Novel Approaches for theTreatment of Alzheimer’s
Disease
Literature Seminar
2020/06/17
M1 Atsushi Iwai

1

Contents

1. Introduction
2. AD pathology
3. AD pathology-based therapeutic target
4. Summary2

Introduction
Alzheimer’s Disease
Symptoms

•
•
•
•
•
•
•
•

Impairment of life functioning
Disorientation
Memory impairment
Speech impediment
Visuospatial cognitive impairment
Executive dysfunction
Psychological symptoms
Aloysius Alois Alzheimer•

Pathology• Degenerative loss of neurons and associated cerebral atrophy• Multiple senile plaques• Multiple neurofibrillary tangles

3Introduction
Social issues of Alzheimer's disease
Number of patients•• 47.5 million (2015)• Over 9.9 million new cases of AD-related dementia are diagnosed every year.

Social costs

•
•
•
•
•
•

Direct medical costs (e.g. nursing home care)
Direct nonmedical costs (e.g. in-home day care)
Indirect costs (e.g. lost productivity of both patient and caregiver)
Dementia costs worldwide have been calculated around $818 billion (2015)
The most costly diseases for society in developed countries

World Alzheimer Report 2015: The Global Impact of Dementia.

4Introduction
Three main hypothesis• Cholinergic hypothesis
AD is caused by reduced synthesis of the neurotransmitter acetylcholine.

• Amyloid hypothesis
Extracellular amyloid beta (Aβ) deposits are the fundamental cause of the disease.

• Tau hypothesis
Tau protein abnormalities initiate the disease cascade.

5

Introduction
Existing drugs• acetylcholinesterase inhibitor
N

O
N

NH2

TacrineN

N

O

Rivastigmine
O

O

O

O

O

N

H

Donepezil•

NMDAR antagonist

HO

Galantamine

NH2

Memantine

6

Introduction
Examples of failed clinical trials of anti-Ab drugs
NameMode of Action

Sponsor Involved in the Clinical Trials

Solanezumab

Monoclonal antibody

Eli Lilly

Crenezumab

Monoclonal antibody

Roche- Genetech

Gantenerumab

Monoclonal antibody

Roche- Genetech

Aducanumab

Monoclonal antibody

BiogenVerubecestat

BACE1 inhibitor

Merck

Lanabecestat

BACE1 inhibitor

Astra, Eli lilly

Atabecestat

BACE1 inhibitor

Janssen

Nurul Husna Ibrahim, Mohamad Fairuz Yahaya, Wael Mohamed, Seong Lin Teoh, Chua Kien and Jaya Kumar,
Front. Pharmacol., 2020, 11, 261.

Contents

1. Introduction
2. AD pathology
3. AD pathology-based therapeutic target
4. Summary8

AD pathology
The general pathways involved in neurodegenerative diseases

Van Bulck M, Sierra-Magro A, Alarcon-Gil J, Perez-Castillo A, Morales-Garcia JA,
Int. J. Mol. Sci., 2019, 20, 719.

9

AD pathology
Aβ changes expression level of mRNA and proteins• mRNA• Proteins

Calkins MJ, Manczak M, Mao P, Shirendeb U, Reddy PH., Hum. Mol. Genet., 2011, 20, 4515.

10AD pathology
Aβ oligomer localizes in mitochondria

Calkins MJ, Manczak M, Mao P, Shirendeb U, Reddy PH., Hum. Mol. Genet., 2011, 20, 4515.

11

AD pathology
Effects on mitochondria

Calkins MJ, Manczak M, Mao P, Shirendeb U, Reddy PH., Hum. Mol. Genet., 2011, 20, 4515.

12

AD pathology
Aβ coordinates metal ions

Barnham, K.J.; Masters, C.L.; Bush, A.I., Nat. Rev. Drug Dis., 2004, 3, 205.

13

AD pathology
Metal ions promote Aβ aggregation

Lovell, M. A., Robertson, J. D., Teesdale, W. J., Campbell, J. L. & Markesbery, W. R., J. Neurol. Sci., 1998, 158, 47.
Atwood, C. S., Moir, R. D., Huang, X., Scarpa, R. C., Bacarra, N. M. E., Romano, D. M., Hartshorn, M. A., Tanzi, R. E., 14
Bush, A. I., J. Biol. Chem., 1998, 273, 12817.

Appendix
Cu & Aβ promote H2O2 production
H2O2cell survival

Bush A. I. et al., J. Biol. Chem., 1999, 274, 37111.
15

AD pathology
Aβ inhibits autophagy

Menzies, F.M.; Fleming, A.; Caricasole, A.; Bento, C.F.; Andrews, S.P.; Ashkenazi, A.; Füllgrabe, J.; Jackson, A.;
Jimenez Sanchez, M.; Karabiyik, C.; et al., Neuron, 2017, 93, 1015.

16

AD pathology
PICALM is low in AD patientsd levels of phosphotau are increased in the total fraction of frontal cortex in AD, Pick disease, FTLD-MAPT P3
17
AndoofK, frontal
Tomimura
K, Sazdovitch
V, et al. Neurobiol
Dis. 2016,
94, 32
fractions from homogenates
cortexfrom control,
Pick Disease,
FTLD-MAPTP301L, FTLD-TDP,LB

AD pathology
Neuro-inflammation

Meraz-Ríos, M. A., Toral-Rios, D., Franco-Bocanegra, D., Villeda-Hernández, J., Campos-Peña, V.,
Front. Integr. Neurosci. , 2013, 7, 59.

18

AD pathology
Aβ increases levels of TNF-α & IL-1βmRNA

Loftis JM, Huckans M, Morasco BJ, Neurobiol Dis., 2010, 37, 519.

protein19

Contents

1. Introduction
2. AD pathology
3. AD pathology-based therapeutic target
4. Summary20

AD pathology-based therapeutic target
Novel treatment strategies for AD
1. Minocycline
2. 4-(1-benzylpiperidin-4-yl)thiosemicarbazone
3. Diethyl (3,4-dihydroxyphenethylamino) (quinolin-4-yl)methylphosphonate
4. Oligomannate

21

Targeting Neuroinflammation
Minocycline
In animal models

OH

O

OH O
OH

O
NH2

N

H

H

OH
N

Minocycline
Tetracycline antibiotics

•
•
•
•
•
•
•
•
•
•

Alzheimers disease (Garcez et al. 2017)
Traumatic brain injury (Hanlon et al. 2016)
Multiple sclerosis (Giuliani et al. 2005)
Cerebral ischemia (Yrjänheikki et al. 1999)
Huntingtons disease (Chen et al. 2000)
Parkinsons disease (Du et al. 2001)
Stroke (Lampl et al. 2007)
Anxiety-related behaviors (Majidi et al. 2016)
Schizophrenia (Chaudhry et al. 2012)
Anti-depressant effect (Amorim et al. 2017)22

Targeting Neuroinflammation
Clinical trial of minocycline
Success example of depression• Mild-to-moderate depression in HIV patients (Emadi-Kouchak et al. 2016)• Major depressive disorder (Dean et al. 2017)• Bipolar depression (Soczynska et al. 2017)

In Alzheimer's disease• “Minocycline did not delay the progress of cognitive or functional impairment
in people with mild AD during a 2-year period.” (Howard et al. 2020)23

Targeting Neuroinflammation
Experimental design of minocycline treatment

Amani,M.;Shokouhi,G.;Salari,A.A., Psychopharmacology, 2019, 236, 1281.

24

Targeting Neuroinflammation
Result of experiment 1

Amani,M.;Shokouhi,G.;Salari,A.A., Psychopharmacology, 2019, 236, 1281.

25

Targeting Neuroinflammation
Result of experiment 2

Amani,M.;Shokouhi,G.;Salari,A.A., Psychopharmacology, 2019, 236, 1281.

26

Targeting Neuroinflammation
The level of Cytokines

Amani,M.;Shokouhi,G.;Salari,A.A., Psychopharmacology, 2019, 236, 1281.

27

Targeting Neuroinflammation
The level of Cytokines

Amani,M.;Shokouhi,G.;Salari,A.A., Psychopharmacology, 2019, 236, 1281.

28

Multi-target drug BPT
Design strategy of multi-target drugOO

thiosemicarbazone
N
N

O

H
N

N

benzylpiperidine

N

N

S

Dp44mT

Donepezil• Chelation of redox-active metals• Induction of autophagy• Inhibition of AChE activity

combine pharmacophores

R

N

H
N

H
N
S

N

4-(1-benzylpiperidin-4-yl)thiosemicarbazone (BPT)
Palanimuthu, D.; Poon, R.; Sahni, S.; Anjum, R.; Hibbs, D.; Lin, H.Y.; Bernhardt, P.V.; Kalinowski, D.S.; Richardson, 29
D.R., Eur. J. Med. Chem. 2017, 139, 612.

Multi-target drug BPT
BPT series
4-(1-benzylpiperidin-4-yl)thiosemicarbazone (BPT)
R

N

H
N

H
N
N

S

1.
2.
3.
4.
5.

Cytotoxicity
Inhibition of AChE activity
Chelation of redox-active metals
Induction of autophagy
BBB permeability

OH

N

OH

OH

SBPT

PBPT

N

OH

NBPT

PCBT

R=
N

N
OH
OH

QBPT

8-OH-QBPT

OH

HO

OH
OH

2,3-OH-BBPT 2,3,4-OH-BBPT

Palanimuthu, D.; Poon, R.; Sahni, S.; Anjum, R.; Hibbs, D.; Lin, H.Y.; Bernhardt, P.V.; Kalinowski, D.S.; Richardson, 30
D.R., Eur. J. Med. Chem. 2017, 139, 612.

Multi-target drug BPT
Anti-proliferative activity

D. Palanimuthu et al. / European Journal of Medi

1
• TheTableanti-proliferative
activity against SK-N-MC neuroepithelioma cells

Anti-Proliferative Activity (IC50 in mM) of the Novel BPT Analogues and their Copperand Iron Complexes Against SK-N-MC Neuroepithelioma Cells, as Determined by the
IC50 (µM)
MTT Assaya.
CompoundLigand (L)CuII(L)FeIII(L2)DFO
Dp44mT
Donepezil

16.81 ± 3.87
0.013 ± 0.01
>100

9.83 ± 0.38
0.014 ± 0.01
83.33 ± 7.60

>25b
2.00 ± 0.49
>100

PBPT
SBPT
NBPT
PCBPT
QBPT
8-OH-QBPT
2,3-OH-BBPT
2,3,4-OH-BBPT>100
34.41 ± 1.24
8.86 ± 0.10
4.23 ± 1.41
17.71 ± 0.70
36.14 ± 3.24
16.85 ± 0.96
79.14 ± 0.36

46.18 ± 7.97
1.30 ± 0.14
1.81 ± 0.53
0.43 ± 0.02
3.86 ± 1.28
1.87 ± 0.50
0.40 ± 0.09
12.22 ± 0.09

>100
77.99 ± 11.58
52.90 ± 8.34
1.33 ± 0.11
1.20 ± 0.26
>100
23.99 ± 4.85
80.06 ± 13.57

aSK-N-MC cells (15,000 cells/well) were seeded in 96-well plates and incubatedfor 24 h/37 " C and the cells then treated with control medium or compounds for"
72 D.;
h/37
are R.;
shown±Bernhardt,
SD (3 experiments).
Palanimuthu,
Poon,C.R.;ICSahni,
S.; Anjum,
Hibbs,asD.;meanLin, H.Y.;
P.V.; Kalinowski, D.S.; Richardson, 31
50 valuesbIII
D.R., Eur. J. Med.ICChem.
2017,
139, 612.for the 1:1 Fe -DFO complex (as dictated by stoichiometry).
measured
50 was

the
of
BP
str
ch

2.6
cel

wh
ag
efﬁ
int
(Fi
(1)
mo
im

Multi-target drug BPT
Inhibition of AChE activity

Palanimuthu, D.; Poon, R.; Sahni, S.; Anjum, R.; Hibbs, D.; Lin, H.Y.; Bernhardt, P.V.; Kalinowski, D.S.; Richardson, 32
D.R., Eur. J. Med. Chem. 2017, 139, 612.

Multi-target drug BPT
Chelation of redox-active metals

alone
Dp44
was
cellu
PBPT
(63%
(Fig.
QBPT
to 91
QBPT
OH-B
ing 5
A
corre
take
(R2 ¼
ising
naliz

59 Fe uptake from 59 Fe-Transferrin•618 59Fe release from prelabeledcellsJournal of Medicinal• Inhibiting
D. Palanimuthu et al. / European
Chemistry 139 (2017) 612e632
and was signiﬁcantly (p < 0.001e0.01) less effective than Dp44mT
or DFO (Fig. 3B).
Overall, the BPT series demonstrated varying ability to inhibit
cellular 59Fe uptake (Fig. 3B). All of the BPT analogues weresigniﬁcantly (p < 0.001e0.01) more effective than control media
alone, but were signiﬁcantly (p < 0.001) less effective thanDp44 mT at inhibiting cellular 59Fe uptake (Fig. 3B). Notably, PBPT
was the most effective agent of the BPT series and decreased
cellular 59Fe uptake to 41% of the control (Fig. 3B). The analogues,
PBPT (41%), SBPT (57%), NBPT (68%), PCBPT (63%) and 2,3-OH-BBPT(63%), were signiﬁcantly (p < 0.001) more effective than DFO(Fig. 3B). The least effective analogues at reducing 59Fe uptake were
QBPT, 8-OH-QBPT and 2,3,4-OH-BBPT, which inhibited 59Fe uptake
to 91%, 78% and 93% of the control, respectively (Fig. 3B). While
QBPT and 8-OH-QBPT showed comparable efﬁcacy to DFO, 2,3,4OH-BBPT showed signiﬁcantly (p < 0.05) lower efﬁcacy at inhibiting 59Fe uptake relative to DFO (Fig. 3B).
As demonstrated with 59Fe mobilization experiments, no strong
correlation was observed between their ability to inhibit 59Fe uptake and either their Log Pcalc values (R2 ¼ 0.2064) or IC50 values(R2 ¼ 0.3094). In conclusion, PBPT demonstrated the most prom59 prelabeledFig. 3. ability(A) The effectof theseries of analogues
on 59Fecellularrelease fromisingof theBPTBPT
analogues
at inhibiting
Fe inter59
SK-N-MC neuroepithelioma
cells. Cells were incubated with 59Fe2-Tf (0.75 mM) for
nalization
from
Fe
-Tf.
2
"
3 h/37 C, washed four times with ice-cold PBS and then re-incubated in the presenceor absence of the control compounds (25 mM) or novel BPT analogues (25 mM) for 3 h/
2.8.
effect ofof 59theironthecomplexesof theBPTagents
seriesonthenascorbate
37 " C. The
The releaseFe fromcells mediatedby thewasassessed.
33(B)oxidation
The effect of the BPT series of analogues on 59Fe uptake from 59Fe-transferrin(59Fe2-Tf) by SK-N-MC neuroepithelioma cells. Cells were incubated with 59Fe2-Tf

Palanimuthu, D.; Poon, R.; Sahni, S.; Anjum, R.; Hibbs, D.; Lin, H.Y.; Bernhardt, P.V.; Kalinowski, D.S.; Richardson,
D.R., Eur. J. Med. Chem. 2017, 139, 612.

2.8.
oxida

A
loadi
from
critic
with
indu
To
BPT
catal
[56,6
of its
prote
the
Dp44
hand
nega
More
ascor

Multi-target drug BPT
Chelation of redox-active metals•

The effect on ascorbate oxidation• Inhibiting copper-mediated Ab1-40 aggregation

Palanimuthu, D.; Poon, R.; Sahni, S.; Anjum, R.; Hibbs, D.; Lin, H.Y.; Bernhardt, P.V.; Kalinowski, D.S.; Richardson, 34
D.R., Eur. J. Med. Chem. 2017, 139, 612.

Multi-target drug BPT
Induction of autophagy

analogues,
PBPT, PCBPT,
8-OH-QBPT,
2,3,4-OH-BBPT,
stimulate
autophagic
ﬂux in SK-N-MC
neuroepithelioma
cells as measured
using
Palanimuthu,
D.; Poon,R.; Sahni,orS.;
Anjum, R.; Hibbs,
D.; Lin,
H.Y.; Bernhardt,
P.V.; Kalinowski,
D.S.; Richardson,
35
mycin
A1 Eur.
(Baf J.
A1).
(A) Schematic
diagramthe mechanisms by which total LC3-II protein levels (a classical marker of autophagosome
D.R.,
Med.
Chem. 2017,
139,demonstrating
612.

Multi-target drug BPT
BBB permeability•

Evaluation of physicochemical parameters to cross the blood brain barrier (BBB)
1. Lipinski's Rule of Five (Oral bioavailability)••••

Hydrogen bond donors (OH and NH)  5
Hydrogen bond acceptors (N and O)  10
Molecular weight  500
LogP  5

2. Successful CNS agents• Topological polar surface area < 90 2• 2 < cLogP <5• LogBB > -1 (LogBB = -0.0148 TPSA + 0.152  cLogP +0.139)

Palanimuthu, D.; Poon, R.; Sahni, S.; Anjum, R.; Hibbs, D.; Lin, H.Y.; Bernhardt, P.V.; Kalinowski, D.S.; Richardson,
D.R., Eur. J. Med. Chem. 2017, 139, 612.
36
D.E. Clark, J. Pharm. Sci., 1999, 88, 815.

Multi-target drug BPT
Summary of Multi-Target Drug Therapy
OH
N
N

H
N

OH

H
N
S

N

pyridoxal 4-(1-benzylpiperidin-4-yl)thiosemicarbazone (PBPT)• PBPT is best of the 8 compounds.
• Low anti-proliferative activity.
• Moderate AChE inhibitory activity.
• Favorable iron chelation properties.
• The inhibition of Fe(III)-mediated ascorbate oxidation.
• The inhibition of Cu(II)-mediated aggregation of Ab1-40.
• Increase autophagic initiation.
• The physicochemical properties of PBPT are favorable for CNS permeation.

Lead compound with promising multi-functional activity
to treat the complex pathology associated with AD.

37

Mitochondrial dysfunction target drug DDQ
Design strategy of Mitochondrial dysfunction target drug
• Existing Drp1 inhibiting drugs
OH

O
N
H

N

Dopamine based structure

OH

Dynasore

OH
O

HO

H
N

O
P

O
O

HO
Cl

O

N
N

N

• New Drp1 inhibiting drugs

SHCl

Mdivi1

Diethyl (3,4-dihydroxyphenethylamino)(quinolin-4-yl)methylphosphonate (DDQ)

OH
O
P

O
OH

MitoQ
Kuruva, C.S.; Manczak, M.; Yin, X.; Ogunmokun, G.; Reddy, A.P.; Reddy, P.H., Hum. Mol. Genet. 2017, 26, 3375.

38

Mitochondrial dysfunction target drug DDQ
Experimental design

Kuruva, C.S.; Manczak, M.; Yin, X.; Ogunmokun, G.; Reddy, A.P.; Reddy, P.H., Hum. Mol. Genet. 2017, 26, 3375.

39

3380

| Human Molecular Genetics, 2017, Vol. 26, No. 17
Mitochondrial
dysfunction target drug DDQTable 2. mRNA fold changes of mitochondrial structural, mitochondrial biogenesis and synaptic genes in Human Neuroblastoma (SHSY5Y)
cells treated with DDQ, Ab, AbþDDQ and DDQ þ Ab relative to the untreated SHSY5Y cells and cells treated with AbþDDQ and DDQ þ Ab relative
to the Ab-treated SHSY5Y cells

mRNA levels

GenesmRNA fold changes compare with untreated cells
DDQ

Mitochondrial Structural genes
Drp1
#2.2*
Fis1
#4.4***
Mfn1
1.7*
Mfn2
2.3**
Synaptic genes
Synaptophysin
1.4*
PSD95
1.4*
Synapsin1
1.0
Synapsin2
1.7*
Synaptobrevin1
1.0
Synaptobrevin2
1.3
Synaptopodin
1.0
GAP43
1.2
Mitochondrial Biogenesis genes
PGC1a
1.5*
Nrf1
1.9*
Nrf2
2.7*
TFAM
1.7*

mRNA fold changes compare with Ab-treated cellsAbAbþDDQDDQþAbAbþDDQDDQþAb2.2**
1.7*#2.3**#2.6**

1.9*
#1.2
1.4
1.3

#2.1*
#1.3
1.2
1.1

#1.7*
#2.0*
3.3**
3.4**

#4.6***
#2.5*
2.9**
2.9**

#3.7***#2.5**#1.9*#1.4*#2.4**#2.3*#2.3**#1.9**

#2.4*
#1.4*
1.1
1.1
#1.1
1.0
#1.1
1.1

#1.6*
#1.2
1.3
1.8*
#1.4
1.0
#1.1
1.1

1.6*
1.8*
2.1*
2.4*
2.0*
2.1*
2.0*
1.7*

2.4*
2.1*
2.5*
2.9**
1.7*
2.3*
2.1*
1.7*

#4.4**#4.1**#2.8*#4.6***

#1.8*
#1.7*
#1.3
#2.0*

#1.1
1.1
#1.1
#1.5*

2.5*
2.6**
2.1*
2.3*

3.9**
4.9***
2.6*
3.2**

cells relative to untreated cells. Decreased levels of Drp1 (P ¼ 0.004)
synaptic gene expression levels were upregulated in cells incuwere found in DDQ þ Ab-treated cells relative to untreated cells
bated with Ab and then treated with DDQ and pretreated with
(Fig. 4A and C). Overall, these findings suggest that DDQ reduces
DDQ and incubated with Ab relative to Ab-treated cells. This
40
Kuruva, C.S.;
Manczak,M.; Yin,synaptic
X.; Ogunmokun,
G.;
Reddy, A.P.; Reddy,
Hum.
Mol.
Genet.fusion
2017,activity
26, 3375.
fissionP.H.,
activity
andenhancesin the presence of Ab.
indicatesthat DDQ
is protecting
genes fromAb.

Mitochondrial dysfunction target drug DDQ
Protein levels

Kuruva, C.S.; Manczak, M.; Yin, X.; Ogunmokun, G.; Reddy, A.P.; Reddy, P.H., Hum. Mol. Genet. 2017, 26, 3375.

41

Mitochondrial dysfunction target drug DDQ
Mitochondrial functional assays

Kuruva, C.S.; Manczak, M.; Yin, X.; Ogunmokun, G.; Reddy, A.P.; Reddy, P.H., Hum. Mol. Genet. 2017, 26, 3375.

42

Mitochondrial dysfunction target drug DDQCell viability analysis

N

reduction

N

N N

N

N

N

S

MTT

NH
N

N

S
Human Molecular Genetics, 2017, Vol. 26, No. 17| 3389gure 12. Cell viability analysis. Cell viability of human neuroblastoma (SHSY5Y) cells, while treated with DDQ, Ab, AbþDDQ and DDQ þ Ab relative to untreated cells.

ell viability of pre-treated DDQ and post treated DDQ in Ab-incubated SHSY5Y cells relative to Ab-treated cells.

Kuruva, C.S.; Manczak, M.; Yin, X.; Ogunmokun, G.; Reddy, A.P.; Reddy, P.H., Hum. Mol. Genet. 2017, 26, 3375.

43Mitochondrial dysfunction target drug DDQTransmission electron microscopy

e 10. Electron microscopy of SH-SY5Y cells. We quantified mitochondrial architectures within the cell in all 5 groups to identify mitochondrial number and m

gy. Average number of mitochondria per cell is shown in graphs. Error bars indicate the standard deviation. Mitochondrial number is significantly decrease
treated SH-SY5Y cells, relative to untreated cells. On the contrary, mitochondrial number is significantly increased Ab-treated SH-SY5Y cells. DDQ-pre and p

44 for all gro
d cells in the presence of Ab showed reduced mitochondrial number compared to cells treated with Ab alone. Mitochondrial length was measured

Kuruva, C.S.; Manczak, M.; Yin, X.; Ogunmokun, G.; Reddy, A.P.; Reddy, P.H., Hum. Mol. Genet. 2017, 26, 3375.

s. Mitochondrial length was significantly reduced Ab-treated SH-SY5Y cells. DDQ-pre and post-treated cells in the presence of Ab showed increased mitoch

Mitochondrial dysfunction target drug DDQ
Interaction between Aβ and Drp1

Kuruva, C.S.; Manczak, M.; Yin, X.; Ogunmokun, G.; Reddy, A.P.; Reddy, P.H., Hum. Mol. Genet. 2017, 26, 3375.

45

Mitochondrial dysfunction target drug DDQ
The localization of Drp1 and Aβ

Kuruva, C.S.; Manczak, M.; Yin, X.; Ogunmokun, G.; Reddy, A.P.; Reddy, P.H., Hum. Mol. Genet. 2017, 26, 3375.

46

Mitochondrial dysfunction target drug DDQ
Summary of DDQ
HO

O

H
N

P

O
O

HO
N

Diethyl (3,4-dihydroxyphenethylamino)(quinolin-4-yl)methylphosphonate (DDQ)• Bound at Ab and Drp1 interacting sites to Inhibit Ab and Drp1 complex formation• showed better docking score.
• Enhanced fusion activity, reduced fission machinery, and increased mitochondrial
biogenesis and synaptic activities.
• Reduces the levels of Ab and Drp1 and interaction between Ab and Drp1.
• Maintains mitochondrial function and cell viability.
• DDQ can reduce the negative effects of Aβ• Prevention is better than treatment.

Promising molecule to treat AD neurons.

47

Approved novel AD treatment
AD & gut microbiota

Wang, X., Sun, G., Feng, T. et al. Cell Res., 2019, 29, 787.

48

Approved novel AD treatment
AD progression is associated with the alteration of gut microbiota• The gut microbiome composition of WT and Tg mice

• Aβ & Tau level of Tg mice

Wang, X., Sun, G., Feng, T. et al. Cell Res., 2019, 29, 787.

49

Approved novel AD treatment
GV-971 exhibits ameliorative effects on cognitive impairment

The escape latency time resultsof the Morris Water Maze

Wang, X., Sun, G., Feng, T. et al. Cell Res., 2019, 29, 787.

The number of platform-sitecrossovers in MWM test50Approved novel AD treatment
GV-971 alleviates neuroinflammation by shaping the gut microbiota
gut microbiota

Wang, X., Sun, G., Feng, T. et al. Cell Res., 2019, 29, 787.

brain

51

Approved novel AD treatment
oligomannate(Shanghai Green Valley Pharmaceuticals)

2003 Memantine was approved
16 years2019 Oligomannate was approved in China
2020 The Phase 3 clinical trial in U.S.A & Europe
52

Contents

1. Introduction
2. AD pathology
3. AD pathology-based therapeutic target
4. Summary

53

Summary• In addition to Aβ and tau, multiple factors are involved in Alzheimer's disease.
• Drug discovery research on various causative factors of Alzheimer's disease isbeing conducted.

54

55

Appendix
Dynamic biomarkers of the Alzheimer’s pathological cascade

John Q Trojanowski, et al., Lancet Neurol. , 2010, 9, 119.
56

Appendix
Proteolytic cleavage of APP

Dileep K. Vijayan, and Remya C, Current Drug Targets, 2019, 20, 148.
PJ Ward, et al., Science, 1990, 248, 1122.
BA Yankner, et al., Proc. Natl. Acad. Sci. U.S.A., 1999, 96, 6959.
BA Yankner, et al., Nat. Natl. Cell Biol., 2000, 2, 463.
57
W Song, et al., FASEB J., 2006, 20, 285.

Appendix
Cu & Aβ generate H2O2
O2–/H2O2

O2
Catechols
Vitamin C
Cholesterol,arachidonic
acidand so on
Drugs

e–

Cu+

Cu2+

Fe2+/Cu+

OH•

Aβ
SOD
α-synuclein
α-crystallin

Lipid peroxidation
Protein oxidation
Protein aggregation
DNA/RNA adducts

Figure 1 | ROS generation by abnormal reaction of O2 with protein-bound Fe or Cu. One of
the consequences of normal aging is that the levels of the redox-active metals copper and iron
Barnham,
K.J.; Masters,
A.I., Nat. Rev. Drug
2004, that
3, 205.
in the brain increase.
This increase
could C.L.;
leadBush,
to hypermetallationofDis.,
proteins
normally bind 58
redox-active metals at shielded sites. Adventitial binding — for example; at a loading site — will

In v
will
tion
tion
nor
the
Dur
(30
exp
in th
T
rela
gen
sou
the
whe
T
hom

Appendix
Aβ Reduces Cu()Bush A. I. et al., J. Biol. Chem., 1999, 274, 37111.

59Appendix
3D and 2D docking diagrams showing the binding mode

Palanimuthu, D.; Poon, R.; Sahni, S.; Anjum, R.;
Hibbs, D.; Lin, H.Y.; Bernhardt, P.V.; Kalinowski,
D.S.; Richardson, D.R., Eur. J. Med. Chem. 2017,
139, 612.

60

Appendix
X-ray crystal structures of [Cu(SBPT)Cl2]

N
OH

H
N

H
N
S

N

SBPT

Palanimuthu, D.; Poon, R.; Sahni, S.; Anjum, R.;
Hibbs, D.; Lin, H.Y.; Bernhardt, P.V.; Kalinowski,
D.S.; Richardson, D.R., Eur. J. Med. Chem. 2017,
139, 612.

61Appendix
Bafilomycin’s effect

Palanimuthu, D.; Poon, R.; Sahni, S.; Anjum, R.; Hibbs, D.; Lin, H.Y.; Bernhardt, P.V.; Kalinowski, D.S.;
Richardson, D.R., Eur. J. Med. Chem. 2017, 139, 612.

62

Appendix
Immunofluorescence analysis (DDQ)

Kuruva, C.S.; Manczak, M.; Yin, X.; Ogunmokun, G.; Reddy, A.P.; Reddy, P.H., Hum. Mol. Genet. 2017, 26, 3375.

63

Appendix
Principal component analysis

https://logics-of-blue.com/principal-components-analysis/64Appendix
iPSC-Based Compound Screening

Anti-Aβ Cocktail of BCroT (bromocriptine, cromolyn, topiramate )Inoue H. et al., Cell Rep. 2017, 21, 2304.

65