RARE Thinking for RARE Solutions - PowerPoint Presentation

Slide1

RARE Thinking for RARE Solutions

Company non-confidential

presentation

January 2021

Slide2

Four key drivers of Zikani’s success

TURBO-ZM platform to design ribosomal modulators

Right leadership, team and advisors

Focused on rare genetic diseases and cancers

Deep pipeline with lead program in clinic by 2022

Slide3

Mission: Transform

human ribosome therapy

with macrolide based novel Ribosome Modulating Agents (RMAs)

Corporate

Founded in 2015; Refocused in 2019

Based in Watertown, MA

14 Full time employees

2022 Goals

Complete Ph1a in RDEB

Advance development candidate in APC mutant FAP

Advance APC mutant CRC program

Expand Oncology pipeline

Funding

Investors: Roche Venture Fund, Gurnet Point Capital and Advent Life Sciences

Technology licensed from Harvard

TURBO-ZM Unlocks Large Market Opportunity

Readthrough to treat rare diseases and rare cancers

Inhibition of proteins in Onco- and mito-ribosomal cancers

TURBO-ZMRARE Thinking for RARE Solutions: TUning the RiBOsome with Zikani Molecules (TURBO-ZM)

Slide4

Large and broad applications for human ribosome targeting

Readthrough = “Functional

Gene Therapy”

Stop codon readthrough in rare diseases and cancer

12% of patients across 1800 rare diseases

6-10% of patients with nonsense tumor suppressor genes in cancer

Neoantigen induction for combo therapy in immuno-oncology

Potential addressable market well in excess of

$5 billion

Protein Translation

Inhibition

Zikani RMA

Zikani RMA

Oncoribosome

inhibition

20-30% of cancer patients with c-

myc

overexpression

Mitoribosome inhibitionPatients with cancer dependent on mitochondrial OXPHOS metabolism

Slide5

Experienced Leadership

Zikani Leadership Team

Sumit Aggarwal

President and CEO

20+ years investing and transforming healthcare companies

Raised >$150M

Biotech Investor

Vijay Modur

CSO and CMO

20+ years in translation and drug development

Led Venglustat rare disease program at Sanofi

30+ years building companies

Closed $2B in equity and debt financings for public and private companies

Daniel Geffken

Chief Financial Officer

Roger Clark

Head of Discovery Sciences

20+ years medicinal chemistry

Architect of Zikani RMAs

Slide6

Strong board, advisors and collaborators supporting plans

Keith Flaherty, Chairman SAB (

Oncology

Loxo

, Harvard Medical School)

Pedro Huertas (

Clinical translation

, ex-CMO Eloxx, Shire)

Dr. David Sidransky (

Johns Hopkins

, Advaxis, Champions Oncology)

Dr. Andrew South (Thomas Jefferson University)

David Bedwell (Univ. Alabama Birmingham)

Cystic Fibrosis Foundation (CF)Hubrecht Institute of Technology (HUB)Rina Arbesfeld (KOL FAP/APC, Tel Aviv University)

Mei Chen (KOL DEB, USC Medical School)

Key AdvisorsKey Collaborators

Dr. Alan Walts (Executive Chairman)

Sumit Aggarwal (President and CEO)

Raj Parekh (Advent Life Sciences)Monique Schiersing (Roche Venture Fund)Christopher Viehbacher (Gurnet Point Capital)Board of Directors

Slide7

TURBO-ZM Platform and RMAs

Slide8

Ribosome modulation can address protein translation defects

Normal vs. defective protein translation

Defect: Genetic Nonsense

Mutations

Normal Ribosome:

“Protein Factory”

Defect: Dysregulated

Ribosomes

Premature

stop codon

Less than full length protein formed

Onco- and

mito

-ribosomes

Selective translation

of oncogenic proteins

Aids hyperproliferative phenotype

Slide9

Strong rationale for macrolides to bind the human ribosome

Nascent peptide exit tunnel in E. coli vs. Human ribosomes

1

1

Nucleic Acids Res. 2019;47(8):4198-4210

H. sapiens

E. coli

Macrolide binding site in Prokaryotic (bacterial) ribosome

75% of binding site conserved in the Human (Eukaryotic) Ribosome

1

First

constriction

site

Second

constriction

site

Slide10

Macrolides can readthrough and inhibit protein translation

Macrolide MOA and effect on protein translation

1

Br J

Pharmacol

. 2017;174(18):2967-2983

2

Antimicrob Agents Chemother. 2004 Dec; 48(12): 4889–4891

3

Nucleic Acids Res. 2017 Sep 19; 45(16): 9573–9582

Ribosome crystal structure

1

Stop codon

readthrough

2

Selective protein synthesis inhibition

3

Slide11

TURBO-ZM platform fully unlocks the potential of macrolides

Macrolide

Ribosome Modulating Agent (RMA)

Gram-positive antibiotics

Favorable PK

Orally delivered

Limited therapeutic index

Gram-negative antibiotics

Eukaryotic ribosome binding

High tissue exposure at low doses

Predictable PK and safety

A

B

C

D

F

E

A

BCDFEConventional Semi-synthetic ApproachNovel Fully-synthetic Approach

TURBO-ZM

Slide12

Growing library of RMAs with drug-like properties

Zikani RMA Library (2000+)

Slide13

Achieved goal of more potent and safe RMAs

Advantages of RMAs over macrolide antibiotics

Long term azithromycin and clarithromycin therapy well established

RMAs have

clean CYP, cardiac and cholestatic risk profile

Safety profile confirmed in vivo studies

Clean non-GLP 7-day rat tox for cUTI candidate

Preserve and improve already clean safety profile

Preserve oral dosing with good exposure

Human ribosome modulating activity

TURBO-ZM advantage represents

>1-year savings

in drug development

RMAs designed for high selectivity to the human ribosome

20%-90% oral bioavailability

50uM tissue exposure with 200-500mg qd dosing (similar to Azithromycin)

Slide14

Target

Indication

Discovery

Early

research

Lead

optimization

IND

enabling

Nonsense Readthrough - Rare Disease

Collagen VII A1

RDEB

LAMB3

JEB

APC

FAP

CFTR

CF

Nonsense Readthrough – Oncology

APC

CRC

p53

Pan cancer

Neoantigen induction

IO combination

Protein translation inhibition

OncoribosomeMyc-driven cancer

Mitoribosome

OXPHOS high cancer

Growing RMA pipeline in rare disease and oncology

Slide15

Readthrough

Pipeline

Slide16

Clinical signal with readthrough using antibiotics fall short of therapeutic goals

Clinically relevant readthrough in selected diseases

1

Kariv, R. Ann. Oncol. 2018, 29, suppl3;

2

Sermet-Gaudelus, I. BMC Med. 2007, 5, 5;

3

Malik, V. Ther. Adv. Neurol. Disord. 2010, 3, 379;

4

Woodley, D. J Clin Invest. 2017;127(8):3028,

5Caspi, M., J Mol Med (Berl). 2016 Apr;94(4):469-82; 6Goldmann, T, Hum Gene Ther. 2011 May;22(5):537-47.

Diseases

Evidence

Readthrough Agent(s) Tested

Familial Adenomatous Polyposis (FAP) and APC driven Colorectal cancer

Clinical1

Erythromycin, Tylosin, Gentamicin

Cystic Fibrosis Class 1

Clinical2Gentamicin, Tylosin, GeneticinDuchenne Muscular DystrophyClinical3GentamicinDystrophic Epidermolysis Bullosa (RDEB)Clinical4Gentamicin, GeneticinLysosomal Storage Disorders, e.g., MPSI (Hurler), cystinosisex vivo5Gentamicin, GeneticinRett Syndromeex vivo5Erythromycin, GentamicinSpinal Muscular Atrophy (SMA)ex vivo5Erythromycin, GentamicinAtaxia-Telangiectasia (ATM)ex vivo5Erythromycin, GentamicinUsher syndrome/retinitis pigmentosa (RP)in vivo Preclinical6Gentamicin, Geneticin

16

Readthrough reported in over 36 different diseases

Slide17

RMAs show superior readthrough than alternatives

Readthrough Emax of selected RMA hits

Relative Luciferase Units compared to DMSO in W134X Nanoluc Reporter Assay

ZKN-3

ZKN-1

ZKN-5

ZKN-2

ZKN-4

ZKN-6

+710%

+333%

ZKN-7

G418 (100µM)

Active control 1

Active control 2

Slide18

Class 1 CF

RDEB and JEB

APC mutant FAP and CRC

Readthrough drug pipeline in three high unmet need diseases; RDEB lead

1

targetedcancercare.massgeneral.org/My-Trial-Guide/Diseases/Colorectal-Cancer/APC.aspx

2

Cabrini, G. Mol Diagn

Ther

2019, 23, 263-279; Schmidt, BZ. Clin

Pharmacol

2016, 8, 127-140

3

Cianfarani, F. Am J

Pathol

2017. 187:1445

Mutations in COL7A1 gene (Collagen) and LAMB3

3Most RDEB patient develop skin cancer by age 35Mutations in the CFTR gene2Acute CF patients die by age of 30yMutations in the APC gene (tumor suppressor gene)FAP patients develop CRC by age 40>20,000 patient WW7,000-9,000 patents in the US/EU

8,000-12,5000 FAP patients in the US/EU;

210,000 CRC patients WW

RDEB: Recessive Dystrophic Epidermolysis Bullosa

JEB: Junctional Epidermolysis Bullosa CF: Cystic Fibrosis FAP: Familial Adenomatous Polyposis

Slide19

RDEB: Efficacy occurs at doses easily achievable through oral dosing

COL7 with 48 hr. exposure in RDEB patient derived

primary

fibroblasts*

* Fibroblasts isolated from patients two and five in gentamicin clinical trial.

J Clin Invest 2017, 127, 3028-3038

** 48 hours treatment with media and compounds replaced and refreshed at 24 hours. Study repeated twice with equivalent results.

Full length protein in Hom R578X COL7A Fibroblasts**

Full length protein in R613X/R1683X COL7A Fibroblasts**

Assay with proven translation to clinic

30-60 day Col7 protein half-life

RMAs compounds exceed clinical efficacy threshold of 10% Gentamicin 845uM

Data generated in collaboration with Academic partner

Slide20

Class 1 CF: Highest ever readthrough in Class 1 CF prompted grant request by CF Foundation

Summary of Class 1 CF data

* Forskolin 10 µM/1µM VX-770 - both chambers

** VX 809 and RMA data averaged from 2 separate Ussing chamber results

Submitted $2.5M grant to CF Foundation to support through development candidate

Data generated at Chantest

Het G542X Human Broncho Epithelial (HBE) cells Ussing Chamber

Steady state modulator response measurement**

RMA (15 uM)

DMSO

RMA (30 uM)

VX-809 (3uM)

G418 (50 uM)

G418 (100 uM)

+47%

“

”

Never seen before impressive single agent activity from non aminoglycoside class– need to advance this program

–

CF Foundation

Encouraged to apply for “Path to Cures”

Isc

Chloride Transport (µA/cm

2

)

Slide21

Oncology –

Readthrough and Ribosome inhibition

Slide22

Targeting key pathways in cancer with RMAs

Invasion and

metastasis

Evading growth suppressors

Evading immune destruction

Enabling replicative immortality

Sustaining proliferative signaling

Reprogramming energy metabolism

Resisting apoptosis

Genomic instability

Tumor promoting inflammation

Inducing angiogenesis

8

Hallmarks of cancer

Tumor suppressor gene restoration

Target protein translation addiction in cancer

c-

myc

overexpression

Neoantigen induction

OXPHOS inhibition

Protein translation inhibition

Nonsense Readthrough

Slide23

Tumor suppressor gene readthrough has transformative potential

1,391,372 cancer samples analyzed (Catalog of Somatic Mutations In Cancer)

1

152 tier 1 tumor suppressor genes identified

High frequency of nonsense mutations are common across tumor suppressor genes

>10% of all cancers can be targeted by tumor suppressor gene readthrough

Enrichment of nonsense mutations in tumor suppressor genes

2

1

Nature reviews. Cancer, 31 Oct 2018, 18(11):696-705

2

BMC Bioinformatics volume 19, Article number: 430 (2018)

Frameshift

Missense

Nonsense

Tumor suppressor gene

Oncogene

Z-score

Slide24

Erythromycin readthrough of APC is basis for clinical efficacy in FAP

Results of Erythromycin treatment of FAP patients with 250mg BID over 4 months (n=9)

Kariv, R. Int J Cancer. 2019 doi: 10.1002/ijc.32557

Change in WNT signaling biomarkers after 4 months of treatment

Clinical trial run by Tel Aviv University

Change in Polyp Number

35%

-42%

Change in Polyp cumulative size

46%

-55%

-1 yr to baseline

12 months post treatment

CYCLIN-D

KI67

C-MYC

AXIN

-51%

-97%

-57%

-64%

Change in Polyp burden at 12 months

Treatment with Erythromycin limited by safety/tolerability issues

Slide25

FAP/CRC: RMAs significantly more potent than active controls in APC readthrough

Scaled readthrough activity of RMAs vs Erythromycin in Dual Reporter Assay

1

RMAs tested at 10 ug/mL and scaled 3x vs Erythromycin at 100 ug/mL*

*Erythromycin demonstrates similar readthrough in E1309X and R1450X nonsense mutations; 7-10% wild-type protein

**Assumes up to 3-fold linear dose response based on

NanoLuc

results 

Ref:

1

Caspi. J Mol Med (

Berl). 2016 94:469-82.

AZM (100 ug/mL)

ZKN-010

ZKN-145

ERY (100ug/ml)

ZKN-157

ERY (300ug/ml)

ZKN-068

ZKN-013

ZKN-155

ZKN-034

ZKN-024

ZKN-074

ZKN-137

ZKN-146

ZKN-149

Activity in R1450X (most prevalent germline mutation)

Fold increase to Erythromycin**

AZM (100 ug/mL)

ERY (100ug/ml)

ZKN-024

ZKN-010

ZKN-0018

ERY (300ug/ml)

ZKN-013

ZKN-034

ZKN-040

ZKN-0012

Activity in E1309X (most prevalent somatic mutation)

Fold increase to Erythromycin**

Data generated in collaboration with Academic partner

Slide26

RMAs can induce activation of APC pathway

β-catenin and cyclin D1 response in cell lines treated with RMAs

48-hour exposure to RMAs vs DMSO in CRC cell lines

RMA 1

% Change

RMA 2

RMA 3

% change in phosphorylated

β

-catenin and Cyclin D1 in CRC cell lines SW1417 (R1450X)

B-catenin

Cyclin D1

RMA 1

RMA 3

RMA 2

Positive control

% change in phosphorylated

β

-catenin in CRC cell line SW403 (S1278X/1197fs)

Data generated in collaboration with Tel Aviv University

Slide27

FAP/CRC: Selective inhibition with RMAs in APC LS411N CRC cell line with driver mutation validates

MoA

Growth inhibition in WT ,missense and LS411N CRC cell line. RMA Concentration that results in inhibiting cell growth by 50% (GI50)

Ki67 normalized fold change

PBS

Placebo

CTNNB1

1.2

1.0

0.8

0.6

0.4

0.2

0.0

***

RMA 3 (APC+)

RMA 1 (APC+)

RMA 2 (APC+)

RMA 4 (APC+)

RMA 5 (CF+)

LS411N (Nonsense APC)

LS 513 (WT APC)

SK-CO-1 (Missense APC)

1. Mol Cancer

Ther

; 15(9); 2143–54

LS411N xenograft growth inhibition by b-catenin siRNA

Slide28

RMAs also active in APC mutant patient TumorGraft models

Growth inhibition ex vivo in patient derived TumorGraft models*

* Data generated at Champions Oncology

RMA 1

RMA 2

RMA 3

Slide29

RMA readthrough in immuno-oncology - Neoantigen induction for checkpoint inhibitor combo therapy

Suboptimal neoantigen expression limits checkpoint inhibitor response

1

1

Lancet

Oncol

2017; 18: 1009–21

Neoantigen

Induction by readthrough with RMA

Neoantigens are mainly expressed

by frameshift mutations

Frameshift Mutation

Checkpoint inhibitor patient response rates

(melanoma)

Frameshift causing immediate nonsense limits IO response

Frameshift causing extensive missense – High IO response

Early stop codon restricts frameshift induced neoantigen expression

Slide30

c-

myc

overexpressing tumors are targets for oncoribosome inhibiting RMAs30-50% of all cancers overexpress c-

mycPanel of cell lines with normal or high MYC selectedScreen of RMAs for selective cell death in high MYC cell linesIdentify best oncoribosome leads

Cell viability of H1112* cells to CX5461 (ribosomal inhibitor)

1

1.

British

Journal

of

Haematology, 2017, 177, 80–94

RMA screening for selective targeting of MYC tumors planned

* Myeloma cell line

Slide31

Mitoribosome

function is essential for OXPHOS: RMAs can inhibit

mitoribosomesMitoribosome is distinct from cytoplasmic ribosome

Mitoribosome produces 13 proteins critical for normal respiration by OXPHOSCertain cancers with high OXPHOS activity can be targetedHepatocellular carcinomaSerous ovarian carcinoma

Mitoribosome

produces OXPHOS proteins

OXPHOS genes are highly expressed in some cancer types

1

RMAs can be designed to selectively inhibit

mitoribosome

RMAs with >>15-fold selectivity for mitochondria identified

Promising SAR to drive lead optimization

1. Invest

Ophthalmol

Vis Sci. 2019 Oct; 60(13): 4187–4195

Slide32

Summary

Slide33

Target

Indication

Discovery

Early

research

Lead

optimization

IND

enabling

Nonsense Readthrough - Rare Disease

Collagen VII A1

RDEB

LAMB3

JEB

APC

FAP

CFTR

CF

Nonsense Readthrough – Oncology

APC

CRC

p53

Pan cancer

Neoantigen induction

IO combination

Protein translation inhibition

OncoribosomeMyc-driven cancer

Mitoribosome

OXPHOS high cancer

Growing RMA pipeline in rare disease and oncology

Slide34

Strong interest in platform and data

Submitted grant to the CF Foundation for identifying development candidate in Class 1 CF as part of “Path to Cures” funding

Actively engaged with DEBRA US and UK – EB patient advocacy group

Non Dilutive Financing

Approached by Memorial Sloan Kettering (MSKCC) with an interest in exploring IO combination therapies

Cystic Fibrosis and RDEB abstracts accepted for the 2020 International Conference on Rare Diseases and Orphan Drugs

External Validation

Seeking to expand syndicate with new lead

Targeting close by January 2021

Slide35

Zikani’s

unique approach in rare diseases and oncology

Targeting human ribosome opens up large rare disease and

oncolog

opportunity

Strong leadership and advisors position Zikani for success

Oral administration and benign safety profile make RMAs uniquely positioned in oncology

Compelling efficacy in validated pre clinical colon cancer models

derisks

pipeline