recently drugs are being tested for the other diseases also the same thing we also had followed and studied about this above given drug

1. Presented by-
ANAND REDDY-18MSB0101
DEEPAK KUMAR-18MSB0100
B.NARESH-18MSBO105
HARSHAVARDHAN18MSB0041
SUBMITED TO
DR.SUDHARAMAIAH
PROFESSER SBST

2.  A genetic disease of Duchenne muscular dystrophy (DMD) is the most
common X-liked disorder
 Muscular dystrophy in children, presenting in early childhood and
characterized by muscle weakness and calf hypertrophy in affected male
children
 Presently there is no permanent treatment available at a commercial level
for this disease.
 DMD affects about one in 5,000 males at birth.

3.  Drug repurposing is re-investigating existing drugs that failed approval for
new therapeutic indications
 Repurposed drugs may have a good effectiveness record with their
primary condition.
 It takes almost 9-12yrs to introduced a new drug for particular disease but
re-purposing impact only take 2-3 yrs.
 Using drug repurposing, pharmaceutical companies have achieved a
number of successes for example pfizer’s viagra(sidenafil) is a medication
used to treat erectile dysfunction and pulmonary arterial hypertension.

4. METHODOLOGY
 For this study on duchenne muscular dystrophy we obtained the mapped
genes list using EBI- GWAS (genome-wide association study).
 It is an observational study of a genome-wide set of genetic variants in
different individuals to see if any variant is associated with a trait.
 GWAS typically focus on associations between single-nucleotide
polymorphisms (SNPs) and traits like major human diseases but can
equally be applied to any other genetic variants and any other organisms.
 The obtained mapped genes which serve as marker for the muscular
dystrophy disease are used to generate whole genomic and proteomic data
associated with those genes using GeneAlaCart.

5.  Data obtained from this genecard is vast and need to be filtered manually
to obtained relation among genes and pathways obtained .
 Adenylate cyclase which is coded by gene ADCY is a membrane bound
enzyme plays a key role in many signaling pathways likes MAPK, G
protein coupled signalling pathways.

6. Activation of signalling pathways due to effect in Dystrophin
Glycoprotein Complex:
Over-expresion of cAMP signalling , MAPK signalling
exacerbate disease progression(DMD)
 Adenylate cyclase which is responsible for conversion of ATP to
cyclicAMP (a secondary messenger) is targeted to inhibit the excess
activation of signalling pathways .


7. Search for ligands:
 Compounds which can act on the target proteins are obtained from zinc
database in sdf format and the suitable ligands are combined with each
target for docking.
 FlexX docking from BioSolveIT is performed to predict the position and
orientation of a ligand to the protein
 Later the score obtained are compared to the ligand which are originally
proposed act against these targets

8. Results

9. Genome wide association studies for Duchenne muscular dystrophy
The mapped genes associated with the disease are obtained using GWAS
catalog : 22 genes
XYLT1 SHKBP1
NUP93 OR5AN2P
TAPT1-AS1 SATB1
MICALL2 MIR138-2
SPEF2 RPL10P1
UNCX SYNGR2P1
SNCA EIF4A1P1
FAM173B NCAPGP2
SLFN12L SLC25A48
RNU6-280P ADCY8
RN7SL435P ADCY1

10. Pathways Genes associated
Metabolism ADCY8,ADCY1,SNCA,RGS6,NU
P93,CYB5A,DGKB
Signaling pathways:
MAPK signaling
Apelin signaling pathway
Ras signaling pathway
Phospholipase D signaling pathway
ADP signalling through P2Y
purinoceptor 12
Activation of cAMP-Dependent PKA
Signaling by GPCR
Signaling by Hedgehog
Calcium signaling pathway
ADCY1,ADCY8
ADCY1, SLC8A1,ADCY8
ADCY1,ADCY8
DGKB,ADCY1,ADCY8,
ADCY1,ADCY8,RGS6
ADCY1,ADCY8,PTPN1,RGS6
ADCY1,ADCY8,RGS6, PTPN1,
DOCK1, DGKB
ADCY1,ADCY8
ADCY1, SLC8A1

11. Secretions:
Pancreatic Secretion
Bile Secretion
Salivary Secretion
Parathyroid Hormone, Secretion And
Action
ADCY1,ADCY8
ADCY1,ADCY8
ADCY1,ADCY8
ADCY1,ADCY8
innate immune system ADCY1,ADCY8,DOCK1,NUP93,PTP
N1
human cytomegalovirus infection ADCY1, SLC8A1,ADCY8,NUP93

12. 2.Duchenne muscular dystrophy disease associated genes and their role
in various functions
 The data obtained in GeneAlaCart is filtered to screen specific relation
among genes and associated pathways.
GENES NUMBER OF PATHWAYS
XYLT1 13
SLC8A1 16
SATB1 30
RGS6 6
PTPN1 14
NUP93 46
JAM2 63
DOCK1 21
DGKB 39
CYB5A 6
ADCY1 175
ADCY8 143

13.  Genes and their pathways associated.
2% 3%
5%
1%
2%
8%
11%
4%
7%
2%
1%
24%
30%
XYLT1 SNCA SLC8A1 SATB1 RGS6 PTPN1 NUP93
JAM2 DOCK1 DGKB CYB5A ADCY8 ADCY1

14. GENES BIOLOGICAL PROCESS
XYTL1 8
SNCA 88
SLC8A1 47
SLC25A48 3
SHKBP1 2
SATB1 12
RGS6 6
PTPN1 27
MIR138-2 19
NUP93 15
MICALL2 7
UNCX 12
FAM173B 3
JAM2 4
DOCK1 14
DGKB 14
CYB5A 4
CLSTN2 4
ADCY8 12

15.  Genes and the number of biological process involved
2%
27%
15%
1%1%
4%
2%
9%
5%
5%
2%
4%
1%
1%
4%
4%
1%
1%
4%
6%
XYLT1 SNCA SLC8A1 SLC25A48 SHKBP1 SATB1 RGS6
PTPN1 MIR138-2 NUP93 MICALL2 UNCX FAM173B JAM2
DOCK1 DGKB CYB5A CLSTN2 ADCY8 ADCY9

16. genes Molecular functions
XYLT1 5
SNCA 29
SLFN12L 1
SLC8A1 10
SLC25A48 8
SHKBP1 1
SATB1 9
RGS6 2
PTPN1 15
MIR138-2 1
NUP93 2
MICALL2 6
UNCX 3
FAM173B 3
JAM2 2
DOCK1 5
DGKB 7
CYB5A 5
CLSTN2 1
ADCY8 6
ADCY1 9

17. 4%
22%
1%
8%
6%
1%7%
2%
12%
1%
2%
5%
2%
2%
2%
4%
5%
4%
1% 5%
7%
Genes and number of molecular functions
XYLT1 SNCA SLFN12L SLC8A1 SLC25A48 SHKBP1
SATB1 RGS6 PTPN1 MIR138-2 NUP93 MICALL2
UNCX FAM173B JAM2 DOCK1 DGKB CYB5A

18. 3.Ligand and the target protein interactions:
 Six compounds are obtained from zinc database along with 2D structure
(sdf format)
 Zinc 504 (mianserin hydrochloride - MHC) – FDA approved drug used for
anti-depression.
 MHC is indirectly associated with adenylate cyclase in provoking
neurotransmitters in brain and hence considered for repurposing for DMD

19. Zinc20245-ADCY1 ZINC504-ADCY1
ZINC2048-ADCY1
ZINC968257-ADCY1 ZINC1530611-ADCY1ZINC1530741-ADCY1
Molecular docking of six compounds with ADCY1protein using
flexX

20. Zinc20245-ADCY8 ZINC504-ADCY8 ZINC20248-ADCY8
ZINC968257-ADCY8 ZINC1530611-ADCY8 ZINC1530741-ADCY8
ZINC2048-ADCY8
Molecualr docking of 6 compounds with ADCY8 protein using
FlexX

21. 3. Mianserin hydrochloride ligand interaction with target
ADCY8 protein
SCORE -7.9292
kcal/mol
MATCH -7.3864
LIPO -6.5316
CLASH -3.30053

22. Binding affinity= -7.9292kcal/mol

23. SCORE -14.8140
MATCH -9.2980
LIPO -16.0042
CLASH 8.5724

24. Binding affinity=-14.8140kcal/mol

25. Conclusion
 Duchenne muscular dystrophy is one of the most lethal genetic disorder in
humans and there is no permanent cure
 In our study we proposed a strategy, to inhibit a crucial protein adenylate
cyclase (ADCY1 and ADCY8) which is over expressed in DMD patients
only
 Its involved in MAPK signaling pathway and cAMP pathway
 The drug that can be repurposed against ADCY1 and ADCY8 for DMD are
MHC and NHC
 Docking studies using FlexX shows that NHC shows high binding affinity
towards ADCY1
 Docking studies using FlexX shows that MHC shows high binding affinity
towards ADCY8
 Further molecular dynamics simulation (MDS), principal component
analysis (PCA) and invitro/invivo studies are required to confirm their
stability and activity against the selected drug targets

26.  R. J. Marshall, Department of Pharmacology, Organon Laboratories Ltd.,
Newhouse, Lanarkshire MLI 5SH, Scotland, UK
 Chiavegatti T, Costa VL Jr, Araújo MS, Godinho RO.Br J Pharmacol. 2008
Mar;153(6):1331-40. Epub 2007 Dec 24. PMID: 18157164
 Beytía Mde L, Vry J, Kirschner J.Acta Myol. 2012 May;31(1):4-8. Review.
PMID: 22655510
 Rodan GA, Rodan SB, Raible DG, Cutler LS, Wacholtz M, Sha'afi RI. Ann
N Y Acad Sci. 1979;317:670-91.PMID: 157710
 Narayanan R. Druggable vitiligo genome: a fast track approach to take the
genome wide association to the clinic. MOJ Proteomics Bioinform.
2015;2(3):102‒110.
 Mullard A. Nat Rev Drug Discov. 2012 Jun 29;11(7):505-6. doi:
10.1038/nrd3776. PMID: 22743966