US 2011 0166063A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0166063 A1 BOSsard et al. (43) Pub. Date: Jul. 7, 2011

(54) POLYMER CONJUGATES OF THERAPEUTIC Related U.S. Application Data PEPTIDES (60) Provisional application No. 61/192,672, filed on Sep. 19, 2008, provisional application No. 61/208,089, (75) Inventors: Mary J. Bossard, Madison, AL filed on Feb. 18, 2009, provisional application No. (US); Steven O. Roczniak, 61/153,966, filed on Feb. 19, 2009. Greensboro, NC (US); Harold Zappe, Harvest, AL (US); Yujun Publication Classification Wang, Madison, AL (US); Ping (51) Int. Cl. Zhang, Madison, AL (US); Dawei A638/28 (2006.01) Sheng, Madison, AL (US); C. A638/16 (2006.01) Simone Jude-Fishburn, Redwood A638/08 (2006.01) City, CA (US); Elizabeth Louise A638/06 (2006.01) Minamitani, Lacey's Spring, AL A638/07 (2006.01) (US); Xiaofeng Liu, Union City, A638/10 (2006.01) CA (US); Haim Moskowitz, San C07K 5/08 (2006.01) Diego, CA (US); Dennis G. Fry, C07K 7/06 (2006.01) Huntsville, AL (US); Cherie F. Ali, C07K 7/08 (2006.01) Burlingame, CA (US); Christine C07K I4/00 (2006.01) Taylor Brew, Pacifica, CA (US) A6IP3/10 (2006.01) (52) U.S. Cl...... 514/5.9; 514/21.3; 514/21.2: 514/21.7; (73) Assignee: Nektar Therapeutics, San 514/21.6; 514/21.9; 514/21.8; 514/21.5; 530/303; Francisco, CA (US) 530/331; 530/330,530/329; 530/328; 530/327; 530/326; 530/324; 530/350 (21) Appl. No.: 13/119,297 (57) ABSTRACT (22) PCT Filed: Sep. 17, 2009 The invention provides peptides that are chemically modified by covalent attachment of a water-soluble oligomer. A con (86). PCT No.: PCT/US09/05.192 jugate of the invention, when administered by any of a num ber of administration routes, exhibits characteristics that are S371 (c)(1), different from the characteristics of the peptide not attached (2), (4) Date: Mar. 16, 2011 to the water-soluble oligomer.

P es Mono 8sa 8. Conjugate s r c h S e 9 O

OO 250 Volume (mL) Patent Application Publication Jul. 7, 2011 Sheet 1 of 84 US 2011/O166063 A1

2 4. O

S 9 O es 5. Mono 8. Conjugate l4O S 2 e 9 O

150 2OO 250 Volume (ml)

Figure KISS1.1

DAD-E. Sig38S, 16 Re-38C, GE LMKISSPEPTIFFGF2308A8-07-23 (3-53-530723.8Aloob)

Figure KISS 1.2 Patent Application Publication Jul. 7, 2011 Sheet 2 of 84 US 2011/0166063 A1

340.25 --- 400

200

1776,028

too

sess 20000 2sode Soo ssood 48-88 also Az Figure KISS 1.3

- UV1280nm - UV2225mm Cond ConC pH 14 13 KP-1 ButALD10K 12 W Coniugate

O

---m Un-reacted KP-10

ml O 2O 40 80 BO 1OO 120 140 18O

Figure KISS2.1 Patent Application Publication Jul. 7, 2011 Sheet 3 of 84 US 2011/O166063 A1

Figure KISS2.2

:

: s

3

5

25

s f SEAllish t ------T t t t

at SN Quality Fac. Res. Entens. Area 2 is 00 0 104. 9.

Figure 2.3 Patent Application Publication Jul. 7, 2011 Sheet 4 of 84 US 2011/O166063 A1

mAU T UV1280nm - UV2 225nm - Cond - Conc - pH - Un-reacted 1 -10 200 \ KP-10/But ALD30K 1000 A. Conjugate

Figure KISS3.1

DAD1E, Sig-280.6 Ref-360,100, TT (11108 PRO.TY 1108 PRODUCT PURITY 2008-11-17-11-3005-050.0) AU

Figure KISS3.2 Patent Application Publication Jul. 7, 2011 Sheet 5 of 84 US 2011/0166063 A1

rtiz SN Quality Fac. Rcs. Intens, Arca 3493.80s 349. s46ss 37.66 SS388 6629.ss 12. 72 4 696 9ss Figure KISS3.3

Figure KISS4.1 Patent Application Publication Jul. 7, 2011 Sheet 6 of 84 US 2011/O166063 A1

DAD1E, Sig=280,16Ref=360,100, TT (111108 PRO.TY111108 PRODUCT PURTY 2008-11-111-1143.008-09010) es s

Figure KISS4.2

42038.83

o o w ". do

raft SN Quality Fac. Res, latens. Aree 203,983 9. 49 to so. Oss

Figure KISS4.3 Patent Application Publication Jul. 7, 2011 Sheet 7 of 84 US 2011/O166063 A1

AA

Unreacted KP-10/SBC30K / / s SBC30K Conjugate N s \ Un-reacted KP-10 2 Unreacted d s O a Ne--- A ° N\1 Nis n-4 s A. A. A. As As A 3AS . S. 2 2s. ... " Figure KISS5.1

Figure KISS5.2 Patent Application Publication Jul. 7, 2011 Sheet 8 of 84 US 2011/O166063 A1

O DAD1E, Sig-280,s Refe380,100, TT (111g08 PRODUCT PURITY 11g08 PRODUCT PURTY 2008-11-1915.140205 na N

2 w |Marks:

Figure KISS5.4 Patent Application Publication Jul. 7, 2011 Sheet 9 of 84 US 2011/O166063 A1

mono-mPEG2-Butyraldehyde 4KHKispeptin-san Un-reacted

Figure KISS6.1

DAD1E, Sig-280.16 Ref=360,100, TT (111908 PRO.TY111908 PRODUCTPURITY 2008-11-1915-140204-0401.D) AU w

25

15

10

2.5 s 5 10 125 15 17.5 20 : Figure KISS6.2 Patent Application Publication Jul. 7, 2011 Sheet 10 of 84 US 2011/0166063 A1

Figure KISS6.3

ordat Yan2008Now OPButyradiok

Corriff KP5By AD40K Coveyda

i.

st sn Quality Fac, Rea has Area 34) d is a a 12 co, was Frt p.r. go yr24.3a is 4

Figure KISS6.4 Patent Application Publication Jul. 7, 2011 Sheet 11 of 84 US 2011/O166063 A1

to KP10 1 O Ald1 OK-KP10 A Ald3OK-KP10 v KP13 0 KP54 O Ald-30K-KP13 O Ald-40K-KP54 A metastin

log Conc M Figure KISS8.1

120 1 O CAC-4OK-KP1O 96.hr CAC-4OK-KP10 48hr CAC-4OK-KP1 O 96ht CAC-4OK-KP10 Ohr metastin

log Conc M

Figure KISS8.2 Patent Application Publication Jul. 7, 2011 Sheet 12 of 84 US 2011/O166063 A1

O SBC-3OK-KP1O 2hr SBC-30K-KP10 Ohr A metastin

log (ConcM

Figure KISS8.3 Patent Application Publication Jul. 7, 2011 Sheet 13 of 84 US 2011/O166063 A1

^ me 215 mm | mus 280 mm ra f D1 and High-conjugate

Mono-conjugate

Volume (mL)

WD assinosis. Soto Figure ZIC2.1 s s W s so

so

Figure Zic2.2 Patent Application Publication Jul. 7, 2011 Sheet 14 of 84 US 2011/O166063 A1

239).54

4000 6000 8. 2000) 2200 24 2s) 2800 300 3abo s4000 re Figure ZIC2.3 "sissiste-2.3TiSEATV ve -acetitectZick is er - gEg is guagers m T ' f T 215 mm

TT- 280 mm t "I Diard High-conjugate E. and free PEG an --" - / W / a Mono-conjugate -/

ge M 1.A. \

Volune (un) Figure ZIC3.1 Patent Application Publication Jul. 7, 2011 Sheet 15 of 84 US 2011/O166063 A1

SS

o

500

3. r

U | /\-

------H------H Figure ZIC3.2 44479.53 30

2

10

r 3000 3SOOO 4000 also 50 550 so so occo Figure ZIC3.3 Patent Application Publication Jul. 7, 2011 Sheet 16 of 84 US 2011/O166063 A1

f 2 1 s 280 mm

D and High-conjugate and free PEG

Volumne in Li Figure ZIC4.1

Figure ZIC4.2 Patent Application Publication Jul. 7, 2011 Sheet 17 of 84 US 2011/0166063 A1

200

34209,596

1CO

50

00 3000 3200 34000 Soo 3800 40000 fe

Patent Application Publication Jul. 7, 2011 Sheet 18 of 84 US 2011/O166063 A1

100 Ziconotide A Mono-mpG-20K-C2-FMOC-Zicono. 80 W Mono-mpEG-30K-SBA-Zicono. 60 0 Mono-mpEG-40K-CAC-FMOC-Zicono.

40

20

10-16 O 10-13 10-12 10-11 10-10 10-9 10-8 10-7 Log Inhibitor) (M)

Ligand): 1.0e-11 Ko: 1.Oe-11

Figure ZIC6.1 Patent Application Publication Jul. 7, 2011 Sheet 19 of 84 US 2011/O166063 A1

Figure BIP1. Biphalin structure

Figure BIP2.1 Patent Application Publication Jul. 7, 2011 Sheet 20 of 84 US 2011/O166063 A1

Figure BIP2.2

so

5291.467 558.75 5423.91

Soo,367 SBB. 92 - 5028,053 SS12.24

4982.4 5558,543

SB00704 sail|| || | 5644,888ye -4849,493 5688.995 4805,95 5733.29s- 4761.110 577,849 4718,828 4627,743

Figure BIP2.3 Patent Application Publication Jul. 7, 2011 Sheet 21 of 84 US 2011/O166063 A1

ec2cc 7Soboe YWoo: uvason is is Yst far. peogfiscegoeywoo; Rasos

O 50 O 15 20

Figure BIP3.1

2, 4, 5, 800 10, 120 140. 6.00 8.0 20, 200 2.0 26.0 28.00 3000 Minutes RT Area 120722 2 2.355 560602

Figure BIP3.2 Patent Application Publication Jul. 7, 2011 Sheet 22 of 84 US 2011/O166063 A1

4358,531

21865,641

Figure BIP4.1 Patent Application Publication Jul. 7, 2011 Sheet 23 of 84 US 2011/0166063 A1

mu

OO

distristirubhirleridis S1) E228Ac. 2S35s CO S. D. S. M

120932 689892 319 4069 2214322092491496,81708a04

Figure BIP4.3 Patent Application Publication Jul. 7, 2011 Sheet 24 of 84 US 2011/O166063 A1

plcohol

Figure BIP5.1 Patent Application Publication Jul. 7, 2011 Sheet 25 of 84 US 2011/O166063 A1

(SBC-30K)-biphalin Free PEG/(SBC-30K)-biphalin

Figure BIP5.2 Patent Application Publication Jul. 7, 2011 Sheet 26 of 84 US 2011/O166063 A1

Opioid Receptor (Human Recombinant) Binding Assay

120 O Biphalin o Biphalin (72hG37°C) iss 100 Di-CAC-20K-biphalin 80 O Di-CAC-20K-biphalin (72hG37°C g 60 3. Z 40 E 20

10-10 10-9 10-8 10-7 10-6 10-5 10-4 log inhibitor (M)

Ligand): 5.0 nM Kid: 4.0 nM

& Opioid Receptor (Human Recombinant) Binding Assay

120 Biphalin

100 Biphalin (72hQ37°C) Di-CAC-20K-biphalin 80 D-CAC-2OK-biphalin (72hG37°C) 60

40

20

H-H--- O 10-10 10-9 10-8 107 10-6 10-5 104 Log Inhibitor (M)

(Ligand): 5.0 nM Ko: 3.9 nM Figure BIP6.1 Patent Application Publication Jul. 7, 2011 Sheet 27 of 84 US 2011/O166063 A1

A. Opioid Receptor (Human Recombinant) Binding Assay

100 O Biphalin A Di-C2-20K-Biphalin 8O A Di-C2-20K-Biphalin (20hG37°C) 60 V Di-SBC-30K-Biphalin (20ha)37°C) 0 Di-SPA-2 K-Biphalin 40

20

10-10 10-9 10-8 10-7 10-6 10-5 10-4 log inhibitor (M)

(Ligand): 5.0 nM Ko: 40 nM Figure BIP6.2A

8 Opioid Receptor (Human Recombinant) Binding Assay

100 Biphalin Di-C2-2OK-Biphalin 80 Di-C2-2OK-Biphalin (20ha)37°C) Di-SBC-30K-Biphalin (20h(Q37°C) 60 Di-SPA-2K-Biphalin 40

20

O 10-10 10-9 10-8 10-7 10-6 10-5 10-4 Log inhibitor (M)

(Ligand): 5.0 nM K: 3.9 nM

Figure BIP6.2B Patent Application Publication Jul. 7, 2011 Sheet 28 of 84 US 2011/O166063 A1

PEGylation rate of BNP-32 with mpFG2-40kDa Butyr-ALD, 2 molecuivalents of PEG

=-0-% peptide -- % mono-PEG -- % di-PEG

O 12C 240 360 48O 5OO 20 84O 96O 18O2OO Time (min) Figure BNP2.1

300

200

di-PEG conjugate Oo

oo 0.0 200 300 400 S0.0 600 n Figure BNP2.2 Patent Application Publication Jul. 7, 2011 Sheet 29 of 84 US 2011/O166063 A1

Figure BNP2.3

3.

2 -

2333.243

Nied 892, 1998

s o 300) SO) 80 100

Figure BNP2.4 Patent Application Publication Jul. 7, 2011 Sheet 30 of 84 US 2011/O166063 A1

kDa Std 1 2 3 4 5 6

200 5 97 66 55

36.5 3. 25.5 4.

3.5 2.5 Figure BNP2.5

nono-PEG-BNP-32

Time (min) Figure BNP4.1 Patent Application Publication Jul. 7, 2011 Sheet 31 of 84 US 2011/O166063 A1

kDa Std 1 2 3 4

15 97 55 55 di-PEG Conjugate 35.5 31. mono-PEG conjugate 25.5 4.4 peptide only

Figure BNP4.2

“"Wiawesignizingostics"""““Wid sengths 2.sassis-oco my g so zooi 150 : to:; : l so l 3 in xxxx-xx-xx-xx-aa-Mow-wow ^:*:::ss.xxxx-xx-a-www.s:ss. wer- ra-taaaar s

t m -- or--or-o- --- *x n- l --

Figure BNP4.3 Patent Application Publication Jul. 7, 2011 Sheet 32 of 84 US 2011/O166063 A1

arraya232,38

00 800 000 20 400 soo n Figure BNP4.4

BNPO2S003:10 UW 2S Ni2SOk: it

Ali a-a-a-a-odayaw------ma-aa-ud-e-et-as-arara-arrara

S00 monokohjugate\ M / \ v

s00

400 y A. free peptide

A 1 2 di-Conjugate / 200 '

o ... N-4------/ 1 30,0 35.0 0.0 SO 500 55.0 60.0 m Figure BNP5.1 Patent Application Publication Jul. 7, 2011 Sheet 33 of 84 US 2011/O166063 A1

Sto 1 2 3.

di-PEG conjugate mono-PEG conjugate

free peptide

Figure BNP5.2

------a-rror------rra-error-ra-re Figure BNP5.3 Patent Application Publication Jul. 7, 2011 Sheet 34 of 84 US 2011/O166063 A1

Figure BNP5.4

BNOOO. W. R. “ NO: i) ?ets: 1A free peptide - O - 00 mono-conjugate SOO

600

:00 di-conjugate

40,0 50.0 60.0 700 80.0 90.0 min Figure BNP6.1 Patent Application Publication Jul. 7, 2011 Sheet 35 of 84 US 2011/0166063 A1

kDa Std 1 2

200 116 97 66 55

36.5 31 25.5 4.4

Figure BNP6.2

i" - " won A Wavelenges mizzo.20soo-ooo - " - “- i i 175

s

25

T is in Figure BNP6.3 Patent Application Publication Jul. 7, 2011 Sheet 36 of 84 US 2011/O166063 A1

200

OO |M "y with "Navy M.M.

0000 20000 30000 Figure BNP64 Scoxo soooo root

10000

O O O

100

-O- BNP 10. -- Metabolite -O- PEG-BNP O 8 16 24 32 40 48 56 64 72 Time Chr) Figure BNP7.1 Patent Application Publication Jul. 7, 2011 Sheet 37 of 84 US 2011/O166063 A1

10000 E 1000 St. S. U E 100 -e- ButyraLD-40K-BNP - ButyraLD-10K-BNP

E 10t A.

O 8 16 24, 32 40 48 S6 (64. 72 Time (hr) Figure BNP7.2 Patent Application Publication Jul. 7, 2011 Sheet 38 of 84 US 2011/O166063 A1

its in anal Azional alonia steal fismissed "s: Allisers: tal

- Unreacted CAC 4OK PEG PG-1 Peptide

1-CAC4OK ugate

- in-a-fittitriti?fiftyligitaticra ea away - O - s 3.

Figure PRO2.1

Figure PRO2.2 Patent Application Publication Jul. 7, 2011 Sheet 39 of 84 US 2011/0166063 A1

DAD1E, S280,46 Ref-300,100 T (01508 PG1.408PG-CACOK-TMECOURSE 2008-09-151637-17033-5001D) y co

Figure PRO2.3

--8540.9.142

Figure PRO2.4 Patent Application Publication Jul. 7, 2011 Sheet 40 of 84 US 2011/O166063 A1

s t s

Figure PRO3.1

Figure PRO3.2 Patent Application Publication Jul. 7, 2011 Sheet 41 of 84 US 2011/O166063 A1

DAD1E, Sig-280,6Re-360,100, TT (11108 PRODUCT PURITY 1108 PRODUCT PURITY 2008-117-143023-1501D)

Figure PRO3.3

Figure PRO3.4 Patent Application Publication Jul. 7, 2011 Sheet 42 of 84 US 2011/O166063 A1

Protegrin-1}-PEG-di Butyraldehyde-5K-Protegrin-l

Figure PRO4.2 Patent Application Publication Jul. 7, 2011 Sheet 43 of 84 US 2011/O166063 A1

DAD1E,Sig280.16 Ref-36,10, TT 12:08 PGoPEGSKREPURRERUN1238PGPEGREPUR228-12.its

5 s s O 25 s 15 mir Figure PRO4.3

a 7172 323 2so . 8: x inss 745. f 8 -----part-Ti-r O occ sco 4. S. Fo so coo o tood m

Figure PRO4.4 Patent Application Publication Jul. 7, 2011 Sheet 44 of 84 US 2011/O166063 A1

Figure PRO5.1

w

<-Free peptide

Figure PRO5.2 Patent Application Publication Jul. 7, 2011 Sheet 45 of 84 US 2011/O166063 A1

karusun 10:W1289 "“isalm 10:1 cord ""trix is as larusiuri. Fackra

Lalil islallel latellulalabdeleteliskollablelabs stadalalelululellslebleakalladislds was so Figure PRO6.1

5.00 . 9. O o 2. Minutes RT Area • Area Height 8O

Figure PRO6.2 Patent Application Publication Jul. 7, 2011 Sheet 46 of 84 US 2011/O166063 A1

Ad2 2K PG1

235,927

343.826

number-average molecular weight: weight-average molecular weight: poly dispersity continuous range (start) continuous range (end) - rumber of peaks continuous Peak f Integral List

Figure PRO6.3 Patent Application Publication Jul. 7, 2011 Sheet 47 of 84 US 2011/0166063 A1

Marus run euv280mt " " " Merial runs; cord "sars: ru is cris: Manual runs;1. Fractions Asrua rury slogbok

| | | | | | | o in

saraining UV128orn As rust run, Gord r: 1: ify a 1 oz., " Manuel ?ung: 1Fractions are ring:tlegsock

(Continued)

300 nM. NaC

5 nM NC

2 MNC

a-a-a-a-a-D --alour t— f 2o.o i - i.

i

| | | || ||: |aliolali:lial alislelullalalalalalulislelelalalellalells als, O 2 30 Ax 50 Figure 7.1.2 Patent Application Publication Jul. 7, 2011 Sheet 48 of 84 US 2011/O166063 A1

fodine stain Coonassie stain

ALD4OK-PG-1

Figure PRO7.2 Patent Application Publication Jul. 7, 2011 Sheet 49 of 84 US 2011/O166063 A1

8. 2. 14. 18 2. 24. Males RT area is area Height 159s 1869 o 12 482

416.103 15513so 962-111953 s22.283 ogg oo, 31s 62585 45518 23: 1423 Figure PRO7.3 Patent Application Publication Jul. 7, 2011 Sheet 50 of 84 US 2011/O166063 A1

O

3.

42693,603

nurbs avei age volecular weight: weight average tolecular weight: Poly dispersity continuous range (start continuous range (end) k numbor of peaks 39 continuous Peak / Integral List Figure PRO7.4 Patent Application Publication Jul. 7, 2011 Sheet 51 of 84 US 2011/O166063 A1

Manual run 6:1. UV-280m Manat rune; cord Misruarrie:tfractions lanuel run 6:1 logbook

G-4)K flow through

al -: : i * | i i i ; 2 waste F2 23 as 878 alchildlisle SO Figure PRO8.1

4. 6. 8. . 2. 4. 6. S. O 2. 2. inites RT Area Area Height 115900 7412 023 839 216.286.317739 997327418 Figure PRO8.2 Patent Application Publication Jul. 7, 2011 Sheet 52 of 84 US 2011/O166063 A1

nurber average ?toletalia. Attigat: 3328 weight-average molecular weight: 89 poly dispersity ... O 97 continuous range at art) ... 9.30 m/z) continuous range (end) . 5597 (m/z) n11thar of peake r continuous Peak f Integral List

Figure PRO8.3 Patent Application Publication Jul. 7, 2011 Sheet 53 of 84 US 2011/O166063 A1

lodine stain Coomassie stain

1 2 3 4

116.3

97.4

66.3

554

36.5

Figure PRO8.4

Hemolysis of Rat RBCs by PG1 conjugates

90 50 -- PG1 40 -- CAC-40K-PG1. Ohr g -- CAC-40K-PG1-48 hr g 30 -- CAC-4OK-PG1-96 ht 2 -6- Dextran-PG1 s -e- 2 mM HCl (buffer control)

10

O O 30 65 90 Incubation time (min) Figure PRO9.1 Patent Application Publication Jul. 7, 2011 Sheet 54 of 84 US 2011/O166063 A1

Hemolysis of Rat RBCs 10035l. 40

30 PG1. Ohr preincubation 3. PG196hr preincubation s CAC 40K-fulvene Ohr incubation 20 CAC 40K-fulvene 96 hr incubation SS PG 1-ButyralD-5K-PG1 1o mPEG2-NHS 20K-gly mPEG-SMB 30K-gly o Buffer control (2 mM HCl) male-la-H Matrix control (RBC) so { Sv v' f f Final concentration in incubation (ug of PG1 equivalent concentration/mL)

Figure PRO9.2

45 40 35 30 25 15 d 10

O

S s S s S s S s 8 S & Sst S S Ns S g S S O S { Sp So S SSis s S S s S s S s S s& c S &QY S S & - S $ s SS Q Sigis’s’o cy & ‘’’$ s

Figure PRO9.3 Patent Application Publication Jul. 7, 2011 Sheet 55 of 84 US 2011/O166063 A1

234. OOO 1. O

Peptide (ig/ml) Figure PRO9.4

10000 E s S

asE 100 A. ale Protegrin in Metabolite -- PEG-conjugate 1 O 4 8 12 16 20 24 Time (hr) Figure PRO10.1 Patent Application Publication Jul. 7, 2011 Sheet 56 of 84 US 2011/O166063 A1

100000

10000

100

9 Protegrin a Metabolite as PEG-conjugate 1 ------O 4 8 12 16 2O 24 Time (hr) Figure PRO10.2

Analyte=Protegrin 100000

10000

Sb * Protegrin-1 s 100 - CAC-PG -- CG-PG1 T A.

O 4 8 12 16 2O 24 Time (h) Figure PRO10.3 Patent Application Publication Jul. 7, 2011 Sheet 57 of 84 US 2011/O166063 A1

Analyte=PEG-conjugate 100000 10000 Sh E. s mPEG-PG-1 100 PG-1PEG-PG-1 PG-1-PEG-PG-1

O 4 8 12 16 20 24 Time (hr) Figure PRO10.4 Patent Application Publication Jul. 7, 2011 Sheet 58 of 84 US 2011/O166063 A1

irst L. Liffiliff, L. KFRSE PEGylated form Free peptide Figure V2.1

ra 9 t 2 tri.2 3,ch- 2 9 al5. as C E - V V s

16- Di-Pogy atcdpaptido Mono-Pegylated peptide

Frce peptide Figure V2.2 Patent Application Publication Jul. 7, 2011 Sheet 59 of 84 US 2011/O166063 A1

BAbi B, Sir280.8 Ref-380,00(68MPEG2NHS2OKC672CAO:00)

Figure V2.4 Patent Application Publication Jul. 7, 2011 Sheet 60 of 84 US 2011/O166063 A1

first if PEGylated form Figure V3.1 A. S. aV C s to C a c e S. C cos 2 co - Di-PEGylated peptide --xx is - Mono-PEGylated peptide

is Free peptide Figure V3.2 Patent Application Publication Jul. 7, 2011 Sheet 61 of 84 US 2011/O166063 A1

DA) 8, S86 Ro:800 (8HPEGS803NAIFOOOL)

" . "alogy Figure V3.4 Patent Application Publication Jul. 7, 2011 Sheet 62 of 84 US 2011/O166063 A1

ea

sE O)

s O (S w G) (9 O V681 (V13AD) O (SMB-30K-V681 (V13AD)) y (NHS-20K-V681 (V13AD)) E 1 O

l

12 Time (hr) Figure V4.1 Patent Application Publication Jul. 7, 2011 Sheet 63 of 84 US 2011/O166063 A1

Hemolysis of Rat RBCs by V681 (V13AD) conjugates

90 12

10 -e- V681 (V13AD) p -- mPEG2-NHS2OK-V681(V13AD) 8 -- mPEG-SMB 30K-V681 (V13AD)for pa 2 -e- V681 (V13AD)desA12 9. 6 -e- 2 mM HCl (buffer control) SS

4.

2

O O 30 65 90 Incubation time (min) Figure V5.1 Patent Application Publication Jul. 7, 2011 Sheet 64 of 84 US 2011/O166063 A1

Figure C-PEP2.1 cant A, gr. 44 Asia total EastEgg E. Patent Application Publication Jul. 7, 2011 Sheet 65 of 84 US 2011/0166063 A1

::::::::::::::::::: as - Patent Application Publication Jul. 7, 2011 Sheet 66 of 84 US 2011/O166063 A1

AE1 A. Sigs 2134 Refuses. 100 CPEPIDE520CBA intact-401)

Figure C-PEP3.2

Figure C-PEP3.3 Patent Application Publication Jul. 7, 2011 Sheet 67 of 84 US 2011/0166063 A1

setse C2 FM994K tet are ce.

Figure C-PEP4.1

ODADA, Sig-2154 Ref-380,100 (C2 RELEASEDMFE.050. D)

Figure C-PEP4.2 Patent Application Publication Jul. 7, 2011 Sheet 68 of 84 US 2011/O166063 A1

------girls retrayerseyss-cis------e.g. rers creasiergaressesser-sis so so 4EO so so s sta Figure C-PEP4.3

Figure C-PEP5.1 Patent Application Publication Jul. 7, 2011 Sheet 69 of 84 US 2011/O166063 A1

O DAD. A. Si-254Rek:38.0) (CPEPTOESOCACA) 28LAE-100)

stru is risis Ery is to

s

e

Figure C-PEP6.1 Patent Application Publication Jul. 7, 2011 Sheet 70 of 84 US 2011/O166063 A1

m.a...... , , it it, si" i. "if" SFS

Figure C-PEP6.2 Patent Application Publication Jul. 7, 2011 Sheet 71 of 84 US 2011/O166063 A1

O CADASI-2154 REO,10(CPEPTIDE DETRAIMctoo.)

drilasaraesarayarreyer-et------essary s & 5. g r r re Figure C-PEP6.4 Patent Application Publication Jul. 7, 2011 Sheet 72 of 84 US 2011/O166063 A1

Unreacted CAC 4OK PEG s

s 4-mono-mPEG2-CAC FMOC-40K)-(OGF)

MWD1B, Sig-280,16Ref=360,100XLOGF CON.JUGATEPURITYO712809.2009-07-2816-383ONS-DEXPEGPH71,D)

Figure OGF2.2 Patent Application Publication Jul. 7, 2011 Sheet 73 of 84 US 2011/O166063 A1

OGF-CAdack-to c. 141 in

Accuisitor Parameter sis alsoulsion 3. atgusor recorers O:sods accrrouxcdsp4kDa par spectrince r Frritories ?craiserspects Focard for selectrowsr?sssss ficero or fear or nisian wave?e Later sists Figure OGF2.3

41 Unreacted C2

4-1 mono-mPEG2-C2-FMOC-40K Unreacted C2

(OGF) 40K PEG

loaded mono-mPEG2

MOC-40K)-(OGF)

Figure OGF3.1 Patent Application Publication Jul. 7, 2011 Sheet 74 of 84 US 2011/O166063 A1

MWD1B, Sig-280.16 Ref-360.100(XLOGFCON.JUGATEPURITY0712809.2009-07-2817-32-49INSDE-PEGPH74.D)

200

00

------5 10 15 20 25 Eli Figure OGF3.2

o:dataxlogF-240 bag

on-cateca 4.11 in

date ocessor Easters;3fes. Acquisitoy freshodnare D:Methods excorrorwethodsraadspar Secr's Tof Parration rissa farrkirspacers corserior sector warrass first soarsey on selector adder tress of ce waverage Nurey sets Figure OGF3.3 Patent Application Publication Jul. 7, 2011 Sheet 75 of 84 US 2011/O166063 A1

silisir Scalaxy. 2n is 333rick caisities tyrata E. & Sirst scussi.itsa actise:Estuax 12

i Sexx i i i ! i s : i i i f 28 1-Unrected C2-PEG : Sk | - mono-mPEG2-C2 Unreacted C2- -FMOC-40K-OGF)

40K PEG i

Figure OGF4.1

Figure OGF4.2 Patent Application Publication Jul. 7, 2011 Sheet 76 of 84 US 2011/O166063 A1

Figure OGF5.1

Figure OGF5.2 Patent Application Publication Jul. 7, 2011 Sheet 77 of 84 US 2011/O166063 A1

ono-mPEG-ButALD-10K)-OGF)

Overloaded mono mPEG-ButaLD 10K-OGF

Figure OGF6.1

DA319. Si2806 Ref:36). COXLOGFBUTAD19K2199 CG7SFRNBE-90 D

Sy 12

Figure OGF6.2 Patent Application Publication Jul. 7, 2011 Sheet 78 of 84 US 2011/O166063 A1

u. Opioid Receptor (Human Recombinant) Binding Assay

120 OGF g 100 OGF (16Ohg37°C, pH7.5) s O OGF (16hG237°C, pH10.0) S 80 60 2 9 4.

E 20

O ------10-11 10-0 0-9 10-s 10-7 10-6 10-6 10-4 Log Inhibitor (M)

Ligand): 5.0 nM Ko: 2.0 nM Figure OGF7.1A

8 Opioid Receptor (Human Recombinant) Binding Assay

120 p OGF s 10 = OGF (16Oh@37°C,0. pH7.5) o OGF (16hg37°C, ph10.0) E 80 E 60 So 40 E. 20

------O 10-11 10-10 10-0 0-6 10-7 0-6 10-5 104 Log Inhibitor (M)

Ligand): 5.0 nM Kid: 3.0 nM Figure OGF7.1B Patent Application Publication Jul. 7, 2011 Sheet 79 of 84 US 2011/O166063 A1

A. u Opioid Receptor (Human Recombinant) Binding Assay

120 OGF CAC-40K-OGF (16Oh@37°C, pH7.5) C2-40K-OGF (68hG)37°C, pH7.5) CAC-40K-OGF (16h337°C, pH10.0) SO C2-4OK-OGF (16hG37°C, pH10.0) 40 CAC-40K-OGF C2-40K-OGF 20

O O 10-11 10-10 10-9 10-s 10-7 10-s 10-5 104 Log inhibitor (M)

(Ligand: 5.0 nM Kd: 2.0 nM Figure OGF7.2A

8 Opioid Receptor (Human Recombinant) Binding Assay

120 OGF 100 A CAC-40K-OGF (16Ohg37°CpH7.5) 80 v C2-40K-OGF (68ha37°CpH7.5) i A CAC-40K-OGF (16hig37°CpH10.0) 60 W C2-40K-OGF (16hG37°CpH10.0) S 40 {0 CAC-40K-OGF

E 2 O

O O 0-1 10-10 10-0 10-s 10-7 10-6 -5 10-4 Log Inhibitor (M)

Ligand): 5.0 nM Kid: 3.OM Figure OGF7.2B Patent Application Publication Jul. 7, 2011 Sheet 80 of 84 US 2011/O166063 A1

Opioid Receptor (Human Recombinant) Binding Assay

20 0 OGF O O 0 CAC-4OK-fulwene (Free PEG) o C2-40K-fulvene (Free PEG) 8O Oo 2 O

O H-H -- O 10-11 10-10 10-9 10-8 1-7 10-6 10-5 10-4 Log Inhibitor (M)

Ligand): 5.0 nM Kd: 2.0 nM Figure OGF7.3A

8 Opioid Receptor (Human Recombinant) Binding Assay

120 OGF 100 0 CAC-40K-fulvene (Free PEG) s o C2-40K-fulvene (Free PEG) 80

i 60

40 E 20

O H-H-I-H O 10-1 10-10 10-9 10-8 0-7 0-6 10-5 10-4 Log Inhibitor (M)

(Ligand): 5.0 nM Kid: 3.0 M Figure OGF7.3B Patent Application Publication Jul. 7, 2011 Sheet 81 of 84 US 2011/O166063 A1

mAU %B 2500 i.e.,als -00 : \ i 2000 A - 80

! 1500 1000 500

O s as onnn ml Figure INS1.1

Figure INS1.2 Patent Application Publication Jul. 7, 2011 Sheet 82 of 84 US 2011/0166063 A1

mAU %B -00

S0 - 80

60 100 y

- 40 50

O 200

s Dextran-butyraLD-40K 97.4/ is insulin

31 21.5 14.4

Figure INS1.4 Patent Application Publication Jul. 7, 2011 Sheet 83 of 84 US 2011/0166063 A1

Dextram mAU i Conjugates %B -100 50.0

400

30.0 A 60

| Free insulin 40

| 20 00 - SF3 waste Was O min

Figure INS1.5

Competition binding assay CHO-hiR (Clone 4.1) O2Sep09

SOOOO m S nSun 3. Dextran-TEG-butyriadehyde-40K d R A Glycine-Dextran s"4000 s 20000 6

O H O -11 -10 -9 -8 - 6 -5 -4 Insulin, nM Figure INS2.1 Patent Application Publication Jul. 7, 2011 Sheet 84 of 84 US 2011/O166063 A1

6SO 600 550 500 450 4OO 3SO 3OO 2SO 200 -- PBS --Dex-InS 250ug 150 -- ins 50Ug 100 -- nS 5ug 50 --Dextran 1.75ng

Time (hr) Figure INS3.1 US 2011/0166063 A1 Jul. 7, 2011

POLYMER CONUGATES OF THERAPEUTIC each and every feature described herein, and each and every PEPTIDES combination of two or more of such features, is included within the scope of the present disclosure provided that the CROSS REFERENCE TO RELATED features included in Such a combination are not mutually APPLICATION inconsistent. In addition, any feature or combination of fea 0001. This application claims the benefit of priority under tures may be specifically excluded from any embodiment of 35 U.S.C. S 119(e) to U.S. Provisional Patent Application Ser. the present invention. Additional aspects and advantages of No. 61/153,966, filed 19 Feb. 2009, to U.S. Provisional Patent the present invention are set forth in the following description Application Ser. No. 61/208,089, filed 18 Feb. 2009, to U.S. and claims, particularly when considered in conjunction with Provisional Patent Application Ser. No. 61/192,672, filed 19 the accompanying examples and drawings. Sep. 2009, the disclosures of which are incorporated herein by reference in their entirety. BRIEF DESCRIPTION OF DRAWINGS 0008 FIG. KISS1.1: Cation exchange purification of the FIELD OF THE INVENTION PEGylation reaction mixture. 0002 Among other things, the present invention relates to 0009 FIG. KISS1.2: RP-HPLC analysis of purified conjugates comprising a therapeutic peptide moiety mono-mPEG-Butyr ALD-30K-Kisspeptin-13. covalently attached to one or more water-soluble polymers. (0010 FIG. KISS1.3 MALDI-TOF spectrum of purified mono-mPEG-Butyr ALD-30K-Kisspeptin-13. BACKGROUND OF THE INVENTION 0011 FIG. KISS2.1. Typical reversed phase purification 0003. In many ways, the chemical and biological proper profile of mono-mPEG-Buty Aldehyde-1OK-Kisspeptin ties of peptides make them very attractive candidates for use 10. as therapeutic agents. Peptides are naturally occurring mol (0012 FIG. KISS2.2 Purity analysis of mono-Butyr Alde ecules made up of amino acid building blocks, and are hyde-10K-Kisspeptin-10 by Reversed Phase HPLC. involved in countless physiological processes. With 20 natu (0013 FIG. KISS2.3. MALDI-TOF spectrum of purified rally occurring amino acids, and any number of non-naturally mono-mPEG-butyraldehyde-10k-Kisspeptin-10. occurring amino acids, a nearly endless variety of peptides (0014 FIG. KISS3.1. Typical reversed phase purification may be generated. Additionally, peptides display a high profile of mono-mPEG-Buty Aldehyde-3OK-Kisspeptin degree of selectivity and potency, and may not suffer from 10. potential adverse drug-drug interactions or other negative (0015 FIG. KISS3.2. Purity analysis of mono-Butyr Alde side effects. Moreover, recent advances in peptide synthesis hyde-30K-Kisspeptin-1 by Reversed Phase HPLC. techniques have made the synthesis of peptides practical and (0016 FIG. KISS3.3. MALDI-TOF spectrum of purified economically viable. Thus peptides hold great promise as a mono-mPEG-Butyraldehyde-30K-Kisspeptin-10. highly diverse, highly potent, and highly selective class of (0017 FIG. KISS4.1. Typical reversed phase purification therapeutic molecules with low toxicity. profile of mono-mPEG2-CAC-FMOC-40K-Kisspeptin 0004. A number of peptides have been identified as thera 10. peutically promising; however in vitro results have often not (0018 FIG. KISS4.2. Purity analysis of mono-ICAC proven to bear out in vivo. Significantly, peptides suffer from PEG2-FOMC-40K-Kisspeptin-10 by Reversed Phase a short in vivo half life, sometimes mere minutes, making HPLC. them generally impractical, in their native form, for therapeu (0019 FIG. KISS4.3. MALDI-TOF spectrum of purified tic administration. Thus there exists a need in the art for mono-ICAC-PEG2-FMOC-40K-Kisspeptin-10. modified therapeutic peptides having an enhanced half-life (0020 FIG. KISS5.1. Typical reversed phase purification and/or reduced clearance as well as additional therapeutic profile of mono-mPEG-SEC-30K-Kisspeptin-10. advantages as compared to the therapeutic peptides in their 0021 FIG. KISS5.2. SDS-PAGE, with Coomassie blue unmodified form. staining) of purified mono-mPEG-SEC-30K-Kisspeptin 10. SUMMARY OF THE INVENTION (0022 FIG. KISS5.3. Purity analysis of mono-mPEG 0005 Accordingly, the present invention provides conju SBC-30K-Kisspeptin-10 by Reversed Phase HPLC. gates comprising a therapeutic peptide moiety covalently (0023 FIG. KISS5.4. MALDI-TOF spectrum of purified attached to one or more water-soluble polymers. The water mono-mPEG-SEBC-30k-Kisspeptin-10. soluble polymer may be stably bound to the therapeutic pep 0024 FIG. KISS6.1 Typical cation exchange purification tide moiety, or it may be releasably attached to the therapeutic profile of mono-mPEG2-Butyr Aldehyde-40K-Kisspep peptide moiety. tin-54. 0006. The invention further provides methods of synthe (0025 FIG. KISS6.2. Purity analysis of mono-mPEG2 sizing such therapeutic peptide polymer conjugates and com Butyraldehyde-40K-Kisspeptin-54 conjugate by positions comprising such conjugates. The invention further Reversed Phase HPLC. provides methods of treating, preventing, or ameliorating a (0026 FIG. KISS6.3. SDS-PAGE with Coomassie staining disease, disorder or condition in a mammal comprising of purified mono-mPEG2-Butyraldehyde-40K administering a therapeutically effective amount of a thera Kisspeptin-54. peutic peptide polymer conjugate of the invention. (0027 FIG. KISS6.4. MALDI-TOF spectrum of purified 0007 Additional embodiments of the present conjugates, mono-mPEG2-Butyraldehyde-40K-Kisspeptin-54. compositions, methods, and the like will be apparent from the (0028 FIG. KISS8.1. Agonist activity at GPR54 for stable following description, examples, and claims. As can be PEG conjugates of Kisspeptin 10, Kisspeptin 13, and appreciated from the foregoing and following description, Kisspeptin 54. US 2011/0166063 A1 Jul. 7, 2011

0029 FIG. KISS8.2. Agonist activity at GPR54 for releas 0057 FIG. BNP2.2. Typical purification profile for the 40 able PEG conjugate of Kisspeptin 10. kDa mPEG2-Butyr-ALD mono-PEG conjugate of BNP-32. 0030 FIG. KISS8.3. Agonist activity at GPR54 for releas 0.058 FIG. BNP2.3. HPLC analysis of the 40 kDa able PEG conjugate of Kisspeptin 10. mPEG2-Butyr-ALD mono-PEG conjugate of BNP-32. 0031 FIG. ZIC2.1: Cation exchange purification of 0059 FIG. BNP2.4. MALDI-TOF analysis of the 40 kDa mono-mPEG-C2-FMOC-20K-Ziconotide from the PEGyla mPEG2-Butyr-ALD mono-PEG conjugate of BNP-32. tion reaction mixture. 0060 FIG. BNP2.5. SDS-PAGE (4-12% Bis-Tris-Nu 0032 FIG. ZIC2.2: RP-HPLC analysis of purified mono PAGE, Invitrogen) analysis of BNP-32 and purified mono mPEG-C2-FMOC-2OK-Ziconotide. mPEG2-Butyr-ALD-40K-BNP-32 conjugate. 0033 FIG. ZIC2.3: MALDI-TOF analysis of purified 0061 FIG. BNP4.1. Typical cation-exchange purification mono-mPEG-C2-FMOC-2OK-Ziconotide. profile of mono-mPEG-Butyr-ALD-10K-BNP-32. 0034 FIG. ZIC3.1: Cation exchange purification of 0062 FIG. BNP4.2. SDS-PAGE analysis of BNP-32 and mono-mPEG-CAC-FMOC-40K-Ziconotide from the PEGy the purified mono-mPEG2-Butyr-ALD-40K-BNP-32 lation reaction mixture. conjugate. 0035 FIG. ZIC3.2: RP-HPLC analysis of purified mono 0063 FIG. BNP4.3. RP-HPLC analysis of the purified mPEG-CAC-FMOC-40K-Ziconotide. mono-mPEG-Butyr-ALD-10K-BNP-32 conjugate. 0036 FIG. ZIC3.3: MALDI-TOF analysis of purified 0064 FIG. BNP4.4. MALDI-TOF analysis of the purified mono-mPEG-CAC-FMOC-40K-Ziconotide. mono-mPEG-Butyr-ALD-10K-BNP-32 conjugate. 0037 FIG. ZIC4.1: Cation exchange purification of 0065 FIG. BNP5.1. Typical first cation-exchange purifi mono-mPEG-SEA-30K-Ziconotide from the PEGylation cation profile for mono-mPEG-SEC-30K-BNP-32. reaction mixture. 0066 FIG. BNP5.2. SDS-PAGE analysis of the purified 0038 FIG. ZIC4.2: RP-HPLC analysis of purified mono mono-mPEG-SEC-30K-BNP-32 conjugate. mPEG-SEBA-3OK-Ziconotide. 0067 FIG. BNP5.3. RP-HPLC analysis of the purified 0039 FIG. ZIC4.3: MALDI-TOF analysis of purified mono-mPEG-SEC-30K-BNP-32 conjugate. mono-mPEG-SEA-30K-Ziconotide. 0068 FIG. BNP5.4. MALDI-TOF analysis of the purified 0040 FIG. ZIC5.1: Cation exchange FPLC chromatogra mono-mPEG-SEC-30K-BNP-32 conjugate. phy of the PEGylation reaction mixture between Ziconotide 0069 FIG. BNP6.1. Typical first cation-exchange purifi and mPEG-SEBC-3OK-NHS. cation profile of mPEG2-C2-fmoc-NHS-40K). 004.1 FIG. ZIC6.1. Mean (ESEM) percent specific bind 0070 FIG. BNP6.2. SDS-PAGE analysis of the purified ing of Ziconotide conjugates to calcium channel, N-type, in mPEG2-C2-fmoc-NHS-40K-BNP-32 conjugate. rat cortical membranes. (0071 FIG. BNP6.3. RP-HPLC analysis of the purified 0042 FIG. BIP2.1: (SPA-2K)2-biphalin purification with mPEG2-C2-fmoc-NHS-40K-BNP-32 conjugate. CG-71S resin. (0072 FIG. BNP6.4. MALDI-TOF analysis of the purified 0043 FIG. BIP2.2: RP-HPLC analysis of reconstituted mPEG2-C2-fmoc-NHS-40K-BNP-32 conjugate. (SPA-2K)2-biphalin. (0073 FIG. BNP7.1 shows the mean plasma concentra 0044 FIG. BIP2.3. MALDITOFMS analysis of reconsti tion-time profiles of for C2-FMOC-PEG2-40K-BNP its cor tuted (SPA-2K)2-biphalin. responding metabolite and released BNP. 004.5 FIG. BIP3.1: (C2-20K)-biphalin purification with 0074 FIG. BNP7.2 shows the non-released PEG-BNP CG-71 S resin. levels after the administration of the two non-releasable PEG 0046 FIG. BIP3.2: RP-HPLC analysis of reconstituted constructs (Butyr ALD-40K-BNP. Butyr ALD-10K-BNP). (C2-20K)-biphalin. (0075 FIG. PRO2.1. Typical cation exchange purification 0047 FIG. BIP3.3 MALDI-TOF analysis of reconstituted profile of mono-mPEG2-CAC-FMOC-40K-PG-1). (C2-20K)-biphalin. 0.076 FIG. PRO2.2. SDS-PAGE of purified monol 0048 FIG. BIP4.1: (CAC-20K)-biphalin purification ICAC-PEG2-FOMC-NHS-40K-Protegrin-1. with CG-71S resin. (0077 FIG. PRO2.3. Purity analysis of mono-ICAC 0049 FIG. BIP4.2: (CAC-20K)-biphalin re-purification PEG2-FOMC-40K-Protegrin-1 by RP-HPLC. with CG-71S resin. (0078 FIG. PRO2.4. MALDI-TOF spectrum of purified 0050 FIG. BIP4.3: RP-HPLC analysis of reconstituted mono-ICAC-PEG2-FMOC-40K-Protegrin-1. (CAC-20K)-biphalin. (0079 FIG. PRO3.1 Typical cation exchange purification 0051 BIP4.4: MALDI-TOF analysis of reconstituted profile of mono-mPEG-SEC-30K-PG-1. (CAC-20K)-biphalin. 0080 FIG. PRO3.2. SDS-PAGE of purified monol 0.052 FIG. BIP5.1: RP-HPLC analysis of SBC-30K and mPEG-SEC-30K--Protegrin-1. biphalin conjugation reaction mixture. I0081 FIG. PRO3.3. Purity analysis of mono-mPEG 0053 FIG. BIP5.2. The purification of (SBC-30K)-bi SBC-30K--Protegrin-1 by RP-HPLC. phalin from the reaction mixture. I0082 FIG. PRO3.4. MALDI-TOF spectrum of purified 0054 FIG. BIP6.1. Competition binding assay of biphalin mono-mPEG-SEBC-30K--Protegrin-1. and di-CAC-20K-biphalin conjugate at human (A) L opioid I0083 FIG. PRO4.1 Typical reversed phase purification and (B) 8 opioid receptors. profile of Protegrin-1-PEG-di-Butyraldehyde-5K-Pro 0055 FIG. BIP6.2. Competition binding assay of biphalin tegrin-1. and di-C2-20K-biphalin, di-SBC-30K-biphalin, and di-SPA I0084 FIG. PRO4.2. SDS-PAGE of purified Protegrin-1- 2K-biphalin conjugate at human (A) Lopioid and (B) 8 opioid PEG-di-butyraldehyde-5K-Protegrin-1. receptors. I0085 FIG. PRO4.3. Purity analysis of Protegrin-1- 0056 FIG. BNP2.1. PEGylation rate of BNP-32 with PEG-di-butyraldehyde-5K-Protegrin-1 by reversed phase mPEG2-40 kDa Butyr-ALD. HPLC. US 2011/0166063 A1 Jul. 7, 2011

I0086 FIG. PRO4.4. MALDI-TOF spectrum of Prote 0115 FIG.V5.1. Hemolysis relative to the 100%hemoly grin-1-PEG-di-butyraldehyde-5K-Protegrin-1. sis produced by 0.25% Triton X-100. 0087 FIG. PRO5.1. Typical cation-exchange chromatog 0116 FIG. C-PEP 2.1. Typical anion-exchange chroma raphy profile of dextran-butryaldehyde-40K-protegrin-1. tography profile of mono-mPEG-ru-MAL-30K-C-pep 0088 FIG. PRO5.2. SDS-PAGE analysis (4-12% gel) of tide(S20C). purified dextran-butryraldehyde-40K-protegrin-1. 0117 FIG. C-PEP 2.2. Purity analysis of mono 0089 FIG. PRO6.1: PG-1 and (ALD)2K conjugates puri mPEG-ru-MAL-30KC-peptide(S20C) by reversed phase fication with CM Sepharose FF resin. HPLC. 0090 FIG. PRO6.2: RP-HPLC analysis of (PG-1)-(ALD) 0118 FIG. C-PEP2.3. MALDI-TOF spectrum for 2K-(PG-1). mono-mPEG-ru-MAL-30K-C-peptide(S20C). 0.091 FIG. PRO6.3: MALDI analysis of (PG-1)-(ALD) 0119 FIG. C-PEP3.1. Typical anion-exchange chroma 2K-(PG-1). tography profile of mono-mPEG-Butyraldehyde-30K 0092 FIG. PRO7.1.1 and 7.1.2: ALD40K-PG-1 purifica C-peptide(S20C). tion with SP Sepharose HP resin. I0120 FIG. C-PEP3.2. Purity analysis of mono-mPEG 0093 FIG. PRO7.2. SDS-PAGE of the purified and con Butyraldehyde-30K-C-peptide(S20C) by reversed phase centrated ALD40K-PG-1. HPLC. 0094 FIG. PRO7.3: RP-HPLC analysis of ALD40K 0121 FIG. C-PEP3.3. MALDI-TOF spectrum for PG-1 (lot #YW-pgALD40K-01). mono-mPEG-Butyraldehyde-30K-C-peptide(S20C). 0.095 FIG. PRO7.4: MALDI analysis of ALD40K-PG-1 0.122 FIG. C-PEP4.1. Typical anion-exchange chroma (lot #YW-pgALD40K-01). tography profile of mono-C2-PEG2-FMOC-40K-C-pep tide(S20C). 0.096 FIG. PRO8.1: CG40K-PG-1 purification with SP (0123 FIG. C-PEP4.2. Purity analysis of mono-C2 Sepharose HP resin. PEG2-FMOC-40KIC-peptide(S20C) by reversed phase 0097 FIG. PRO8.2: RP-HPLC analysis of purified HPLC. CG4OK-PG-1. 0.124 FIG. C-PEP4.3. MALDI-TOF spectrum for 0.098 FIG. PRO8.3: MALDI-TOF analysis of purified mono-C2-PEG2-FMOC-40K-C-peptide(S20C). CG4OK-PG-1. (0.125 FIG. C-PEP5.1. Typical anion-exchange purifica 0099 FIG. PRO9.1. Hemolysis relative to the 100% tion profile of mono-ICAC-PEG2-FMOC-40K-C-pep hemolysis produced by 0.25% Triton X-100. tide(S20C). 0100 FIG. PRO9.1. Hemolysis by PEG reagent controls. (0.126 FIG. C-PEP5.2. Purity analysis of mono-ICAC 0101 FIG. PRO9.3. Hemolysis at the maximum concen PEG2-FMOC-40K-C-peptide(S20C) by reversed phase tration. HPLC. 0102 FIG. PRO9.4. Hemolytic activities of PG-1 I0127 FIG. C-PEP6.1 Typical anion-exchange chromatog (0103 FIG. PRO10.1 and PRO10.2 show the mean plasma raphy profile of dextran-butryaldehyde-40K-C-peptide concentration-time profiles for CG-PEG-FMOC-40K-PG-1 (S20C). and CAC-PEG-FMOC-40K-PG-1, their corresponding 0128 FIG. C-PEP6.2. Concentration of fraction II from PEG-metabolite and released Protegrin-1. the anion-exchange chromatogram shown in FIG. c-pep6.1 0104 FIG. PRO10.3 shows the released Protegrin-1 levels by a second anion-exchange chromatography run. after the administration of the two releasable PEG constructs I0129 FIG. C-PEP6.3. Purity analysis of mono-Dext versus the level of Protegrin-1 given as native protein at the ran-40K-C-peptide(S20C) by reversed phase HPLC. same dose (mg/kg). 0.130 FIG. C-PEP6.4. MALDI-TOF spectrum for 0105 FIG. PRO10.4 shows the mean plasma concentra mono-Dextran-40K-C-peptide(S20C). tion-time profiles for mPEG-PG-1, PG-1PEG-PG-1, I0131 FIG. OGF2.1. Typical CG71 S reversed phase puri PG-1-PEG-PG-1. fication profile of mono-mPEG2-CAC-FMOC-40K 0106 FIG. V2.1. Typical cation-exchange purification OGF. profile of mPEG2-NHS-20K-V681 (V13AD). (0132 FIG. OGF2.2. Purity analysis of mono-ICAC 0107 FIG. V2.2. SDS-PAGE analysis of V681 (V13 AD) PEG2-FOMC-40K-OGF by reversed phase HPLC. PEGylation. (0.133 FIG. OGF2.3. MALDI-TOF spectrum of purified 0108 FIG. V2.3. Purity analysis of mono-mPEG2 mono-mPEG2-FMOC-CAC-40K-OGF. NHS 20K-V681 (V13 AD) conjugate by reverse phase I0134 FIG. OGF3.1. Typical CG71S reverse phase purifi HPLC. cation profile of mono-mPEG2-C2-FMOC-40K-OGF). 0109 FIG. V2.4. MALDI-TOF spectra for mono I0135 FIG. OGF3.2. Purity analysis of mono-mPEG2 mPEG2-NHS 20K-V681 (V13 AD). FMOC-C2-40K-OGF by reversed phase HPLC. 0110 FIG. V3.1. Typical cation-exchange purification (0.136 FIG. OGF3.3. MALDI-TOF spectrum of purified profile of mPEG-SMB-301 (I-IV681 (V13 AD). mono-mPEG2-FMOC-C2-40K-OGF. 0111 FIG. V3.2. SDS-PAGE analysis of V681 (V13 AD) I0137 FIG. OGF4.1. Typical CG71S reversed phase puri PEGylation and purification on the SP ion-exchange column. fication profile of mono-mPEG-Butyraldehyde-30K 0112 FIG.V3.3. Purity analysis of mono-mPEG-SMB OGF. 30K-V681 (V13 AD) conjugate by reverse phase HPLC. I0138 FIG. OGF4.2. Purity analysis of mono-mPEG-Bu 0113 FIG. V3.4. MALDI-TOF spectra for mono tyr Aldehyde-30K-OGF by reversed phase HPLC. mPEG-SMB 30K-V681 (V13 AD). I0139 FIG. OGF5.1. Typical CG71S reversed phase puri 0114 FIG. V.4.1 shows the mean plasma concentration fication profile of mono-mPEG-epoxide-5K-OGF. time profiles for V681 (V13 AD), SMB-30K-V681 (V13AD), (O140 FIG. OGF5.2. Purity analysis of mono-mPEG-ep and NHS-20K-V681 (V13 AD). oxide-5K-OGF by reversed phase HPLC. US 2011/0166063 A1 Jul. 7, 2011

0141 FIG. OGF6.1. Typical CG71 S reversed phase puri may be made that result in an increase in therapeutic activi fication profile of mono-mPEG-Butyraldehyde-1OK ties. Thus, in the spirit of the invention, the terms “therapeutic OGF. peptide' or “therapeutic peptides are meant to encompass 0142 FIG. OGF6.2. Purity analysis of mono-mPEG-Bu modifications to the therapeutic peptides defined and/or dis tyr Aldehyde-10K-OGF by reversed phase HPLC. closed herein that do not alter, only partially abrogate, or 0143 FIG.OGF7.1. Competition binding assay of OGF at increase the therapeutic activities of the parent peptide. human (A) u opioid and (B) 8 opioid receptors: effects of 0156 The term “therapeutic activity” as used herein refers incubation treatment conditions. to a demonstrated or potential biological activity whose effect 014.4 FIG. OGF7.2. Competition binding assay of OGF is consistent with a desirable therapeutic outcome in humans, and PEG-OGF conjugates (released and unreleased) at or to desired effects in non-human mammals or in other human (A) L opioid and (B) 8 opioid receptors. species or organisms. A given therapeutic peptide may have (0145 FIG. OGF7.3. Competition binding assay of OGF one or more therapeutic activities, however the term “thera and free PEGs at human (A) u opioid and (B) 8 opioid recep peutic activities' as used herein may refer to a single thera tOrS. peutic activity or multiple therapeutic activites. “Therapeutic 0146 FIG. INS1.1 Typical anion-exchange chromatogra activity” includes the ability to induce a response in vitro, and phy profile of the conjugation reaction mixture with partially may be measured in vivo or in vitro. For example, a desirable acetylated insulin. effect may be assayed in cell culture, or by clinical evaluation, 0147 FIG. INS1.2 SDS-PAGE analysis of fractions con ECso assays, ICso assays, or dose response curves. In vitro or taining dextran-butyraLD-40K-insulin collected from cell culture assays, for example, are commonly available and anion-exchange chromatography. known to one of skill in the art for many therapeutic peptides 0148 FIG. INS1.3 Concentration of purified dextran-bu as defined and/or disclosed herein. Therapeutic activity tyr ALD-40K-insulin by anion-exchange chromatography. includes treatment, which may be prophylactic or ameliora 0149 FIG. INS1.4. SDS-PAGE analysis of purified dext tive, or prevention of a disease, disorder, or condition. Treat ran-butyr ALD-40K-insulin. ment of a disease, disorder or condition can include improve 0150 FIG. INS1.5 Typical anion-exchange chromatogra ment of a disease, disorder or condition by any amount, phy profile of the conjugation reaction mixture with non including elimination of a disease, disorder or condition. acetylated insulin. 0157. As used herein, the terms “peptide.” “polypeptide.” 0151. In vitro binding of the Insulin-dextran conjugate. and “protein, refer to polymers comprised of amino acid 0152 FIG. INS3.1. Glucose levels after compound admin monomers linked by amide bonds. Peptides may include the istration (0-8 hr). standard 20 C-amino acids that are used in protein synthesis by cells (i.e. natural amino acids), as well as non-natural DETAILED DESCRIPTION amino acids (non-natural amino acids may be found in nature, 0153. As used in this specification and the intended but not used in protein synthesis by cells, e.g., ornithine, claims, the singular forms “a,” “an,” and “the include plural citrulline, and sarcosine, or may be chemically synthesized), referents unless the context clearly dictates otherwise. Thus, amino acid analogs, and peptidomimetics. Spatola, (1983) in for example, reference to “a polymer includes a single poly Chemistry and Biochemistry of Amino Acids, Peptides, and meras well as two or more of the same or different polymers: Proteins, Weinstein, ed., Marcel Dekker, New York, p. 267. reference to “an optional excipient’ or to “a pharmaceutically The amino acids may be D- or L-optical isomers. Peptides acceptable excipient” refers to a single optional excipient as may be formed by a condensation or coupling reaction well as two or more of the same or different optional excipi between the C-carbon carboxyl group of one amino acid and ents, and the like. the amino group of another amino acid. The terminal amino 0154) In describing and claiming one or more embodi acid at one end of the chain (amino terminal) therefore has a ments of the present invention, the following terminology free amino group, while the terminal amino acid at the other will be used in accordance with the definitions described end of the chain (carboxy terminal) has a free carboxyl group. below. Alternatively, the peptides may be non-linear, branched pep 0155 As used herein, the terms “therapeutic peptide' and tides or cyclic peptides. Moreover, the peptides may option “therapeutic peptides' mean one or more peptides having ally be modified or protected with a variety of functional demonstrated or potential use in treating, preventing, orame groups or protecting groups, including on the amino and/or liorating one or more diseases, disorders, or conditions in a carboxy terminus. subject in need thereof, as well as related peptides. These 0158 Amino acid residues in peptides are abbreviated as terms may be used to refer to therapeutic peptides prior to follows: Phenylalanine is Phe or F: Leucine is Leu or L; conjugation to a water-soluble polymer as well as following Isoleucine is Ile or I; Methionine is Metor M. Valine is Val or the conjugation. Therapeutic peptides include, but are not V: Serine is Ser or S. Proline is Pro or P; Threonine is Thr or limited to, those disclosed herein, including in Table 1. Thera T; Alanine is Ala or A: Tyrosine is Tyr orY. Histidine is His or peutic peptides include peptides found to have use in treating, H. Glutamine is Glin or Q: Asparagine is ASn or N. Lysine is preventing, or ameliorating one or more diseases, disorders, Lys or K; Aspartic Acid is Asp or D: Glutamic Acid is Glu or or conditions after the time of filing of this application. E: Cysteine is Cys or C: Tryptophan is Trp or W: Arginine is Related peptides include fragments of therapeutic peptides, Arg or R; and Glycine is Gly or G. therapeutic peptide variants, and therapeutic peptide deriva 0159. The terms “therapeutic peptide fragment' or “frag tives that retain some or all of the therapeutic activities of the ments of therapeutic peptides’ refer to a polypeptide that therapeutic peptide. As will be known to one of skill in the art, comprises a truncation at the amino-terminus and/or a trun as a general principle, modifications may be made to peptides cation at the carboxyl-terminus of a therapeutic peptide as that do not alter, or only partially abrogate, the properties and defined herein. The terms “therapeutic peptide fragment” or activities of those peptides. In some instances, modifications "fragments of therapeutic peptides' also encompasses US 2011/0166063 A1 Jul. 7, 2011 amino-terminal and/or carboxyl-terminal truncations of alkoxy (e.g., methoxy, ethoxy and benzyloxy), as well as aryl, therapeutic peptide variants and therapeutic peptide deriva heteroaryl, cyclo, heterocyclo, and the like. It must be remem tives. Therapeutic peptide fragments may be produced by bered that the end-capping moiety may include one or more synthetic techniques known in the art or may arise from in atoms of the terminal monomer in the polymer e.g., the Vivo protease activity on longer peptide sequences. It will be end-capping moiety “methoxy” in CH O—(CH2CH2O) understood that therapeutic peptide fragments retain some or — and CH(OCH2CH) . In addition, Saturated, unsat all of the therapeutic activities of the therapeutic peptides. urated, substituted and unsubstituted forms of each of the 0160. As used herein, the terms “therapeutic peptide vari foregoing are envisioned. Moreover, the end-capping group ants' or “variants of therapeutic peptides’ refer to therapeutic can also be a silane. The end-capping group can also advan peptides having one or more amino acid Substitutions, includ tageously comprise a detectable label. When the polymer has ing conservative Substitutions and non-conservative Substitu an end-capping group comprising a detectable label, the tions, amino aciddeletions (eitherinternal deletions and/or C amount or location of the polymer and/or the moiety (e.g., and/or N-terminal truncations), amino acid additions (either active agent) to which the polymer is coupled can be deter internal additions and/or C- and/or N-terminal additions, e.g., mined by using a suitable detector. Such labels include, with fusion peptides), or any combination thereof. Variants may be out limitation, fluorescers, chemiluminescers, moieties used naturally occurring (e.g. homologs or orthologs), or non in enzyme labeling, colorimetric (e.g., dyes), metal ions, natural in origin. The term “therapeutic peptide variants' may radioactive moieties, gold particles, quantum dots, and the also be used to refer to therapeutic peptides incorporating one like. Suitable detectors include photometers, films, spectrom or more non-natural amino acids, amino acid analogs, and eters, and the like. The end-capping group can also advanta peptidomimetics. It will be understood that, in accordance geously comprise a phospholipid. When the polymer has an with the invention, therapeutic peptide fragments retain some end-capping group comprising a phospholipid, unique prop or all of the therapeutic activities of the therapeutic peptides. erties are imparted to the polymerand the resulting conjugate. 0161 The terms “therapeutic peptide derivatives” or Exemplary phospholipids include, without limitation, those "derivatives of therapeutic peptides' as used herein refer to selected from the class of phospholipids called phosphatidyl therapeutic peptides, therapeutic peptide fragments, and cholines. Specific phospholipids include, without limitation, therapeutic peptide variants that have been chemically altered those selected from the group consisting of dilauroylphos other than through covalent attachment of a water-soluble phatidylcholine, dioleylphosphatidylcholine, dipalmi polymer. It will be understood that, in accordance with the toylphosphatidylcholine, disteroylphosphatidylcholine, invention, therapeutic peptide derivatives retain some or all of behenoylphosphatidylcholine, arachidoylphosphatidylcho the therapeutic activities of the therapeutic peptides. line, and lecithin. 0162. As used herein, the terms “amino terminus protect 0.165. The term “targeting moiety' is used herein to refer ing group' or “N-terminal protecting group.” “carboxy ter to a molecular structure that helps the conjugates of the inven minus protecting group' or "C-terminal protecting group:” or tion to localize to a targeting area, e.g., help enter a cell, or 'side chain protecting group' refer to any chemical moiety bind a receptor. Preferably, the targeting moiety comprises of capable of addition to and optionally removal from a func vitamin, antibody, antigen, receptor, DNA, RNA, sialyl tional group on a peptide (e.g., the N-terminus, the C-termi Lewis Xantigen, hyaluronic acid, Sugars, cell specific lectins, nus, or a functional group associated with the side chain of an steroid or steroid derivative, RGD peptide, ligand for a cell amino acid located within the peptide) to allow for chemical Surface receptor, serum component, or combinatorial mol manipulation of the peptide. ecule directed against various intra- or extracellular recep (0163 “PEG.,” “polyethylene glycol” and “poly(ethylene tors. The targeting moiety may also comprise a lipid or a glycol)” as used herein, are interchangeable and encompass phospholipid. Exemplary phospholipids include, without any nonpeptidic water-soluble poly(ethylene oxide). Typi limitation, phosphatidylcholines, phospatidylserine, cally, PEGs for use inaccordance with the invention comprise phospatidylinositol, phospatidylglycerol, and phospatidyle the following structure"—(OCH2CH), where (n) is 2 to thanolamine. These lipids may be in the form of micelles or 4000. As used herein, PEG also includes “ CHCH O liposomes and the like. The targeting moiety may further (CH2CH2O), CHCH and “ (OCH2CH), O—.” comprise a detectable label or alternately a detectable label depending upon whether or not the terminal oxygens have may serve as a targeting moiety. When the conjugate has a been displaced. Throughout the specification and claims, it targeting group comprising a detectable label, the amount should be remembered that the term "PEG' includes struc and/or distribution/location of the polymer and/or the moiety tures having various terminal or “end capping groups and so (e.g., active agent) to which the polymer is coupled can be forth. The term “PEG” also means a polymer that contains a determined by using a suitable detector. Such labels include, majority, that is to say, greater than 50%, of —OCH2CH2— without limitation, fluorescers, chemiluminescers, moieties repeating subunits. With respect to specific forms, the PEG used in enzyme labeling, colorimetric (e.g., dyes), metalions, can take any number of a variety of molecular weights, as well radioactive moieties, gold particles, quantum dots, and the as structures or geometries such as “branched.” “linear.” like. "forked,” “multifunctional, and the like, to be described in 0166 “Non-naturally occurring with respect to a poly greater detail below. meras described herein, means a polymer that in its entirety 0164. The terms "end-capped' and “terminally capped' is not found in nature. A non-naturally occurring polymer of are interchangeably used herein to refer to a terminal or the invention may, however, contain one or more monomers endpoint of a polymer having an end-capping moiety. Typi or segments of monomers that are naturally occurring, so long cally, although not necessarily, the end-capping moiety com as the overall polymer structure is not found in nature. prises a hydroxy or Co alkoxy group, more preferably a (0167. The term “water soluble' as in a “water-soluble Coalkoxy group, and still more preferably a Cls alkoxy polymer is any polymer that is soluble in water at room group. Thus, examples of end-capping moieties include temperature. Typically, a water-soluble polymer will transmit US 2011/0166063 A1 Jul. 7, 2011

at least about 75%, more preferably at least about 95%, of due of a therapeutic peptide and a water-soluble polymer that light transmitted by the same solution after filtering. On a can be attached directly or indirectly through a spacer moi weight basis, a water-soluble polymer will preferably be at ety). least about 35% (by weight) soluble in water, more preferably 0173 A "monomer' or “mono-conjugate.” in reference to at least about 50% (by weight) soluble in water, still more a polymer conjugate of a therapeutic peptide, refers to a preferably about 70% (by weight) soluble in water, and still therapeutic peptide having only one water-soluble polymer more preferably about 85% (by weight) soluble in water. It is molecule covalently attached thereto, whereas a therapeutic most preferred, however, that the water-soluble polymer is peptide “dimer' or “di-conjugate' is a polymer conjugate of about 95% (by weight) soluble in water or completely soluble atherapeutic peptide having two water-soluble polymer mol in water. ecules covalently attached thereto, and so forth. 0168 “Hydrophilic, e.g., in reference to a “hydrophilic 0.174 Alkyl refers to a hydrocarbon, typically ranging polymer.” refers to a polymer that is characterized by its from about 1 to 15 atoms in length. Such hydrocarbons are solubility in and compatibility with water. In non-cross linked preferably but not necessarily saturated and may be branched form, a hydrophilic polymer is able to dissolve in, or be or straight chain, although typically straight chain is pre dispersed in water. Typically, a hydrophilic polymer pos ferred. Exemplary alkyl groups include methyl, ethyl, propyl. sesses a polymer backbone composed of carbon and hydro butyl, pentyl, 2-methylbutyl, 2-ethylpropyl, 3-methylpentyl, gen, and generally possesses a high percentage of oxygen in and the like. As used herein, “alkyl includes cycloalkyl as either the main polymer backbone or in pendent groups Sub well as cycloalkylene-containing alkyl. stituted along the polymer backbone, thereby leading to its 0.175 “Lower alkyl refers to an alkyl group containing “water-loving nature. The water-soluble polymers of the from 1 to 6 carbon atoms, and may be straight chain or present invention are typically hydrophilic, e.g., non-natu branched, as exemplified by methyl, ethyl, n-butyl, i-butyl, rally occurring hydrophilic. and t-butyl. 0169 Molecular weight in the context of a water-soluble 0176 “Cycloalkyl refers to a saturated or unsaturated polymer, such as PEG, can be expressed as either a number cyclic hydrocarbon chain, including bridged, fused, or spiro average molecular weight or a weight average molecular cyclic compounds, preferably made up of 3 to about 12 car weight. Unless otherwise indicated, all references to molecu bon atoms, more preferably 3 to about 8 carbon atoms. lar weight herein refer to the weight average molecular “Cycloalkylene' refers to a cycloalkyl group that is inserted weight. Both molecular weight determinations, number aver into an alkyl chain by bonding of the chain at any two carbons age and weight average, can be measured using gel perme in the cyclic ring system. ation chromatography or other liquid chromatography tech niques. Other methods for measuring molecular weight 0177 "Alkoxy' refers to an —O R group, wherein R is values can also be used. Such as the use of end-group analysis alkyl or Substituted alkyl, preferably Ce alkyl (e.g., meth or the measurement of colligative properties (e.g., freezing oxy, ethoxy, propyloxy, and so forth). point depression, boiling-point elevation, and osmotic pres 0.178 The term “substituted as in, for example, “substi Sure) to determine number average molecular weight, or the tuted alkyl refers to a moiety (e.g., an alkyl group) Substi use of light scattering techniques, ultracentrifugation or vis tuted with one or more noninterfering Substituents, such as, cometry to determine weight average molecular weight. The but not limited to: alkyl, C.s cycloalkyl, e.g., cyclopropyl. polymers of the invention are typically polydisperse (i.e., cyclobutyl, and the like; halo, e.g., fluoro, chloro, bromo, and number average molecular weight and weight average iodo; cyano; alkoxy, lower phenyl; Substituted phenyl; and molecular weight of the polymers are not equal), possessing the like. “Substituted awl' is aryl having one or more nonin low polydispersity values of preferably less than about 1.2, terfering groups as a Substituent. For Substitutions on a phenyl more preferably less than about 1.15, still more preferably ring, the Substituents may be in any orientation (i.e., ortho, less than about 1.10, yet still more preferably less than about meta, or para). 1.05, and most preferably less than about 1.03. 0179. “Noninterfering substituents' are those groups that, (0170 The term “active' or “activated when used in con when present in a molecule, are typically nonreactive with junction with a particular functional group refers to a reactive other functional groups contained within the molecule. functional group that reacts readily with an electrophile or a 0180 Aryl means one or more aromatic rings, each of 5 nucleophile on another molecule. This is in contrast to those or 6 core carbon atoms. Aryl includes multiple aryl rings that groups that require strong catalysts or highly impractical may be fused, as in naphthyl or unfused, as in biphenyl. Aryl reaction conditions in order to react (i.e., a “non-reactive' or rings may also be fused or unfused with one or more cyclic “inert' group). hydrocarbon, heteroaryl, or heterocyclic rings. As used 0171 As used herein, the term “functional group’ or any herein, “aryl' includes heteroaryl. synonym thereof is meant to encompass protected forms 0181 “Heteroaryl' is an aryl group containing from one to thereofas well as unprotected forms. four heteroatoms, preferably Sulfur, oxygen, or nitrogen, or a (0172. The terms “spacer moiety,” “linkage” and “linker” combination thereof. Heteroaryl rings may also be fused with are used herein to refer to an atom or a collection of atoms one or more cyclic hydrocarbon, heterocyclic, aryl, or het optionally used to link interconnecting moieties such as a eroaryl rings. terminus of a polymer segmentandatherapeutic peptide oran 0182 “Heterocycle' or "heterocyclic' means one or more electrophile or nucleophile of a therapeutic peptide. The rings of 5-12 atoms, preferably 5-7 atoms, with or without spacer moiety may be hydrolytically stable or may include a unsaturation or aromatic character and having at least one physiologically hydrolyzable or enzymatically degradable ring atom that is not a carbon. Preferred heteroatoms include linkage. Unless the context clearly dictates otherwise, a Sulfur, oxygen, and nitrogen. spacer moiety optionally exists between any two elements of 0183 “Substituted heteroaryl' is heteroaryl having one or a compound (e.g., the provided conjugates comprising a resi more noninterfering groups as Substituents. US 2011/0166063 A1 Jul. 7, 2011

0184 “Substituted heterocycle' is a heterocycle having ent that may optionally be included in the compositions of the one or more side chains formed from noninterfering Substitu invention and that causes no significant adverse toxicological entS. effects to the patient. 0185. An “organic radical as used herein shall include 0193 “Pharmacologically effective amount,”99 “physi&g alkyl, Substituted alkyl, alkenyl, Substituted alkenyl, alkynyl, ologically effective amount, and “therapeutically effective Substituted alkynyl, aryl, and Substituted aryl. amount are used interchangeably herein to mean the amount 0186 “Electrophile” and “electrophilic group” refer to an of a polymer-(therapeutic peptide) conjugate that is needed to ion or atom or collection of atoms, that may be ionic, having provide a desired level of the conjugate (or corresponding an electrophilic center, i.e., a center that is electron seeking, unconjugated therapeutic peptide) in the bloodstream or in capable of reacting with a nucleophile. the target tissue. The precise amount will depend upon numerous factors, e.g., the particular therapeutic peptide, the 0187. “Nucleophile' and “nucleophilic group' refers to an components and physical characteristics of the therapeutic ion or atom or collection of atoms that may be ionic having a composition, intended patient population, individual patient nucleophilic center, i.e., a center that is seeking an electro considerations, and the like, and can readily be determined by philic center or with an electrophile. one skilled in the art, based upon the information provided 0188 A “physiologically cleavable' or “hydrolyzable” or herein. “degradable bond is a bond that reacts with water (i.e., is 0194 “Multi-functional' means a polymer having three or hydrolyzed) under physiological conditions. The tendency of more functional groups contained therein, where the func a bond to hydrolyze in water will depend not only on the tional groups may be the same or different. Multi-functional general type of linkage connecting two central atoms but also polymeric reagents of the invention will typically contain on the Substituents attached to these central atoms. Appropri from about 3-100 functional groups, or from 3-50 functional ate hydrolytically unstable or weak linkages include but are groups, or from 3-25 functional groups, or from 3-15 func not limited to carboxylate ester, phosphate ester, anhydrides, tional groups, or from 3 to 10 functional groups, or will acetals, ketals, acyloxyalkyl ether, imines, orthoesters, pep contain 3, 4, 5, 6, 7, 8, 9 or 10 functional groups within the tides and oligonucleotides. polymer backbone. A "difunctional polymer means a poly 0189 "Releasably attached, e.g., in reference to a thera mer having two functional groups contained therein, either peutic peptide releasably attached to a water-soluble polymer, the same (i.e., homodifunctional) or different (i.e., heterodi refers to a therapeutic peptide that is covalently attached via a functional). linker that includes a degradable linkage as disclosed herein, (0195 The terms “subject.” “individual,” or “patient” are wherein upon degradation (e.g., hydrolysis), the therapeutic used interchangeably herein and refer to a vertebrate, prefer peptide is released. The therapeutic peptide thus released will ably a mammal. Mammals include, but are not limited to, typically correspond to the unmodified parent or native thera murines, rodents, simians, humans, farm animals, sport ani peutic peptide, or may be slightly altered, e.g., possessing a mals, and pets. short organic tag. Preferably, the unmodified parent therapeu 0196) “Optional or “optionally’ means that the subse tic peptide is released. quently described circumstance may or may not occur, so that 0190. An “enzymatically degradable linkage” means a the description includes instances where the circumstance linkage that is Subject to degradation by one or more occurs and instances where it does not. enzymes. (0197) “Substantially” (unless specifically defined for a 0191 A“hydrolytically stable' linkage or bond refers to a particular context elsewhere or the context clearly dictates chemical bond, typically a covalent bond, that is Substantially otherwise) means nearly totally or completely, for instance, stable in water, that is to say, does not undergo hydrolysis satisfying one or more of the following: greater than 50%, under physiological conditions to any appreciable extent over 51% or greater, 75% or greater, 80% or greater, 90% or an extended period of time. Examples of hydrolytically stable greater, and 95% or greater of the condition. linkages include, but are not limited to, the following: carbon 0198 Unless the context clearly dictates otherwise, when carbon bonds (e.g., in aliphatic chains), ethers, amides, ure the term “about precedes a numerical value, the numerical thanes, and the like. Generally, a hydrolytically stable linkage value is understood to mean the stated numerical value and is one that exhibits a rate of hydrolysis of less than about 1-2% also it 10% of the stated numerical value. per day under physiological conditions. Hydrolysis rates of 0199 Turning now to one or more aspects of the invention, representative chemical bonds can be found in most standard conjugates are provided, the conjugates comprising a thera chemistry textbooks. It must be pointed out that some link peutic peptide covalently attached (either directly or through ages can be hydrolytically stable or hydrolyzable, depending a spacer moiety or linker) to a water-soluble polymer. The upon (for example) adjacent and neighboring atoms and conjugates generally have the following formula: ambient conditions. One of ordinary skill in the art can deter PEP-I-X-POLY mine whether a given linkage or bond is hydrolytically stable or hydrolyzable in a given contextby, for example, placing a wherein PEP is a therapeutic peptide as defined herein, X is a linkage-containing molecule of interest under conditions of covalent bond or is a spacer moiety or linker, POLY is a water interest and testing for evidence of hydrolysis (e.g., the pres soluble polymer, and k in an integer ranging from 1-10, pref ence and amount of two molecules resulting from the cleav erably 1-5, and more preferably 1-3. age of a single molecule). Other approaches known to those of ordinary skill in the art for determining whether a given Therapeutic Peptides linkage or bond is hydrolytically stable or hydrolyzable can 0200. As previously stated, the conjugates of the invention also be used. comprise a therapeutic peptide as disclosed and/or defined 0.192 The terms “pharmaceutically acceptable excipient’ herein. Therapeutic peptides include those currently known and “pharmaceutically acceptable carrier refer to an excipi to have demonstrated or potential use in treating, preventing, US 2011/0166063 A1 Jul. 7, 2011

orameliorating one or more diseases, disorders, or conditions dotin hydrochloride; ceruletide diethylamine; ceruletide, in a subject in need thereofas well as those discovered after Fukuoka; cetrorelix acetate; chimaeric peptides, NIH; chole the filing of this application. Therapeutic peptides also cystokinin, Ferring; collagenase IV inhibitors; collamers; include related peptides. contraceptive vaccine, Cephalo; contraceptive vaccine, 0201 In some embodiments of the invention, PEP is a Novarti; corplatin S compounds; corticoliberin, Pharma Bis therapeutic peptide selected from the group consisting of send; corticoliberin, Salk; corticoliberin, Unigene; corti carperitide; alpha-neoendorphin; 348U87; A-3847; A-41 14: coliberin, Vanderbilt: D-21775; D-22213: Demegel; DAP A-68552; A-75998; A-84861; AN-1792: AAMP-1: inhibitors: DP-640; DP-107; DSIP; DU-728; Dynorphin A; exenatide; AC-625; ACE-inhibitors, Aventis; ACE-inhibitors, daniplestim; defensins, LSB; desirudin; detirelix; dialytic oli SRI; ACTH, Amgen; ruprintrivir; AI-102: AI-202; Neu gopeptides; disagregin; E-2078; ECE inhibitor, SmithKline: roVax: AI-402: AI-502; AIDS therapeutic vaccine, Repl; ELS-1; EMD-73495; Enhancins; ecallantide: ES-1005; AIDS therapy, Inst Pasteur: AIDS vaccine, J&J: AIDS vac ES-305; echistatin; efegatran; eglin derivatives; elafin deriva cine, Liposome Co: AIDS vaccine, Arana: AIDS vaccine, tives; elcatonin; eledoisin; encapsulated insulin, INSERM; Peptimmune: AIDS vaccine, Sanofi Past-3: AIDS vaccine, endorphin, B-, Antigenics; endorphin, pancreatic; endorphin, Protherics: AIDS vaccine, SSVI: AIDS vaccine, SWFBR: B-, Mitsubishi; endorphin, B-, Amgen; endothelial cell AIDS vaccine, United-1, AIDS vaccine, United-2: AIDS vac growth factor, endothelin antagonists, ResCo. eptifibatide; cine-2, Yokohama: AIDS vaccine-3, NIH AIDS vaccine-4, examorelin; Factor VIII fragments, Pharma; FG-002: NIH: AIT-083; tediuglutide: Skelite: Allotrap-2702; Alzhe FG-003; FG-004; FG-005; FR-113680; FTS-Zn; fibrin-bind imer's imaging agent, Dia; AM-425; AN-238; AnergiX.RA; ing peptides, ISIS; fibronectin inhibitors, AstraZ; fibronectin AnervaX.RA; AS-109; AV-9; AZM-134; addressin, Lilly: related peptide; follicular regulatory protein: G-4120; GAG allergy vaccine, BioResearch; ambamustine; amylin antago V3-VDP vaccine, Vern; GDL-peptides, Cytogen; EP-51216: nists, Amylin; anaritide analogues, Bio-Mega; anaritide, GLP-1+exendin-4, NIH: GLP-1, Amylin; GLP-1, TheraT Bayer, anaritide, Bristol; anaritide, Aventis-2; anaritide, ech; GM-1986: GM-CSF blocker, Hospira; GnRH-associ Astellas; anaritide, GlaxoSmithKline-2; anaritide, Aventis-1, ated peptide: GPCR antagonists, NIH; GPIb/IIIa antags, anaritide, Mitsubishi Tanabe; anaritide, Novartis; anaritide, Selectide; GRF1-44; GRF, Lilly; GT2342; GT2501: GYKI OmniGene; anaritide, Sankyo; anaritide, Scios; angiotensin 14451; galanin; gastrin antagonists; gastrin, Novo; glaspi II antagonists; anti-inflammatories, Affymax; anti-inflamma mod; glicenting glucagon antagonists, SynVista; glucagon, tory peptide, BTG.; anti-integrin peptides, Burnha; anti-TCR Lilly; glucagon, ZymoGenetics; glucagon-121; glycoprotein vaccines; antiallergy peptides, Ajin; antiallergy vaccine, 1 balpha fragments; gonadorelin analogues, Syntex; gona Acambis-1, anticancer matrix, Telios; anticancer peptides, dorelin antagonist, Ortho; gonadorelin preparations; gona Micrologix; antiflammins; antifungal peptides, BTG.; anti dorelin, Arana; gonadorelin-MDP vaccine; goralatide; fungal tripeptides, BTG.; antiGnRH immunogen, Aphton; gp120-V3 peptides; growth factor peptides, Biothe: ENMD Gastrimmune; antirenin vaccine; antirheumatic peptides, 0996; H-142, AstraZeneca; Her-2/Neupeptides, GSK; herpes Acambis; antithrombin polypeptides; antiviral peptide, Bio simplex vaccine, Wistar: AIDS vaccine, Cel-Sci; HP-101; Mega; antiviral peptides, Non-indust; antiviral peptides, vitespen; HSV vaccine, Cel-Sci; HSV-1 g/vaccinia vaccine; Yeda; apolipoprotein, NeuroSearch; apoptosis technology, heparin binding peptides, NC1; hepatitis-B receptor; hepati Receptag. BCH-143; arthritis antigen; atrial natriuretic pep tis-B Vaccine, Tokyo; hepatitis-B Vaccine, Protherics; hepati tide, Ph; atrial natriuretic peptide, Ra; avorelin; B-956; BCH tis-B Vaccine-2, BTG; hirugen; I5B2; iseganan hydrochlo 2687: BCH-2763; frakefamide; BIM-22015; BIM-26028; ride; IgE peptides; IgG binding factor, Hoechst Ma: BIM-44002: BIO-1006: BIO-1211: Bio-Flow; BPC-15; Brit netarmftide; Insulin Aspart; Zorcell; icrocaptide; icatibant: istatin; BST-2001; bivalirudin; bombesin antagonist; brain immunomodulating peptides, Bio; infertility, E-TRANS: natriuretic peptide; brain natriuretic peptide, Phar; C-peptide influenza vaccine, GSK-1; influenza vaccine, Yeda; instimu analogues, UCB: C5a antagonist, Abbott; C68-22; Casocidin, lin; insulin analogue, Lilly; insulin analogues, Lilly; insulin Pharis; CBT-101; CCK(27-32), Organon: CD4, Genelabs; analogues, Scios; insulin formulation, Pasteur, insulin CD4-liposome conjugate, Sumito; CEE-04-420; CEP-079; glargine; insulin, Nektar, inhaled; insulin molecules, Novo; CEP-903; CETP vaccine, Avant; mi?amurtide; CGRP ana insulin oral, Inovax; insulin transdermal; insulin, Organon; logues, Asahi Chemical; CGRP. CSL; CGRP Celltech: insulin, ocular; insulin, AERX; insulin, AutoImmune; insulin, CGRP, Novartis; CGRP, Asahi Kasei: CGRP, SmithKline BEODAS; insulin, Biobras; insulin, Ferring Pharma; insulin, Beecham; CGRP Unigene; rusalatide acetate; CI-782. CKS CJ Corp; insulin, Chiron; insulin, Chong Kun Dang; insulin, 17: CMV peptides, City of Hope; CNTF, Fidia; CP-95253; Sanofi Pasteur; insulin, Di-Arg, Hoechst Mario; insulin, corticorelin acetate; CT-112, BTG: CT-1508: CTAP-III, Cre E-TRANS: insulin, Forest; insulin, Hoechst, semisynth; insu ative; CTP-37; PMD-2850; CVFM; CVT-857; CY-725: lin, Lilly, iodinated; insulin, Genentech, recombi; insulin, CY-726; CYC101: CYC103: CYC102; calcitonin, Peptitrol; Provalis; insulin, Novartis; insulin, nasal; insulin, Ohio; insu calcitonin, Rockefeller, calciseptine; calcitonin analogues, lin, Nazdel; insulin, Novo, synthetic; insulin, nasal, Novo SB; calcitonin, Amgen, calcitonin, Armour, calcitonin, Beau Nordisk; insulin, oral; insulin, buccal, Generex; insulin, four, calcitonin, Inhale; calcitonin, Bridgelock, calcitonin, Arana; insulin, Anesiva; insulin, Procter & Gamble; insulin, microspheres; calcitonin, NaZdel; calcitonin, Novartis; calci Qmax; insulin, Innovata; insulin, Roche; insulin, recombi tonin, nasal, Novartis; calcitonin, oral, Mannkind; calcitonin, nant, Aventis; insulin, Shionogi; insulin, Shire; insulin, Panoderm; calcitonin, Pharma Bissendorf, calcitonin, Phar Spiros; insulin, SRI; insulin, Structured Biological; insulin, mos; calcitonin, Anesiva, calcitonin, Aventis; calcitonin, semisynthetic, Biobra; insulin, synthetic, Powerpatch; insu Teijin; calcitonin, Teikoku; calcitonin, TheraTech; calcitonin, lin, Zymo, recombinant; insulin, monocomponent, Novo; Yissum; calf thymus derived peptides; calpain inhibitors, IL-1 receptor antagonist, Affym; interleukin-1B, Sclavo; ResCo., calphobindin I; cancer vaccines, Argonex; carguto interleukin-8 antags, Select; J015X; J018X; AG-1776; KNI cin; casokefamide; cekropin-P; chemokines, Dompe; tasi 549; prailmorelin; KPI-022; katacalcin; ketomethylureas: US 2011/0166063 A1 Jul. 7, 2011

L-34.6670; L-364210; L-659837; L-693549; L-709049; Centocor, somatoliberin, Takeda; PTR-3173; somatostatin L-75; L-761191; L-histidyl peptides; LDV-containing pep analogue, Shira; somatostatin analogues, Merck; somatosta tides, Antiso: LEAPS-101; LHRH antagonists, Abbott; tin analogues, Tulane; somatostatin derivatives; Somatostatin, PD-6735; Lys-Phe; hLF1-11; lagatide; laminin A peptides, Merck Serono. Somatostatin, Ferring; Somatostatin, Arana; NIH; laminin technology, NIH; lanreotide; leuprolide somatostatin, Sanofi-Synthelabo; somatostatin, BayerScher acetate, Atrigel; leuprorelin, Takeda, leuprorelin, Merck ingPhar, T-205, Streptococcus A vaccine, Active; Sulglic Serono; leuprorelin, DUROS; leuprorelin, Powerpatch; lipid otide; syndyphalin; synthetic p16, Dundee; synthetic peptide linked anchor technology; lysozyme metabolites, SPA; MCI BPC, Pliva; synthetic peptides, ICRT; T cell receptor peptide 826; omiganan pentahydrochloride; MBP ImmuLogic: MCI vaccin; T-118; T-786; T-cell receptor peptides. Xoma; T22; 536; MDL-104238; MDL-28050; Metascan; MMP TA-3712: TASP inhibitors; TCMP-80; Tc-99m P215: inhibitors, NIH, MN-10006; MOL-376; MR-988: MSH Tc-99m P4.83H; Tc-99m P773; Tc-99m depreotide; Tc-99m derivatives: MUC-1 vaccine, Pittsburgh; malaria vaccine, P280; TEI-1345; THF, Pfizer; Theradigm-HBV: Theradigm Axis; malaria vaccine, Vernalis; malaria vaccine, Cel-Sci; HIV: Theratides; Stimuvax; ThCRF 1-29; tesarnorelin malaria vaccine, Roche; melanoma vaccine, Nobilon; men acetate; ThromboScan: TIMP, Creative BioMolecules: TIMP. ingitis vaccine, Acambis; mertiatide; metkephamide; metor Sanofi-Aventis: TJN-135; TNF inhibitor, Genelabs; phamide; monocyte chemotactic factor, montirelin hydrate; TP-9201; TRH analogues, Roche: TRH. Daiichi: TRH, Japan motyline; murabutide; muramyl dipeptide derivatives; Tobacco; TRH, Medicis: TRH, Arana; TRH-R, Medical myelopid; N-acetyl Leu-28Leu-31NPY24-36: N-carboben Research Counci; TT-235; tabilautide; tendamistat; terlipres Zoxy peptides; NAGA; tiplimotide; opebecan; insulin sin; terlipressin, Nordic, teverelix; INKP-2001; thymic pep detemir, liraglutide: Nona CCK; NP-06: NPC-18545; Nva tide; thymoleptic peptides; thymopentin; thymopentin ana FMDP: nacartocin, natural peptide BPC, Pliva; nerve growth logues; thymosin alpha-2; thymosin 134, thymosin fraction factor, Synergen; nesiritide citrate; neuropeptides, Prother 5; tolerizing peptide, Acambis; trefoil peptides, ICRT; tri ics; neuropeptides, Pfizer; neurotensin, Merck; neurotrophic letide; tuftsin, Abic; tuftsin, Sclavo; Type I diabetes vaccine, factors, CereMedix; nifalatide; CL22, Innovata; nootropic, RCT tyrosine kinase antags, ICRT; tyrosine-containing Yakult; nociceptin, Euroscreen; Org-2766: Org-30035: OSA dipeptides; UA 1041; UA 1155. UA 1248; Uroguanylin, peptides, Osteopharm; octreotide; opioid peptides, Unigene; Pharis; urodilatin; V.F.; VIC, Astellas; VIP analogues, osteogenic growth peptide; osteoporosis peptides, Telios; TRION: VIP derivative, Eisai; VIP fusion protein, Kabi: vap oxyntomodulin; P-113, Demegen: PACAP 27: PAPP; reotide, immediate-release; varicella Vaccine, ResCo.; Vit PD-83176:PD-122264; PD-132002; PEP-F; Penetratin: Pep ronectin receptor antag; vicalcins: Mycoprex: YIGSR tigen agents; Phe-X-Gly, ResCo; PL-030; PN1 antagonists, Stealth; Yissum Project No. 11607; Pharmaprojects No. Allelix: POL-443; POL-509; PPA, ResCo; PR-39; Prodap 1088; Pharmaprojects No. 1113: Pharmaprojects No. 1269; tin-M technology; PSP; tigapotide triflutate; PT-14; PT-5: Pharmaprojects No. 1448; Pharmaprojects No. 1507; Phar semparatide; PTL-78968; parathyroid hormone fragments: maprojects No. 1573: Pharmaprojects No. 1583; Phar pancreastatin: papillomavirus Vaccine constru; parathyroid maprojects No. 1626; Pharmaprojects No. 1779; Phar antagonist, Merck; enfuvirtide; peptide heterodimers, maprojects No. 1797; Pharmaprojects No. 1843; Cortech; peptide imaging, Diatide; pentapeptide 6A, pen Pharmaprojects No. 1876; Pharmaprojects No. 1913; Phar tigetide; peptide analogues, ResCo. peptide 6, NY Medical maprojects No. 1939: Pharmaprojects No. 1994; Phar College; peptide G, Arana; peptide inhibitors, ICRT; peptide maprojects No. 2043; Pharmaprojects No. 2044; Phar T analogue, Carl; peptide T analogues; peptide T. Arana; maprojects No. 2063; Pharmaprojects No. 2100; peptide? drug vehicle, BTG; peptides, Sanofi-Aventis; pep Pharmaprojects No. 2122; Pharmaprojects No. 2202: Phar tides, Scios; peptides, Yeda; peptomers, NIH; pertussis vac maprojects No. 2363; Pharmaprojects No. 2388; Phar cine-1, TRION: ph-914; ph-921; ph-9313; phospholipase maprojects No. 2425; Pharmaprojects No. 2476; Phar inhibitors, Poli; prolactin, Genzyme; pramlintide; pranlukast; maprojects No. 2527; Pharmaprojects No. 2560; proinsulin, Lilly; proinsulin-2, Novartis; progenitor cell Pharmaprojects No. 2571; Pharmaprojects No. 2825: Phar inhibitor, RCT proinsulin fragments, Lilly; proinsulin ana maprojects No. 2866; C-type natriuretic peptide, Sun: Phar logues, Lilly; proinsulin, Genentech; prostate cancer Vaccine, maprojects No. 2909; Pharmaprojects No. 2912; Phar United; prostate cancer vaccine, GSK; protirelin; protirelin, maprojects No. 2913; Pharmaprojects No. 3009: Takeda: Pseudomonas elastase inhibitor; QRS-10-001; QRS Pharmaprojects No. 3020; Pharmaprojects No. 3051; Phar 5-005; Quilimmune-M: Retropep; RGD peptides: RHAMM maprojects No. 3127; Pharmaprojects No. 3284; Phar targeting peptides, Cange: Ro-25-1553; RP-128; RSV vac maprojects No. 3341; Pharmaprojects No. 3392: Phar cine, Avant; RSV vaccine, Acambis; RWJ-51438: TRH, Fer maprojects No. 3393; Pharmaprojects No. 3400; ring; renin inhibitors, Pfizer-2; relaxin, Novartis; renin inhibi Pharmaprojects No. 3415: Pharmaprojects No. 3472: Phar tors, INSERM; romurtide; rubella vaccine, Protherics; maprojects No. 3503; Pharmaprojects No. 3581; Phar S-17162: S-2441; SC-40476; SC-44900; SDZ-CO-611; maprojects No. 3597; Pharmaprojects No. 3654; Phar SIDR-1204; SK&F-101926; SK&F-1 10679; SLPI, Syner maprojects No. 3667; Pharmaprojects No. 3777: gen; edotreotide: SP-1; SPAAT: SR-41476; SR-42128; Pharmaprojects No. 3862: Pharmaprojects No. 3863; Phar SR-42654; SRIF-C. Streptococcus A vaccine, ID: Strepto maprojects No. 3891; Pharmaprojects No. 3903; Phar coccus A vaccine, SIGA; calcitonin, PPL: salmon calcitonin, maprojects No. 3939: Pharmaprojects No. 3963; Phar Therapicon; sermorelin, Kabi: Saralasin acetate; secretin, maprojects No. 3989; Pharmaprojects No. 4004: Eisai; Secretin, Ferring; secretin, Wakunaga; sermorelin, Pharmaprojects No. 4093: Pharmaprojects No. 4098: Phar Novartis; sermorelin peptides, Sanofi-Ave; sermorelin, Anti maprojects No. 4113: Pharmaprojects No. 4182; Phar genics; sermorelin, Molecular Genetics; sermorelin acetate, maprojects No. 4209: Pharmaprojects No. 4246; Phar Merck Ser; sermorelin, Sanofi-Aventis; sermorelin, Unigene: maprojects No. 4251; Pharmaprojects No. 4300: Sinapultide; sleep inducing peptide, Bissen; Small peptides, Pharmaprojects No. 4323; Pharmaprojects No. 4347: Phar US 2011/0166063 A1 Jul. 7, 2011 maprojects No. 4367; Pharmaprojects No. 4385; Phar AG-702; insulin, AeroDose; anti-inflammatory, Taplmmune; maprojects No. 44.02: Pharmaprojects No. 4445; Phar insulinglulisine: GPG-NH; hepatitis-B therapy, Tripep; Sta maprojects No. 4544: Pharmaprojects No. 4625; phylococcus therapy, Tripep; angiogenesis inhibitor, Tripep; Pharmaprojects No. 4626; Pharmaprojects No. 4643; Phar bone marrow inhibitor, Tripep: melanoma vaccine, BioVec maprojects No. 4705; Pharmaprojects No. 4708; Phar tor; lipopeptides, Cubist; ABT-510; parathyroid analogue, maprojects No. 4766; GHRP-1, QLT; Pharmaprojects No. Unigene; Adageon-E: A-443654; CJC-1131; FE200 665; 4865; Pharmaprojects No. 491; Pharmaprojects No. 4915; insulin, TranXenoGen; Gilatide; TFPI, EntreMed; desmo Pharmaprojects No. 4936; Pharmaprojects No. 494; Hema pressin, Unigene; leuprorelin, oral, Unigene; antimicrobials, tide; Pharmaprojects No. 4975; Pharmaprojects No. 5048; Isogenica; insulin, oral, Unigene; metastin, TRI-1144; DEBI Pharmaprojects No. 5055; Pharmaprojects No. 5076; anti 4022; HM-9239; insulin, Bentley, intranasal; F-992: ZP-10; HER2/neu mimetic, Cyclacel; Pharmaprojects No. 5131; E1-INT: DEBIO-0513; spinal cord injury vacc, Weizrn; Pharmaprojects No. 5173: Pharmaprojects No. 5181; Phar DAC:GLP-2: uPAR inhibitors, Message: MBP-8298; maprojects No. 5200; Pharmaprojects No. 5216; Phar PL-14736; anaritide peptides, BTG: SP-1000, Samaritan: maprojects No. 5292; Pharmaprojects No. 5348; Phar leuprorelin, Ardana; melanocyte modulators, IsoTis; maprojects No. 5356; Pharmaprojects No. 5412: DMP-444; HF-1020; leucocyte immobilizing peptide; Dentonin; MET Pharmaprojects No. 5657; Pharmaprojects No. 5728; Phar 1000; SGS-111: 5-Helix. HPV vaccine, Ludwig: caries vac maprojects No. 5839: Pharmaprojects No. 5910; TGF-f cine, Forsyth; taltobulin; ATN-161; T05; LY-307161; Spneu antagonists, Inspiraplex: Pharmaprojects No. 5961; Phar moniae vaccine, Milleniu; Alphastatin; anticancer peptides, maprojects No. 5991; Pharmaprojects No. 6021; Phar Wockhardt; PGN-0052: INNO-201; leuprolide, Nektar; insu maprojects No. 6063: Pharmaprojects No. 6083; PI-0824; lin, BioSante, oral; ADD-9903; viral vaccines, Bio-Virus; RIP-3, Rigel; NBI-6024; Pharmaprojects No. 892: Phar AOD-9604; calcitonin, oral, Pfizer; insulin, INJEX; ETD maprojects No. 955; IR-501; A6, Angstrom; leuprolide, Pro XXXX; analgesic, Sigyn; anti-infectives, AM-Pharma: Maxx: Orolip DP; edratide; 131-I-TM-601; Prosaptide TX14 human AMPs, AM-Pharma; INGAP peptide; osteomyelitis (A), Savient; insulin, Flamel: p1025; NIH; protein kinase R peptides, AM-Phar: XOMA-629; XMP-293 derivatives: antags, NIH; GLP-1, Daiichi: EMD-249590; secretin, Repli BlockAide/VP: Eradic Aide: BlockAide/CR; VAC-12; leu Gen: RANTES inhibitor, Milan; Pharmaprojects No. 6236: prolide, oral, DOR BioPharm; synthetic erythropoiesis pro; NY ESO-1/CAG-3 antigen, NIH; BILN-504 SE; NIPs, RCT; B-amyloid vaccine, Intellect; CEL-1000; sincalide; Panko insulin, Biphasix; ZRXL peptides, Novartis; BIM-23190; Pep; albiglutide; insulin, Bharat; leuprorelin, Norwood; leuprorelin, TheriForm; B-amyloid peptides, CeNeS; oglu Reversin 121, Solvo; SB-144; SB-29, STIL; cancer vaccine, fanide disodium; amyloid inhibiting peptides, AX: iprP13; Sedac: SDT-021; malaria vaccine, Sedac, ther; malaria vac PN-277; differentiation inducers, Topo; immune privilege cine, Seda, prophyl; hepatitis-C cellular ther, Seda; Factor factor, Proneu; TASP-V: anticancer vaccine, NIH, Phar XIIIa inhib. Curacyte; insulin, Micronix; AIDS vaccine, Anti maprojects No. 6281; HAV peptide matrix, Adherex: calcito gen Express-1; exenatide LAR: AIDS vaccine, Bionor nin, oral, Biocon; analgesic, Nobex; PTH 1-34, Biocon; insu Immuno-1; GV-1002; GV-1001; MSI vaccine, GemVax: lin, oral, Biocon-2: BLS-0597; leuprorelin, Depocore; IDPS: PEP-14; PV-267; antibacterials, Provid; hepatitis-B Vaccine, AIDS vaccine, Hollis-Eden; insulin, Nova)el; insulin, Ora Innovata; BA-058; BIM-51077; malaria vaccine, Immuno some; Pharmaprojects No. 6310: TRP-2. NIH; Phar genics; TM-701; VG-104; AC-162352; antivirals, Genencor; maprojects No. 6320: Re-188 P2045; calcitonin, Inovio: leuprolide acetate, Voyager, calcitonin, nasal, Archimedes; golotimod, angiotensin-II, topical, Trine; ETRX-101; anti insulin, nasal, West, calcitonin, oral, Unigene; calcitonin, allergy vaccine, Acambis-2: Tc-99m-P424; Tc-99m-P1666: nasal, Unigene. IMX-735: IMX-775; PPI-01; anti-IgE pep insulin, Transfersome; Yissum Project No. 11649; SP(V5.2) tide, Allergy Ther. BZK-1 11: TH-0318; Enkastim; antibiot C. melanoma vaccine, Therion-2; insulin Aspart, biphasic, ics, Bayer; Cerebrolysin; colorectal cancer therapy, IDM: Novo; Tat peptide analogues, NIH; Pharmaprojects No. 6365; wound growth factor, NephRx; JPD-105; osteoporosis drugs, Pharmaprojects No. 6373; Ramot project No. 981; ESP Ferring: PN-951; CZEN-002: ZP-120; pasireotide; HerVac: 24218: Pharmaprojects No. 6395; calcitonin, oral, Emi CTT, LLG peptide, CTT; Pharmaprojects No. 6779; mep sphere; omiganan, topical: AIDS vaccine, United-3; leupro tides, Senexis; Q-8008; FX-06; PhG-alpha-1; insulin, oral, relin, Archimedes; HPV 16 E6+E7 vaccine, NIH; peptide Biocon; PP-0102; GTP-010: PAR-2 antagonists, EntreMed: vaccine, NC1; Chlamydia vaccines, Argonex; delimitide parathyroid analogue, Zelos: K-1020; CTCE-9908; CTCE acetate; RSV vaccine, Pierre Fabre-2; F-50040; CPI-1500; 0214; urocortin-II, Neurocrine; telomerase vaccine, Den AIDS vaccine, BioOuest; insulin, BioSante, inhaled; antian dreon: AKL-0707: PYY3-36, Nastech; prostate cancer vac giogenics, GPC; TNF degradation product, Oncot; insulin, cine, Pepsca: AEZS-130; LYN-001: CUV-1647: AL-108; Emisphere; ozarelix; bremelanotide; Pseudomonas Vaccine, AL-309; HNTP-15; BIM-28131; CSF-Gagonists, Affymax: Millenium: AIDS vaccine, CIBG: AIDS vaccine, Wyeth Vac IL-5 antagonists, Affymax; TRAIL agonists, Affymax; IgE cines-3; HCV serine protease inhib, BI; insulin, Wockhardt; inhibitors, Affymax; TM-801; TM-901; BN-054: APTA-01; cat PAD, Circassia:NOV-002: PPI-3088; insulin 24 hr, Altea; HB-107; AVE cancer vaccine: PxSR; STD peptides, Helix: AP-811; hNLP, Pharis: ANUP-1, Pharis; serine protease CFanti-infectives, Helix; HB-50; Homspera; S-0373: PYY3 inhibs, Pharis; Pharmaprojects No. 6523; respiratory mucus 36, oral, Emisphere; XG-101; XG-201CS: XG-102; insulin, inhibitor, Em; CLX-0100; AIDS vaccine, Panacos; SPHERE oral, Coremed; Alzheimer's vaccine, Prana; AIDS vaccine, peptide vaccine, Genzyme: P-16 peptide, Transition; Bionor Immuno; leuprolide acetate, ALZAmer; AUX-202; EP-51389; insulin, ProMaxx: ET-642; P-50 peptide, Transi AR-HO44.178; PYY3-36, Thiakis; lanreotide SR; malaria tion Ther; Famoxin; insulin, Alkermes, inhaled; GPCR pep vaccine, Pevion; Alzheimer's vaccine, Pevion; melanoma tide ligand, Synaptic; DiaPep227; alpha-1-antitrypsin, vaccine, Antigen Expr; melanoma vaccine, Pevion; OGP Cortech; IC-41; tuberculosis vaccine, Intercell; immunosup (10-14)-L: ABS-13; ABS-17; cancer therapeutics, Argolyn; pressant vaccine, Aixl; malaria vaccine, NYU-2; netupitant; substance P-saporin; diabetes therapeutic, Thera; CGX US 2011/0166063 A1 Jul. 7, 2011

1051; OTS-102; Xen-2174; insulin, inhaled, Coremed; inflammatory, Forbes; labour inhibitor. Theratechnolo: glau WP9QY: osteoporosis treatment, Fulcr; AHNP. Fulcrum; coma therapy, Theratechnolo; AG-EM-0040: MS therapy, insulin, Technosphere, Mannkind; FX-07: CBP-501; E7 vac AplaCen; interleukin-2 mimetic, AplaGen; CNS therapy, cine, Neovacs: LSI-518.P. aviptadil, Mondobiotech; antican AplaCen; Mesd-based peptides, Raptor, paratohormone, cer peptide, OrthoLogic; AL-209; OP-145: AT-001: AT-008: Sidus; asthma therapy, Synairgen: dekafin-1, anticancer vac CHP-105: AMEP. BioAlliance; cardiovascular ther, Argolyn; cine, Ulm; BT-15; cancer imaging agent, Speci; cardiovascu TEIPP-03; mental retardation ther, Argol; IMX-002: IMX lar imaging, Speci; E-75; Prothyx; anticancer, Prothyx, 942: NLC-001; octreotide, Indevus: DRF-7295; opioid pep Stealthyx; IL12-NGR; allergy vaccine, China Bio; amylin tide derivatives, Ka; CDX-110; ALT-212; desmopressin, mimetic, 2nd-gen, Amylin; influenza vaccines, Variation; Orexo; IMA-901; obinepitide; TM-30335; HIV therapy, VLP-0012M; PLT-101; AL-408; anticancers, Aileron; antivi OyaGen-1; calcitonin, oral. ThioMatrix; insulin, oral. Thi rals, Ambrx; hSPN-16; HDL, Cerenis; enterostatin: BSc oMatrix; BRX-00585; Insulin Aspart, biphasic-2. No: 2118; SB-006; antimicrobials, Spider Biotech; peptide CG-55069-11; GLP-1, Emisphere; linaclotide acetate: NPT therapy, Angioblast; octreotide, Ambrilia; GAP-134; Alzhe 002; terlipressin, Orphan Therapeut; ZT-153; SciClone: imer's therapy, Il Dong; BL-4020; von Willebrand factor, FGLL; Syn-1002: MIP-160; PI-2301: PI-3101; BDM-E: Baxter; IL-1 aOb; POT4: gamma-secretase inhibitors, BMS; insulin, Medtronic; ST-03: TH-0312; hepatitis-C vaccines, ISCOMATRIX; enfuvirtide, needle-free; connexin modula Kochi; cetrorelix acetate, once-weekly; RPI-MN; neurode tors, NeuroSol; BT-25; BT-20; AmpTide: HepTide: antimi generative ther, Recepto; RPI-78M; B-amyloid inhibitor, crobial peptides, Helix; NPYagonists, Bayer; ragweed PAD, Alzhyme: DMI-3798; DMI-4983; ruzam: CT-319; Circassia; dust mite PAD, Circassia; grass PAD, Circassia: EN-122004; glyponectin: EN-122001; EN-122002: KAI transplant rejection PAD: insulin, oral, Oramed; cardiac 9803; insulin, Advancell; larazotide acetate; calcitonin, oral, ischaemia therapy, Phy: PYC-18; antidiabetics, Phylogica; Bone Medical; parathyroid hormone, Bone Medi; calcitonin, PEP-35; ACE-041: ACE-031; ovarian cancer vaccine, Merrion; desmopressin, Merrion; acyline, Merrion: IMX Generex: ATX-MS-1467; iATX FVIII; diabetes vaccine, 503; AP-214; Streptococcus vaccine, Vaccine; cytomegalovi Apitope; allergy vaccine, Apitope; FX-06 analogue: PR-22G: rus vaccine, Vacc: RHS-08: AG-707; antiallergics, Phylogica; PR-21, Pharmaxon; LT-1945; LT-1942; XG-414; XG-517; PYC-36S: anticancers, Phylogica; Glypromate; NNZ-4945: AC-163794: MDPTO; B27PD; AC-2307; sedatives, Pro calcitonin, intranasal, ITI; Peptide T. Advanced Immuni T. Therapeutics; L-Type Ca channel blocker, Pro; phospholi APTA-O; CGRP Akela: TKS-1225; GalR2 peptide agonist, pase A2 inhibitor, Pro; PGL-3001: PGL-1001; influenza vac NeuroTa; botulinum vaccine, Emergent; HIV fusion inhibi cine, Variation-2; Homspera nanoparticle. Immune: CVX tors, Sequoia: AL-208: APP-018; BKT-RP3; smallpox vac 096; COR-1; Survivin-2B: imMucin, GLP-1, PharmaiN: cine, Antigen Expr: CMLVAX-100; INNO-105; insulin, atherosclerosis vaccine, Affir, adeptide; Somatostatin antago Intravail; leptin, Intravail; calcitonin, Intravail; Somatropin, nists, Preg: Casimax; CD-NP, Nile; PRX-111; ACT1-C; Intravail; heparin, Intravail; erythropoietin, Intravail; PRX-102; ACT1-G: AIDS vaccine, ITS; influenza vaccine, CT-201; telomerase variants, GemVax; INT, transplantation; ITS; hepatitis-C vaccine, ITS; ALTY-0601; BGLP-40; soma INT-3; SPI-1620: BIO-037; anticancers, Bracco: BIO-023; tropin, INB: trypansomal vaccine, INB; RU-COH, Pantarhei; ZT-100; MC-4Ragonists, Lilly; LT-1951; PTH (1-34), IGI; LH-COH, Pantarhei; GLP-1 analogues, Unigene; Poly CGRP, VasoGenix; BIO-145; BIO-142; stem cell factor, fensin; VIR-576; Xen-0568; Xen-0495: Xen-0468; LEKTI Affymax: VEGFR-2 antagonist, Affymax; KGF receptor 6; leukaemia vaccine, MD Anderson; Met receptor agonists, agonist, Affymax: YM-216391; AT-007: AT-011; EK2700; MRCT; insulin HDV, short-acting, Dia; glucagon antago EK900-1800; EK900-12: FGLm: ABS-201: Mdbt-12; nists, CoGene: GLP-1 agonists, CoGenesys; insulin HDV, autoimmune therapy, Antigen; VX-001; IPP-102199; IPP oral, Diasome; insulin HDV, long-acting, Dia; glucagon, Par 201101: CTA1-DD; Factor VIIa inhibitor, ProTher; antian ticle Therapeutics; GLP-1. Mannkind; insulin, next-genera giogenic, ProTherapeutic; IMT-1012; colon cancer vaccine, tion, Flamel; Ostabolin-C, topical; DAC: HIV; antiviral, Hep Immunoto; prohanin, ProTherapeutics; Smallpox vaccine, Tide: Insulin Aspart, biphasic-3, No: Innotide; influenza BioDefense; heart failure therapy, ElaCor; PA-401; 802-2: vaccine, Bionor; HPV vaccine, Bionor Immuno; hepatitis-C insulin, nasal, Nastech: SEN-304; IMA-920; IMA-940; vaccine, Bionor; Affitope AD-02: Affitope AD-03: RHS-02: IMA-910; influenza vaccine, Antigen, HN; Primacoll; oct RHS-03; insulin, Access; inherbins, Enkam; Dekafin-2: reotide, PR Pharmaceuticals; female infertility th, Vyteris; BL-4050; ALS vaccine, Amorfix; cancer vaccine, Canopus: FAR-404; athlete's foot therapy, Helix; leishmaniasis ther, relaxin, Corthera; rhNRG-1: rhErbB3-f; hepatitis-C vacc, Helix; INNO-305; ALS-O; sNN-0465; N,N-5401; TRI-999; Green Cross-3; androgen receptor antag, CRT, GLP-1 ana Org-214444; Org-33409; IMA-930: YH-APC: PYC-35B; logue CR, OctoPlus: AIDS vaccine, Sanofi Past-12; insulin, Rev-D4F; insulin, Phosphagenics; coeliac disease ther, Nex Diabetology; Combulin; AIDS vaccine, Sanofi Past-11; pep; coeliac disease therapy, BTG.; exendin-4, PC-DAC: AnergiX.MG: AnergiX.MS: insulin, CritiTech:YP-20; NDR/ exenatide, nasal spray; CAP-232; ACE-011; Cardeva; NCE-18; CLT-002: CLT-007: CLT-008, Charlesson; CLT BL-3020; FM-TP-2000; GGTI-2418; TM-30339; DP-74; 009: PYC-38: AIM-101; AIM-102: AIM-501: APL-180; DP-68; PPH ther, GeoPharma; MPL-TLB100: AZX-100: metabolic disease therapy, Xen; NP-213: NP-339; antimicro Alloferon: S2: S3; S4; PAC-G31P: PAC-745; PAC-525; PAC bial peptides, NovaB: lung anti-infectives, NovaBiot; c-pep 113: VEBV; lipopeptide, Combinature: mondopeptide-1; tide analogue, Diabetology; CGEN-855; NN-1250: N,N- mondopeptide-2; mondopeptide-2+mondopeptide-3; mon 9535; insulin, rectal, Oramed; insulin, 12 hr. Altea; pancreatic dopeptide-4; MLIF; carfilzomib: Affitope AD-01; LT-ZP001; cancer vaccine. Onco; SB-101; L-glutamine, Emmaus; glu LT-ZMP001; CGX-1204; C3d, Enkam: C5a antagonist, cagon antagonists, Kisspeptin-54, Kisspeptin-14: Kisspep Eucodis; adenocarcinoma vacc, ImmvaRX; insulin, oral, tin-13; Kisspeptin-10; Ziconotide; Biphalin; Nesiritide; Pro Apollo; renin inhibitors, Servier; Factor VIIa, GTC; ABS tegrin-1; Protegrin-2: Protegrin-3: Protegrin-4: Protegrin-5; 212: NAFB001: NAFB002; insulin, MediVas; ZT-181; anti Preprotegrin: V681; V681 (V13A); GLP-2: GLP-2 (A2G): US 2011/0166063 A1 Jul. 7, 2011

GLP-2 (A2G/C34); AOD-9604: Ac-AOD-9604(S8K); cation. In some instances, these peptides contain features that Ac-AOD-96.04(K17); C-peptide: CR845; and Marcadia. are either inconsistent with or not amenable to inclusion in the 0202. In certain embodiments of the invention, PEP is a Sequence Listing. For example, a sequence with less than therapeutic peptide selected from the therapeutic peptides four-amino acids; a sequence with a D-amino acid; or certain listed in Table 1. modification that cannot be described in the Sequence Listing presently, and therefore are not provided in the Sequence Table 1 Listing. However, for the ease of use and description, a SEQ ID NO. has been provided to these peptides (i.e., SEQ ID 0203 This table lists the SEQID NOS., names, sequences, NOs: 302-469). and known or Suspected therapeutic activities of various pep (—NH indicates amidation at the C-terminus; Ac indicates tides described herein. The SEQ ID NOS. 1-301 describe acetylation; other modifications areas described herein and in sequences that are required to be provided with the Sequence the specification; SIN indicates Sequence Identification Listing and are therefore appended with the instant Specifi Number)

SEQ ID Sequence and/or other Identifying Therapeutic NO: Name Family Information Activity

1. carperitide ANP SLRRSSCFGGRMDRIGAOSGLGCNSFRY, Cardio stimulant human alpha-atrial natriuretic peptide; Respiratory Atriopeptin-28 (human); 2 alpha- Endorphin H-Tyr-Gly-Gly-Phe-Leu-Arg-Lys-Tyr-Pro-Lys-OH Analgesic, other neoendorphin

3 A-3847 Insulin gi|386828|gb|AAA59172.1|insulin Homo Antidiabetic sapiens MALWMRLLPLLALLALWGPDPAAAFVNOHLCGSHLVEALYLVCG ERGFFYTPKTRREAEDLOVGOVELGGGPGAGSLOPLALEGSLOK RGIVEOCCTSICSLYOLENYCN

4. A-4 114 Insulin MALWMRLLPLLALLALWGPDPAAAFVNOHLCGSHLVEALYLVCG Antidiabetic ERGFFYTPKTRREAEDLOVGOVELGG

5 A- 68552 GPGAGSLOPLALEGSLQKRGIVEOCCTSICSLYOLENYCN Anorectic/ Antiobesity

3 O2 and A-75998 Ac-D-2Nal1-D-4ClPhe2-D-3 Pal3-NMeTyr5-D- Releasing 3O3 Lys (Nic) 6-Lys (Isp) 8-D-Ala1OGnRH; N-acetyl- hormone D-2-naphthylalanyl-D-4-chlorophenylalanyl- Reproductive/ D-3-pyridylalanyl-seryl-N-methyltyrosyl- gonadal, general D-N (epsilon) - nicotinyllysyl-leucyl N(epsilon) - isopropyllysyl-prolyl-alaninamide acetate

6 AN-1792 beta- gi|8176733|gb|AAB26264. 21 beta-amyloid Cognition amyloid peptide precursor; beta APP Homo sapiens enhancer peptide GSGLTNIKTEEISEVKMDAEFRHDSGYEVHHOKLVFFAEDWGSN KGAIIGLMWGGVWIATVIIITLVMLKKOYTSNHHGVVE

7 AAMP-1 MESESESGAAADTPPLETLSFHGDEEIIEWWELDPGPPDPDDLA Anticoagulant OEMEDVDFEEEEEEEGNEEGWVLEPOEGVWGSMEGPDDSEWTFA Anti LHSASWFCWSLDPKTNTLAWTGGEDDKAFWWRLSDGESSFECAG inflammatory HKDSVTCAGFSHDSTLVATGDMSGLLKVWOVDTKEEVWSFEAGD Immunological LEWMEWHPRAPWLLAGTADGNTWMWKVPANGDCKTFOGPNCPAT Anticancer, CGRVLPDGKRAVWGYEDGTIRIWDLKOGSPIHVLKGTEGHOGPL other TCVAANODGSLILTGSVDCOAKLVSATTGKVVGVFRPETVASOP Vulnerary SLGEGEESESNSWESLGFCSWMPLAAWGYLDGTLAIYDLATOTL RHOCOHOSGIVOLLWEAGTAVWYTCSLDGIWRLWDARTGRLLTD YRGHTAEILDFALSKDASLVVTTSGDHKAKVFCVORPDR

8 Exenatide GLP-1 HGEGTFTSDLSKOMEEEAVRLFIEWLKNGGPSSGAPPPS Antidiabetic Anorectic/ Antiobesity

9 AC- 625 Acetyl-ATORLANELVRLQTYPRTNVGSNTY-NH, Anti hypertensive, renin system Symptomatic antidiabetic

1O ACTH gi|80861463|ref|NP 001030333.1| Adrenal and proopiomelanocortin preproprotein pituitary Homo sapiens disorders

US 2011/0166063 A1 Jul. 7, 2011 14

- Continued

SEQ ID Sequence and/or other Identifying Therapeutic NO : Name Family Information Activity

3 O 6 ambamustine L-Methionine, N-3-bis(2-chloroethyl)amino Anticancer, N-(4-fluoro-L-phenylalanyl)-L-phenylalanyl-, alkylating ethyl ester Anticancer, antimetabolite

19 amylin DTTVSEPAPSCVTLYOSWRYSOADNGCAETWTVKVVYEDDTEGL Antidiabetic antagonists CYAVAPGOITTVGDGYIGSHGHARYLARCL

anaritide ANP gil 178638|gb|AAA35529. 11 atrial Anti analogues natriuretic peptide hypertensive, MSSFSTTTVSFLLLLAFOLLGOTRANPMYNAVSNADLMDFKNLL diuretic DHLEEKMPLEDEVWPPOVLSDPNEEAGAALSPLPEVPPWTGEVS PAORDGGALGRGPWDSSDRSALLKSKLRALLTAPRSLRRSSCFG GRMDRIGAOSGLGCNSFRY

21-28 anti As disclosed in U.S. Pat. No. 5, 470, 831: Anti inflammatory Thr-Thr-Ser-Gln-Val-Arg-Pro-Arg inflammatory peptide Val-Lys-Thr-Thr-Ser-Gln-Wal-Arg-Pro-Arg. Immuno Ser-Gln-Wal-Arg-Pro-Arg suppressant Wal-Arg-Pro-Arg Multiple Thr-Thr-Ser-Gln-Val-Arg-Pro-Arg-His-Ile-Thr. Sclerosis Thir-Thr-Ser-Gln-Wall treatlet Thr-Ser-Gln-Val-Arg Antiarthritic, Thr-Thr-Ser-Gly-Ile-His-Pro-Lys other Stomatological Dermatological

3. Of antiflammins L-Leucine, N-N-N-N-N2-(N2-N-(N-L- Anti histidyl-L-alpha-aspartyl)-L-methionyl-L- inflammatory asparaginyl-L-lysyl-L-valyl-L-leucyl-L- alpha-aspartyl antifungal tripeptides of N3 - 4-methoxyfumanyl and di Antifungal tripeptides and tripeptides of N3-D-trans 2, 3 epoxy succinamoyl-L-2, 3-diaminopropanoic acid

29 Gastrimmune G17-DT; G17DT (vaccine) ; Gastrimmune; Anticancer, Glu-Gly-Pro-Trp-Leu-Glu-Glu-Glu-Glu immunological diphtheria toxoid; anti-gastrin 17 immunogen; gastrin 17 vaccine; gastrin-17-diphtheria toxoid conjugate

3 O antithrombin gi|312673|emb|CAA51292.1| Hirudin Antithrombotic polypeptides Hirudinaria manillens is Anticoagulant MFSLKLFVVFLAVCICVSOAVSYTDCTESGONYCLCVGGNLCG GGKHCEMDGSGNKCVDGEGTPKPKSOTEGDFEEIPDEDILN

31 antiviral NH-Tyr-Ala-Gly-Ala-Val-Val-Asn-Asp-Leu-COOH Antiviral, other peptides

32 apolipoprotein gil 671882emb|CAA28583. 1 apolipoprotein Hypolipaemic / Homo sapiens Antilathero MKLLAATWLLLTICSLEGALVRROAKEPCVESLWSOYFOTWTDY Sclerosis GKDLMEKVKSPELOAEAKSYFEKSKEOLTPLIKKAGTELVNFLS YFVELGTHPATO

33 arthritis gi|463696O3|gb|AAS89650. 11 secreted antigen Recombinant, antigen 85A precursor Mycobacterium bovis BCG other MOLVDRVRGAVTGMSRRLVVGAVGAALVSGLVGAVGGTATAGAF Antiarthritic, SRPGLPWEYLOWPSPSMGRDIKVOFOSGGANSPALYLLDGLRAQ immunological DDFSGWDINTPAFEWYDOSGLSVVMPVGGOSSFYSDWYOPACGK Immuno AGCOTYKWETFLTSELPGWLOANRHVKPTGSAVWGLSMAASSAL suppressant TLAIYHPOOFWYAGAMSGLLDPSQAMGPTLIGLAMGDAGGYKAS DMWGPKEDPAWORNDPLLNVGKLIANNTRWWVYCGNGKPSDLGG NNLPAKFLEGFVRTSNIKFODAYNAGGGHNGVFDFPDSGTHSWE YWGAOLNAMKPDLORAL.GATPNTGPAPOGA

309 Avorelin 5-Oxo-L-prolyl-L-histidyl-L-tryptophyl-L- Releasing seryl-L-tyrosyl-2-methyl-D-tryptophyl-L- hormone leucyl-L-arginyl-N-ethyl-L-prolinamide Anticancer, hormonal