From: David Shayne <ShayneDav@Katori.

org>
Subject: Reevaluation of Protein Assay
Date: 25 May 2013
To: DMPK <Drug Metabolism and Pharmokinetics>
(1 Attachment, 347 KB)
Dear All,
This morning we will be starting the reevaluation of KT-TTRsc, a subcutaneously
administered RNAi therapeutic targeting TTR for the treatment of TTR-mediated
amyloidosis. Hopefully, this is will help us start our first phase in clinical trials by
the end of June.
We are excited to be advancing what we believe to be the industry leading effort in
ATTR, which includes KT-TTR02 for the treatment of FAP and now KT-TTRsc for the
treatment of FAC.
Attached is the data that was shown at our last group meeting. Please review it
before meeting again on May 27th, for the reevaluation.
-David
David Shayne, PhD,
Senior Director of DMPK,
Katori Pharmaceuticals Inc.

(Attachment)
Male (30 mg/kg)
<LLOQ (20.0)
3911
3137
2343
2156
231
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)

Male (100 mg/kg)

<LLOQ (20.0)
16867
24467
11167
14340
2534
534
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)

Male (300 mg/kg)

<LLOQ (20.0)
70989
43828
54254
13498
45434
3483
64.0
32.1
<LLOQ
<LLOQ
<LLOQ
<LLOQ

Male (30 mg/kg)

Male (100 mg/kg)
Male (300 mg/kg)

100000
Plasma ALN-51547 (ng/mL)

Time
Hours
0
0.5
1
2
4
8
24
48
120
168
336
504
672

10000
1000
100
10
1
0

12

24

36

48

60 Time
72 (h) 84

96

108

120

132

144

Time
Hours
0
0.5
1
2
4
8
24
48
120
168
336
504
672

Female (30
mg/kg)
<LLOQ (20.0)
5442
3435
2333
1627
434
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)

Female (100
mg/kg)
<LLOQ (20.0)
45345
34523
45345
23534
3433
101.4333333
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)

Female (300
mg/kg)
<LLOQ (20.0)
23423
34234
12434
34234
23433
4343
43
32
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)

Plasma ALN-51547 (ng/mL)

100000
10000

Female (30 mg/kg)

Female (100 mg/kg)

1000
100
10
1
0

12

24

36

48

60

72
84
Time (h)

96

108

120

132

144

From: Jane Bowe <BoweJa@Katori.org>

Subject: Group Meeting
Date: 27 May 2013
To: DMPK <Drug Metabolism and Pharmokinetics>
CC: David Shayne <ShayneDav@Katori.org>
Hi All,
Just reminding you all that we have our reevaluation of TTRsc meeting today. Please
meet in Hystor Conference Room at 1:00 pm.
Peter, please give an update on the invitro metabolism on the AAT.
Ju, please give an update on the NHP AT3 protein data.
Heiral, please give an update on the mice AT3 ELISA.
Vince, please give an update on the PEG antibody assay interference assay with KTTTR02 in rat.
Thanks,
Jane
Jane Bowe
Senior Scientist of DMPK,
Katori Pharmaceuticals Inc.

From: David Shayne <ShayneDav@Katori.org>

Subject: Reevaluation of Protein Assay
Date: 27 May 2013
To: DMPK <Drug Metabolism and Pharmokinetics>
(1 Attachment, 590 KB)
Dear All,
Thank you for attending the group meeting this afternoon for the reevaluation of
ALN-TTR02. There will be a few changes made in the previous protocol to eliminate
three values that read <LLOQ under the female category with a concentration of 300
mg/kg.
As we discussed, we will be retesting TTR02 from mice plasma. We will follow the
attached protocol instead of the previous one. Please respond if you have any
questions about it.
David
David Shayne, PhD,
Senior Director of DMPK,
Katori Pharmaceuticals Inc.

(Attached)
KT17128_TTR02-RS11-002_Mouse PK_Plamsa_20130527
Preparation of Solution
Proteinase
.K solution

Vol._each
sample

Vol._total
samples

Cat. #

Lot #

Brand

PBS

90

6120

AM9624

906010

Ambion

Proteinase
K

408

AM2547

0902010

Ambion

Lysis
Buffer

30

2040

MTC096H

TC-90329

Epicentre

Total

126

8568

A-probe
(1uM)
A-94151

Vol._each
sample

Vol._total
samples

N-probe
(10 uM)

Vol._each
sample(ul)

Vol._total
samples(ul)

408

A-93254

408

Hypbribuffer
Total

36

2448

36

2448

42

2856

Hybribuffer
Total

42

2856

# of Std, QC,
SST, Blank

24

# of samples

26

# of total
samples

50

Hypridization:
1 Add 3 ul of stock solution of standard, SST and QC into 27 ul of blank matrix (250 ul PCR plate). The total volume is 30 ul
2 Add 30 ul of unknown samples into 250 ul PCR plate and the position are sown in the plate map below
3 Add 126 ul of proteinase K solution into each standard, QC, SST samples and unknown samples
4 Incubate all samples at 60C for 45 min
5 Add 42 ul of atto probe solution into each samples and incubate at 95C for 15 min
6 Put the plate of wet ice for 1 min, and keep all samples at 20C (r.t.) for 30 min
7 Add 42 ul of anti-probe solution to each sample while still at 20C and continue to incubate at 20C for 15 min
8 Transfer all samples into Agilent 96 well plate and load on HPLC for analysis
Plate 1

10

11

12

STD1-0.2

STD1-0.5

STD1-1

STD1-2

STD1-5

STD1-10

STD1-20

STD1-50

STD1-100

STD1-200

STD1-500

STD1-1000

DB

SST200

QC1

QC5

QC100

QC500

DB

SST200

QC1

QC5

QC100

QC500

1001/10

1002/10

1003/10

1004/10

1005/10

1006/10

1007/10

1008/10

1009

1010

1011

1012

1013

1014

1015

1016

1017

1018

1019

1020

1021

1022

1023

1024

1001/100

1002/100

1003/100

1004/100

F
G
H

From: Jane Bowe <BoweJa@Katori.org>

Subject: New Protocol
Date: 27 May 2013
To: Francine Chen <ChenFra@Katori.org>, Vince Reye <ReyeVin@Katori.org>
CC: David Shayne <ShayneDav@Katori.org>
Vince,
Please work with Francine on the new protocol David just attached in the previous
e-mail tomorrow. Hopefully this will be completed by the end of the next two weeks.
Thanks,
Jane
From: Vince Reye <ReyeVin@Katori.org>
Subject: TTR02
Date: 28 May 2013
To: Francine Chen <ChenFra@Katori.org>
(1 Attachment, 200 KB)
Francine,
When you arrive to work today, please see me to discuss about the new protocol
David just sent out and about the dilution table attached. We will be doing three of
the same tests to verify the results. Attached below is the analytical method
development for the mouse plasma ID: KT-17128.
Vince

(Attached)
The preparation of stock solution of standard
Conc. (ng/ml)

Total_Vol. (uL)

Additive

Vol._PBS
(uL)

Vol._Additive(uL)

Stock 1

1000000

121.2

Formulation

48.5

72.7

Stock 2

100000

211.8

Stock 1

21.2

190.6

Stock 3

200000

159

Stock 2

31.8

127.2

DB

200

PBS

180

WS-A

80

200

WS-F

20

180

WS-B

100

200

WS-G

20

180

WS-C

150

200

WS-H

20

180

WS-D

200

200

WS-I

20

180

WS-E

400

200

WS-J

20

180

WS-F

800

200

WS-K

20

180

WS-G

1000

200

WS-L

20

180

WS-H

1500

200

Stock 3

15

185

WS-I

2000

200

Stock 3

20

180

WS-J

4000

200

Stock 2

192

WS-K

8000

200

Stock 2

16

184

WS-L

10000

200

Stock 2

20

180

From: Francine Chen <ChenFra@Katori.org>

Subject: RE: TTR02
Date: 29 May 2013
To: Vince Reye <ReyeVin@Katori.org>
Vince,
I have just completed the dilution task. The plates are being refrigerated in Fridge 2,
Shelf A. They are labeled Dilution Stocks. When you want to start on the new
protocol please let me know.
Thanks,
Francine
From: Vince Reye <ReyeVin@Katori.org>
Subject: RE: TTR02
Date: 29 May 2013
To: Francine Chen <ChenFra@Katori.org>
Francine,
Please start on the protocol without me. I am in a conference meeting on the 4th
floor for another hour. I should be done in time to help you to set up the HPLC
machine to start the test.
Thanks,
Vince
From: Vince Reye <ReyeVin@Anlam.org>; Francine Chen <ChenFra@Katori.org>
Subject: TTR02 new data
Date: 5 June 2013
To: Jane Bowe <BoweJa@Katori.org>
CC: David Shayne <ShayneDav@Katori.org>
(Attachment, 550 KB)
David and Jane,
Francine and I completed the new experiments for TTR02. Attached is the analyzed
data. We will plot them in graphs to present during our scheduled group meeting
this Friday.
(Attachment Below)

Subject
Group

Subject

Hour
Nominal

Concentration
(ng/mL)

1/2
1/2
1/2
1/2
1/2
1/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2

B41371
B41372
B41373
B41451
B41452
B41453
B41384
B41384
B41384
B41384
B41384
B41385
B41385
B41385
B41385
B41385
B41386
B41386
B41386
B41386
B41386
B41387
B41387
B41387
B41387
B41388
B41388
B41388
B41388
B41389
B41389
B41389
B41389
B41390
B41390
B41390
B41390
B41391
B41391
B41391
B41391
B41392
B41392
B41392
B41392
B41464
B41464
B41464
B41464
B41464
B41465
B41465

1
1
1
1
1
1
0
2
24
168
672
0
2
24
168
672
0
2
24
168
672
0.5
4
48
336
0.5
4
48
336
0.5
4
48
336
1
8
120
504
1
8
120
504
1
8
120
504
0
2
24
168
672
0
2

<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
2650
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
2210
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
1990
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
5510
2090
<LLOQ (20.0)
<LLOQ (20.0)
3900
2020
<LLOQ (20.0)
<LLOQ (20.0)
2510
1900
<LLOQ (20.0)
<LLOQ (20.0)
4870
595
<LLOQ (20.0)
<LLOQ (20.0)
3610
299
<LLOQ (20.0)
<LLOQ (20.0)
3840
430
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
1990
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
1970

Comments

Assay
Date/Time

Custom
ID

01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
04-Jun-2013
01-Jun-2013
02-Jun-2013
03-Jun-2013
04-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
04-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
03-Jun-2013
03-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
04-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
04-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
04-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013

20121320001
20121320002
20121320003
20121320094
20121320095
20121320096
20121320004
20121320005
20121320006
20121320007
20121320187
20121320008
20121320009
20121320010
20121320011
20121320188
20121320012
20121320013
20121320014
20121320015
20121320189
20121320016
20121320017
20121320018
20121320190
20121320019
20121320020
20121320021
20121320191
20121320022
20121320023
20121320024
20121320192
20121320025
20121320026
20121320027
20121320193
20121320028
20121320029
20121320030
20121320194
20121320031
20121320032
20121320033
20121320195
20121320097
20121320098
20121320099
20121320100
20121320214
20121320101
20121320102

2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2
2/2

B41465
B41465
B41465
B41466
B41466
B41466
B41466
B41466
B41467
B41467
B41467
B41467
B41468
B41468
B41468
B41468
B41469
B41469
B41469
B41469
B41470
B41470
B41470
B41470
B41471
B41471
B41471
B41471
B41472
B41472
B41472
B41472

24
168
672
0
2
24
168
672
0.5
4
48
336
0.5
4
48
336
0.5
4
48
336
1
8
120
504
1
8
120
504
1
8
120
504

<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
2830
<LLOQ (20.0)
<LLOQ (20.0)
<LLOQ (20.0)
2620
1340
<LLOQ (20.0)
<LLOQ (20.0)
2790
1440
<LLOQ (20.0)
<LLOQ (20.0)
2920
1390
<LLOQ (20.0)
<LLOQ (20.0)
3210
236
<LLOQ (20.0)
<LLOQ (20.0)
4120
322
<LLOQ (20.0)
<LLOQ (20.0)
2880
183
<LLOQ (20.0)
<LLOQ (20.0)

01-Jun-2013
01-Jun-2013
04-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
04-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
04-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
04-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
01-Jun-2013
04-Jun-2013
01-Jun-2013

20121320103
20121320104
20121320215
20121320105
20121320106
20121320107
20121320108
20121320216
20121320109
20121320110
20121320111
20121320217
20121320112
20121320113
20121320114
20121320218
20121320115
20121320116
20121320117
20121320219
20121320118
20121320119
20121320120
20121320220
20121320121
20121320122
20121320123
20121320221
20121320124
20121320125
20121320126
20121320222

From: David Shayne <ShayneDav@Katori.org>

Subject: RE: TTR02 new data
Date: 5 June 2013
To: Vince Reye <ReyeVin@Katori.org>; Francine Chen <ChenFra@Katori.org>; Jane
Bowe <BoweJa@Katori.org>
Thank you for all your hard work. I am looking forward to seeing yours and
Francines presentation at our group meeting. Glad that the data has a better
concentration than our last data.
David
From: David Shayne <ShayneDav@Katori.org>
Subject: Phase 1 Clinical Trials
Date: 6 June 2013
To: DMPK <Drug Metabolism and Pharmokinetics>
Hi All,
From the desired data received from our latest experiments, we are now sending
that set information for approval by the health authority and ethics committee to
initiate Phase 1 in clinical trials.
I just want to congratulate you all beforehand. Im hoping to hear good news in a
few weeks.
David

From: David Shayne <ShayneDav@Katori.org>

Subject: Phase 1 Clinical Trials
Date: 20 June 2013
To: DMPK <Drug Metabolic and Pharmokinetic>; Katori <124+.>
Dear All,
Congratulations!
This morning we announced that we have initiated dosing in a Phase I clinical trial
with KT-TTR02, a subcutaneously administered RNAi therapeutic targeting TTR for
the treatment of TTR-mediated amyloidosis. This is an exciting development in our
company - one that represents an important milestone.
We are excited to be advancing what we believe to be the industry leading effort in
ATTR, which includes this KT-TTR02 for the treatment of familial amyloidotic
polyneuropathy (FAP) and also KT-TTRsc for the treatment of familial amyloidotic
cardiomyopathy (FAC). With the start of this Phase I trial, KT-TTR02 becomes the
first RNAi therapeutic that utilizes our proprietary GalNAc conjugate delivery
platform to enter the clinic.
Furthermore, it is the industry's first systemic RNAi therapeutic to be delivered
subcutaneously. We expect to present data from this Phase I trial in the middle of
this year, and, upon completion of the trial, plan to start a Phase II study in FAC
patients by the end of this year.
Congratulations again to all on reaching this important milestone!
Best,
David
David Shayne, PhD,
Senior Director of DMPK,
Katori Pharmaceuticals Inc.