Pharmacological Mode of Action
EDDITED BY JIN-TAE KIM, AGRANCO N.E ASIA COUNTRY MANAGER
AGRAZINC 100
AGRAZINC 100
 Zinc, especially, ZnO which is one of inorganic
zinc products is very effective to prevent and cure
the digestive disorder such as ‘diarrhea’ in young
animal (piglet, calf etc.), even in matured animal
 Also, Zinc is very important trace mineral involved
in natural immune system
Bivalent cation minerals and the
mucus on intestinal surface
 Bivalent cation: Zinc(Zn++), Copper(Cu++),
Magnesium(Mg++) etc.
 On the other hand, Mucus/intestinal surface is
consist of ‘glycoprotein’ and ‘glycolipid’. And, all of
them show a negative charge
 Bivalent cation compounds become free ions in
GIT and bind to mucus through the ionic bond
 To cope with this barrier in digestion of mineral,
‘Organic minerals’ were developed
 Differences between ZnO and Organic Zinc
 ZnO: can be free ion easily (Zn++)
 organic Znic: strong bond
 To get any effects such as anti-viral, antibiotic and
reduction in excessive immune response (allergy
etc) in GIT, Zinc complex should release free ions
easily.
Preventing pathogenic bacteria
[GIT; gastro intestinal tract]
EDDITED BY JIN-TAE KIM, AGRANCO N.E ASIA COUNTRY MANAGER
AGRAZINC 100
E. coli and GIT dysfunction
 Most important reason for GIT dysfunction like a
diarrhea is E. coli; K88, K99 etc.
 E. coli bind to specific receptors consist of
‘Glycoprotein’ and ‘Glycolipid’ on intestinal surface
and exert toxicity on the epithelial cell in typical
condition.
Effects of ZnO and CuSO4
 In effects of ZnO and CuSO4 on GIT…
 ZnO and CuSO4 increased the performance in
production and growth rate, when it’s inclusion rate
was increased.
 Both of them suppressed the activity of E. coli in
pharmacological level in diets
Intestinal
epithelial
cell
Mucus layer: the 1st
barrier to the pathogen
Depiction of normal condition in GIT and
settle down on mucus layer
[Mathew. 2001]
Effects of some countermeasures
for E. coli
 High inclusion level of Zinc(ZnO) could reduce the
activity of beneficial bacteria [Ole Hojberg et al, 2005].
 AGRAZINC 100 reduced the activity and number of
E. coli, but increased the activity and number of
beneficial bacteria in GIT.
 Lactobacilli can also bind to specific receptors of
E. coli on intestinal surface competitively. So, it
prevents excessive proliferation of E. coli in GIT.
[Mathew, 2001]
AGRAZINC 100
: Effects on E. coli
8
7
E. coli, log10
AGRAZINC 100 showed
reduction in number of
E. coli significantly
[China, 2007]. From
AGRANCO’s China trial,
Zinc ion seems to have
an affinity for specific
receptors for E. coli on
the intestinal surface
than beneficial bacteria
6
AGZ 300g/t
AGZ 100g/t
5
ZnO 3000ppm
4
3
Duodenum
Jejunum
Ileum
Caecum
9
Lactobacillus
acid., log10

8
7
6
5
4
3
Duodenum
Jejunum
Ileum
Caecum
[Dalian Polytechnic Univ. China, 2007]
Logical approach to MOA of
Zinc Ion(Zn++)


Zinc ion, bivalent cation, seems to have an
affinity for E. coli’s binding sites on intestine
more than other beneficial microorganisms.
With this hypothesis, Zinc ion seems to be
bound to ‘glycoprotein’ and ‘glycolipid’ on
intestinal surface preferentially that are the
binding sites of E. coli.

Then, E. coli cannot find any binding sites to
be adhered and channel to penetrate and
exert toxicity in target tissue. And, move
along with excreta.

As we have seen in many other trial, if ZnO
was supplied more than specific level
(unknown), it could suppress not only E. coli,
but also beneficial bacteria in GIT.
Zn++
Zn++
E. coli
First is E. coli,
second is
Beneficial
Beneficial bacteria
bacteria
• Graph 1
60
• Graph 2
8
7
50
6
40
5
100ppm Zn
2500ppm Zn
30
20
10
0
pH value
Lactic acid conc., umol/g wet digesta
Logical approach to MOA of
Zinc Ion(Zn++)
100ppm Zn
2500ppm Zn
4
3
2
1
intestine
caecum
0
intestine
caecum
• ZnO which have bypassed stomach would be ionized gradually and Zinc ions would be attached to
receptors consist of glycoprotein/glycolipid on intestinal surface or ‘Cell adhesion molecules’ on
epithelial cells.
• Meanwhile, Zinc ions would be attached to specific binding sites for E. coli competitively. However,
too much higher level of zinc ions flux could block the docking sites for other beneficial bacteria, and
then, it could suppress the activity of these beneficial bacteria such as Lactobacilli (Graph 1)
• After then, the amount of intact ZnO, which could be ionized, would be reduced to the minimum
level to suppress E. coli gradually. Then, the activity of beneficial bacteria would be increased (Lactic
acid, it’s by-product, would be increased)
[Ole Hojberg et al, AEM.71.5.2267-2277. 2005]
The limit of ZnO and effects of
AGRAZINC 100
 The effect of pharmacological level of ZnO is well known.
However, it also effects on beneficial bacteria.
 Still, there need to study what incorporation level (a guide
line) do not depress the activity of beneficial bacteria in
certain conditions, while it depress typical pathogen such as
E. coli.
 AGRAZINC 100 release Zn ion where it should be released in
order to protect gut health effectively from the pathogen.
Then, AGRAZINC 100 can also promote any beneficial
bacteria.
How AGRAZINC100 works in GIT?
Lactobacilli and factors produced by lactobacilli also
can block E. coli attachment sites
K88
E. coli
lactobacillus
Zn+
+
Zn+
+
E. coli attachment to
intestinal surface
glycoproteins
Free Zn++ released from AGRAZINC 100
attach to E. coli binding sites competitively.
Then, AGRAZINC 100 block E. coli
attachment on intestinal surface
Preventing Virus
EDDITED BY JIN-TAE KIM, AGRANCO N.E ASIA COUNTRY MANAGER
AGRAZINC 100
Infection routes of Virus
 The viral infection through the gut and respiratory
system(throat, lungs etc.) is commenced from the
binding (penetration) between Virus and the host
cell.
 Virus can get into the host cell through CAMs (Cell
Adhesion Molecules). There are IgSF CAMs(ICAM1, VCAM-1, etc), Selectins, Integrins and
Cadherins
Infection routes of Virus
 Nucleic acid of Virus is covered with a certain
protein called ‘Capsid’ seems like a “canyons’” or
“pockets” pattern.
 This capsid will be attached to a certain receptor
(ICAM-1, Integrins, etc.) just like a ‘key and lock’
formation
Capsid of Virus
CAMs of
Host cell
Infection routes of Virus,
CAMs
 CAMs (Cell Adhesion Molecules)

CAMs are proteins located on the cell surface involved with the binding with other
cells or with the extracellular matrix (ECM) in the process called cell adhesion
-Wikipedia-

Rotavirus utilize different integrins (glycoproteins) as cell receptors.

The outer end of ICAM-1 have a narrow, wedge-shaped segment that can reach
and bind with side chains on the canyon floor of rhinovirus.

CAMs allow the virus to penetrate the cell and replicate.
Inside of
Host cell
Capsid of Virus
CAMs of
Host cell
channel
Zinc and cell adhesion molecules
 In common cold virus(rhinovirus), Zinc ion is a
competitive inhibitor of ICAM-1 in both rhinovirus
particles and nasal epithelium.
[Darrell Hulisz, J Am Pharm Assoc 44(5):594-603, 2004]
 Zinc ions are small, positively charged spheres
with an affinity for CAMs that can easily reach the
virus canyon floor
 Zinc ions may block the binding between Virus and
CAMs on the cell membrane. Therefore, it may
reduce viral infection of intestinal epithelial cells.
Development of ICAM (Intercellular Adhesion
Molecule)
Development of ICAM-1
Cell membrane without
ICAM-1 development
Cell membrane with
ICAM-1 development
Flourescent-dyed ICAM-1 and ‘wedge’ liked micro projection
Normal cell surface with ICAM-1 (wt-IC1) has micro projections, but
other cell surface without inner cell domain(ICAM-1 [IC1ᅀCTD])has
no projections
TOP VIEW
A micro projection in
ICAM-1 developed cell
Surface of ICAM-1
developed cell
SIDE VIEW
Jun, Chang-Duk et al, Molecular Biology of the
Cell Vol. 18, Issue 6, 2322-2335, June 2007
CAMs, Infection, Immune and
Zinc ion (Zn++)
 CAMs provide a channel to immune cells and pathogen in
blood to adhere them on epithelial cell line. Therefore, these
immune cells can move into target tissue.
 This kind of method can be applied for both of virus and
bacteria or immune cell
 It means; if these kind of channel were blocked, we could
reduce the rate of viral infection in host cells and excessive
immune cells flux by increment of the permeability of the cell
into the target area/tissue that can lead ‘inflammation’
 Zinc ions may attach glycoproteins/glycolipids on the cell
membrane, such as CAMs. Then, it may prevent the binding
among epithelial cells and other cells or extracellular matrix
Zn++ : preventing allergy in GIT
EDDITED BY JIN-TAE KIM, AGRANCO N.E ASIA COUNTRY MANAGER
AGRAZINC 100
Immune response..
 In young animals, allergenic factors (ANFs in soy,
etc) can induce allergenic reaction in GIT, even in
matured animal.
 These induce excessive immune reaction on
intestinal epithelium, such as ‘hemorrhage’,
‘inflammation’
The basic of inflammatory
response (immune)
 Mast cell
 A mast cell is a resident cell of areolar connective tissue
(loose connective tissue) and contains many granules
rich in histamine and heparin
 Once allergen is bound with IgE on a mast cell, ‘mast cell’
is activated and release ‘histamine’, ‘leukotriene’ and
‘Prostaglandine(PG)’
 Histamine increase a permeability of blood vessel. Then,
it makes immune cells move into target tissue.
 At the same time, ‘mast cell’ release Zn++ as a natural
“circuit breaker” to prevent a excessive immune response;
regulate the amount of immune cell movement.
A permeability of tissue and
immune response
Zn ions attach to CAMs
and regulate the amount
of immune cell
movement into tissue
Michael J. Holtzman et al, Washington Univ. School of Medicine, 2001
AGRAZINC 100 : reduction in
inflammation and a excessive immune
response
 Zinc ions released from AGRAZINC 100 are...
 acting right on time and place: Once zinc ions
are absorbed at intestinal epithelium, they
regulate immune cell flux into intestinal
epithelium to prevent a allergy-like excessive
immune response.
 How?
 Once Zn ++ of AGRAZINC 100 attach to CAMs
(Cell Adhesion Molecules), they can block the
binding between CAMs and immune cells that
lead inflammation
Differences in types of zinc complex
EDDITED BY JIN-TAE KIM, AGRANCO N.E ASIA COUNTRY MANAGER
AGRAZINC 100
Types and effects of zinc complex
 Until now, we have studied ‘mode of action’ are
related to AGRAZINC 100
 However, all of zinc complexes are performing
same effects were shown in previous slides
effectively.
 To bring such effects in GIT, zinc ion must be
released easily right on time and place. It means
too tight bond in a zinc complex is not effective
such as chelated minerals.
Types and effects of zinc complex
benefits
AGRAZINC 100
ZnO
2500ppm
Organic
Zinc
weakness
Reduction in E. coli’s activity
Reduction in viral activity
Reduction in allergy in GIT
Immune enhancer (ingredient for
specific enzyme using in
metabolism/immune response)
Supply zinc ions
Etc.
advantage
Eco-friendly
Minor loss in GIT(100%
bypass in stomach)
Minor dosage
Effective for diarrhea
Distal effect
Reduction in E. coli’s activity
Reduction in viral activity
Reduction in allergy in GIT
Immune enhancer (ingredient for
specific enzyme using in
metabolism/immune response)
Supply zinc ions
Etc.
Environmental pollution
Huge loss in stomach &
intestine
Require higher inclusion
rate
Reduction in palatability
Effective for diarrhea
Supply zinc ions
Immune enhancer (ingredient for
specific enzyme using in
metabolism/immune response
Cannot be ionized easily
in GIT
No effects on diarrhea
May increase absorption
rate
Minor dosage
AGRAZINC 100 summery
 Glycoproteins, Glycolipids and CAMs are very important in
AGRAZINC 100
 AGRAZINC 100 attaches to these receptors competitively to
prevent pathogen exert their own toxicity on epithelium
 AGRAZINC 100 reduces the activity of E. coli as a pathogen,
but increase the activity of beneficial bacteria in GIT.
 AGRAZINC 100 may reduce a excessive immune response
like a ‘allergy’ induced by ANFs (anti nutritional factors).
 AGRAZINC 100 bypasses ‘stomach’ by 100% and commence
releasing of zinc ions from the beginning of intestine
(duodenum).