Peptides as a therapeutic method attract much attention due to the synthetic accessibility, high degree of specific binding, and the ability to target protein surfaces traditionally considered "undruggable". Macrocyclic peptides possess a lot of pharmacological characteristics distinct from other well-established therapeutic molecular classes, resulting in a versatile drug modality with a unique profile of advantages. https://www.medicilon.com/services/biology-services/

1. MEDICILON
Peptides as a therapeutic method attract much attention due to the synthetic accessibility, high degree of
specific binding, and the ability to target protein surfaces traditionally considered "undruggable". Macrocyclic
peptides possess a lot of pharmacological characteristics distinct from other well-established therapeutic
molecular classes, resulting in a versatile drug modality with a unique profile of advantages.
Peptide Drug Advantages
Synthetic accessibility
Low immunogenicity and toxicity
High binding affinity and selectivity
The ability to target protein surfaces traditionally considered “undruggable”
Macrocyclic peptide: Compared to the linear peptide precursor
Advances in high-throughput in vitro screening techniques
more stable
more selective
more potent
increased membrane permeability
Phage display
mRNA display
Flexizymetechnology

2. MEDICILON
Medicilon Peptide Drug Discovery Service
Full Time Equivalent (FTE)
Dedicated R&D and Management team: standard 10 FTE team: 1 senior group leader (8-10 years) + 1
senior chemist + 2 subgroup leader (3-7 years), senior scientist, middle level scientist, junior scientist (3
member subgroup)
Flexibility in
Fee For Service (FFS)
Others
Project priorities
Staffing
Protocol
Services provided for specific needs/projects. Fixed fees and delivery timeframe.
Customization of needs
mg to kg custom synthesis
chiral separation
impurity separation
CMC work for IND application and IND investigation
Non-GMP and GMP scale-up, commercial production of API and intermediates
Customized synthesis: mg-100 g scale
Linear peptides (up to 50 AAs via one-by one coupling; >70 AAs via ligation strategy)
Cyclic peptides (S-S bridge, lactam/lactone, thioether, olefin metathesis, click reaction etc.)
Modifiedpeptides (pegylation, DOTA, phosphorylation, thiopeptide etc.)
Peptidomimetics
Peptides containing special amino acid: containing isotope atom amino acid, D-amino-acid, other unnatural
amino acid etc.
Macro cyclic peptide library synthesis
Unnatural AAs Synthesis –Chemical and Enzymatic
ADC-like compounds
Saltexchange: TFA/HCl,TFA/acetate

3. MEDICILON
Instruments for Peptide Synthesis & Purification
Medicilon Peptide Drug Discovery Service Team
> 60 chemists
> 15 biologists
Peptide Team
Solid Phase
Synthesis
Solution
Phase
Synthesis
Synthetic
Biology
Synthesis
Purification
& QC
Solid phase synthesis on resin by Fmocor other chemistry
Support various modifications on peptides
Synthesis by both auto-synthesizer and manual apparatus
Unnatural amino acid synthesis
Solution/solid combination synthesis for special peptides
ADC-like compound synthesis
mRNA Display technology for coded macro cyclic peptide library synthesis
Enzymatic catalysis for unnatural amino acid synthesis
Enzymatic peptide ligation and cyclization
Various prep-HPLC equipped with up to 20 cm columns
Multiple types of columns: C18, C8, C4….
Up to 60L tray-lyophilizer
OEB4 lab for ADC project Automated small scale purification system HRMS
>60 Chemists
>40% MS/Ph.D.
Customized synthesis
mg to 100 g scale
Unnatural AA
ADC-like compounds
Customized manual peptide synthesis system CsBioauto-synthesizer

4. Peptide Synthesis
Linear/Modified Peptides
(S)
N
H (R)
O
HN
(Z)
HN O
(S)
O
N
(S) N
O
(S)
NH
O
(S)
O
NH
HN
(Z)
H2N
Fixed peptidomimetics
WO2008092281
AAsc
(S)
N
H
O
HN
(S)
R1
O
HN
(S)
O R3
O
(R)
NH
N
H
H2N
NH
(S)
O
R6
NH
O
(R)
NH
R5
O
(S)
H
N
R4
HN
(R)
O
R2
CPPs
WO2022/241408
NH2
(R)
HN
(S)
NH
(S)
N
H
(R)
HN
(R)
O
O
O
O
R2
H
N (S)
O
O
HN
(S)
R6
HN O
(R)
OH
O
S
S
S
S
Bicyclic disulfide bridge
H2N (S)
H
N (S)
N
H
H
N (S)
N
H
(S)
O
O
O
O
R1
R2 R4
H
N (S)
O
R5 O
N
H
R6
H
N
O
(S)
OH
O
R8
(Z)
RCM monocyclic peptide
N
(S)
O NH
(S)
O
N
(S)
O
(S)
N
O
(S)
(R)
O
O
(S)
O
NH
F
F
O
(E)
ADEP4
J. Med. Chem.2006, 689
Cyclic peptides (S-S bridge, lactam/lactone, thioether, olefin metathesis, click reaction etc.)
Cyclic Peptides
H2N
(S)
H
N (S)
N
H
(S)
H
N (S)
N
H
(S)
O
O
O
O
R1
R2 H
N (S)
O
R3
O
P
HO OH
O
O
P
OH
O
OH
O
N
H
(S)
R4
OH
O
R5
Phospho-peptide
H2N (S)
H
N
O
H
N (S)
N
H
(S)
O O
O
R1
R2
H
N (S)
O
R3 O
N
H
(S)
R4
OH
O
R5
n
PEGylated peptide
N
H
(S)
O
NH2
H
N
O
(S)
NH2
N
H
O
(S)
NH2
H
N
O
(S)
HN NH2
NH
N
H
(S)
O
NH2
H
N
O
(S)
N
H
O
(S)
O
HO
H
N
(R)
R
R = SH or aliphatic
amine, or other groups
used for modification
N
H
(S)
H
N (S)
N
H
(S)
H
N (S)
N
H
(S)
O
O
O
O
R1
R2
R3
R4
H
N (S)
O
R5 O
N
H
(S)
R6
OH
O
R7
Dye
Dye modified peptide
H2N (S)
H
N (S)
N
H
(S)
H
N (S)
N
H
(S)
S
S
S
S
R1
R2
R3
R4
H
N (S)
S
R5 S
N
H
(S)
R6
OH
O
R7
Thio-peptide
Linear/Modified Peptide synthesis in solution phase, on solid phase and automatic synthesizer
Peptide modification (N-terminal or side chain modification, C-terminal modification, peptide phosphorylation
labeling, methylation and other alkylation modification, isotope labeling, fluorescence modification, PEG modifi-
cation, peptide protein coupling, etc.)
MEDICILON

5. Unnatural Amino Acids
NHFmoc
OH
O
OH
O
NHFmoc
F3C
OH
O
S
AcHN
NHFmoc
MeO
OH
OH
NH2
O
CO2H
NH2
NH2
HO O
NH2
HN
O
OH
O
F3C
NH
Boc
O
OH
NHFmoc
N
NH O
OH
Boc
HN
N
HO2C
Boc
R
CO2H
BocHN
NHFmoc
CO2H
→Enantioselective chemical synthesis
Unnatural α-amino acids Unnatural β/γ-amino acids
Rich experience to prepare various unnatural amino acids in-house
Help client to build up amino acids library
Explore the synthesis of special AAs upon request
→Enzyme Catalyzed unnatural amino acid synthesis
→Enzyme evolution was applied to improve amino acid C-N lyase
R1
COOH
+ R2
NH2
C-N lyase
R1
COOH
or
R1
COOH
*
NHR2 *
NHR2
R1 COOH
NH2
+
R2 COOH
O
amino-acid
aminotransferase
R2 COOH
NH2
+
R1 COOH
O
R3
NH2 +
R2 COOH
O
aminotransferase R2 COOH
NH
R3
or
O
NH
OH
L-Pro
trans-4-hydroxylase
Fe2+ + O2
2-oxoglutarate CO2
+ succinate
O
NH
OH
HO
COOH COOH
NH3.H2O
amino acid lyase
NH2
MEDICILON

6. Enzymatic Peptide Ligation & Cyclization
Peptide ligases: Sortase A (SrtA) Butelase-1 Trypsiligase ……
mRNA Display Technology for Macrocyclic Peptide Library
mRNA display technology for cyclic peptide library construction
Library size exceeding 1013
Unnatural amino acids will be incorporated into the peptides through PURE cell-free translation system
A typical round of an mRNA display selection takes 2-3 days
Hits will be easily identified through sequencing of the cDNA linked with the peptide
O
HO
NH
O
NH2
O
OH
N
N
N
N
N
Puromycin
O
O
O OH
N
N
N
N
NH2
P
O
O
-
O
O
NH3
+
AA
tRNA
Aminoacyl-tRNA
5`
AUG
mRNA
Ribosome
P A
Peptide
Linker
Puromycin
A) B)
C)
MEDICILON
S
O
LPXT
Sortase A
Ca2+
HS
Sortase A
Ca2+
LPXT
H2O
LPXTG XX
G XX
LPXTG motif
(acyl donor)
(G)n
amino nucleophile
(acyl acceptor)
sortase A catalyzed
ligation product or
cyclic peptides
LPXT(G)n
X = any amino acid
LPXT(G)n
LPXT(G)n
Ligation Product
OR
Cyclic peptides

7. MeO
HO
O C
O
H
N
1) Fmoc-Pro-OH, DIC
DMAP, DMF, rt
2) 20% 2-MP in DMF, rt
HN
O
O linker
Fmoc SPPS
N
O
O linker
O
N
O
H
N
O
N
O
N
O
N
O
N
O
1) 20% HFIP in CH2Cl2
2)
HN
O
O
N
N
O
N
O
H
N
O
N
O
N
O
N
O
N
O
O
Target
Medicilon successfully made the reference compound in 2 weeks
The purity of the desired compound is >98%
The amount of the final peptide >500 mg
Case Study
Medicilon Case: Peptide Synthesis
N
H
N
N
N
NH
O
O
O O
O
O OH
O
O
HN
O
NH
NH2
O
N
H
O
N
H
O
N
O
O
N
H
N
O
O
O
O
HN
O
N
N
H
O
H2N
R
R
N
H
N
O
O
O
O
HN
O
R
N
H
O
H2N
N
H
N
O
O
O
O
R
HN
O
N
N
H
O
H2N
R
R
N
H
N
R
O
O
O
O
HN
O
N
N
H
O
H2N
R
R
NH2
HN
O
O
HN O
NH
O
O
O
N
+
H
N
HN
N
F
O
S
N
N
N
S
HN
N
O
O
O
N O
NH
O
O
N
H
HO
O
N
H
Fmoc
OH
O
Fmoc
N
O
OH
O
Fmoc
N
O
OH
O
R
N
H
Fmoc
OH
O
R
Amino acid modification and application in ADC drug Amino acid modification in PCSK9 peptide inhibitor synthesis
> 10 novel amino acid were made in Medicilon > 20 novel amino acid were made in Medicilon
Medicilon Case: Solid-Phase Synthesis of Peptide
Peptide synthesizer Peptide HPLC purification system
MEDICILON

8. MEDICILON
Email: marketing@medicilon.com Website: www.medicilon.com Tel: +1(626)986-9880
Global Headquarters: 585 Chuanda Road, Pudong, Shanghai, 201299, China
US Laboratory: 50 Soldiers Field Place, Boston, MA 02135, United States
N
O
N
H
O
O
P
OH
O
O
5’
3‘
NH2
O
P
OH
O
O
5’
3‘
N
O
O
O
F
F
F
F
F
100-150nt
100-150nt
EEV-2-linker
O
N
H
O
O
P
OH
O
O
5’
3‘
N
N
N
N
(S) N
H
NH
H
N
O
O
(S)
H
N
O
N
H
(S)
HN
(S)
O
HN
HN NH
O
N
H
(S)
HN
H
N (S)
O
NH
O
N
H
(S)
HN
H
N (S)
O
N
O
O
(S)
H
N
O
NH
(S)
O NH
(S)
O
O
(R)
HN
H
N H
N
HN
(S)
O
HN
HN
NH
HN
O
(R)
HN
N
H
HN
NH
O
(S)
N
H
NH
HN NH
NH
(R)
O
Boc
Boc
Boc
Boc
Pbf
Pbf
Pbf
Pbf
Pbf
Medicilon Case: Synthesis of AOC
5 mg, 95% purity
MEDICILON
Peptide Bio-analysis
LC-MS/MS and High Resolution MS (GLP and non-GLP)：all Waters
ACQUITY PREMIER UPLC Triple Quad 6500+ XevoTQ-XS