter 
(time-savings analysis)  
• Percentage of subjects with unchanged accommodation from baseline (-1 hour) at 
0 minutes, 90 minutes, 2 hours, 3 hours, and 6 hours  
• Change (in diopters) in accommodation from max pupil dilation (0 minutes) at 
90 minutes, 2 hours, 3 hours, and 6 hours 
• 
Change (in mm) in the pupillary light reflex from max pupil dilation (0 minutes) at 
90 minutes, 3 hours, and 6 hours (study eye) 
• 
Change (in letters) in BCDVA under normal photopic lighting versus glare conditions 
(using Brightness Acuity Tester [BAT]) from baseline (-1 hour) at 0 minutes, 60 minutes, 
2 hours, and 6 hours (study eye) 
• 
Percentage of subjects with unchanged (within ± 4 letters) BCDVA under glare 
conditions (BAT) from baseline (-1 hour) at 0 minutes, 60 minutes, 2 hours, and 6 hours 
(study eye) 
• Change in glare discomfort from max pupil dilation (0 minutes) at 60 minutes, 2 hours, 
and 6 hours (study eye) 
Measurements: 
• Pupil diameter and pupillary light reflex will be measured with a NeurOptics VIP-300 
pupillometer (mm) 
• Accommodation will be measured by the Royal Air Force (RAF) Near Point Rule 
(measured in cm and then converted to diopters). Unchanged accommodation from 
baseline (-1 hour) is defined as a change from baseline value ≥ -1, as measured in 
diopters 
• Glare testing will be performed using the Marco BAT 2000 
• Glare discomfort will be measured on a 4-point scale from 0 (none) to 3 (severe) 
Some of the efficacy endpoints will be analyzed overall, by mydriatic agent, and by light/dark 
irides at all time points. Each mydriatic agent will be analyzed individually, and an additional 
analysis combining 1% tropicamide and Paremyd subjects into a “tropicamide or Paremyd” 
group will be performed. Additional analyses may be performed to compare efficacy endpoints 


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between the study eye and fellow eye within the same subject as well as the age given the 
pediatric population. 
Safety: 
The primary safety measures are conjunctival hyperemia, impairment in visual acuity 
(BCDVA and distance-corrected near visual acuity [DCNVA]), subjective ocular 
tolerability, and AEs. Other safety measures include intraocular pressure (IOP), subject 
questionnaire, and systemic safety as measured by HR and BP.  
Safety analyses will include: 
• Change from baseline (-1 hour) in the conjunctival hyperemia grading (Cornea and 
Contact Lens Research Unit [CCLRU] images) at each time point (0 minutes,  
30 minutes, 60 minutes, 90 minutes, 2 hours, 3 hours, 4 hours, 6 hours, and 24 hours) 
for the study eye and fellow eye   
• 
Change from baseline (-1 hour) in BCDVA at 0 minutes, 60 minutes, 2 hours, 6 hours, 
and 24 hours for the study eye, fellow eye, and binocular 
• 
Change from baseline (-1 hour) in DCNVA at 0 minutes, 90 minutes, 3 hours, 6 hours, 
and 24 hours for the study eye, fellow eye, and binocular 
• 
Subjective ocular tolerability at 0 minutes after administration of study medication for 
the study eye and fellow eye 
• 
Change from screening in IOP at 6 hours for the study eye and fellow eye 
• 
Change from baseline (-1 hour) in subject questionnaire responses at 0 minutes, 
60 minutes, 2 hours, 4 hours, and 24 hours 
• 
Change from screening in vital signs (HR and BP) at 6 hours and 24 hours  
Measurements: 
• Conjunctival hyperemia will be assessed visually with a 4-point grading scale (0-3) using 
CCLRU images (Appendix 2)  
• Best-corrected distance visual acuity will be measured in photopic conditions by a 
standard Early Treatment Diabetic Retinopathy Study (ETDRS) illuminated chart (on 
wall or stand) at 4 m (letters recorded) 
• Distance-corrected near visual acuity will be measured in photopic conditions by Near 
Visual Acuity Chart in the Precision Vision Small 914 Illuminator Cabinet (light box) at 
16 inches (~40 cm) (letters recorded)  
• Subjective ocular tolerability will be measured on a 4-point scale (0-3) 
• Intraocular pressure will be measured with the Tono-Pen or Goldmann applanation 
tonometer 
• Subject questionnaire will be a brief symptom survey (Appendix 3) 
4.2. Description and schedule of visits and procedures 
A sample size of approximately 330 healthy subjects ≥ 12 years of age will be randomized in a 
2:1 ratio to 1 of 2 treatment arms (Nyxol or placebo, respectively). Randomization will be 


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stratified 1:1 by iris color (light and dark irides) and 3:1:1 by mydriatic agent 
(2.5% phenylephrine, 1% tropicamide, or Paremyd, respectively). Subjects will receive their 
mydriatic agent OU 1 hour before treatment with study medication (Nyxol or placebo). Study 
procedures are shown in detail in Table 1. For this study, the following time points apply: 
• Baseline (-1 hour): just prior to administration of mydriatic agent at -1 hour and time 
point at which PD measurement is considered normal 
• Max pupil dilation (0 minutes): just prior to administration of study medication and time 
point at which maximum PD is expected   


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Table 1: Screening and Mydriatic/Treatment Schedule 
 
Screening [a]
Mydriasis/Treatment 
Follow-Up 
Day 
1 
1 
2 
Visit 
1 
1 
2 
Start time [b] 
 
-1 hour 
(Baseline) 
0 ±5 min 
(Max) 
30 min 
±5 min 
60 min 
±5 min 
90 min 
±5 min 
2 hours 
±5 min 
3 hours 
±10 min 
4 hours  
±10 min 
6 hours 
±15 min 
24 hours      
+6 hours 
Informed consent/assent 
X 
 
 
 
 
 
 
 
 
 
 
Subject identification number assigned 
X 
 
 
 
 
 
 
 
 
 
 
Medical/Ophthalmic history 
X 
 
 
 
 
 
 
 
 
 
 
Demographics 
X 
 
 
 
 
 
 
 
 
 
 
Prior/Concomitant medications [c] 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Urine pregnancy test [d] 
X 
 
 
 
 
 
 
 
 
 
 
HR/BP 
X 
 
 
 
 
 
 
 
 
X 
X 
Pupil diameter 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
BCDVA 
 
X 
X 
 
X 
 
X 
 
 
X 
X 
DCNVA 
 
X 
X 
 
 
X 
 
X 
 
X 
X 
Accommodation 
 
X 
X 
 
 
X 
X 
X 
 
X 
 
Pupillary light reflex [e] 
 
X 
X 
 
 
X 
 
X 
 
X 
 
BAT – glare test (BCDVA) [e] 
 
X 
X 
 
X 
 
X 
 
 
X 
 
Glare discomfort 
 
X 
X 
 
X 
 
X 
 
 
X 
 
Conjunctival hyperemia  
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Biomicroscopy 
X 
 
 
 
 
 
 
 
 
 
 
IOP [f] 
X 
 
 
 
 
 
 
 
 
X 
 
Ophthalmoscopy (undilated) 
X 
 
 
 
 
 
 
 
 
 
 
Subject questionnaire 
 
X 
X 
 
X 
 
X 
 
X 
 
X 
Mydriatic agent and kit number assigned 
(randomization) 
X 
  
 
 
 
 
 
 
 
 
 
Mydriatic agent administration 
 
X 
 
 
 
 
 
 
 
 
 
Treatment: Nyxol/Placebo administration 
 
 
X 
 
 
 
 
 
 
 
 
Ocular tolerability 
 
 
X 
 
 
 
 
 
 
 
 
PK sampling [g] 
 
 
X[h] 
 
X 
 
 
X 
 
 
 
AEs [c] 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
AE, adverse event; BAT, Brightness Acuity Tester; BCDVA, best-corrected distance visual acuity; BP, blood pressure; DCNVA, distance-corrected near visual acuity; HR, 
heart rate; IOP, intraocular pressure; PK, pharmacokinetic. 
[a] Screening Visit: if subject qualifies, this becomes Treatment Visit 1. The assigned mydriatic agent will be given at -1 hour. 


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[b] Start time refers to the time that first assessment is performed. Assessments at each time point are shown in the order performed.  
[c] Investigators to note changes to concomitant medications and AEs at any time throughout the visit. 
[d] Urine pregnancy test is for females of childbearing potential only. 
[e] Study eye only. 
[f] IOP with Tono-Pen or Goldmann applanation tonometer.  
[g] Blood sampling for Nyxol PK measurements will be performed at approximately 2 select study sites in approximately 30 adult subjects.  
[h] PK sampling will be performed at 15 minutes + 5 minutes, 60 minutes ± 10 minutes, and 3 hours ± 10 minutes post dose. 


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4.3. Measures taken to minimize/avoid bias 
This is a placebo-controlled, double-masked, 2:1 randomized (Nyxol or placebo, respectively), 
2-arm, Phase 3 study. Nyxol vehicle is used as placebo. 
4.4. Study medications 
Study Medication Identification 
Established name 
Phentolamine mesylate – parent phentolamine 
CAS registry number 
65-28-1 – parent 50-60-2 
Chemical class 
An α-adrenergic antagonist, it is a member of 
the following classes: imidazoles, of phenols, 
is a tertiary amino compound and a substituted 
aniline. 
Chemical name 
3-[N-(4,5-dihydro-1H-imidazol-2-ylmethyl)-4-
methylanilino]phenol;methanesulfonic acid 
Molecular formula 
C18H23N3O4S– parent C17H19N3O 
Molecular weight 
377.140– parent 281.352 
Drug name/formulation 
Nyxol / aqueous, sterile, non-preserved, 
isotonic, ophthalmic solution  
Concentration active 
1% – parent 0.75% (free acid) 
Manufacturer drug substance 
Industriale Chimica s.r.l. (Italy) 
Manufacturer drug product, placebo Woodstock Sterile Solutions, Inc. 
Storage requirements 
Shipped and stored refrigerated (2°C to 8°C; 
36°F to 46°F). Stored at the site at room 
temperature (15ºC to 25ºC; 59ºF to 77ºF) in a 
secured location (locked) with no access for 
unauthorized personnel per ICH GCP 
Guidelines.  
Formulation 
Nyxol (0.75% Phentolamine Ophthalmic Solution) is a clear, colorless to slightly brown, sterile, 
non-preserved, isotonic, buffered aqueous solution containing 1% phentolamine mesylate 
(equivalent to 0.75% phentolamine free base), mannitol, and sodium acetate. Placebo for Nyxol 
is a clear, colorless, sterile, non-preserved, isotonic, buffered aqueous solution containing 
mannitol and sodium acetate. The pH of the study medications may be adjusted with NaOH 
(United States Pharmacopeia [USP]) and/or HCl (USP) to pH 4.8.  


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4.4.1. Packaging and labeling 
The investigational products, active and placebo, are packaged in a 0.5-mL low-density 
polyethylene Blow-Fill-Seal vial containing 0.3 mL solution for single-dose use. Five individual 
vials are included in a molded strip. Each strip containing 5 vials is wrapped with a multicolor 
“rainbow” aluminum foil overwrap that has been purged with nitrogen. The foil is impermeable 
to water and oxygen and will be labeled with an investigational label showing the study protocol 
number and other relevant information, including a statement “Caution – New Drug – Limited 
by Federal (US) Law to Investigational Use”.  
4.4.2. Storage of study medication and dispensing 
Prior to dispensing, all investigational material must be stored in a secure locked location with 
limited access documented by signature of authorized persons who may dispense investigational 
materials. The products should be shipped refrigerated (2°C to 8°C; 36°F to 46°F) and stored at 
the site at room temperature (15ºC to 25ºC; 59ºF to 77ºF). Study medication must not be frozen 
and must be protected from light.  
4.4.3. Study medication administration 
Study medication will be administered by the Investigator or designee at the site on Treatment 
Visit 1. To administer the study medication, the foil pouch is opened and the strip of 5 vials 
removed from the pouch. A single vial is removed from the strip by twisting the vial, which 
exposes the dropper tip and contents of the vial. The vial should be removed immediately prior 
to dosing to protect the contents of the vial. After administration, all used (1 vial) and unused 
vials (4 vials [5 vials total]) should be returned to the labeled foil pouch and the original kit box 
and retained at site, for proper accountability and reconciliation.   
4.4.4. Study medication accountability 
4.4.4.1. Receipt and disposition of study medication 
The Investigator or designee (eg, study coordinator or pharmacist) will maintain a full 
accountability record for the study medication and will be responsible for recording the receipt, 
dispensing, and return of all supplies of the study medication using the inventories supplied for 
the study. The Investigator or designee will account for all study medication. The monitor will 
review dispensing and study medication accountability records during study visits and at the 
completion of the study and note any discrepancies.  
4.4.4.2. Return of study medication 
When the study is completed or is terminated by Ocuphire, all study material, including used and 
unused study medication vials and strips in the original kits, will be returned to Ocuphire (or its 
designee) or destroyed under the specific direction of same. All study medication accounting 
procedures must be completed before the study is considered completed. A final study 
medication disposition will be completed by the study coordinator. 
4.5. Expected duration of subject participation 
The total length of subject participation is 2 days, with 2 clinic visits, as summarized below: 
• Screening Visit Day 1/Treatment Visit 1/Day 1  
• Follow-Up Visit 2/Day 2 


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The execution of the entire study (first subject screened through last randomized subject 
completed) is expected to be approximately 4 to 6 months. 
4.6. Randomization and procedure for breaking the code 
A randomization code for allocating subjects to treatment will be prepared by an unmasked 
biostatistician not directly connected with the study. Randomization to study medication 2:1 
(Nyxol or placebo, respectively) will be stratified 1:1 by light/dark color irides and further 
randomized 3:1:1 to mydriatic agent (2.5% phenylephrine, 1% tropicamide, or Paremyd, 
respectively) (Appendix 4). Note that the mydriatic agent administered is unmasked. 
At the initiation of study-related procedures, every potential subject will be assigned a subject 
identification number.  
The study medication will be masked to both Investigator and study subjects, as well as 
Ocuphire. Only in case of medical emergency or occurrence of SAEs will the code be unmasked 
by the Medical Monitor and made available to the Investigator, Ocuphire, and/or other personnel 
involved in the monitoring or conduct of this study. 
4.7. Collection of data 
Study-specific data that have been outlined in the protocol will be entered into the clinical 
database by individual(s) designated by the Investigator. Data will be verified electronically 
using a series of online programmed edit checks that have been created by the Clinical Data 
Manager and programmed by the Clinical Data Programmer or designee. Data discrepancies will 
be brought to the attention of the clinical team and investigated by the study monitor and site 
staff. Study monitors will review and verify all data collected in the electronic Case Report Form 
(eCRF) against any applicable source documentation during remote review or scheduled 
monitoring visits. The study monitor will work closely with the site staff to address any 
discrepancies found so that proper resolutions can be made and documented in the clinical 
database. An audit trail within the system will track all changes made to the data. 
4.8. Study medication discontinuation 
The study medication may be discontinued for the following reasons: 
• Adverse events: AEs include clinically significant laboratory abnormalities and 
intercurrent diseases reported by the subject or observed by the Investigator with 
documentation on the eCRF 
• Death: If a subject dies, the AE that caused the death should be documented on the eCRF 
and be noted as serious and fatal 
• Disallowed concurrent medication: Any medication not allowed by the protocol would 
be a protocol violation 
• Lack of efficacy: A subject may elect to discontinue participation in the study for a 
perceived lack of efficacy 
• Investigator decision: A subject may be discontinued for reasons other than those 
bulleted previously if the Investigator thinks it is not in the best interest of the subject to 
continue 
• Other: Any other reason for subject discontinuation should be noted on the eCRF 


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The reason for premature study medication discontinuation should be entered onto the 
appropriate eCRF.  
4.8.1. Reasons for withdrawal from study 
• Subject withdraws consent 
• Subject is lost to follow-up 
• Subject withdraws for other reason 
4.8.2. Entire study terminated 
The entire study may be terminated by Investigators or Ocuphire. Prompt, written notice of 
reasonable cause to the other party (Ocuphire or Investigators, respectively) is required. Prompt 
notice to the Institutional Review Board (IRB) and to regulatory authorities is also required. 
4.8.3. Actions after discontinuation 
All subjects who discontinue study medication due to a report of an AE must be followed-up and 
provided appropriate medical care until their signs and symptoms have remitted or stabilized or 
until medical assessments have returned to acceptable or pre-study limits. 
For any subject who chooses to withdraw consent or who is noncompliant, every possible effort 
should be made by the Investigator to assure the 6-hour measurements are assessed prior to 
discontinuation, in addition to a follow-up telephone call that includes assessments for AEs, 
concomitant medications, and subject-evaluated conjunctival hyperemia.  
4.9. Completed study 
The study is completed when all randomized subjects have completed the study, all eCRFs have 
been completed, and all eCRF data have been entered into the database. Final database lock will 
occur after the last randomized subject completes last visit, all data have been entered, and all 
queries have been resolved.  
5. SUBJECT INCLUSION AND EXCLUSION CRITERIA 
5.1. Subject inclusion criteria 
Included in the study will be individuals with the following characteristics: 
1. Males or females ≥ 12 years of age 
2. Ability to comply with all protocol-mandated procedures independently and to attend all 
scheduled office visits  
3. Adults (≥ 18 years of age) willing to give written informed consent to participate in this 
study. Children aged 12-17 years to provide signed assent form, as well as a separate 
parental/Legal Guardian consent 
5.2. Subject exclusion criteria 
Excluded from the study will be individuals with the following characteristics: 
Ophthalmic (in either eye):  


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1. 
Clinically significant ocular disease as deemed by the Investigator (eg, glaucoma, 
corneal edema, uveitis, severe keratoconjunctivitis sicca) that might interfere with the 
study 
2. 
Unwilling or unable to discontinue use of contact lenses at screening until study 
completion 
3. 
Unwilling or unable to suspend use of topical medication at screening until study 
completion 
4. 
Ocular trauma, ocular surgery, or non-refractive laser treatment within the 6 months 
prior to screening 
5. 
Use of any topical prescription or over-the-counter (OTC) ophthalmic medications of 
any kind within 7 days of screening, with the exception of lid scrubs with OTC 
products (eg, OCuSOFT® lid scrub, SteriLid®, baby shampoo, etc.)  
6. 
Recent or current evidence of ocular infection or inflammation in either eye (such as 
current evidence of clinically significant blepharitis, conjunctivitis, or keratitis). Subjects 
must be symptom free for at least 7 days prior to screening 
7. 
History of herpes simplex or herpes zoster keratitis 
8. 
Closed or very narrow angle that in the Investigator’s opinion is potentially occludable 
if the subject’s pupil is dilated  
9. 
History of any traumatic (surgical or nonsurgical) or nontraumatic condition affecting 
the pupil or iris  
10. History of cauterization of the punctum or punctal plug (silicone or collagen) insertion 
or removal 
11. Known allergy, hypersensitivity, or contraindication to any component of the 
phentolamine ophthalmic solution or to any component of the mydriatic agents or 
vehicle formulation 
12. Prior participation in a study involving the use of Nyxol for the reversal of mydriasis 
Systemic: 
13. Known hypersensitivity or contraindication to α- and/or β-adrenoceptor antagonists 
14. Clinically significant systemic disease (eg, uncontrolled diabetes, myasthenia gravis, 
cancer, hepatic, renal, endocrine, or cardiovascular disorders) that might interfere with 
the study 
15. Initiation of treatment with or any changes to the current dosage, drug, or regimen of 
any systemic adrenergic or cholinergic drugs within 7 days prior to screening or during 
the study (Appendix 5) 
16. Participation in any investigational study within 30 days prior to screening 
17. Females of childbearing potential who are pregnant, nursing, planning a pregnancy, or 
not using a medically acceptable form of birth control. Acceptable methods include the 
use of at least one of the following: intrauterine device (IUD), hormonal (oral, 
injection, patch, implant, ring), barrier with spermicide (condom, diaphragm), or 


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abstinence. A female is considered to be of childbearing potential unless she is 
premenstrual, 1 year postmenopausal, or 3 months post-surgical sterilization. All 
females of childbearing potential, including those with post-tubal ligation, must have a 
negative urine pregnancy test result at Visit 1/Screening and must intend to not become 
pregnant during the study 
18. Resting HR outside the normal range (50-110 beats per minute) at the Screening Visit. 
Heart rate may be repeated only once if outside the normal range following at least a 5-
minute rest period in the sitting position 
19. Hypertension with resting diastolic BP > 105 mmHg or systolic BP > 160 mmHg at the 
Screening Visit. Blood pressure may be repeated only once if outside the specified 
range following at least a 5-minute rest period in the sitting position 
6. TREATMENT OF SUBJECTS 
Approximately 330 healthy subjects ≥ 12 years of age will be randomized in a 2:1 ratio to 1 of 
2 treatment arms (Nyxol or placebo, respectively). Randomization will be stratified 1:1 by 
light/dark color irides and further randomized 3:1:1 to mydriatic agent (2.5% phenylephrine, 
1% tropicamide, or Paremyd, respectively). Subjects will receive their mydriatic agent OU 
1 hour before treatment with study medication on Treatment Visit 1/Day 1. There will be no 
treatment administered on Follow-Up Visit/Day 2. 
6.1. Treatment adherence 
All subjects will be treated by the Investigator or designee at the study clinic on Visit 1.  
6.2. Concomitant medications 
As noted in the exclusion criteria (Section 5.2), use of any topical prescription or OTC 
ophthalmic medications of any kind within 7 days of screening is prohibited, with the exception 
of lid scrubs with OTC products (eg, OCuSOFT lid scrub, SteriLid, baby shampoo, etc.) which 
may have been used prior to but not at screening until study completion.  
Additionally, initiation of treatment with or any changes to the current dosage, drug, or regimen 
of any systemic adrenergic or cholinergic drugs (Appendix 5) within 7 days prior to screening or 
during the study is prohibited. However, a subject can be treated with a systemic adrenoceptor 
antagonist, for example, as long as the particular agent and its dose and regimen had been 
consistent for the 7 days prior to screening, and there was no reason to believe that alteration 
would be necessary at some point later during the study. 
If there is any question about whether a medication is acceptable, the Medical Monitor should 
be consulted before proceeding. 
Use of all medications should be documented on the appropriate eCRF. Investigators are 
encouraged to contact the Medical Monitor for any questions regarding allowed medications. 
Judgment of continued study participation by the subject, and inclusion of this subject’s 
subsequent visits in the safety and efficacy analysis will be made by Ocuphire. 
All medications taken by the subject within 30 days prior to the Screening Visit and during the 
study will be recorded in the eCRF. The name of the drug, dose, route of administration, duration 
of treatment, and indication will be recorded for each medication. For combination products (eg, 
Contac®, Cosopt®), the brand name is required. For non-combination products, the generic name 


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is desired. The use of routine ophthalmic diagnostic pharmaceutical agents (eg, fluorescein and 
local anesthetic) will be allowed and should be documented. Any change in dosing parameters 
should also be recorded in the eCRF. 
7. ASSESSMENT OF EFFICACY 
7.1. Specification of the efficacy parameters 
The primary efficacy endpoint is the percentage of subjects’ study eyes with 2 drops of treatment 
returning to ≤ 0.2 mm baseline (-1 hour) photopic pupil diameter at 90 minutes. 
The study eye is defined as the right eye (OD). The fellow eye is defined as the left eye (OS). 
Unless specified, the study eye and fellow eye will both be evaluated at all assessments. 
Specified endpoints will also be analyzed for both eyes. Secondary efficacy endpoints can be 
found in Section 4.1.  
The mITT Population will be used for the primary endpoint analysis and to analyze selected 
secondary efficacy endpoints. The PP Population will be used to analyze selected secondary 
efficacy endpoints. Some of the efficacy endpoints will be analyzed at all time points overall, by 
mydriatic agent, and by light/dark irides.  
7.2. Assessing, recording, and analyzing efficacy parameters 
Pupil diameter will be measured at Treatment Visit 1 and at Follow-Up Visit 2. Accommodation, 
pupillary light reflex, glare testing (BAT), and glare discomfort will all be measured only at 
Treatment Visit 1. 
• Pupil diameter and pupil reactivity will be measured with the NeurOptics VIP-300 
pupillometer (mm) 
• Accommodation will be measured by the RAF Near Point Rule (measured in cm and then 
converted to diopters). Unchanged accommodation from baseline (-1 hour) is defined as a 
change from baseline value ≥ -1, as measured in diopters 
• Glare testing will be performed using the Marco BAT 2000 
• Glare discomfort will be measured on a 4-point scale from 0 (none) to 3 (severe) 
The photopic lighting conditions in the room will be defined as lights off with ambient light 
coming from the distance ETDRS illuminated chart (on wall or stand) 4 m away at a luminance 
level of 85 cd/m2 (or 85 lux). Subjects will be allowed to acclimate to these lighting conditions 
(with the eyes open normally for a minimum of 2 minutes) prior to the PD, BCDVA, and 
DCNVA (safety measures) measurements at all scheduled time points. Subjects will sit in the 
exam chair facing the illuminated chart during the acclimation period and for all assessments. 
Scotopic PD will be measured OD with room lights and illuminated chart off and OS occluded. 
BAT testing will be performed OD with OS occluded. Room lights will be off with the distance 
ETDRS chart illuminated. Room lights should be turned on for accommodation measurements 
and for the remaining safety assessments (eg, conjunctival hyperemia, AEs, brief subject 
questionnaire, etc.). The subject will be in the same room for all assessments, and every effort 
will be made to have the same person perform the measurements at all time points. 
Some of the efficacy endpoints will be analyzed overall, by mydriatic agent, and by light/dark 
irides at all time points. Each mydriatic agent will be analyzed individually, and an additional 
analysis combining 1% tropicamide and Paremyd subjects into a “tropicamide or Paremyd” 


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group will be performed. Exploratory analyses may be performed to compare efficacy endpoints 
between the study eye and fellow eye within the same subject. 
7.2.1. Screening/Day 1 
Individuals who are potential subjects are identified by the study center to schedule the 
Screening Visit. The Screening Visit should occur the same day as Treatment Visit 1, when the 
dose of study treatment is given.  
Once subjects arrive at the study center, a member of the study center staff will interview the 
individual as to their qualifications for participation in the study, and if the subject wishes to 
continue, the informed consent form is signed, and a subject identification number is assigned. 
Children aged 12 to 17 years must provide assent. Screening includes an explanation of the 
study, a medical and ophthalmic history, demographics, a review of prior/concomitant 
medications, a urine pregnancy test (for females of childbearing potential), and HR/BP 
assessment. The second step in screening includes procedures such as IOP assessment and an 
ophthalmic examination consisting of biomicroscopy and direct or indirect ophthalmoscopy 
without dilation.  
Investigators are cautioned to appropriately note all observations of conjunctival hyperemia (also 
called conjunctival erythema) on the biomicroscopy eCRF at screening. 
7.2.2. Treatment Visit 1/Day 1 
Treatment Visit 1 should be the same day as the Screening Visit. Once the subject has completed 
the screening assessments part of the visit and it is confirmed that he/she meets all of the 
inclusion criteria but none of the exclusion criteria, the visit will then transition to the Treatment 
Visit 1 assessments. The study eye is OD and the fellow eye is OS.  
As part of the Treatment Visit 1, the subject:   
• Will be randomized to 1 of 2 treatment arms, with further randomization by mydriatic agent 
and stratification by iris color. 
• Will receive 1 of 3 approved mydriatic agents (2.5% phenylephrine, 1% tropicamide, or 
Paremyd) at -1 hour (baseline). The mydriatic agent will be administered as a single drop in 
each eye. If a drop of mydriatic agent is missed, the Investigator should give the drop again. 
Note: Mydriatic agent is administered after assessments conducted at -1 hour (baseline).  
• Will receive the masked study medication (Nyxol or placebo), based on his/her randomized 
treatment arm (kit assignment) at 0 minutes (max pupil dilation). Note: Study medication is 
administered after assessments conducted at 0 minutes (max pupil dilation). Study medication 
will be administered OU 1 hour after mydriatic drug instillation. Subjects will have a total of 
2 drops (1 drop each 5 minutes apart) in the study eye (OD) and 1 drop in the fellow eye (OS). 
If a drop of study medication is missed, the Investigator should give the drop again.  
• Will be assessed for ocular tolerability immediately after instillation of study medication at 
0 minutes (max pupil dilation).  
• Will be assessed at -1 hour (baseline), 0 minutes (max pupil dilation), 30 minutes, 60 minutes, 
90 minutes, 2 hours, 3 hours, 4 hours, and 6 hours relative to study treatment for the following: 
PD (OD, OS) and conjunctival hyperemia (OD, OS), as well as concomitant medications and 
any AEs. 


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• Will be assessed at -1 hour (baseline), 0 minutes (max pupil dilation), 60 minutes, 2 hours, and 
6 hours relative to study treatment for the following: BCDVA (OD, OS, and OU), BAT ‒ glare 
test BCDVA (OD), and glare discomfort (OD). 
• Will be assessed at -1 hour (baseline), 0 minutes (max pupil dilation), 90 minutes, 2 hours, 
3 hours, and 6 hours relative to study treatment for the following: accommodation (OD, OS, 
and OU). 
• Will be assessed at -1 hour (baseline), 0 minutes (max pupil dilation), 90 minutes, 3 hours, and 
6 hours relative to study treatment for the following: DCNVA (OD, OS, and OU) and pupillary 
light reflex (OD). 
• Will be assessed at -1 hour (baseline), 0 minutes (max pupil dilation), 60 minutes, 2 hours, and 
4 hours relative to study treatment for the following: subject questionnaire. 
• Will have PK blood samples taken at 15 minutes, 60 minutes, and 3 hours relative to study 
treatment (for approximately 30 adult subjects).  
7.2.3.Follow-Up Visit/Day 2  
On Day 2, the subject will return to the clinic for a Follow-Up Visit.  
As part of the Follow-Up Visit, the subject will be assessed at 24 hours + 6 hours relative to 
study treatment administration during Treatment Visit 1 for the following: concomitant 
medications, HR/BR, PD, BCDVA, DCNVA, conjunctival hyperemia, subject questionnaire, and 
AEs. 
7.2.4. Unscheduled Visits 
An Unscheduled Visit may be any visit to the Investigator other than the specific visits requested 
in the protocol as possibly required for the subject’s ophthalmic condition. The Investigator will 
perform all procedures necessary to evaluate the subject at these visits and record any AEs in the 
eCRF. 
As noted in Section 4.8.3, every possible effort should be made by Investigators to assure that 
subjects who discontinue early from the study have a telephone follow-up that includes 
assessments of AEs, concomitant medications, and subject-evaluated conjunctival hyperemia. 
7.2.5. Visit Variation 
Visits on Day 2 may occur between 24 to 30 hours after the baseline (-1 hour time point on 
Treatment Day 1). 
8. PHARMACOKINETIC ASSESSMENTS 
For Nyxol PK analysis at Treatment Visit 1, blood samples will be collected to establish drug 
levels of Nyxol from approximately 30 adult subjects at approximately 2 select study sites. 
Six mL of blood will be drawn 15 minutes, 60 minutes, and 3 hours post dose. Analysis of 
plasma samples for Nyxol concentration determinations will be performed by a central PK 
laboratory using a validated liquid chromatography-mass spectrometry and liquid 
chromatography-tandem mass spectrometry (LC-MS/MS) method. 


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9. ASSESSMENT OF SAFETY 
9.1. Specification of safety parameters 
The assessment of safety and tolerability is a secondary objective of this study. The assessment 
of safety will be performed by: 
• Conjunctival hyperemia measured with CCLRU images using a 4-point scale (0-3) 
(Appendix 2). 
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 
• Best-corrected distance visual acuity will be measured in photopic conditions by a 
standard ETDRS illuminated chart (on wall or stand) at 4 m (letters recorded)  
• Distance-corrected near visual acuity will be measured binocularly in photopic conditions 
by Near Visual Acuity Chart in the Precision Vision Small 914 Illuminator Cabinet (light 
box) at 16 inches (~40 cm) (letters recorded) 
• Intraocular pressure will be measured with the Tono-Pen or Goldmann applanation 
tonometer 
• Heart rate and BP (as per the site’s normal equipment and procedures) 
• Adverse events 
• Brief subject questionnaire (Appendix 3) 
9.2. Assessing, recording, and analyzing safety parameters 
The timing for recording safety parameters may be found in Section 4.2. 
9.3. Adverse events and serious adverse events 
All AEs and SAEs that occur following consent and until the final study visit should be collected 
and recorded on the AE or SAE eCRF page. Only treatment-emergent adverse events 
(TEAEs)/adverse reactions will be summarized (Section 10.3.6). 
All AEs/adverse reactions occurring during the study (ie, once the subject has signed the 
informed consent/assent) must be documented, regardless of the assumption of causal 


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relationship, on the respective eCRF. All TEAEs/adverse reactions must be documented from the 
time the subject receives the study medication until the subject’s participation in the study has 
been completed. If a subject has ongoing TEAEs/adverse reactions at the time of study 
completion or discontinuation from the study, the ongoing TEAEs/adverse reactions are to be 
followed and provided appropriate medical care until the signs and symptoms have remitted or 
stabilized or until medical assessments have returned to acceptable or pre-study limits.  
Documentation of AEs/adverse reactions includes start date and end date, severity, relationship 
to study medications, action(s) taken, seriousness, and outcome.  
9.3.1. Adverse event definitions 
The following definitions of terms apply to this section: 
Adverse event. An AE is any untoward medical occurrence associated with the use of a study 
medication in humans, whether or not considered drug related. An AE can therefore be any 
unfavorable and unintended sign (including abnormal laboratory findings), symptom, or disease 
temporarily associated with the use of the study medication, whether or not related to the study 
medication. Study medication includes the investigational drug under evaluation and the 
comparator product or vehicle placebo that is given or administered during any phase of the 
study.  
Medical conditions/diseases present before starting the investigational treatment are only 
considered AEs if they worsen after starting the investigational treatment. Abnormal test results 
constitute AEs only if they induce clinical signs or symptoms, are considered clinically 
significant, or require therapy. 
The occurrence of AEs should be sought by open-ended questioning of the subject at each visit 
during the study. At each clinic assessment/visit, study personnel should ask the following 
question: “Have you had any problems since your last assessment/visit?”. Adverse events also 
may be detected when they are volunteered by the subject during or between visits or through 
study assessments. 
Life-threatening adverse event or life-threatening suspected adverse reaction. An AE or 
suspected adverse reaction is considered “life-threatening” if, in the view of either the 
Investigator or Ocuphire, its occurrence places the subject at immediate risk of death. It does not 
include an AE or suspected adverse reaction that, had it occurred in a more severe form, might 
have caused death.  
Serious adverse event or serious suspected adverse reaction. An AE or suspected adverse 
reaction is considered ‘‘serious’’ if, in the view of either the Investigator or Ocuphire, it results 
in any of the following outcomes:  
• Death 
• Life-threatening AE 
• Inpatient hospitalization or prolongation of existing hospitalization 
• Persistent or significant incapacity or substantial disruption of the ability to conduct normal 
life functions  
• Congenital anomaly/birth defect  


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• Other medically important serious event 
Important medical events that may not result in death, be life-threatening, or require 
hospitalization may be considered serious when, based upon appropriate medical judgment, they 
may jeopardize the patient or subject and may require medical or surgical intervention to prevent 
one of the outcomes listed in this definition.  
Treatment on an outpatient emergency basis that does not result in hospital admission, or a 
hospitalization that is elective or is a preplanned treatment for a pre-existing condition that is 
unrelated to the medication under study and has not worsened since the start of the study, is not 
considered an SAE. 
Suspected adverse reaction means any AE for which there is a reasonable possibility that the 
drug caused the AE. For the purposes of Investigational New Drug Application safety reporting, 
“reasonable possibility” means there is evidence to suggest a causal relationship between the 
drug and the AE. Suspected adverse reaction implies a lesser degree of certainty about causality 
than adverse reaction, which means any AE caused by a drug.  
Unexpected adverse event or unexpected suspected adverse reaction. An AE or suspected 
adverse reaction is considered “unexpected” if it is not listed in the IB or is not listed at the 
specificity or severity that has been observed; or, if an IB is not required or available, is not 
consistent with the risk information described in the general investigational plan or elsewhere in 
the current application, as amended. “Unexpected,” as used in this definition, also refers to AEs 
or suspected adverse reactions that are mentioned in the IB as occurring with a class of drugs or 
as anticipated from the pharmacological properties of the drug but are not specifically mentioned 
as occurring with the particular drug under investigation. 
The study medication relationship for each AE/adverse reaction should be determined by the 
Investigator using these explanations: 
• Not related 
• Unlikely related 
• Possibly related 
• Probably related 
• Definitely related 
• Unknown 
Unless the relationship is considered to be “Not related” or “Unlikely related” and there is any 
valid reason, even if undetermined, for suspecting a possible cause-and-effect relationship 
between the study medication and the occurrence of the AE, then the AE should be considered 
“related”.  
If the relationship between the AE/SAE and the study medication is determined by Ocuphire to 
be anything other than “Not related” or “Unlikely related” the event will be considered to be 
related to the study medication for the purposes of expedited regulatory reporting. 
Severity of an AE is defined as a qualitative assessment of the level of discomfort of an AE as is 
determined by the Investigator or reported to him/her by the subject. The assessment of severity 


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is made irrespective of study medication relationship or seriousness of the event and should be 
evaluated according to the following scale: 
• 1 = Mild: present, but not distressing, and no disruption of normal daily activity 
• 2 = Moderate: discomfort sufficient to reduce or affect normal daily activity 
• 3 = Severe: incapacitating, with inability to work or perform normal daily activity 
A change in severity for a reported AE will require an end date for the previous severity and a 
new start and end date for the new severity. For example, a change in severity may go from mild 
to severe or from severe to moderate. In either case the start or end times/dates should be 
recorded.  
The term “severe” is used to describe the intensity of an event/reaction; the event/reaction itself 
may be of relatively minor medical significance (such as a severe headache). This is not the same 
as a “serious” AE, which is based on a subject/event outcome or action criteria usually associated 
with events that pose a threat to the subject’s life or vital functions. “Seriousness” (NOT 
severity) serves as a guide for defining regulatory reporting obligations. 
Action taken in response to an AE is coded as:  
• Dose increased: An indication that a medication schedule was modified by addition; 
either by changing the frequency, strength, or amount 
• Dose not changed: An indication that a medication schedule was maintained 
• Dose reduced: An indication that a medication schedule was modified by subtraction, 
either by changing the frequency, strength, or amount 
• Dose interrupted: An indication that a medication schedule was modified by temporarily 
terminating a prescribed regimen of medication 
• Drug withdrawn: An indication that a medication schedule was modified through 
termination of a prescribed regimen of medication 
• Not applicable: Determination of a value is not relevant in the current context 
• Unknown: Not known, not observed, not recorded, or refused 
Additional other action taken: 
• Concomitant medication 
• Hospitalization 
Outcome of an AE is coded as: 
• Fatal: The termination of life as a result of an AE 
• Not recovered/not resolved: One of the possible results of an AE outcome that indicates 
that the event has not improved or recuperated 
• Recovered/resolved: One of the possible results of an AE outcome that indicates that the 
event has improved or recuperated 


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• Recovered/resolved with sequelae: One of the possible results of an AE outcome where 
the subject recuperated but retained pathological conditions resulting from the prior 
disease or injury 
• Recovering/resolving: One of the possible results of an AE outcome that indicates that 
the event is improving 
• Unknown: Not known, not observed, not recorded, or refused 
In previous clinical studies of Nyxol, the most frequently reported AE was conjunctival 
hyperemia. 
Investigators are cautioned to use the appropriate verbatim term on the AE form to 
describe this observation:  
• Redness related to instillation that is transient (ie, is no longer present 2 hours after 
instillation) = “conjunctival erythema upon instillation” 
• Redness that is NOT transient (ie, is present 2 hours after instillation) = 
“conjunctival hyperemia” 
Expedited reporting of Serious and Unexpected Adverse Events: All SAEs (related and 
unrelated) will be recorded following subject signature of the informed consent/assent and until 
the Follow-Up Visit (Day 2). Any SAEs “suspected” to be related to the study medication and 
discovered by the Investigator at any time after the study should be reported. 
Any SAE that occurs must be reported to the clinical research organization (CRO) within 
24 hours of its occurrence or within 24 hours of learning of its occurrence. Recurrent episodes, 
complications, or progression of the initial SAE must be reported to the CRO as follow-up to the 
original episode within 24 hours of the Investigator receiving the information. Information about 
all SAEs will be collected and recorded on the SAE form. All pertinent medical records and 
information collected during the treatment and follow-up of the subject should be maintained at 
the site with a copy emailed to MIRA3_Safety@Oculoscr.com. The Investigator must assess the 
SAE relationship and complete the SAE form. The CRO may request additional information. 
Follow-up information (eg, discharge summary) will be retained in the subject’s chart and a copy 
will be emailed to MIRA3_Safety@Oculoscr.com. In addition, all SAEs should be recorded on 
the AE eCRF page with the serious question marked “Yes”. 
It is the Investigator’s responsibility to notify the approving IRB of any SAEs on a timely basis 
as instructed by Ocuphire following Ocuphire’s determination of causality. All subjects who 
experience an SAE should be followed clinically and undergo the appropriate diagnostic 
evaluations until stabilization or resolution of the event. Ocuphire will report all SAEs to the 
FDA on the appropriate schedule depending if the event is drug related, not drug related, 
expected, or unexpected (based on the available information in the IB). 
Any death occurring during the study and follow-up period must be reported as an SAE. For any 
death occurring through the end of the study, regardless of the degree of relationship to study 
medication, the SAE resulting in the death must be reported to the CRO. A death occurring after 
completion of the study including the Safety Follow-up Visits, that is not reasonably associated 
with study medication administration, does not require completion of the SAE form. 


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9.3.2. Follow-up of subjects after adverse events 
If an AE/adverse reaction occurs, the Investigator will institute support and/or treatment as 
deemed appropriate. All SAEs ongoing at the time of the last visit or discontinuation from the 
study will be followed up until the AE/adverse reaction is resolved or stabilized, the subject is 
lost to follow-up, or there is other resolution to the event.  
10. STATISTICS 
A detailed presentation of the statistical approach is outlined in the Statistical Analysis Plan. 
10.1. Sample size 
A sample size of approximately 330 subjects (approximately 220 treated with Nyxol) in this 
study will result in a total of > 300 subjects treated with Nyxol in the reversal of mydriasis 
program (including prior studies); this number of subjects is needed to meet the minimum 
number of subjects exposed to Nyxol to assess safety in this population. The primary efficacy 
endpoint will be met if subjects show a positive effect for Nyxol (α = 0.05 significance, two-
tailed). A total of 330 subjects (Nyxol, n=220; placebo, n=110) will provide > 90% power to 
detect a difference of 25% between the Nyxol and placebo arms in percentage of subjects 
returning to ≤ 0.2 mm from baseline PD at 90 minutes. This calculation is based on a 20% 
placebo effect and a 45% Nyxol treatment effect. The assumptions for this power calculation are 
estimated from the MIRA-2 Phase 3 results.  
All subjects will be randomized into the study in a 2:1 ratio to 1 of the 2 treatment arms (Nyxol 
or placebo, respectively), with a 1:1 stratification by light/dark irides (equal number of light and 
dark irides stratified across treatment groups). Furthermore, subjects will be randomized into the 
study at a ratio of 3:1:1 to mydriatic agent (2.5% phenylephrine, 1% tropicamide, or Paremyd, 
respectively). Therefore, if 198 subjects are randomized to 2.5% phenylephrine, then 66 subjects 
will be assigned to 1% tropicamide and 66 subjects to Paremyd, resulting in 330 total subjects.  
10.2. Analysis Populations 
Modified Intent-to-Treat Population: The mITT Population will include all randomized 
subjects who received 2 drops of study treatment in the study eye and had at least 1 scheduled 
post-treatment PD measurement during Visit 1. The mITT Population 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. 
Per Protocol Population: The PP Population includes all subjects in the mITT Population who 
had 2 drops of study treatment in the study eye, had all scheduled PD measurements during Visit 
1, had an increase of > 0.2 mm in PD in the study eye at 0 minutes compared to baseline 
(-1 hour), and had 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.  
All Randomized Population (ARP): The ARP will include all randomized subjects. This 
population is also known as the Intent-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. 


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Safety Population (SP): The SP will include all randomized subjects who received at least 
1 drop of study treatment. The SP will be used to summarize safety variables, using the treatment 
they actually received. 
Pharmacokinetic Population: The PK Population will include all subjects who had at least one 
PK sample taken at any post-treatment timepoint. 
10.3. Statistical methods  
10.3.1. General considerations 
All continuous variables will be summarized by treatment and time point (as applicable) using 
descriptive statistics (n, mean, median, standard deviation, minimum, and maximum). All 
categorical variables will be summarized by treatment and time point (as applicable) using 
frequency counts and percentages.  
All study data will be listed by treatment, subject, and time point (as applicable). 
All statistical tests will be performed using a significance level of 5% (two-tailed). The p-values 
for the analysis of secondary efficacy endpoints will be considered descriptive. 
10.3.2. Demographic and baseline characteristics 
Demographic and baseline characteristics such as age, race, and sex will be summarized by 
treatment group using the mITT Population, PP Population, SP, PK Population, and the ARP. 
These data will also be provided in by-subject listings. 
10.3.3. Subject disposition 
Subject disposition, including randomization, completion, and withdrawal from the study, will be 
summarized using the ARP. These data will also be provided in by-subject listings. 
10.3.4. Medical history and prior/concomitant medications 
Medical history will be coded using the latest version of Medical Dictionary for Regulatory 
Activities (MedDRA) and will be summarized by treatment group using the SP.  
Prior medications (medications with an end date prior to the date of randomization) and 
concomitant medications (medications with a start or end date after the date of randomization) 
will be coded using WHODrug and will both be summarized by treatment group using the SP. 
Medical history and prior and concomitant medications will also be provided in by-subject 
listings.  
10.3.5. Analysis of efficacy 
Efficacy will be assessed using the mITT and PP Populations, with subjects included in the 
treatment arm into which they were randomized. For the analysis of the primary efficacy 
endpoint, imputation will be performed for missing data as described in the Statistical Analysis 
Plan. 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 analyses 
may be performed using the ARP, with imputation performed for missing data. For the analysis 
of the secondary efficacy endpoints, only observed case data will be used.  
For all efficacy endpoints, baseline is defined as -1 hour prior to treatment on Visit 1. Max pupil 
dilation is defined as 0 minutes, during which maximum PD is expected.  


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All efficacy data will be summarized by treatment group, study day, and time point (-1 hour 
[baseline], 0 minutes, 30 minutes, 60 minutes, 90 minutes, 2 hours, 3 hours, 4 hours, 6 hours, and 
24 hours), as appropriate. 
The primary efficacy endpoint is the percentage of subjects returning to ≤ 0.2 mm from baseline 
(-1 hour) photopic pupil diameter at 90 minutes post treatment in the study eye. Subjects who did 
not have an increase of > 0.2 mm in pupil diameter in the study eye at the max pupil dilation 
(0 minutes) will not be counted in the PP Population as returning to ≤ 0.2 mm from baseline 
pupil diameter. The primary efficacy endpoint will be analyzed using a logistic regression model 
with treatment, mydriatic agent, and light/dark irides as factors and the baseline pupil diameter as 
a covariate. The percentage of subjects in each treatment group meeting the criteria and the odds 
ratio (OR) with 95% confidence interval (CI) and p-value will be provided. The analysis will be 
performed using the mITT and PP Populations, with subjects included in their randomized 
treatment regardless of the treatment they actually received.  
A sensitivity analysis of the primary efficacy endpoint will be completed using a logistic 
regression model with treatment as a factor and the baseline pupil diameter as a covariate; the 
mydriatic agent and irides type are excluded from this model. 
Secondary efficacy endpoints are indicated in Section 4.1.  
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 (-1 hour) value 
included as the covariate. Note that most secondary efficacy endpoints are in relation to baseline 
(-1 hour), whereas some endpoints are in relation to max pupil dilation (0 minutes). Each 
ANCOVA will be performed using the mITT or PP Populations (as specified in the Statistical 
Analysis Plan), with subjects included in their randomized treatment regardless of the treatment 
they actually received. The output from each ANCOVA will include the least squares mean 
(LSM) and standard error for both treatment groups, along with the placebo-corrected LSM, its 
95% CI, and associated p-value.  
A comparison of the study eye and fellow eye for each subject will be completed for the primary 
efficacy endpoint, as well as by mydriatic agent and light/dark irides.  
For each of the secondary endpoints related to percentage of subjects achieving certain criteria, 
the analysis will be performed using a logistic regression model with treatment, mydriatic agent, 
light/dark irides as factors, and the respective baseline as a covariate. For each analysis, the 
percentage of subjects in each treatment group meeting the criteria and the OR with 95% CI and 
p-value will be provided. For these endpoints, the mITT and PP Populations will be used, with 
subjects included in their randomized treatment regardless of the treatment they actually 
received. 
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 treatment, mydriatic agent, light/dark irides as factors, and the baseline pupil diameter as a 
covariate. 
In addition, secondary efficacy endpoints will be analyzed by light/dark irides and by mydriatic 
agent using the same model indicated above but without irides or mydriatic agent as a factor, as 
appropriate. Each mydriatic agent will be analyzed individually, and an additional analysis 


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combining 1% tropicamide and Paremyd subjects into a “tropicamide or Paremyd” group will be 
performed. Other subgroups, such as age, sex, and race, may be analyzed as well. If there is 
sufficient sample, analysis of selected efficacy endpoints will be completed for the subgroup of 
pediatric subjects.  
10.3.6. Analysis of safety 
Safety will be assessed using the SP, with subjects included in the treatment group they actually 
received regardless of their randomized treatment. Observed case data will be used; no 
imputation will be performed for missing safety data. 
Safety endpoints are indicated in Section 4.1.  
For HR and BP, baseline is defined as the screening value. Heart rate and BP values and change 
from baseline in the values will be summarized by treatment group and time point (screening, 
6 hours, and 24 hours).  
Observed values and change from baseline (-1 hour) in conjunctival hyperemia at each time 
point (0 minutes, 30 minutes, 60 minutes, 90 minutes, 2 hours, 3 hours, 4 hours, 6 hours, and 
24 hours) will be summarized for the study eye and the fellow eye. Treatments will be compared 
using the same ANCOVA model proposed for the continuous secondary efficacy endpoints. 
Conjunctival hyperemia will also be summarized categorically. 
Visual acuity assessments (BCDVA and DCNVA) will also be summarized at selected time 
points (BCDVA at 0 minutes, 60 minutes, 2 hours, 6 hours, and 24 hours; DCNVA at 0 minutes, 
90 minutes, 3 hours, 6 hours, and 24 hours) using the number of letters correctly identified. 
Letters will be recorded and may later be converted to logMAR by programming for such 
analyses. One letter is equivalent to 0.02 logMAR. As a reference five letters is represented by 
1 line. Treatments will be compared using the same ANCOVA model proposed for the 
continuous secondary efficacy endpoints. Distance-corrected near visual acuity will also be 
analyzed by mydriatic agent using a model that does not include the mydriatic agent as a factor. 
For IOP, baseline is defined as the screening value. Observed values and change from baseline in 
IOP at 6 hours will be summarized for the study eye and the fellow eye. Treatments will be 
compared using the same ANCOVA model proposed for the continuous secondary efficacy 
endpoints. 
Ocular tolerability values will be summarized by treatment group at max pupil dilation 
(0 minutes). Additionally, the categories “No Discomfort” and “Mild Discomfort” will be pooled 
into a single category and summarized descriptively, as will the categories “Moderate 
Discomfort” and “Severe Discomfort”. Treatments will be compared for the 2 pooled categories 
using a Fisher’s exact test. 
Subject questionnaire values will be summarized by treatment group (overall and by mydriatic 
agent) and time point (-1 hour, 0 minutes, 60 minutes, 2 hours, 4 hours, and 24 hours). 
Verbatim descriptions of AEs will be coded using MedDRA. Only TEAEs (those that occur after 
the first dose of study medication or increasing in severity after initiation of study medication) 
will be summarized. Treatment-emergent AEs and SAEs will be summarized by treatment group, 
by system organ class (SOC), severity, and relationship to study medication. Deaths, withdrawal 
from study medication due to AEs, and withdrawal from the study due to AEs will each be 
summarized by treatment group. Note that in MedDRA, ocular events are coded to the SOC of 


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“special senses”. Thus, using SOC in the summaries will provide a separation of ocular and non-
ocular AEs. 
All safety data will be provided in by-subject listings. Safety tables and listings for pediatric 
subjects may be provided. 
10.3.7. Analysis of Pharmacokinetics 
Nyxol plasma concentrations will be summarized by nominal sampling time using descriptive 
statistics. Individual subject PK data will be listed.  
10.4. Procedure for accounting for missing, unused, or spurious data 
For the summarization and analysis of the primary efficacy endpoint, imputation will be 
performed for missing data as described in the Statistical Analysis Plan. For the summarization 
and analysis of secondary efficacy endpoints and safety data, observed case data only will be 
used. 
10.5. Procedure for reporting deviations from the statistical plan 
Any deviations from the Statistical Analysis Plan will be described and a justification given in 
the final Clinical Study Report.  
11. DIRECT ACCESS TO SOURCE DATA AND DOCUMENTS 
The Investigator will permit study-related monitoring visits, audits, IRB review, and regulatory 
inspection(s) by providing direct access to source data and documents. 
12.  QUALITY CONTROL AND QUALITY ASSURANCE 
The progress of the study will be monitored by on-site, written, and telephone communications 
between personnel at the Investigator’s site and the Medical Monitor. Should the COVID-19 
pandemic restrict monitors from traveling to a site, remote review will be conducted to the extent 
possible, while still ensuring the study is monitored appropriately per applicable regulations and 
guidelines. The Investigator will allow Ocuphire, the study monitor, and the Medical Monitor to 
inspect all eCRFs, subject records (source documents), signed consent/assent forms, records of 
study medication receipt, storage, preparation, and disposition, and regulatory files related to this 
study.  
13.  ETHICAL CONSIDERATIONS AND GOOD CLINICAL PRACTICE 
COMPLIANCE  
13.1. Good Clinical Practice compliance 
The proposed study is subject to all applicable governmental rules and regulations concerning 
the conduct of clinical trials on human subjects. This includes, but is not necessarily limited to, 
the approval of IRBs, the Helsinki Declaration, US FDA law, ICH GCP Guidelines, obtaining 
prospective informed consent, monitoring of the conduct of the study and the completeness of 
the eCRFs by Ocuphire or its designee(s), and appropriate record retention by the Investigator.  
13.2. Institutional Review Board 
This protocol, materials used to recruit subjects, and materials used to document consent/assent 
must be approved by the IRB prior to initiation of the study. Written IRB approval must 
adequately identify the protocol and informed consent/assent. In addition to approving the 
protocol, the IRB must also approve the Subject Information and Consent/Assent/Parental 


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Consent Form, as well as any advertising tools that will be used for the study. Copies of all 
approved materials, all correspondence with the IRB, and written approval from the IRB must be 
made available to Ocuphire, prior to the start of subject enrollment into the study. 
13.3. Protocol deviations/violations 
The Investigator should not deviate from the requirements of this protocol without prior written 
approval of the Medical Monitor or Sponsor except in the event of a medical emergency. 
A reportable protocol deviation is defined as nonadherence to the protocol that involves 
inclusion/exclusion criteria, affects subject safety, rights, or welfare, or has the potential to affect 
the integrity of the data. Examples of major protocol deviations include study enrollment by 
ineligible subject, loss of key data such as equipment malfunction (eg, pupillometer), and/or use 
of a prohibited medication during the study.  
All protocol deviations will be reported by entering the event in the appropriate eCRF page. 
Protocol deviations should be reported to the IRB in accordance with IRB-specific guidelines. If 
there is any question as to whether the deviation is reportable, Ocuphire or its designee and the 
IRB should be contacted. 
All changes to the protocol will be made by the Sponsor or designee as an approved amendment 
to the protocol, submitted to the FDA, and approved by the IRB prior to implementation.  
Changes implemented without prior approval will be considered protocol violations. 
13.4.  Informed consent and assent requirements 
Written informed consent will be obtained from each adult subject. A signed assent form will be 
obtained for all minors ages 12 to 17, as well as a separate parental/Legal Guardian consent. A 
copy of the signed and dated consent/assent document will be given to each subject or parent 
guardian. The original signed and dated informed consent/assent document must be maintained 
in the study files at the Investigator’s site.  
The Investigator is responsible for ensuring that no subject is subject to any study-related 
examination or activity before that subject has given informed consent/assent. The subject must 
give written consent/assent after the receipt of detailed information. The verbal explanation will 
cover all the elements specified in the written information provided to the subject. 
It should be emphasized that the subject is at liberty to withdraw consent/assent to participate at 
any time, without penalty or loss of benefits to which the subject is otherwise entitled. Subjects 
who refuse to give or withdraw written informed consent/assent may not be included or 
continued in this study, but this will not impact on their subsequent care.  
The Investigator will inform the subject of the aims, methods, anticipated benefits, and potential 
hazards of the study, including any discomfort it may entail. The subject must be given every 
opportunity to clarify any points he/she does not understand and, if necessary, ask for more 
information. At the end of the interview, the subject may be given time to reflect if this is 
required, or if the subject requests more time. Subjects and/or legal guardian will be required to 
sign and date the informed consent form.  
A copy of the signed and dated consent/assent document will be given to each subject. The 
original signed and dated informed consent/assent document must be maintained in the study 


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files at the Investigator’s site. Signed informed consent/assent must be obtained prior to the 
conductance of any study procedures. 
14.  DATA HANDLING AND RECORD KEEPING 
All procedures for the handling and analysis of data will be conducted using good computing 
practices meeting ICH and US FDA guidelines for the handling and analysis of data for clinical 
trials. 
14.1.  Data entry 
Study-specific data that have been outlined in the protocol will be entered into the clinical 
database by individual(s) designated by the Investigator.  
14.2. Data quality control and reporting 
Data are verified electronically using a series of programmed edit checks that have been created 
by the Clinical Data Manager and programmed by the Clinical Data Programmer or designee. 
Data discrepancies will be brought to the attention of the clinical team and investigated by the 
clinical research associate (CRA) and site staff. Clinical research associates will review and 
verify all data collected in the eCRF against source documentation during scheduled monitoring 
visits. The CRA will work closely with the site staff to address any discrepancies that have been 
found so that proper resolutions can be made and documented in the clinical database. An audit 
trail within the system will track all changes made to the data. 
14.3.  Archiving of data 
Archived versions of the database will be saved by Ocuphire consistent with ICH GCP 
Guidelines, complying with whichever of the requirements is longer. Ocuphire will notify the 
Investigator when documents should be returned. 
14.4.  Records retention 
The Investigator’s site and clinical laboratory will retain all records related to the study in 
compliance with ICH GCP Guidelines. 
14.5.  Amendments to the protocol 
Modifications of the signed protocol are only possible by approved protocol amendments and 
with the agreement of all responsible persons. The procedure for approval of a protocol 
amendment is identical to that for approval of the protocol. The IRB must be informed of all 
protocol amendments and should be asked for its opinion as to whether a full re-evaluation of the 
ethical aspects of the study is necessary by the committee. This should be fully documented. 
The Investigator must not implement any deviation from or change to the protocol, without 
discussion with and agreement by Ocuphire and prior review and documented 
approval/favorable opinion of the amendment from the relevant ethics committee, except where 
it is necessary to eliminate an immediate hazard to study subjects, or where the change(s) 
involves only logistical or administrative aspects of the study (eg, change in monitor, change of 
telephone number). 
Protocol amendments will be submitted to the appropriate authority(ies) as required by the 
applicable regulatory requirement(s). 
 
 


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15. BIBLIOGRAPHY 
1. Hogan TS, McDaniel DD, Bartlett JD, Hart KK, Paggiarino DA. Dose-response study of 
dapiprazole HCl in the reversal of mydriasis induced by 2.5% phenylephrine. J Ocul 
Pharmacol Ther. 1997;13(4):297–302. 
2. Holladay J, Meyer A, Pitlick B. Phentolamine mesylate ophthalmic solution 1% once daily 
reduces pupil diameter and improves night vision disturbances. American Academy of 
Ophthalmology 2018 annual meeting. Presented at Chicago, IL; October 27-30, 2018. 
3. Karpecki PM, Foster SA, Montaquila SM, et al. Phentolamine eye drops reverse 
pharmacologically induced mydriasis in a randomized Phase 2b trial. Optom Vis Sci. 
20211;98(3):234‒242.  
4. McDonald M. Phentolamine mesylate treatment of severe night vision complaints. American 
Society of Cataract and Refractive Surgery 2011 annual meeting. Presented at San Diego, 
CA; March 25-29, 2011. 
5. OP-NYX-01a2 CSR. Ocuphire Pharma, Inc. 24 May 2013. 
6. OP-NYX-SNV CSR. Ocuphire Pharma, Inc. 07 February 2008. 
7. OPI-NYXG-201 CSR. Ocuphire Pharma, Inc. 23 April 2020. 
8. OPI-NYXRM-201 CSR. Ocuphire Pharma, Inc. 25 April 2020. 
9. Pepose JS, Hartman PJ, DuBiner HB, et al. Phentolamine mesylate ophthalmic solution 
provides lasting pupil modulation and improves near visual acuity in presbyopic glaucoma 
patients in a randomized Phase 2b clinical trial. Clin Ophthalmol. 2021;15:79–91. 
10. Pepose JS, Sooch MP, Charizanis K, et al. Phase 3 clinical trial to evaluate the efficacy of 
phentolamine ophthalmic solution on the reversal of pharmacologically induced mydriasis. 
ASCRS ASOA 2021 annual meeting. Presented at Las Vegas, NV; July 23-27, 2021.  
11. Pepose JS, Sooch MP, Charizanis K, et al. Phase 2 clinical trial to evaluate the efficacy of 
phentolamine ophthalmic solution and low-dose pilocarpine for the treatment of presbyopia. 
ASCRS ASOA 2021 annual meeting. Presented at Las Vegas, NV; July 23-27, 2021.  
12. Steinhauer SR, Siegle GJ, Condray R, Pless M. Sympathetic and parasympathetic innervation 
of pupillary dilation during sustained processing. Int J Psychophysiol. 2004;52(1):77–86. 
13. Yoshitomi T, Ito Y, Inomata H. Adrenergic excitatory and cholinergic inhibitory innervations 
in the human iris dilator. Exp Eye Res. 1985;40(3):453–9. 
14. Yu Y, Koss MC. alpha(1A)-adrenoceptors mediate sympathetically evoked pupillary dilation 
in rats. J Pharmacol Exp Ther. 2002;300(2):521–5. 
 


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APPENDIX 1: IRIS COLOR CHART 
Study enrollment includes both light- and dark-colored eyes. Examples of light irides (pictures 
A, B, C, D, and E) and dark irides (pictures F, G, and H) for the purposes of this study are 
detailed in the chart below.  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
A = Light blue 
B = Dark blue 
C = Blue with peripupillary brown 
D = Uniform green 
E = Green with brown iris ring 
F= Central brown and peripheral green 
G = Brown with some peripheral green 
H = Brown 


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APPENDIX 2: CONJUNCTIVAL HYPEREMIA GRADING SCALE USING IMAGES 
FROM CCLRU 
 
None (0) 
Mild (+1) 
Moderate (+2) 
Severe (+3) 
Normal; appears 
white with a small 
number of 
conjunctival blood 
vessel easily 
observed 
Prominent, pinkish-
red color of both the 
bulbar and palpebral 
conjunctiva 
Bright, scarlet red 
color of the bulbar 
and palpebral 
conjunctiva 
Beefy red with 
petechiae, dark red 
bulbar and palpebral 
conjunctiva with 
evidence of 
subconjunctival 
hemorrhage  
 
 
 


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APPENDIX 3: SUBJECT QUESTIONNAIRE 
 
Study: OPI-NYXRM-302 (MIRA-3) 
Subject Questionnaire 
Subject Number: _____  
 
 
                        
 Visit#___, Date_____  
 
 
 
 
 
 
 
 
Assessment Time point _______ 
 
For this questionnaire, if you normally wear glasses to read, please put them on now. 
Please read the following paragraph without squinting, and then answer the following questions.  
   
 
 
Sunset is the time of day when our sky meets the end of the horizon. 
There are blue, pink, and purple swirls spinning and twisting in the sky. 
There is a coolness, a calmness, when the sun does set. 
 
  
1. Circle the number below to describe the severity of your symptoms while reading the above 
paragraph.  
  
None  
Mild  
Moderate  
Severe  
A. Unable to read the text  
0  
1  
2  
3  
B. Blurry text  
0  
1  
2  
3  
C. Words going in and out of focus  
0  
1  
2  
3  
 
 
2. For the following statement, circle the number to indicate whether you agree or disagree.  
  
 
3. For the following statement, circle the number to indicate whether you agree or disagree. 
 
 
4. For the following question, circle the number to indicate how you would respond. 
 
 
 
  
Strongly 
Disagree 
Disagree 
Neither 
Agree/ 
Disagree 
Agree 
Strongly 
Agree 
My distance vision is blurrier than 
normal 
1 
2 
3 
4 
5 
  
Strongly 
Disagree 
Disagree 
Neither 
Agree/ 
Disagree 
Agree 
Strongly 
Agree 
I am satisfied with my vision at the 
moment.  
1 
2 
3 
4 
5 
  
Extremely 
Unlikely 
Somewhat 
Unlikely 
No 
Preference 
Somewhat 
Likely 
Extremely 
Likely 
How likely would you be to take this 
eye drop the next time your eyes are 
dilated? 
1 
2 
3 
4 
5 


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APPENDIX 4: RANDOMIZATION SCHEMA BY INVESTIGATIONAL TREATMENT, 
MYDRIATIC AGENT, AND IRIS COLOR 
All subjects 
1:1 iris color 
3:1:1 mydriatic agent 
2:1 treatment 
330 
165 Light 
99 Phenylephrine 
66 Nyxol 
33 Placebo 
33 Tropicamide 
22 Nyxol 
11 Placebo 
33 Paremyd 
22 Nyxol 
11 Placebo 
165 Dark 
99 Phenylephrine 
66 Nyxol 
33 Placebo 
33 Tropicamide 
22 Nyxol 
11 Placebo 
33 Paremyd 
22 Nyxol 
11 Placebo 
 
 
 


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APPENDIX 5: ADRENERGIC AND CHOLINERGIC DRUGS 
The following drugs are examples of drugs which cannot be used within 7 days prior to 
screening, or during the study unless the drug, dose and regimen has been consistent for the 7 
days prior to screening. This list is not inclusive of all drugs in these classes. If there is any 
doubt, please consult with the Medical Monitor. 
Alpha-1-agonists 
Methyl norepinephrine 
Naphazoline 
Oxymetazoline 
Tetrahydrozoline 
Phenylephrine 
Xylometazoline 
 
Alpha-2-agonists 
Brimonidine 
Clonidine 
Guanfacine 
Guanabenz 
Guanoxabenz 
Guanethidine 
Xylazine 
Tizanidine 
Methyldopa 
Nonselective alpha-
antagonists 
Phenoxybenzamine 
Tolazoline 
Labetalol 
Carvedilol 
 
Alpha-1-
antagonists  
Alfuzosin 
Prazosin 
Doxazosin 
Tamsulosin 
Terazosin 
 
Alpha-2-
antagonists 
Atipamezole 
Idazoxan 
Yohimbine  
Acetylcholine 
receptor agonists 
Pilocarpine (M3 
receptors) 
 
Acetylcholine 
receptor antagonists 
Scopolamine 
Dicycloverine 
Tolterodine 
Oxybutynin 
Ipratropium 
Mamba toxin (MT7) 
Pirenzepine 
Telenzepine 
 
Antivertigo 
Meclizine  
Scopolamine 
Gastrointestinal 
Atropine  
Belladonna 
 
Parkinsonism 
Amantadine 
Benztropine 
Biperiden 
Trihexyphenidyl