Applications of Plant Cell Cultures from Bench to

Applications of Plant Cell Cultures – from Bench to Commercial Scale – www. phytonbiotech. com Development. Solutions@phytonbiotech. com

Leader in Plant Cell Fermentation (PCF®) Technology Plant cell culture is a controlled biotechnological process that provides a sustainable and scalable supply solution for plant-made actives 2

Who we are Phyton Biotech® is … • … a global supplier of high-quality paclitaxel and docetaxel API via plant cell fermentation (PCF®) • … the only company with the know-how, expertise, and infrastructure to develop, scale-up, and commercially manufacture plant-based actives via PCF® • … a provider of development services in the field of plant based actives through PCF® and value-added API services • … a creator of proprietary and partnered products and platforms 3

Phyton – From Integrated Development Solutions to API Supply Upstream (Germany) Cryobank Plant sourcing Implementation of suspension cultures Lab to pilot scale Scale-up Validation PCF®-based production of actives Regulatory support: in house QA, QC Plant raw material PCF®-based starting material Interface DSP Extraction Chemical processing Purification Pilot batch production Scale-up / Validation Product Downstream (Canada) 4

PCF® - Characteristics • For every batch, the plant cell culture is recovered from a cryopreserved cell bank • The medium is fully defined, protein-free and does not contain any animalderived component and … it is cheap • The process is operated in aerated stirred tanks with proven scale-up technology – no light is needed for the heterotrophic growth of cells • Fermentation product can be directly purified to an API => e. g. paclitaxel • Fermentation product can be semi-synthetically processed => e. g. docetaxel • The entire process complies with GMP requirements 5

Plant Cell Culture Selected strains Cell suspension Amplification of cell material Cryopreservation Homogeneity of cell material Transfer to liquid medium Stability Growth Expression Master cell bank Working cell bank The best of the best Controlled conditions Reliable strain Almost unlimited supply source 6

Why Plant Cell Fermentation (PCF®)? Sustainability - Reliability - Quantity Destructive plant harvest Phyton provides favorable supply solution for plant actives • Environmentally responsible and sustainable • Not subject to climate conditions • Allows for growth optimization • Allows for optimization of expression of active • Allows for optimization of active/impurities ratio Clean PCF® technology • Controlled supply / entire production chain fully GMP • Flexible in commercial scale to address market-driven demand 7

Case Study Production The Paclitaxel and Docetaxel experience Processing PCF® broth to complex finished API’s 8

From Plant to Cell Culture to Product 9

Production Cascade – Overview > 200, 000 L Bioreactor Volume Pre-Fermentation Fermenter cascade Harve st 75 cbm 45 cbm Seed tanks GMP Produ ction tanks

Large Scale Fermentation Environment 11

Recovery of Paclitaxel from Fermentation Broth - Paclitaxel Concentrate Transfer broth to adsorption tanks Adsorption to resin Adsorption tanks Separation of loaded resin from biomass and spent medium 75, 000 L fermentor Recycling of desorbed resin Concentration, precipitation, filtration, drying, packaging 200 kg of 40 -50% paclitaxel concentrate Move to desorption Solvent based desorption of active from resin Loaded resin in transport containers 12

Straightforward Processing of Paclitaxel Concentrate 1 2 Stable Paclitaxel Polymorph 13

Commercial Production Finished API Production - Canada 14

Green Chemistry – Case Study Paclitaxel Organic solvent usage for Paclitaxel manufacturing 92% less No Hazardous Solvents for PCF® 80% less PCF® Semi-synthesis from 10 -DAB CONFIDENTIAL Direct extraction from plants 15

Capabilities - Quality • Entire process strictly GMP compliant • FDA, EMA, EDQM, TGA, KFDA, Health Canada inspected • All API are USP/EP • Full internal QA/QC support • Strong internal regulatory support • Broad regulatory approvals for finished API: • • • CEP and US DMF for paclitaxel from PCF CEP and US DMF for docetaxel Japan approval for paclitaxel PCF and docetaxel Korea approval for paclitaxel PCF China registration in process for paclitaxel and docetaxel 16

Case Study cell culture generation Development of Ingenol Mebutate producing cell cultures 17

Ingenol Mebutate … is a diterpenoid ester which is a powerful protein kinase C agonist providing pro-apoptotic effects. Ingenol Mebutate gel is approved for the topical treatment of Actinic Keratosis 18

Ingenol Mebutate • Ingenol - the precursor of Ingenol Mebutate - has been synthesized chemically in a 14 step synthesis starting from 3 -carene Jörgensen et al. 2013, Science • Ingenol Mebutate is found at concentrations of only 1. 1 mg/kg in the sap of Euphorbiaceae Hohmann et al. 2000, Planta Med • Why not combining nature’s famous and straightforward chemical expertise with human knowledge Þ Plant Cell Culture Phyton PCT/EP 2016/055989 19

Plant Cell Culture Derived Ingenol Mebutate 20

Plant Cell Culture Derived Ingenol Mebutate • Euphorbiaceae cells in plant cell culture produce Ingenol Mebutate, Ingenol and other Ingenol esters • Euphorbiaceae cells are adaptable to bioreactor environment • Euphorbiaceae cells in culture produce higher concentrations of Ingenol Mebutate than found in the sap of plants • Euphorbiaceae cells can be cryopreserved and recovered • Ingenol Mebutate is found in the biomass as well as in the surrounding medium => secreted • Ingenol Mebutate can be captured/recovered from cell culture by in-situ resin adsorption without cell disruption 21

Case Study high producing cell lines Screening for high Thapsigargin-producing Thapsia garganica cell cultures -funded by Inspyr (former Gen. Spera)- 22

Thapsigargin … is a sesquiterpene lactone which is a powerful cytotoxin – a sarco-endoplasmic reticulum calcium-ATPase (SERCA) inhibitor – and the precursor for the derivative ADT, the key ingredient of Mipsagargin (G-202). 23

Initiation of Hundreds of Suspension Cultures 24

High Producing Cell Lines – the Best From the Best Pre-Screening High producing cell line 98. 5 mg/kg Phyton WO 2015082978 25

Thapsigargin / Thapsigargicin Ratio in Seeds 26

Improved Ratio in the Cell Culture Seeds Cell culture #8 27

PCF® - the Ideal Choice for many Plant-derived Drugs • Expression of HPAPI through plant cells cultivated in well controlled bioreactor environment • Huge enrichment of compound of interest • Reduction of impurity profiles • Demand-driven scalability • Competitive Technology • Comply with GMP requirements 28

PCF® Project Pipeline Identification of high producing cell lines and optimized culture conditions Evaluation of applicability Implementation of reliable process at pilot scale Scale-up Internal Client Inspyr Client Co-dev. Client 29

Contact Dr. Gilbert Gorr gilbert. gorr@phytonbiotech. com Development. Solutions@phytonbiotech. com