Codexis : TIDES Europe 2023 Presentation -November 02, 2023 at 09:17 pm EDT

ECO Synthesis™ Platform: Positioned to Deliver in RNAi Market

RNAi Demand is Coming

Chemical synthesis (phosphoramidite chemistry) alone cannot meet

projected future wave of demand for RNAi therapeutics

Customers are asking us for a scalable, more sustainable enzymatic solution to complement chemical synthesis

1 kg of siRNA requires ~ 1000 kg of MeCN (BioSpring)

PMI per nucleotide added ~200 kg/kg ( Org. Proc. Dev, 2021, 86, 1, 49-61)

Codexis positioned to deliver based on 20-yearhistory of enzyme

engineering and directly relevant Pharmaceutical Manufacturing commercial expertise

Codexis ECO Synthesis™ Technology

Enzyme Catalyzed Oligonucleotide Synthesis

Core process:

Oligonucleotide synthesis by iterative, single nucleotide extension

• Extend: Controlled, enzymatic addition of modified, 3′-phosphate blocked ribonucleotides

• Deblock: Enzymatic cleavage of 3′-phosphate blocking group & excess ribonucleotides

… Repeat

Supply processes:

• Enzyme cascade for synthesis of modified, 3′- phosphate blocked ribonucleotides (NQPs)

• Enzymatic synthesis of starter oligonucleotide

Codexis ECO Synthesis™ Technology – Key Platform Traits

Enzyme Performance

• High incorporation efficiency

• Robustness & manufacturability

Scalable & Economical Manufacturing

• High volumetric productivity

• “Oligo in solution, enzyme immobilized”

• Controlled addition of monomers

• Low impurity production

• Smaller infrastructure/facilities footprint

• Established reagents supply

Supplying Critical NQP Reagents For ECO Synthesis™ Platform

Building a “one-pot,two-step” enzyme cascade with engineered kinases

Targeted Key Performance Indicators

Robustness/Manufacturability – soluble expression & stability

Productivity – >98% conversion at millimolar substrate concentration

Substrate Tolerance – accepts ribonucleotides with 2′ modified sugars & phosphorothioate backbone modifications

Supplying Critical NQP Reagents For ECO Synthesis™ Platform

Building a “one-pot,two-step” enzyme cascade with engineered kinases

Step 1: N→NTP conversion via three consecutive phosphorylation steps, using three kinases

Percent conversion for individual nucleosides to the corresponding NTPs

Supplying Critical NQP Reagents For ECO Synthesis™ Platform

Building a “one-pot,two-step” enzyme cascade with engineered kinases

Step 1: N→NTP conversion via three consecutive phosphorylation steps, using three kinases

Status: NTP Forming Cascade

• 
  Full base promiscuity & emerging activity for 2′-modified nucleosides
• Operational at process-relevant substrate concentration
• Engineering on track to deliver full base and 2′-modification promiscuity for NTP formation

Supplying Critical NQP Reagents For ECO Synthesis™ Platform

Building a “one-pot,two-step” enzyme cascade with engineered kinases

Step 2: NTP→NQP percent conversion with current 3′-O kinase

Percent conversion for individual NTPs to the corresponding NQPs

Supplying Critical NQP Reagents For ECO Synthesis™ Platform

Building a “one-pot,two-step” enzyme cascade with engineered kinases

Step 2: NTP→NQP percent conversion with current 3′-O kinase

Status: 3′-O-kinase engineering

• 
  Engineering challenge as desired activity is non-natural (& potentially cytotoxic to expression host)
• Break through from initial strict A selectivity
• 
  Ongoing engineering of 3’O-kinase starting to deliver activity on 2′-modification for NQP formation

Terminal Deoxynucleotidyl Transferase (TdT)

A high engineered enzyme for catalyzing the extension reaction