About OMICS GroupAbout OMICS Group

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

About OMICS Group About OMICS Group ConferencesConferences

OMICS Group International is a pioneer and leading science event OMICS Group International is a pioneer and leading science event organizer, which publishes around 400 open access journals and organizer, which publishes around 400 open access journals and conducts over 300 Medical, Clinical, Engineering, Life Sciences, conducts over 300 Medical, Clinical, Engineering, Life Sciences, Phrama scientific conferences all over the globe annually with the Phrama scientific conferences all over the globe annually with the support of more than 1000 scientific associations and 30,000 editorial support of more than 1000 scientific associations and 30,000 editorial board members and 3.5 million followers to its credit.board members and 3.5 million followers to its credit.

OMICS Group has organized 500 conferences, workshops and national OMICS Group has organized 500 conferences, workshops and national symposiums across the major cities including San Francisco, Las Vegas, symposiums across the major cities including San Francisco, Las Vegas, San Antonio, Omaha, Orlando, Raleigh, Santa Clara, Chicago, San Antonio, Omaha, Orlando, Raleigh, Santa Clara, Chicago, Philadelphia, Baltimore, United Kingdom, Valencia, Dubai, Beijing, Philadelphia, Baltimore, United Kingdom, Valencia, Dubai, Beijing, Hyderabad, Bengaluru and Mumbai.Hyderabad, Bengaluru and Mumbai.

Neurotrophic Compounds of Javanese Ginger, Zingiber purpureum

Yoshiyasu Fukuyama

Pharmaceutical Sciences, Tokushima Bunri UniversityTokushima, Japan

Pharmacognosy 2014, BeijingTrack 9, 15:00-15:20Aug., 26, 2014

Drugs for Treatment of Alzheimer’s Disease

Neurotrophins (NGF, BDNF, NT-3, NT-4/5 )

Ageing

Astrocyte

Oligodendrocyte

Neurons

Death

MaturationDevelopment

Differentiation

Neuronal precursor cells

Neuronal stem cells

Effects of Neurotrophins

1. 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

Neurotrophic factors

Nonpeptidyl small molecule neurotrophic compounds

PC12 cells used for Screening neurotrophic/neuroprotective/cytotoxic compounds

Cytotoxic

Neurotrophic

Neuroprotective

after NGF withdrawal

“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.” Science, 296(5573):1648 (2002)

The Principles for Screening Neurotrophic/Neuroprotective 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

Rat pheochromocytomacell line (PC12 cells)

Primary cultured rat neurons (Cortical neurons)

Search for Neurotrophic Mimic Natural Products

Neurotrophic Compounds Library

Purpose:Rhizome: Fever, headache Cough, expectorate Stomach ache Constipation Jaundice Rheumatism The ingredients of herbal medicine for women after childbirth.

Leaf: Revitalizer Digestive trouble

BANGLE (Zingiber purpureum)

This plant is used as a spice and also used for traditional Indonesian medicine ‘jamu’.

NGF-like Activity of BANGLE in PC12 Cells

Ctrl ( 0.5% EtOH )

MeOH ext (25 μg/mL)

Absence of NGF

BANGLE (Zingiber purpureum)

PC12 Cells

: Neurotrophic activity (+)

Fr.4 ( 12.5 μg/mL ) Fr.5 ( 12.5 μg/mL )Ctrl ( 0.5% EtOH )

Screening Result of Compounds from BANGLE by PC12 Cells

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. ## P<0.01 vs control; by Chi-square test

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 < 0.01 vs control; Dunnett’s t test.

Ave

rage

of

neu

rite

len

gth

(μ

m

)

Neurite Length of NGF-differentiated PC12 Cells Effected by 1

1

Neurite outgrowth- promoting activity

Protocol for screening

Seeding1 day 6 day

Fixed by PFA

overnight

Change mediumin the absence or presence

of sample compounds

Stained withanti-MAP2

DMEM/10 % FBS NB/2 % B27

Neuron density : 5 x103 cells/cm2

control

bFGF (10 ng/mL)

1 (3 μmol/L)

2 (3 μmol/L)

Neurite Outgrowth of Cultured Rat Cortical Neurons Promoted by 1 and 2

Neuronal cells were first cultured in 24-well plate in DMEM/10% FBS for 1 day at the density of 5000 cells/cm 2, 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.

Quantitative Analysis of the Neurite Outgrowth of 1, 2, (+)-1, and (-)-1

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. (n=140); Dunnett's t-test vs. control, **p<0.01.

1(μM)

2(μM)

(+)-1(μM)

(-)-1(μM)

Neurite Number from Cell Body

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. (n=60); Dunnett's t-test vs. control, **p<0.01.

1(μM)

2(μM)

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.

14 28 day

AnimalsddY (8W)

Schedule

0 217

BrdU (50mg/kg I.p.)OBX

IHC (BrdU / NeuN)

IHC : ImmunohistochemistryNeuN: Neuron specific nuclear proteinBrdU: 5-Bromo-2’-deoxyuridine

Comp.1 (50mg/kg/day p.o.)Comp.2 (50mg/kg/day p.o.)fluoxetine (10mg/kg/day i.p.)

Neurogenesis of Phenylbutenoid Dimers in OBX Mice

Olfactory Bulbectomized (OBX )Mice

OBX

olfactory bulb

memory impairmentdeppression

neurodegenerative model mice

SSRI : Selective Serotonin Reuptake Inhibitors

Protocols

Double-labeled for NeuN and BrdU

Confocal laser-scanning microscope

Brain hippocampal

BrdU(green) NeuN(red) Merged (yellow)

immunohistochemistry

Confocal microscopy images of double staining for BrdU and NeuN in DG regions of the hippocampus.

BrdU NeuN Merged

OBXFLU. (i.p.)

OBXVeh. (p.o.)

OBX1 (p.o.)

OBX2 (p.o.)

non sham OBX OBXVeh. (i.p.)

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

1H NMR spectrum (600 MHz, C6D6) of 3

1) Tom J. Mabry, Tetrahedron Letters, 35, No. 7, pp. 981-984, 19941) Tom J. Mabry, Tetrahedron Letters, 35, No. 7, pp. 981-984, 1994

Cassumunarin A (7)1)Cassumunarin A (7)1)

[α]D = + 71.3 (c 0.1, MeOH)IR ν max : 3389 (OH), 1658 (C=O), 1579 (arom.) cm-1 UV λ max (ε) : 205 (44360) nm FAB-MS : m/z 609[M+K]+

HR-FAB-MS : found 609.1819 calcd 609.1819 for C34H34O8K

[α]D = + 71.3 (c 0.1, MeOH)IR ν max : 3389 (OH), 1658 (C=O), 1579 (arom.) cm-1 UV λ max (ε) : 205 (44360) nm FAB-MS : m/z 609[M+K]+

HR-FAB-MS : found 609.1819 calcd 609.1819 for C34H34O8K

2828

ATRATR

MeOHMeOH

HMBC of 3Cassumunarin A

δC = 43.6 ppmCassumunarin A

δC = 43.6 ppm

Plausible biosynthesis of compound 3

Neurite Outgrowth of PC12 Cells Promoted by 3

1 μM + NGF (2 ng/mL)1 μM + NGF (2 ng/mL) 10 μM + NGF (2 ng/mL)10 μM + NGF (2 ng/mL)

NGF (2 ng/mL)NGF (2 ng/mL)Ctrl (0.5% EtOH)Ctrl (0.5% EtOH)

Neuronal stem cellsNeurons

Dead neurons

NeurodifferentiationNeuroprotective activity

Neurite outgrwth-promoting activity

from BANGLE

Neurotrophic Compounds

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