L’uso dell’IL10 nelle malattie reumatiche e gastroenterologiche

Prof. Mauro Galeazzi

Congresso Nazionale della Società Italiana di GastroReumatologia Roma 24-26 giugno 2015

Interleuchina 10 e malattie infiammatorie croniche intestinali In PubMed 1997-2015

332 Lavori sul Morbo di Crohn 276 Lavori sulla Colite Ulcerosa

Evidenza del coinvolgimento della IL-10 nella patogenesi delle MICI

Modelle animali: topi KO (IL10-/Il10-) sviluppano spontaneamete varie forme di MICI Mutazioni dei geni IL-10 e IL-10 R favoriscono comparsa di MCI nell’uomo Trial clinici con IL-10 ricombinante iniziati alla fine degli anni ’90

Immunomodulation of Crohn's disease by interleukin-10. Narula SK1,. Agents Actions Suppl. 1998; inhibiting the production of pro-inflammatory cytokines such TNF-alpha, Interleukin-1, Interleukin-6 and antigen presentation by these professional antigen presenting cells. Interleukin-10 also regulate the function of other immunocompetent cells. Multiple doses of intravenous interleukin 10 in steroid-refractory Crohn's disease. Crohn's Disease Study Group. van Deventer SJ1, Gastroenterology. 1997 Defective IL10 signaling defining a subgroup of patients with inflammatory bowel disease. Begue B, Am J Gastroenterol. 2011 Recombinant human interleukin 10 in the treatment of patients with mild to moderately active Crohn's disease. The Interleukin 10 Inflammatory Bowel Disease Cooperative Study Group. Fedorak RN,.Gastroenterology. 2000

Clinical features of interleukin 10 receptor gene mutations in children with very early-onset inflammatory bowel disease. Beser OF1, J Pediatr Gastroenterol Nutr. 2015 Anti-mouse CD52 monoclonal antibody ameliorates intestinal epithelial barrier function in interleukin-10 knockout mice with spontaneous chronic colitis. Wang H1, Dong J, Immunology. 2015

DEKAVIL

Product Target ACR 20 ACR 50

INFLIXIMAB TNF 64 57

ETANERCEPT TNF 74 41

ADALIMUMAB TNF 67 55

ABATACEPT B7 73 48

TOCILIZUMAB IL6-R 89 70

KINERET IL1-R 38 17

TENOVIL IL 10 63 13-25

*values in combination with MTX

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rhIL10 (Tenovil) has been tested in a variety of clinical trials for the treatment of chronic inflammation

Opportunity to improve Tenovil ACR50 by selectively delivering IL10 to the

inflammation sites

IL10 IN RA TREATMENT

Weinblatt et al. (1999) ACR 63rd meeting. Abstract 598

A PHASE 1B CLINICAL TRIAL WITH DEKAVIL (F8-IL10)

AN IMMUNOREGULATORY 'ARMED ANTIBODY’

FOR THE TREATMENT OF RHEUMATOID ARTHRITIS,

USED IN COMBINATION WITH METHOTREXATE

Prof. Mauro Galeazzi EUlAR 2015

DEKAVIL IS A FULLY HUMAN IMMUNOCYTOKINE

Human IL10

Human IgG

F8 VH

VL

Villa A. et all. Int. J. Cancer: 122, 2405–2413 (2008)

F8-IL10 consists of the vascular targeting antibody F8 in scFv format fused to the anti-inflammatory cytokine interleukin 10 (IL10).

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A MODULAR APPROACH FOR PRODUCT DESIGN

EDA – fibronectin F8 IL10

Vascular target Antibody Payload Final product

N H 2

EDB EDA I I I CS oncofetal fibronectin

F8

COOH

Antibody F8, is specific to the extra-domain A of Fibronectin

DEKAVIL: F8IL10 HUMAN IMMUNOCYTOKINE

Human IgG 150 kDa

Diabody Immunocytokine F8-IL10 86 kDa

F8-IL10 is a fully human immunocytokine consisting of the vascular targeting antibody F8 fused to the anti-inflammatory cytokine interleukin 10 (IL10).

VH

VH VL

VL

NH2

NH2

IL10

IL10

VH

VH VL

VL

NH2

NH2

Villa A. et all. Int. J. Cancer: 122, 2405–2413 (2008) 10

Villa A. et all. Int. J. Cancer: 122, 2405–2413 (2008)

DEKAVIL: F8-IL10 TARGETING THERAPY

The antibody F8 targets to the ED-A of fibronectin, a marker of neoangiogenesis (oncophetal F.)

F8IL10

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ED-A fibronectin has a restricted expression in normal tissue (only in sites of physiological angiogenesis: plancenta and endometrium in the proliferative phase)

Schwager et al. (2009) Arthritis Res. Ther., 11, R142

placenta uterus pancreas lung

spleen bone marrow cervix prostate

breast

muscle

oesophagus colon heart brain liver

TARGETING CHARACTERIZATION ED-A FIBIRONECTIN TARGET IN RA

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TARGETING CHARACTERIZATION ED-A FIBIRONECTIN TARGET IN RA

Schwager et al. (2009) Artritis Res. Ther. 13

Schwager et al. (2009) Arthritis Res. Ther., 11, R142 Trachsel et al. (2007) Arthritis Res. Ther., 9, R9

grade 1 swelling

grade 2 swelling

healthy control paw

Targeting: selective uptake of F8-IL10 at the site of inflammation in the collagen-induced arthritis mouse model.

Phosphorimaging: In vivo targeting of 125I-F8-IL10. Near-infrared imaging: In vivo

targeting of Alexa750- F8-IL10

DEKAVIL PRECLINICAL STUDIES

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DEKAVIL PRECLINICAL STUDIES

0

1

2

3

4

5

6

0 5 10 15 20

salineMTXTNF-blockerF8-IL10 + MTX

days after arthritis onset

arth

ritic

sco

re

Dekavil inhibits disease progression in the collagen-induced arthritis mouse model and shows synergy with methotrexate

Schwager et al. (2009) Arthritis Res. Ther., 11, R142 Trachsel et al. (2007) Arthritis Res. Ther., 9, R9;

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Acute intravenous toxicity study in mice (RTC S.p.A, Study No 74250) - 20 mg/kg i.v. , single dose, termination on Day 15. - All animals survived to scheduled sacrifice. -No abnormalities observed at the injection sites, clinical or macroscopic observations. - Histopathology was not performed. -Slight (22- 26%) reduction in absolute and relative kidney weights of female mice was not considered toxicologically significant. 8-week comparative toxicity study in cynomolgus monkeys (CIT, Study No 34975TSP) - NOAEL, 180 µg/kg administered Dekavil s.c., 3 x /week for 8 weeks. - A transient regenerative anemia was observed. - Minimally increased liver weights. - Increased axillary lymph node weights

DEKAVIL PRECLINICAL STUDIES

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DEKAVIL PHASE I CLINICAL TRIAL IN RA

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Pavia

Siena

Rome

Pisa

1. Siena, Dpt. of Rheumatology, University Hospital.

P.I. and Head of Department Prof. Mauro Galeazzi,

Coordinating Center 2. Pavia, Dpt. of Rheumatology, S.Matteo

University Hospital. P.I. Prof. Roberto Caporali, Head of Department: Prof. Maurizio

Montecucco 3. Pisa, Dpt. of Rheumatology, S.Chiara

University Hospital. P.I. and Head of Department Prof. Stefano

Bombardieri 4. Rome, U.O. of Rheumatology, S.Camillo

Hospital. P.I. Dr. Giandomenico Sebastiani Head of U.O.: Prof. Giovanni Minisola

Primary Study Objectives: • To establish Maximum Tolerated Dose (MTD) and

Recommended Dose (RD) of F8-IL10 when administered with MTX

• To study safety and tolerability of F8-IL10 when administered with MTX

Study Secondary Objectives: • To study the drug Pharmaco Kinatic • To study immunogenicity (HAFA) • To assess preliminary anti-arthritic effect, in terms of ACR

and DAS 28 scores Study design • Treatment with escalating doses of F8-IL10 + fixed dose

of MTX

DEKAVIL PHASE I CLINICAL TRIAL IN RA

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Up to 8 weekly administrations of F8IL10 and MTX, response is assessed after 4 weeks (4 cycles). Patients who tolerate the treatment and are willing to continue, may receive additional 4 treatment cycles. After treatment follow-up visit will be performed monthly. Patients needing new treatment to control rheumatoid arthritis activity will discontinue the study.

SC W1 W2 W3 W4 W5* W6 W7 W8

W5 assessment or

EOT*

W9

Screening Evaluation

EoT assessment

(W9) F.U. 1 (W14) F.U. 2 (W18) Safety

evaluation for DLT

SC: Screening evaluation : DEKAVIL + MTX administration

* EOT for patients who do not continue with the 4 additional weeks of treatment

Mandatory part of the study Optional part of the study

DEKAVIL PHASE I CLINICAL TRIAL IN RA: treatment schedule

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Immunogenicity of F8-IL10: potential development of antibodies against F8-IL10 (HAFA). ELISA and BIAcore on collected samples at baseline, week 4 and week 9. At date 15 patients analyzed.

DEKAVIL PHASE I CLINICAL TRIAL IN RA: HAFA

Results Baseline and week 4 results: negative. Week 9 results: very low signal of HAFA detected in 2 patients.

DEKAVIL PHASE I CLINICAL TRIAL IN RA: CONCLUSIONS

Preliminary safety data suggest that targeted IL-10 treatment (s.c. F8-IL10), in combination with MTX is safe and well tolerated.

At all dose levels explored so far at least one patient experienced a clinical response as measured by ACR and/or EULAR criteria. However, no clear cut dose-response has emerged across the dose levels tested; this may be due to the study design, which is intended to test primarily safety and tolerability of F8-IL10.

Dose escalation of Dekavil in combination with MTX is on-going in order to achieve the Maximum Tolerated Dose.

Phase II trial in order to formally demonstrate Dekavil superiority versus placebo has been approved

The targeted delivery of IL10 to the site of inflammation may be a

promising therapeutic approach for RA patients.

Many thanks for your attention