Liu php min dch dng khng th

IgY kim sot nhim Helicobacter

pylori - nhng kt qu nghin cu ti
Nht Bn

Sa V. Nguyen, Ph. D
Vin NC Min dch Gifu (IRIG)

Cng trnh do chnh ph Nht ti tr

1. Phng nga chng nhim H. pylori bng
khng th khng H. pylori ( H. pylori
). Tin hnh chung vi H Keio
University v hng Ortho Corporation (20032004).
2. "NC pht trin thc phm chc nng nhm
phng nga v gim thiu bnh d dy do
H. pylori (
). Tin hnh chung vi H Keio
University v hng Nisshin Pharma (2006-2008)

Bng sng ch lin quan n thc phm

chc nng khng HP
Major Patent List
Inhibitor of Helicobacter pylori colonization
USA: 6235709 / Canada: 2292012 / Japan: 3255161 / etc.

Glycoprotein having inhibitor activity against

Helicobacter pylori colonization
Japan: Appl.No.2000-113913 / USA: Appl. No.09/833637 / Canada:
Appl.No.2344183 / etc.

Specific antibodies for use in preparation of

pharmaceutical compositions useful in the
prevention or treatment of gastritis, gastric ulcers
and duodenal ulcers
USA: 6419926 / Australia: 735418 / EU: Appl.No.98400874.8 / Japan: Appl.No.199794159 / etc.

Pharmaceutical composition useful in the

prevention or treatment of peptic ulcers
Japan: Appl.No.2000-214835 / USA: Appl. No.09/903734 / EC: Appl.No.98400874.8 /
etc.

Ni dung

H. pylori urease v IgY (Ovalgen-HP)

Nghin cu trong phng th nghim
Mt s kt qu th nghim lm sng

H. pylori urease and IgY

pH: 2-3
Mi trng d dy vn khng
thun cho vi sinh vt pht
trin:
- Axt
- Vn ng co bp lin tc

Vai tr ca men urease i vi H. pylori

Phn hy urea trong thc n to thnh NH 3 v

phn ng ny lm tng pH trong d dy

Urea

Urease

NH3 + CO2

Gip Hp bm vo mng nhy ca d dy

H. pylori urease: thng tin khc

L mt protein ch yu ca H. pylori (chim 15%
tng s protein v)
Cn thit cho Hp lp qun th trong d dy (chng
Hp bin d men urease khng pht trin tt trong d
dy)
Kch thch to cytokines gy phn ng vim

Quy trnh sn xut IgY c hiu vi Urease (Ovalgen HP)

H. pylori NSP335 trong

mi trng BHI , 72 h
nhit 37oC

Thu thp khi t bo

Tch urease tinh

khit

Tinh ch qua 2 bc

Thu thp lng

trng ly IgY
(Ovalgen-HP)

IgY c hiu vi ureasetch ly trong trng ca

g c min dch

Trn urease vi CFA ri

gy min dch cho g mi

u vit ca khng th IgY so vi

IgG t V nui con bng sa
C i lc vi khng nguyn ln hn nhiu so vi IgG
(hiu qu phng tr ln hn).
Khng bm vo Fc receptor v khng phn ng vi h
b th ca ngi nn khng gy d ng (non-specific
inflammation).
C trong khu phn n hng ngy ca ngi nn rt an
ton (IgY lun c trong trng g).
iu kin v sinh nghim ngt trong chn nui g
(cm dng khng sinh).
9

C ch hot ng ca IgY

1. Gy bt hot urease : IgY c ch hot tnh urease dn ti

c ch s tng trng ca vi khun.
2. c ch bm dnh: IgY ngn cn H. pylori bm vo nim
mc d dy.
3. Ngng kt vi trng Hp to iu kin cho macrophage v
cc t bo min dch tiu dit
4. Gy ra cc tn thng cu trc t bo vi khun khin cho
Hp d nhy cm hn vi thuc ( c tc dng hip ng khi
kt hp vi cc thuc )

10

In-vitro test: IgY ngn c n Hp

urease bm vo mng nh y d dy
heo

11

Ngng kt
Bacteria

Bacteria and IgY

12

Macrophage tiu dit vi trng

T cell

B cell

IgG

13

Tc dng hip ng ca IgY v probiotic

H. pylori

Hp + IgY

Hp + L. gasseri

Hp + IgY + L.gasseri

14

Khi nim c gii thiu

Urease ng vai tr quan trng i vi s sinh trng ca
HP trong d dy
IgY t trng g u vit hn so vi IgG t V nui con
bng sa
IgY khng urease ca HP c 4 c ch tc dng chnh

15

Cc thc nghim ti phng th

nghim

16

Thc nghim 1: hiu qu ca IgY trn m hnh

chut nhim bnh
(Icoff 2007)
ng vt:

NS:Hr/ICR hairless mice, 8 tun tui

Tc nhn gy bnh:

chng H. pylori NSP335, 109 CFU/chut

Cc nhm:

cho n thc n trn Ovalgen HP theo t l

0.25, 2.5, 25% v nhm i chng

Thi gian:

10 tun

nh gi:

s lng vi khun trong m d dy

F: 2-day fasting
F

1 TUN

Gy nhim Hp

10 TUN

Cho n IgY

autopsy
17

Kt qu
Nhm

Log10 CFU / 0.1g m d

dy/chut

S chut m
tnh/tng s chut

i chng khng gy
nhim bnh

0.000.00

6/6

i chng c b nhim
bnh

3.640.39

0 / 10

0.25% Ovalgen HP

3.080.79

0 / 10

2.5% Ovalgen HP

1.261.47**

5 / 10#

25% Ovalgen HP

0.941.53**

7 / 10##

**; p<0.01 compared to positive control group value, one-way Anova test
, ; p<0.05 and 0.01 compared to positive control group, chi-square test

# ##

18

Thc nghim 2: tc dng hip ng ca IgY v

probiotic trn chut
(The 15th Annual Meeting of The Japanese Society for Helicobacter Research, 2009 )

Animals: Germfree BALB/C mice

Challenge : 1 109CFU/0.5mL/mouse/day 4days
IgY 1.0mg/mouse/day

4 weeks

8 weeks

H.pylori 1x109/mouse/day

L.gasseri 1x109/mouse/week
Autopsy

Autopsy

19

No. of H.pylori (Log10 CFU / g tissue)

Kt qu

4.30.6
(n=10)

* 3.11.2

**

** p<0.01
* p<0.05
3.01.4
(n=10)

(n=10)

2.31.7
(n=10)

5
4
3
2
1
0
Control IgY

Anti H.p urease IgY

L.gasseri 752

Treatment

Anti H.p urease IgY

+
L.gasseri 752

20

Thc nghim 3: hiu qu ca IgY trn m hnh chut

nhy Mng c
4
Nhm i chng

1 tun
-

F + IgY

8 tun

Tui ( tun)
13

F + IgY

Gy nhim H.pylori

Nhm nhim
H.pylori

F + IgY

IgY: 25 mg/g
F: Famotidine 0.16 mg/g

F + IgY

(Helicobacter 10:43-52, 2005 )

21

Kt qu

F+IgY
Control

105

H.pylori

0.2
0.15
0.1
0.05

Cut off
value

IgG c hiu khng H.pylori

trong huyt
thanh

(CFU/g gastric tissue)

anti H. -pylori IgG

0.25

Control

H.pylori

104
103
102
10
n.d.

n.d.

n.d.

Lng H. pylori
trong d dy

(Helicobacter 10:43-52, 2005

)

22

Vim nim mc d dy

Nhm i chng
khng gy nhim bnh

Nhim bnh v
iu tr F+IgY
Nhm nhim bnh
nhng khng iu tr
(Helicobacter 10:43-52, 2005 )

23

Khi nim c gii thiu

IgY khng urease (Ovalgen-HP) lm gim s xm nhim ca
H. pylori trong d dy chut v chut nhy Mng c.
IgY kt hp vi probiotic hoc famotidine lm tng o thi
H. pylori v gim vim d dy trn chut nhy Mng c.

24

Cc th nghim trn ngi tnh

nguyn

25

Cc th nghim tin
hnh
No

Nm
(ni)

Sn
phm

Thi
gian

nh
gi

2002
(Taiwan)

23

sa chua

8 tun

UBT

Test 24
Plac 24

2002
(Korea)

2004
(Japan)

2004
(Japan)

16

2004
(Japan)

17

2009
(Spain)

Test 27
Plac 24

Bt

2011
(Japan)

Test 21
Plac 21

sa chua

22

Kt qu

Average UBT decreased after 4 & 8 w

Test: average UBT decreased after 2, 4 & 8
w
Placebo: No change in UBT

Bt

8t

UBT

Bt
(capsule)

4t

UBT

8t

UBT &
HpSA

4t

UBT

13/17 subjects show decreased UBT

8t

UBT

Test: average UBT decreased significantly

Placebo: no significant change in UBT

8t

HpSA

sa chua

Bt

Average UBT decreased significantly after 4

w
Both parameters decreased significantly
after 2, 4 & 8 w

Test: average HP decreased significantly

Placebo: tends to show increased HP in stool
26

Th nghim 3
J Dairy Sci. 2004 Dec;87(12):4073-9

S ngi tham gia:

22 ngi tnh nguyn c

H. pylori dng tnh

Sn phm th

Ovalgen HP (vin capsule

1.5 g 3 l/ngy x 4 tun)

nh gi

UBT ( thi im 0, 1 thng )

27

Kt qu
J Dairy Sci. 2004 Dec;87(12):4073-9

P < 0.01
28

Th nghim 4
(Food and Development vol 38 No 11, 2004)

S ngi :

16 ngi tnh nguyn c H. pylori dng tnh

Sn phm:

sa chua cha 2.0 g of Ovalgen HP

Liu:

2 chn/ngy

Thi gian:

2 thng

nh gi :

UBT & HpSA ( ti thi im 0, 1, 2, 3 thng)

29

Kt qu
(Food and Development vol 38 No 11, 2004)

30

Th nghim 5
(Suzuki et al. Aliment Pharmacol Ther 2004, 20: 185-192)

Thit k nghin cu
i tng:

17 ngi tnh nguyn c Hp dng tnh

iu tr:

Ovalgen HP 0.9 g/ngy x 4 tun.

nh gi:

UBT test

31

Kt qu
(Suzuki et al. Aliment Pharmacol Ther 2004, 20: 185-192)

S th t

Trc

Sau

66.4

49.9

62.9

46.3

61.2

5.3

57.7

19.9

35.0

46.6

29.9

44.2

32.4

30.0

43.3

34.3

40.8

19.6

20.0

36.4

28.8

10

28.9

46.7

15.0

11

26.6

28.8

12

23.1

9.6

5.0

13

22.0

25.4

14

19.2

13.0

0.0

15

19.1

11.0

16

9.0

4.4

17

6.9

9.1

45.0
40.0

**

25.0

10.0

Trc

sau

** p<0.01
32

Th nghim 6: Spain
(By Leche Pascual, Spain)

Tham gia:
Nhm TN: s ngi tham gia ban u 27, cui cng cn li 15
Nhm C: s ngi tham gia ban u 24, cui cng cn li 21
Sn phm: Bt Ovalgen HP (YP) 1g trn vo cornflake, dng
n sng ngy 1 ln (Nhm C dng 1 g bt khng
th C). Thi gian 8 tun
nh gi:

UBT ( thi im 0, 8 tun )

33

Kt qu

P=0.543

P=0.028

34

Th nghim 7

Tham gia: 42 ngi tnh nguyn c HP dng tnh chia thnh

2 nhm TN v placebo (n =21)
Sn phm: Nhm TN dng sa chua c Ovalgen HP (3
ln / ngy)
Nhm placebo dng sa chua c khng th C (3
ln / ngy)
(Sa chua c sn xut bi hng Namyang, Hn Quc)
nh gi:

UBT ( thi im 0, 1, 2 thng )

35

Kt qu

Placebo

Test
36

Kt lun
Ovalgen-HP c hiu qu lm gim s xm nhim
ca H. pylori trn ng vt b nhim bnh v trn
ngi tnh nguyn c Hp dng tnh.
Ovalgen-HP c th hu ch trong iu tr nhim Hp
khng khng sinh.

37

38

Acknowledgement
Dr. H. Suzuki, Keio University
Dr. T. Yamane, Matsushita Memorial Hospital
Dr. Koga, Tokai University
Dr. N. Kimura and research staff of Health care Research center, Nisshin Pharma Inc.
Project conducted under grant of Japanese Government

39

Products using Ovalgen HP

Pylorigen: capsules

Dr. Piro yogurt

Containing IgY and bifidobacteria

Pyloridasu: tablets
40