Case Study: The experience of the industrialization for ...c.ymcdn.com/sites/casss.site-ym.com/resource/resmgr/CMC_Japan... · Fingerprint Authentication Centralized ... Educational

Copyright Japan Tissue Engineering Co., Ltd. All Rights Reserved.

CMC Strategy Forum Japan 2014

Japan Tissue Engineering Co., Ltd.,

Ken-ichiro Hata D.D.S. Ph.D.,

Definition and Regulation on Regenerative Medicine (or Cell Therapy Product)

Case Study: The experience of the industrialization for autologous cell based products in JAPAN.


Japan Tissue Engineering Co., Ltd. (J-TEC)

OSAKA

NAGOYA TOKYO

GAMAGORI

Foundation: Feb. 1, 1999

HQ location: Gamagori

Capital: 7,785 mil yen

Employee: 218

Only J-TEC has approved QMS/GMP facilities in Japan, among biotech companies.

QMS/GMP facilities


Regenerative Medicine for skin


Cultured epidermis


Chukyo Hospital burn center

After 4 weeks

Clinical Research :BURNS

Left thigh

Right thigh


[Category] Medical device

[Indication] Severe burn (DDB + DB ≧ 30％)

[Regulatory Status] Approved : Oct. 2007 [Reimbursement price] JPY 314,000 / sheet ($3,140 / sheet)

8 cm

10 cm 80 cm2

The 1st Tissue-Engineered Medical Product Approved in Japan


Severe burn patient

Healthy skin

From a stamp-sized sample of a patient’s own healthy skin, J-TEC cultures enough epidermis to cover the patient’s entire body within 3 weeks.

Incoming inspection

Culture

Shipping inspection

Packaging

Grafting

Medical Institution

JACE

JACE Business Model


Bone

Cartilage Articular cavity

Regenerative Medicine for Cartilage

Cartilage

Bone


Tissue-Engineered Cartilage

3-D Cultivation in

Collagen Gel

Implantation

Improve inadequacies of ACI by 3-D

Cultivation

(prevent de-differentiation of chondrocytes

and retain in implant site after operation )

Existence of chondrocytes having

proliferation potency

Production of cartilage matrices

Prof. Ochi (Hiroshima University)

Isolate

chondrocytes

Covered with

Periosteum

Marketing approval in JAPAN

2012. 7. 27


Transplantation of Tissue-engineered Cartilage

Uchio Y. and Ochi M,

3875 Nihon Iji-Shinpou 1998.


Ochi M. et al., JBJS [Br] 84B, 571-8, 2002.

Lysholm score： pre-operation 70.6 → post-operation 96.7

Arthroscopic assessment：excellent / good 96％

pre-operation 2 yrs after operation

13-yr-old boy, a cartilage defect in the medial femoral condyle

Case Report

Clinical Research :Damaged knee cartilage


Evaluation by MRI

Pre-operative finding 12 months after ope.

Clinical Trial


About 40 patients participated in the clinical trial. J-TEC monitored their results after surgery for 1 year.

J-TEC submitted an application for mfg & sales in Aug. 2009, but excluded the elbow from its indication.

[Patient] [Result]

– Elbow) Chef

– Elbow) Building constructor

– Knee) Physical education teacher

– Knee) Farmer

– Knee) 68 year-old senior

– Knee) 120kg weight female

– Knee) Aerobics instructor

– Knee) Nurse

Excellent

Good

No improvement

NA

Efficacy

Safe

ty

JACC – Result of Clinical Trial


Long term follow up results

Modified MOCART Score

0

5

10

15

20

25

跛行

ロ

ッキ

ング

膝

の不安定性

階段昇降

支持装具

腫脹

疼痛

膝

の屈伸

スコア（最

小値、中央値、最大値）移植前

12ヶ月後

追加検査

**

*

** **

*

*** ***

**

0

10

20

30

40

50

60

70

80

90

100

8項目

の合計

*** ***

Lysholm Score （*p<0.05, **p<0.01, ***p<0.0001 vs Pre operation, Wilcoxon）

According to these results, the efficacy of cultured cartilage grafting

lasted during extra follow up period (1-5 years).

Pre 1Y >5Y


2.5 cm

(4.9 cm2)

[Category] Medical device

[Regulatory Status] Approved : July 2012

[Indication] Traumatic cartilage defects and osteochondritis dissecans (OCD)

exclu. osteoarthritis (OA) for knee joints. Defect area: 4 cm2 +

2nd generation cartilage

[Reimbursement price] JPY 2,130,000 / knee ($21,300 / knee)

2nd RM Product Approved in Japan


Evaluation of Safety and Efficacy

15

How do you know it’s safety?

How do you know it’s efficacy?

・No infectious factor such as pathogenic organism

・No anticipated immune reaction such as allergy

・No risk of transformation after grafting

・Efficacy of the treatment

・Efficacy of the product

Objective evaluation of TEMPs


To develop tissue-engineered regenerative medicine products using autologous cells

1. Establishment of cell banks for research and development.

※Skin 249 (2) Cartilage 295 (6) Cornea 128 (2)

2. Standardization of cell culture procedure for production.

/Age /Biopsy site /Doctors technique /Uncontrolled etc.

3. Evaluate product characterization.

/cell survival rate /cell content /impurities etc.

4. Establishment of standards for product safety and efficacy

5. Development of product packages and delivery system

/avoiding reduce(increase) cell viability /easy to handle etc.

6. Construction of GMP compliant facilities

We have a lot of work to achieve ‘from Bench to Bedside”


Ensuring safety for cell based products

Implemented for ensuring the product safety.

Choice of raw materials Process of manufacture

Animal experiments Clinical trials

Pathogenic organisms

•Microbe clearance •Aseptic manipulation

Evaluation pathogen infectious disease

Evaluation pathogen infectious disease (Long term follow up in some cases)

Immune reaction (allergy)

•Immunogenicity of raw materials •Removal by washing

Evaluation of immunological reaction

Evaluation of immunological reaction

Tumorigenecity (Carcinogenicity)

•Soft-ager colony formation assay •karyotype analysis •Carcinogenicity test for medium.

Evaluation of neoplasia Evaluation of neoplasia (Long term follow up in some cases)

Othres ・・・・・・・・・


A Ki67 / TO-PRO®-3

④ Final Product

③ Confluent ② Subconfluent ①Initial Colony

Growth Potential Cells in Cultured Epidermis


The analysis for the cell contents in cultured epidermis

【Main active component】・Keratinocyte 【Process-Related Impurities】・Feeder cell （3T3-J2）【Raw material-Related Impurities】 Epidermis ・Melanocyte ・Langerhans cell ・Merkel cell Dermis ・Fibroblast Subcutaneous tissue ・Adipocyte

Dermis（papillary）

Dermis （reticular）

Epidermis

Langerhans cell

Glandulae sebaceae

Subcutaneous tissue

Fat Hair follicles Sweat grand

fibroblast

]Melanocyte Basal L. prickle cell L. Granular L.

Merkel cell

H.Green: SCIENTIFIC AMERICAN Nov (1991)

Keratinocyte

capillary

Cornified L.

Keratinocyte Nuclear stein Melanocyte

Langerhans

cell Fibroblast Merkel cell Adipocyte



N=4：Ave. ± SD

Langerhans cell

0

1

2

3

4

5

P0 P1 P2 P3 FP

Co

nte

nt

rate

（%）

Merkel cell

0

1

2

3

4

5

P0 P1 P2 P3 FP

Co

nte

nt

rate

（%）

Melanocyte

0

1

2

3

4

5

P0 P1 P2 P3 FP

Co

nte

nt

rate

（%）

Adipocyte

0

1

2

3

4

5

P0 P1 P2 P3 FP

Co

nte

nt

rate

（%）

Fibroblast

0

1

2

3

4

5

P0 P1 P2 P3 FP

Co

nte

nt

rate

（%）

Feeder cell

0

2

4

6

8

10

P0 P1 P2 P3 FP

Co

nte

nt

rate

（%）

0

92

94

96

98

100

P0 P1 P2 P3 FP

Co

nte

nt

rate

（%）

Keratinocyte



Keratinocyte

「Content rate of keratinocyte

in final product」

Feeder cells

「Residual ratio of feeder cells

in final product」

Shipping inspection

Feeder cells

Adipocyte

fibroblast

Merkel cell

Langerhans cell

Melanocyte

Keratinocyte

98.3

97.2

95.3

94.7

98.0

1.0

0.5

0.9

0.8

1.4

0.9

0.6

0.4

1.6

2.7 3.5

0.4

P0 P1 P2 P3 Final Product

Co

nte

nt

rate（％）

100


Detachment

by dispase Cultured

epidermis

Digestion

Inoculation

Isolated

cell

Feeder Layer

（3T3-J2）

37℃ 10% CO2

Passage

culture

Stratification

Full thickness

Skin

4. Establishment of standards for product safety and efficacy (JACE)

■Process inspection #1 Morphology of cultured 3T3-J2

#2 Morphology of 3T3-J2 feeder

#3 Morphology of cultured keratinocyte

#4 Intensity of sheet by detachment and washing

#5 Proliferative potential of cultured keratinocyte

#6 Viability of frozen-thawed keratinocyte

#7 Number of collected cells on 3T3-J2 subculture

#8 Preservation period of biopsy tissue

■Shipping inspection ; judge until shipping

#1 Sterility 5 days before shipping

#2 Mycoplasma-negative [PCR] 2 days before shipping

#3 Appearance and morphology of cultured epidermis

#4 Endotoxin

#5 Appearance of final product package

#6 Intensity of keratinocyte sheet

#7 Viable cell density of keratinocyte sheet

#8 Viability of keratinocyte sheet

#9 Residual bovine-serum albumin in keratinocyte sheet

#10 Content rate of feeder cell in keratinocyte sheet

#11 Content rate of keratinocyte in keratinocyte sheet

■shipping inspection ; judge after shipping

#1 Mycoplasma-negative [DNA-statining]; judge 5 days after shipping

#2 Sterility of final product; judge 14 days after shipping

■Receiving inspection #1 Transport condition of biopsy tissue

#2 Appearance of biopsy tissue


5. Development of product packages and delivery system

Product packaging assures the preservation stability. Designing logistics, with temperature control and maintaining expiration date, is very important.

°

Expiration:

56 hrs

Storage temperature:

10 – 25 C °


The environment within our production facilities is controlled by air conditioning and room pressure control systems.

The flow of people and objects is also controlled to maintain the environment in its highly clean state.

Safety Cabinet Entry Control by Fingerprint

Authentication

Centralized Monitoring System

6.Construction of GMP compliant facilities


■ Optimization of culture conditions

(Medium preparation, Culture environment, Container design, etc.)

■ Combined culture with scaffolding

■ Cell freezing technology

(Cell banking, Cell freezing in ultracold storage, Quality control)

■ Quality inspection

- Setting cell quality endpoint and standard value

- Setting safety evaluation items

(Microbial contamination examination, Cell character evaluation,

Residual test, etc.)

Productization and optimization of TEMPs

Technology and Know-How (1)


■ Managing schedule in cooperation with Hospital

(Planning- Receiving order-Tissue biopsy-Culture-Shipping-Logistics -Grafting)

■ Raw material management system

(Selection, Procurement, Storage, Information-gathering)

■ Packaging cell culture product

- Defining storage conditions (temperature range, preservative solution)

- Designing product container, Selecting material, Conditions for sterilization

■ Information control

(Identification management, Prevention of misidentification, Traceability)

■ Training system

(Education of operatives, Recognition of skills)

■ Developing logistics for order-made cell product

(Special logistics system with temperature control)

Systematization of production and logistics of TEMPs

Technology and Know-How (2)


Key point to achieve TE Industry

写真 SOP

GMP Facility

HQ

Educational system

for Employee

More! ・Delivery

(Package, Logistics etc.)

・Post-marketing

Surveillance

・Training system for Drs.

etc. SOP and documentation


The training system for Doctors

• What is JACC®（configuration, indication, clinical efficacy and safety etc）

• How to use（pre-operative screening, biopsy, graft procedure, Rehabilitations etc）

• Account for patient and informed consent • About post marketing surveillance • Hand’s on course (Training by Simulation kit)

【Contents】

Simulation Kit

Text for

-Doctor

-Comedical staff

-Patient



Thank you for your attention.

[email protected]

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Case Study: The experience of the industrialization for ...c.ymcdn.com/sites/casss.site-ym.com/resource/resmgr/CMC_Japan... · Fingerprint Authentication Centralized ... Educational