About OMICS Group

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About OMICS Group

• OMICS Group International is an
  amalgamation of Open Access publications and
  worldwide international science conferences and
  events. Established in the year 2007 with the
  sole aim of making the information on Sciences
  and technology Open Access, OMICS Group
  publishes 400 online open access scholarly
  journals in all aspects of Science, Engineering,
  Management and Technology journals. OMICS Group
  has been instrumental in taking the knowledge on
  Science technology to the doorsteps of ordinary
  men and women. Research Scholars, Students,
  Libraries, Educational Institutions, Research
  centers and the industry are main stakeholders
  that benefitted greatly from this knowledge
  dissemination. OMICS Group also organizes
  300 International conferences annually across the
  globe, where knowledge transfer takes place
  through debates, round table discussions, poster
  presentations, workshops, symposia and
  exhibitions.

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About OMICS Group Conferences

• OMICS Group International is a pioneer and
  leading science event organizer, which publishes
  around 400 open access journals and conducts over
  300 Medical, Clinical, Engineering, Life
  Sciences, Phrama scientific conferences all over
  the globe annually with the support of more than
  1000 scientific associations and 30,000 editorial
  board members and 3.5 million followers to its
  credit.
• OMICS Group has organized 500 conferences,
  workshops and national symposiums across the
  major cities including San Francisco, Las Vegas,
  San Antonio, Omaha, Orlando, Raleigh, Santa
  Clara, Chicago, Philadelphia, Baltimore, United
  Kingdom, Valencia, Dubai, Beijing, Hyderabad,
  Bengaluru and Mumbai.

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Track 9, 1500-1520 Aug., 26, 2014
Pharmacognosy 2014, Beijing
Neurotrophic Compounds of Javanese Ginger,
Zingiber purpureum
Yoshiyasu Fukuyama Pharmaceutical Sciences,
Tokushima Bunri University Tokushima, Japan
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Drugs for Treatment of Alzheimers Disease
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Effects of Neurotrophins
Neurotrophins (NGF, BDNF, NT-3, NT-4/5 )
Neuronal stem cells

Differentiation
Neuronal precursor cells

Maturation
Development
Ageing
Death


Astrocyte


Neurons
Oligodendrocyte
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Neurotrophic factors

• Representative neurotrophic factors
• NGF
• BDNF
• NT-3, NT-4/5
• Problems due to large peptide molecule
• No penetration of B.B.B.
• Poor oral pharmacokinetic profiles
• Immune reactions

Nonpeptidyl small molecule neurotrophic compounds
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PC12 cells used for Screening neurotrophic/neurop
rotective/cytotoxic compounds
The PC12 cell line was derived from rat
pheochromocytoma, a tumor arising from chromaffin
cells of the adrenal medulla. It is a useful
model for studying cell signaling for at least
two reasons (i) There are few growth factors,
neurotrophins, and hormones to which it does not
respond and (ii) distinct responses of
differentiation, proliferation, and survival can
all be assessed independently.
Neuroprotectiveafter NGF withdrawal
Science, 296(5573)1648 (2002)
Neurotrophic
Cytotoxic
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The Principles for Screening Neurotrophic/Neuropro
tective Compounds in Rat Cortical Neurons
Neurotrophic compounds can promote neurite
outgrowth, which improves the status of neurons.
Neuroprotective compounds can retard toxic
factor-induced cell death in primary cultured
neurons.
Neurotrophic
toxic
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Search for Neurotrophic Mimic Natural Products
Rat pheochromocytoma cell line (PC12 cells)
Primary cultured rat neurons (Cortical neurons)
Neurotrophic Compounds Library
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BANGLE (Zingiber purpureum)
This plant is used as a spice and also used for
traditional Indonesian medicine jamu.
Purpose Rhizome Fever, headache Cough,
expectorate Stomach ache Constipation
Jaundice Rheumatism The ingredients of herbal
medicine for women after childbirth. Leaf
Revitalizer  Digestive trouble
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BANGLE (Zingiber purpureum)
NGF-like Activity of BANGLE in PC12 Cells
PC12 Cells
MeOH ext (25 µg/mL)
Absence of NGF
Ctrl ( 0.5 EtOH )
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Neurotrophic activity ()
Ctrl ( 0.5 EtOH )
Fr.4 ( 12.5 µg/mL )
Fr.5 ( 12.5 µg/mL )
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Screening Result of Compounds from BANGLE by PC12
Cells
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Differentiation of PC12 Cells Induced by 1 and 2
NGF (10ng/mL)
Control (0.5 EtOH)
1 (30 µM)
2 (30 µM)
PC12 cells were cultured in 24-well plates in
DMEM 10 HS and 5 FBS for 24 h at a density of
2x103 cells/cm2, and then medium was changed to
DMEM 2 HS and 1 FBS containing 1 and 2.
After 4 days cells with neurites were counted.
Plt0.01 vs control by Chi-square test
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Neurite Length of NGF-differentiated PC12 Cells
Effected by 1
Average of neurite length (µm )
1
PC12 cells were cultured in two 24-well plates in
DMEM 10 HS and 5 FBS for 24 h at a density of
2 x103 cells/cm2, and then medium was changed to
DMEM 2 HS and 1 FBS with NGF (10 ng/mL)
containing 19 respectively. After 4 days neurite
lengths of PC12 cells were quantified. Data are
expressed as the mean SE (n 150). P lt 0.01
vs control Dunnetts t test.
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Protocol for screening
Neurite outgrowth- promoting activity
Change medium in the absence or presence of
sample compounds
Seeding
1 day
6 day
Fixed by PFA
DMEM/10 FBS
NB/2 B27
overnight
Neuron density 5 x103 cells/cm2
Stained with anti-MAP2
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Neurite Outgrowth of Cultured Rat Cortical
Neurons Promoted by 1 and 2
control
1 (3 µmol/L)
bFGF (10 ng/mL)
2 (3 µmol/L)
Neuronal cells were first cultured in 24-well
plate in DMEM/10 FBS for 1 day at the density of
5000 cells/cm2, and then medium was changed to
serum-free Neurobasal medium plus 2 B27
supplement with or without 1, 2, ()-1, (-)-1,
and bFGF (10 ng/mL). After 6-days treatment of 1
and 2 neurons were fixed and stained with
anti-MAP2 immunohistochemical method.
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Quantitative Analysis of the Neurite Outgrowth of
1, 2, ()-1, and (-)-1
1 (µM)
2 (µM)
()-1 (µM)
(-)-1 (µM)
After the neuronal cells (5000 cell cm-2)
cultured for 7d in the presence of 0.5 EtOH,
bFGF, 1, 2, ()-1, and (-)-1 were fixed with 4
paraformaldehyde, morphometric analysis was
carried out on these neurons according to the
criteria described. The data are expressed S.E.
(n140) Dunnett's t-test vs. control, plt0.01.
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Neurite Number from Cell Body
2 (µM)
1 (µM)
After the neuronal cells (5000 cell cm-2)
cultured for 7d in the presence of 0.5 EtOH,
bFGF, 1, and 2 were fixed with 4
paraformaldehyde, morphometric analysis was
carried out on these neurons according to the
criteria described. The data are expressed S.E.
(n60) Dunnett's t-test vs. control, plt0.01.
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Synthesis of Phenylbutenoid Dimers
1. Pittaya Tuntiwachwuttikul, Brin Limchawfar,
Vichai Reutrakul, Orasa Pancharoen, Kosan
Kusamran Lindsay T. Byrne, Aust. J. Chem. 1980,
33, 913-916.
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Neurogenesis of Phenylbutenoid Dimers in OBX Mice
memory impairment deppression
Olfactory Bulbectomized (OBX )Mice
neurodegenerative model mice
olfactory bulb
OBX
Animals
ddY (8W)
Schedule
14
28
0
21
7
day
SSRI Selective Serotonin Reuptake Inhibitors
OBX
BrdU (50mg/kg I.p.)
IHC (BrdU / NeuN)
Comp.1 (50mg/kg/day p.o.) Comp.2 (50mg/kg/day
p.o.) fluoxetine (10mg/kg/day i.p.)
IHC Immunohistochemistry NeuN Neuron specific
nuclear protein BrdU 5-Bromo-2-deoxyuridine
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Protocols
Brain
hippocampal
immunohistochemistry
Confocal laser-scanning microscope
Double-labeled for NeuN and BrdU
BrdU(green)
NeuN(red)
Merged (yellow)
100µm
100µm
100µm
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Confocal microscopy images of double staining for
BrdU and NeuN in DG regions of the hippocampus.
non
sham
OBX
OBX Veh. (i.p.)
OBX FLU. (i.p.)
OBX Veh. (p.o.)
OBX 1 (p.o.)
OBX 2 (p.o.)
BrdU NeuN Merged
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Quantitative analysis of the number of BrdU and
NeuN coexpressing cells in DG regions of the
hippocampus
Comp.1 (50mg/kg/day p.o.) Comp.2 (50mg/kg/day
p.o.) fluoxetine (10mg/kg/day i.p.)
non non-operated age-mached mouse
sham operated mouse with olfactory bulb
veh 0.5 CMC
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1H NMR spectrum (600 MHz, C6D6) of 3
Cassumunarin A (7)1)
aD 71.3 (c 0.1, MeOH) IR ? max 3389
(OH), 1658 (CO), 1579 (arom.) cm-1 UV ? max
(e) 205 (44360) nm FAB-MS m/z
609MK HR-FAB-MS found 609.1819
calcd
609.1819 for C34H34O8K
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ATR
MeOH
1) Tom J. Mabry, Tetrahedron Letters, 35, No. 7,
pp. 981-984, 1994
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Cassumunarin A dC 43.6 ppm
HMBC of 3
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Plausible biosynthesis of compound 3
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Neurite Outgrowth of PC12 Cells Promoted by 3
NGF (2 ng/mL)
Ctrl (0.5 EtOH)
1 µM NGF (2 ng/mL)
10 µM NGF (2 ng/mL)
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Neurons
Dead neurons
Neuronal stem cells
Neurodifferentiation
Neuroprotective activity
Neurite outgrwth-promoting activity
Neurotrophic Compounds
from BANGLE
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Lets Meet again at Pharmacognosy-2015

• 3rd International Conference and Exhibition on
  Pharmacognosy, Phytochemistry and Natural
  Products
• October 26-28, 2015 Hyderabad, India
• Theme Advanced trends for the future of Herbal
  Drugs and Products
• Website http//pharmacognosy-phytochemistry-natur
  al-products.pharmaceuticalconferences.com/