Drug discovery

Role and significance of academia

Toshio MIYATA

Tohoku University Graduate School of Medicine

United Centers for Advanced Research

and Translational Medicine (ART)

The 13th Kitasato University - Harvard School of Public Health Symposium

Inhibitors to plasminogen activator inhibitor (PAI)-1

Example

What can academia contribute ？

Our strategy, goal, framework, status…

95 compounds

Hit compounds TM5007

3000 compounds

Docking simulation

28 compounds

Biological screening

Filter selection (size/charge)

Lead compound TM5275

75 compoundsHit-to-lead synthesis

Virtual screening

Chemical synthesis

In silico discovery of PAI-1 inhibitors2,240,000 compounds

0.3 mg/kgIn vivo efficacy

(anti-thrombosis)

Cmax：17.0 μM

Tmax：2 hr

T1/2：3.9 hr

PK (rat, 5mg/kg)

Tmax：4 hr

T1/2：8.2 hr

Cmax：106.5 μM

PK (monkey, 3mg/kg)

IC50

2.82 μM

0.3 mg/kg

Cmax：5.7 μM

Tmax：18 hrT1/2：>198 hr

PK (rat, 50mg/kg)

10 mg/kg

Cmax：34.2 μMTmax：2 hrT1/2：25 hr

PK (rat, 50mg/kg)

Cmax：17.9 μM

Tmax：1 hrT1/2：2.3 hr

PK (rat, 5mg/kg)

1 mg/kg

In vivo efficacy

(anti-thrombosis)

IC50 IC50 IC50

7.94 μM6.47 μM 3.58 μM

Hit TM5007 Lead TM5275 TM5441

In vivo efficacy

(anti-thrombosis)

In vivo efficacy

(anti-thrombosis)

TM5509

300 mg/kg

O

NH

O

NHSS

S S

OOH OH

O

NH

O O

O N

N

Cl

OH

O

75NCEs 166NCEs 207NCEs

Clinical candidate

Hit to lead, lead optimization to a clinical candidate

Newly synthesized over 500 compounds

A panel of GLP pre-clinical studies

A manufacture of the GMP-grade investigational drug (tablet)

Leaching test：Good dissolutionStability test：Unwrapped tablet was stable for 3 months in accelerating test of 40℃ and 75 % relative humidity.

Pathophysiologicalresearch

Target validation

Hit compound(HTS, in silico)

Academia can undertake up to Ph-IIa…

Lead optimization

GMP synthesisformulation

GLPPre-clinical

Phase I (PK, PD) and II (POC)

これまでの実績（個人シーズ）

PMDA戦略相談・事前面談 ３件

PMDA戦略相談・対面助言 ３件

医師主導治験（未承認薬）Ph-1,2a ３件

治験外臨床研究（未承認薬）Ph-2a １件

オーファン申請（共同） １件

企業導出 ２件

厚生労働省 先駆けパッケージ戦略 2014 6月17日

What is the advantage and significance of drug discovery in

academia ？

Open innovation…

A PAI-1 inhibitor regenerates bone marrow and vascular endotheliumBlood 119, 2012

PAI-1 inhibitors improved the mortality and

promoted rapid hematopoietic recovery

C57BL6/J recipient mice, 9Gy TBI, WBM transplantation

0d 7d 21d 28d14d

Treatment Evaluation Evaluation

0

10

20

30

40

Cont 10 100 1 3 10

WBC count (x102/µl)

0

20

40

60

80

100

Cont 10 100 1 3 10

PLT count (x105/µl)

0

10

20

30

40

Cont 10 100 1 3 10

BM MNC (x106/µl)

0.0

0.3

0.6

0.9

1.2

Cont 10 100 1 3 10

plasma active tPA (ng/ml)

0

20

40

60

80

Cont 10 100 1 3 10

plasma total MMP9 (μg/ml)

0

100

200

300

400

Cont 10 100 1 3 10

plasma c-KitLigand (pg/ml)

tPA TM5275 TM5509 (mg/kg) tPA TM5275 TM5509 (mg/kg) tPA TM5275 TM5509 (mg/kg)

tPA TM5275 TM5509 (mg/kg) tPA TM5275 TM5509 (mg/kg) tPA TM5275 TM5509 (mg/kg)

Stem Cell, 2013PAI-1 inhibitor

Irradiation

Blood count Blood count

A PAI-1 inhibitor retards the development of senescence and protects organ

structure and function while prolonging the lifespan in the klotho mouse.

Proc Natl Acad Sci USA, in press

PAI-1 is a critical up-stream factor of

senescence-messaging secretome

(SMS)

Proc Natl Acad Sci USA 2012

Nature Reviews Cancer 2009

Cell regeneration

PAI-1

inhibition

Fibrinolysis

Inhibition of plasminogen activator inhibitor 1 (PAI-1)

Anti-senescence

Anti-thrombosis

Anti-aging

Anti-fibrosis

Kidney disease

Cellregeneration

Anti-inflammation

PAI-1inhibitors

Nature Review Nephrology 2014

Potential applications of our PAI-1 inhibitors

ATVB 2007

ATVB 2007

JCBMF 2010

Circulation 2013

PNAS 2014Blood 2012

Stem Cell 2013

ARCMB 2012

ATVB 2013

ATVB 2013

Open resources

What can academia contributein the futute？

PK, Pharmacokinetics (what the body does to the drug)

Microdosing (phase 0)

Efficiently develop FIH-class drug with lower costs and greater probabilities…?

Significance of ‘exploratory early clinical studies’ in humans !

PD, Pharmacodynamics (what the drug does to the body)

Biomarkers, molecular imaging

Tox, Toxicity

Renal biomarkers for drug toxicities

POC, Proof of concept (efficacy in humans)

Pharmacogenomics (PGx), surrogate end points

Miyata et al. Nature Review Nephrology, 2014

Phase I

Phase 0

Phase I

Phase IIa

A total dose of 100μg as a single dose or divided doses in any subject including patients.

The requirements of pharmaco-toxicological studies are minimum, depending upon the amount or duration of exposure, e.g., a single-dose toxicity study in rodents.

For the conventional phase I, a full program of pharmaco-toxicological studies under the GLP regulation is required

Microdosing test

ICH Harmonized Tripartate Guideline M3(R2)

PK, Pharmacokinetics (what the body does to the drug)

Microdosing (phase 0)Phase 0

Phase I pharmacokinetics of a PAI-1 inhibitor TM5509M

icro

dosi

ng

Q

Is it possible to evaluate early a drug’s pharmacodynamics from analyses of blood or urine specimens? Often NO

Can we directly assess pathological events in situ in the tissue (e.g., inflammation, fibrosis, hypoxia, oxidative stress)? Potentially Yes

How can we evaluate a drug’s pharmacodynamics as early as possible?

Oxygen status is visible non-invasively in human kidney

by blood oxygen dependent level (BOLD)-MRI

Control Water load

Miyata et al, Nature Review Nephrology, 2011

PD, Pharmacodynamics (what the drug does to the body)

Biomarkers, molecular imagingPhase I

POC, Proof of concept (efficacy in humans)

Pharmacogenomics (PGx)Phase IIa

This is not the case with human studies: age, nationality, lifestyle, food …

Pharmacogenomics (PGx)

Normal responder

Weak responder

No responder

Potential side effect

Experimental mice are homogeneous:age, gender, genotype, food …

PAI-1 polymorphism

Does this genetic difference affect the efficacy, optimal

dose, and toxicity of a PAI-1 inhibitor?

PAI-1 GGGG

4G-type

Gene Higher PAI-1 Conc.

TM5509Partial Inhibition

PAI-1 GGGGG

5G-type

Gene Lower PAI-1 Conc.

TM5509Complete Inhibition

PGx-based dose adjustment

PAI-1 Genotypes Frequency in Japanese Plasma PAI-1 conc. Dose of TM5509

4G/4G 40 % 87 ng/mL 100 mg ?

4G/5G 50 % 62 ng/mL 75 mg ?

5G/5G 10 % 42 ng/mL 50 mg ?

Q

Does the genetic difference of metabolizing enzymes of a

PAI-1 inhibitor TM5509 affect the efficacy or side effect?Q

PAI-1 inhibitorTM5509

Patients withCYP2C9 3*/3*

Low activity mutant

・Normal metabolizer・Relatively low blood concentration?

・Poor metabolizer・Relatively high blood concentration?

Prediction of PK, tox, and efficacyGenetic analyses of target molecule

or metabolizing enzymes

Patients withCYP2C9 1*/1*

Wild Type

Efficient clinical trials for POC with pharmacogenomics (PGx)

PAI-1 inhibitorTM5509

(FIH class)

Metabolizing enzymeCYP2C9

Normal responder without side effect

No responder

Potential side effect

Pharmacogenomics (PGx)

Target protein

Phase 0 Phase I Phase IIa Phase IIb Phase III

Towards a more efficient strategy and framework

‘From Serendipity to Rationality’

Biomarker and molecular imaging (PD or

Tox)

Validated surrogate endpoints (Big data)

Pharmacogenomics (PGx)

Collaboration of regulatory authorities

Global networking of clinical trials

Microdosing test (PK)