November 14, 2023
PeptiGrowth Inc.

PeptiGrowth Inc. is Launching a Novel Synthetic Peptide
Wnt3a alternative peptide (β-catenin pathway agonist)
PeptiGrowth Inc. (Headquarters: Chiyoda-ku, Tokyo, President: Jiro Sugimoto) has successfully
developed a novel synthetic peptide called “Wnt3a alternative peptide (β-catenin pathway agonist)”
which has equivalent function to recombinant Wnt3a as well as GSK-3β inhibitors, such as
CHIR99021(CHIR)). This product will be available in the middle of November 2023.

Development of Synthetic Peptide Growth Factors by PeptiGrowth
Conventional growth factors and cytokines used in the manufacturing of regenerative medicine and celltherapy products face various quality challenges such as batch-to-batch variation, potential contamination
with biological impurities, low stability, and high cost. PeptiGrowth has been working on the developmentof a series of synthetic peptides that can address these challenges while maintaining equivalent function
to the conventional growth factors and cytokines in the market. Our peptides are completely chemicallysynthesized and animal component-free, enabling Xeno-free and chemically defined cell culture media.

About Wnt3a alternative peptide (β-catenin pathway agonist) [Product code: PG-008]
The activation mechanism of the β-catenin pathway by Wnt3a alternative peptide
The Wnt3a alternative peptide (PG008) is composed of two cyclic peptides thathave specific and strong affinity to thereceptors of Wnt3a, FZD (Frizzled), andLRP (Low-density lipoprotein receptorrelated protein) 5/6. Similar to Wnt3a, PG008 binds to FZD and LRP5/6, activating theβ-catenin pathway in the Wnt signalingpathway (see the figure on the right)1Superior agonist activity compared to recombinant Wnt3a.
We performed a functional comparative test
between PG-008 and commercially available
recombinant Wnt3a. Specifically, we used the TCFLEF reporter assay in HEK293 cells, which
evaluates the activation of luciferase genes. This
assay allowed us to compare the agonist activity ofPG-008 and recombinant Wnt3a on the β-catenin
pathway. As a result, PG-008 was shown to activate
the reporter gene at lower concentrations andexhibit superior agonist activity compared torecombinant Wnt3a (see the figure on the right).

Achieving cell differentiation at an exceptionally low concentration compared to CHIR99021
Our collaborator conducted an experiment to compare the differentiation efficiency from iPSCs todefinitive endoderm (DE) comparing PG-008, CHIR, and recombinant Wnt3a, following the scheme
outlined below (Dr Nick Hannan, Univ of Nottingham, unpublished data). It is important to note that CHIR
is a small molecule that functions as a GSK-3β inhibitor and commonly used as an indirect activator of theβ-catenin pathway in Wnt3a signaling.

When using recombinant Wnt3a (50 ng/mL, approx.1.3 nM), a significant number of undifferentiatedcells were observed on Day 4 (the left image below), and the induction efficiency of DE was approx. 60%(the right image below). In contrast, when PG-008 and CHIR were used, a uniform differentiation into DE

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was observed, and a differentiation efficiency of over 98% was achieved. These results demonstrate that
only 1 nM of PG-008 exhibits a comparable differentiation induction efficiency to 3 µM of CHIR.
Furthermore, we confirmed that PG-008 does not exhibit cytotoxicity even at a high concentration of 10µM.
We anticipate that PG-008 will be used for various applications, including induction ofdifferentiation from iPSCs and adult stem cells to endodermal lineage cells and maintenance ofvarious organoids.
Product Overview
Product name:Wnt3a alternative peptide (β-catenin pathway agonist)

Product Code:

PG-008

Product form:

Lyophilized

Storage conditions:-20°C or lessPurity:≥95% by HPLC

Molecular weight:

5099.63(Acetate)Size:

10 μg (volume per a glass vial)*This product can be provided with an Animal Component Free (ACF) certificate.
*This product is for testing and research use only (RUO).
*There is a possibility that the specification would be changed.

Purchase Our Product
If you would like to purchase our products, please contact us at the information below.
For customers who would like to purchase GMP-grade products or RUO-grade products in bulk scale,please contact us at the Contact Information below.

Development of other peptides from PeptiGrowth
We have developed alternative peptides targeting numerous growth factors and cytokines. To date, wehave launched eight products, and we also plan to launch several growth factor alternative peptides
targeting PDGF-AA, TPO, FGF2, etc. from the end of 2023 to early 2024. Details will be updated on ourwebsite. If you are interested, please contact us at the following Contact Information.
Contact Information

Contact for US Market

Contact for European Market

PeptiGrowth Inc.

MIFI Bioceuticals

IVICT Europe

Tel : + 81-(0)70-4503-1497

Tel : + 1 -(201)-242-5104

Tel : +49-(0)172-204-4403

E-mail: contact@peptigrowth.com

E-mail: team@mifibioceuticals.com

E-mail: koji.okada@ivic-t.com

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