Synthesis and biological investigation of pyrimido 1,2 a 1,3,5 triazine and its analogues 2

12 Dihydroorotate dehydrogenase, mitochondrial Nucleotide transport and metabolism 2fpy, 1d3h, 2prl, 1tv5, 3i6r 13 Cell division protein kinase 2 Involved in protein kinase activity 1p

Table XVIII Retrieval of top targets of 149b by 3 approaches:

Pharm Mapper (fit score)

Reverse Screen 3d score

1 Epidermal growth factor

3.473 0.9375

1odj

5 Leukotriene A-4 hydrolase Hydrolyzes an epoxide moiety of leukotriene A4

(LTA-4) to form leukotriene B4 (LTB-4) 1gw6, 3fum 3.413 0.4483

6 Corticosteroid

11-beta-dehydrogenase isozyme 1

Catalyzes reversibly the conversion of cortisol to

the inactive metabolite cortisone

3czr, 3czr,

7 Aspartate aminotransferase Amino acid transport and metabolism 1c9c, 2z9v, 3.131 0.6842

8 Thymidylate synthase Nucleotide transport and metabolism

1tls,1j3j, 1tlc, 3jsu, 2blc, 3clb, 3k2h, 2aaz, 1sej, 1ju6, 1qzf, 1f28, 1jtq, 1f4d,

9 Mitogen-activated protein

kinase 14 Involved in MAP kinase activity

3e92, 3fc1, 1ouy,3new, 3fl4, 1w82

3.024 0.4324

11 Androgen receptor

Steroid hormone receptors are ligand-activated transcription factors that regulate eukaryotic gene expression and affect cellular proliferation

1z95, 2ax8, 2piq, 3b66 2.994 0.281

12

Dihydroorotate dehydrogenase, mitochondrial

Nucleotide transport and metabolism

2fpy, 1d3h, 2prl, 1tv5, 3i6r

13 Cell division protein kinase

2 Involved in protein kinase activity

1p2a, 3eoc, 2v22, 1vop 2.979 0.36111

14 Serine/threonine-protein

kinase Chk1

Required for checkpoint mediated cell cycle arrest in response to DNA damage or the presence of unreplicated DNA May also negatively regulate cell cycle progression

Recognizes the substrate consensus sequence

2.963 0.3871

1nhu, 2f9v, 1yvx, 1nhv, 2a4g, 2pxa

2.942 0.3333

17

Tyrosine-protein phosphatase non-receptor

type 1

May play an important role in CKII- and src-induced signal transduction cascades

p60c-1no6, 1nny, 3o5x, 1c85, 2shp

2.936 0.2917

18 Dipeptidyl peptidase 4 Removes N-terminal dipeptides sequentially

from polypeptides having unsubstituted

N-1rwq, 2gbi, 2iit, 1rwq,

2.931 0.5667

termini provided that the penultimate residue is proline Plays a role in T-cell activation

2aj8

19

2-C-methyl-D-erythritol

2,4-cyclodiphosphate synthase

phosphorus-oxygen lyase participates in

biosynthesis of steroids 1h48, 3ke1 2.927 0.3636

20 Retinoic acid receptor

unstressed cells

23 Collagenase 3 Degrades collagen type I Does not act on gelatin

or casein Could have a role in tumoral process 1xud, 3kek 2.916 0.359

25

methylglutaryl-coenzyme

3-hydroxy-3-A reductase

This transmembrane glycoprotein is the limiting enzyme of sterol biosynthesis

rate-2q1l, 3cd0, 3cd7, 3cd5, 3cct

2.855 0.4412

31

3-phosphoinositide-dependent protein kinase 1

Phosphorylates and activates not only PKB/AKT, but also PKA, PKC-zeta, RPS6KA1 and RPS6KB1 May play a general role in signaling processes and in development

1uu3, 2xck, 3nax, 2v4l, 3oaw, 3ml8, 3dpd, 3ml9

2.852 0.3409

1e7v, 2chz, 2chx, 2chw, 1e8x, 3ls8

32 Signal recognition particle

34 Pantothenate synthetase Coenzyme transport and metabolism 1n2h, 3ivc 3.106 0.4571

35 UDP-glucose 4-epimerase Cell wall/membrane/envelope biogenesis 1lrj, 1lrl,

36

dimethylbenzimidazolepho

Nicotinate-nucleotide-sphoribosyltransferase

37 Kinesin like protein kif11

Motor protein required for establishing a bipolar spindle Blocking of KIF11 prevents centrosome migration and arrest cells in mitosis

3k5e, 2gm1, 2uym, 3cjo 2.992 0.491

38 Lactoylglutathione lyase Catalyzes the conversion of hemimercaptal,

formed from methylglyoxal and glutathione, to

1fro, 2za0 2.908 0.5517

S-lactoylglutathione

39 Hepatocyte growth factor

receptor Involved in hepatocyte growth factor activity 3cd8, 3c1x 2.901 0.3478

41 TGF-beta receptor type-1

On ligand binding, forms a receptor complex consisting of two type II and two type I transmembrane serine/threonine kinases It then bind and activate SMAD transcriptionalcomplex, enters the nucleus where it binds transcription cofactors and causes the

2.948 0.5862

43

1-aminocyclopropane-1-carboxylate deaminase Amino acid transport and metabolism 1j0e, 1j0b 2.867 0.5

44 Cell division protein kinase

Table XIX Retrieval of top targets of 149f by 3 different VS approaches

Pharm Mapper (fit score)

Reverse Screen 3d score

1 Nucleoside diphosphate

kinase

enzymes that catalyze the exchange of phosphate groups between different nucleoside diphosphates

1s59, 1mn9 3.907 0.2703

2 Purine nucleoside

phosphorylase Nucleotide transport and metabolism

1lv8, 1ovg, 1rfg, 1odj, 3iom, 2ai1

3

Phosphoenolpyruvate carboxykinase, cytosolic

[GTP]

Energy production and conversion

1m51, 1nhx 1khb

3.569 0.3429

4 UDP-glucose 4-epimerase Cell wall/membrane/envelope biogenesis

1lrj, 1hzj 2p5y, 2c20, 3ehe, 3enk

3.397 0.2917

5 Hypothetical protein -

1vhn, 2q6i, 3kyf, 3kyg, 1ofu, 2e87, 2yqz, 2hek, 2ogi, 1yre, 2d1y, 1p0h, 1q6y, 2gf6

3.358 0.2799

6,7-dimethyl-8-ribityllumazine synthase Coenzyme transport and metabolism

1nqu, 2c9b, 1t13, 2f59, 2obx, 1nqu, 1kyy

10

3-hydroxy-3-methylglutaryl-coenzyme A reductase

This transmembrane glycoprotein is involved in the control of cholesterol biosynthesis It is the rate-limiting enzyme

of sterol biosynthesis

2q1l, 3cd7, 3ccw, 1qax, 3cd5, 1dqa, 1r31

13 Enoyl-[acyl-carrier-protein]

reductase [NADH] Lipid transport and metabolism

1jvf, 1d8a, 3gr6, 3oig, 1qsg, 2h7i, 2h9i, 2pd4

permeability

2oh4, 1ywn, 3c7q, 3c7q 2.959 0.3288

15 2-C-methyl-D-erythritol

3q8h

Involved in the activation cascade of caspases; responsible for apoptosis execution It proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at

implicated in the pathogenesis of chronic

myelogenous leukemia 1o4r, 3el8 3.157 0.2201

20

dimethylbenzimidazole phosphoribosyltransferase

Nicotinate-nucleotide-plays a central role in the synthesis of ribazole, which is a key component of the

α-lower ligand of cobalamin

1l5l, 1l5m 3.12 0.3667

21 Phosphopantetheine

22 Beta-secretase 1

Involved in aspartic-type signal peptidase

Table XX Retrieval of top targets of 173b by the ReverseScreen 2D and 3D Methods, PharmMapper and Binding DB

S No Protein name (target

Pharm Mapper (fit score)*

Reverse Screen 3d score*

1 Adenylate cyclase

2

Vascular endothelial growth factor receptor

2

Has a tyrosine-protein kinase activity The VEGF-kinase ligand/receptor signaling system plays a key role in vascular development and regulation of vascular

permeability

3dtw, 3cpc, 1ywn, 3be2, 3cpb, 3oh4, 3ewh, 1y69, 3c7q

ligand-cellular proliferation

1gs4, 3b67, 3b68, 1t7t, 1i37, 2piq, 3b68

Serine/threonine-Signal transduction mechanisms 1it6, 1u32 3.079 0.2369

9

protein] reductase [NADH]

10

hydroxymethyl dihydropteridine pyrophosphokinase

2-amino-4-hydroxy-6-Coenzyme transport and metabolism 1cbk, 1ex8 3.129 0.15

11 Purine nucleoside

phosphorylase Nucleotide transport and metabolism

2ai1, 1a9t, 1ovg, 1z36,

1z37, 1a9p, 1rfg, 3iom, 1k9s, 1odj, 1rhw, 2ac7, 2ai1

12

Serine/threonine-protein kinase Chk1

Required for checkpoint mediated cell cycle arrest in response to DNA damage or the presence of unreplicated DNA May also negatively regulate cell cycle progression during unperturbed cell cycles

Recognizes the substrate consensus sequence [R-X-X- S/T]

2bro, 2c3k, 3jvr, 2e9p, 2e9u

13

Proto-oncogene tyrosine-protein kinase

Src

-

3el7, 3g6h, 2h8h, 3f3t, 2qq7, 1o4r

3k5e, 2gm1, 2uym, 3cjo, 2fme

16

methylglutaryl-coenzyme A reductase

3-hydroxy-3-This transmembrane glycoprotein is involved in the control of cholesterol biosynthesis It is the rate-limiting enzyme

of sterol biosynthesis

1hw8, 3cct, 3cd0, 3cd5, 3ccw, 3cdb

17

hydroxymethyldihydro

2-amino-4-hydroxy-6-pteridine pyrophosphokinase

Coenzyme transport and metabolism 1ex8, 1cbk 3.12 0.15

Post-translational modification, protein

20 Scytalone dehydratase Involved in hydro-lyase activity 6std, 7std,

22

Dihydrofolate reductase /Thymidylate synthase

Nucleotide trans port and metabolism

1tls, 1ju6, 1qzf, 3k2h, 3jsu, 3cl9, 2aaz, 2blc,

1j3j, 1tvv, 1hvy, 3nrr, 1tlc, 1f28, 1sej, 1tsd, 1o26, 2cfa, 3fnn, 3gwc

3i65, 1tv5 3.327 0.3714

25

Proto-oncogene tyrosine-protein kinase

LCK

Tyrosine kinase that plays an essential role for the selection and maturation of developing T-cell in the thymus and in

mature T-cell function

2og8, 3byu, 3byo, 2ofv 3.463 0.3171

26 Mitogen-activated

protein kinase 14 Involved in MAP kinase activity

3e92, 3d83, 3e93, 3fc1, 3huc, 2zb0,

3.136 0.3514

27 Beta-secretase 1 Involved in aspartic-type signal peptidase 3duy, 2qky,

3f3e, 1ymz,

activity 1ym4, 1xs7,

3hvg

28 Cell division protein

kinase 2 Involved in protein kinase activity

1p2a, 1fvt, 1r78, 2vtp, 3bhv

29

Tyrosine-protein phosphatase non-receptor type 1

May play an important role in CKII- and p60c-src-induced signal transduction cascades (By similarity)

1q6s, 1ecv, 1nwe, 1no6, 1c83

31

Phosphatidylinositol 3-kinase regulatory subunit alpha

Binds to phosphorylated protein-Tyr kinases, through its SH2 domain, and acts

as an adapter, mediating the association of the p110 catalytic unit to the plasma membrane Necessary for the insulin-stimulated increase in glucose uptake

Table XXI Retrieval of top targets of 173c by the ReverseScreen 2D and 3D Methods, PharmMapper and Binding DB

S No

Protein name (target name)

Pharm Mapper (fit score)*

Reverse Screen 3d score*

1

Phosphatidylinositol-4,5-bisphosphate-3-kinase

Involved in phosphotransferase activity,

alcohol group as acceptor 2a4z, 2a5u 4.223 0.731

2 Epidermal growth factor

receptor

Cell-surface receptor for members of the epidermal growth factor family (EGF-family) of extracellular protein

ligands

2gs7, 1mox, 1ivo 3.915 0.436

3

Phosphoenolpyruvate carboxykinase, cytosolic

3.815 0.3462

5 Scytalone dehydratase Involved in hydro-lyase activity 6std, 7std,

5std, 4std 3.654 0.275

6 Pyruvate oxidase Amino acid transport and metabolism 1pox, 1pow,

4.2659 0.371

10 Dihydrofolate reductase

/Thymidylate synthase Nucleotide transport and metabolism

1tls, 1ju6, 1qzf, 3k2h, 3jsu, 3cl9, 2aaz, 2blc, 1j3j, 1tvv, 1hvy, 3nrr, 1tlc, 1f28, 1sej, 1tsd, 1o26, 2cfa,

3.561 0.375

3fnn, 3gwc

11 Mast/stem cell growth factor

receptor

It has a tyrosine-protein kinase activity

Binding of the ligands leads to the autophosphorylation of KIT and its association with substrates such as phosphatidylinositol 3-kinase (Pi3K)

1t46, 1t46 3.5339 0.25

12 Angiopoietin-1 receptor

This protein is a protein tyrosine-kinase transmembrane receptor for angiopoietin 1 Regulates endothelial cell proliferation, differentiation and guides the proper pattern

vascular permeability

3dtw, 2p2h 3.4957 0.3095

14

Proto-oncogene serine/threonine-protein kinase

16 Mitogen-activated protein

kinase 14 Involved in MAP kinase activity 3e92, 3d83, 3e93, 3fc1, 3.441 0.3514

3huc

17 Beta-secretase 1 Involved in aspartic-type signal

peptidase activity

3duy, 2qky, 3f3e, 1ymz, 1ym4, 1xs7

mature T-cell function

2og8, 3byo, 3byu, 2ofv 3.463 0.3404

19 Tyrosine-protein phosphatase

non-receptor type 1

May play an important role in CKII- and p60c-src-induced signal transduction cascades (By similarity)

1q6s, 1ecv, 1nwe, 1no6, 1c83

3.565 0.2812

Biologically important in the destruction of opioid peptides such as Met- and Leu-enkephalins by cleavage

of a Gly-Phe bond

1r1i, 1y8j 3.298 0.3548

21 Pyruvate oxidase Amino acid transport and metabolism

1pow, 2ez9, 1pox, 1v5e

3.644 0.2258

22 DNA polymerase I,

thermostable Replication, recombination and repair 5ktq, 1qss 3.271 0.2326

endopeptidase, whose function is to

1bim, 1bil,

generate angiotensin I from angiotensinogen in the plasma, initiating

a cascade of reactions that produce an elevation of blood pressure and increased sodium retention by kidney

24 Ribonuclease Involved in nucleic acid binding

1goy, 1rca, 1rga, 2rok, 1iyb, 1j1f, 3d45, 1rds

3.17 0.3125

25 Cell division protein kinase 2 Involved in protein kinase activity

1p2a, 1fvt, 1r78, 2vtp, 3bhv

2x24

30 Poly [ADP-ribose] polymerase

31 NADPH-flavin oxidoreductase Involved in oxidoreductase activity 2bkj, 2h0u,

32 Tyrosine-protein kinase JAK2

Plays a role in leptin signaling and control of body weight Tyrosine kinase

of the non-receptor type, involved in interleukin-3 and interleukin-23 signal

37 Major NAD(P)H-flavin Energy production and conversion 1v5z, 1vfr, 2.972 0.286

3.743 0.2773

39 Matrix metalloproteinase-9

May play an essential role in local proteolysis of the extracellular matrix and in leukocyte migration Could play

a role in bone osteoclastic resorption

plays an important role in transcriptional regulation, cell cycle

reticulum

44 Purine nucleoside

phosphorylase Nucleotide transport and metabolism

1a9t, 1a9p, 1z36, 1rfg, 3iom, 1k9s, 1odj

3.048 0.68

45 Proto-oncogene

3g6h, 1o4r, 3f3t, 2h8h 2.998, 0.432

46 Retinoic acid receptor

ligand-3b67, 2pip,

48 TGF-beta receptor type-1

On ligand binding, forms a receptor complex consisting of two type II and two type I transmembrane serine/threonine kinases Type II receptors phosphorylate and activate

type I receptors which autophosphorylate, binds and activate

SMAD transcriptional

1vjy, 3gxl 2.967 0.4705

49 Serine/threonine-protein kinase

Chk1

Required for checkpoint mediated cell cycle arrest in response to DNA damage Recognizes the substrate consensus sequence [R-X-X- S/T]

2e9p, 2c3k,

50 ATP-dependent molecular

chaperone HSP82

Post-translational modification, protein

turnover, chaperones 1bgq, 2bre 3.26 0.4118

51

dimethylbenzimidazole phosphoribosyltransferase

Table XXII Retrieval of top targets of 173f by the ReverseScreen 3D Method and PharmMapper

S No

Protein name (target name)

Pharm Mapper (fit score)*

Reverse Screen 3d score*

2gs7, 1mox, 1ivo, 2itt 3.641 0.392

3 Protein kinase C beta II - 2i0e

4

dependent protein kinase

3-phosphoinositide-1

Phosphorylates and activates not only PKB/AKT, but also PKA, PKC-zeta, RPS6KA1 and RPS6KB1 May play a general role in signaling processes and in

development

3iop, 3nax, 1okz, 2pe1 3.5 0.463

3.602 0.36

7 Dihydrofolate reductase Coenzyme transport and metabolism

1cd2, 1m4h, 1hfq, 1dyh, 1vif, 1dr1, 3nrr, 1sej, 3jwf, 1ly4, 2w9s, 1qzf, 3jsu, 1d1g, 1u70, 3frd, 1ia2, 3k2h,

3.414 0.41

3ix9, 2w3w, 1bzf, 2zza, 2blc, 1dg5

Participates in the Wnt signaling pathway

Implicated in the hormonal control of several regulatory proteins including glycogen synthase, MYB and the transcription factor

JUN

1j1b, 3f88 3.305 0.353

3.21 0.371

15 Androgen receptor

Steroid hormone receptors are activated transcription factors that regulate eukaryotic gene expression and affect cellular proliferation in target tissues

ligand-1gs4, 2pip, 2piq, 2hvc 3.174 0.4

16 DNA ligase Involved in DNA ligase (NAD+) activity 1ta8, 3bac,

3ba9, 3bab 3.114 0.31

17 Ribonuclease Involved in nucleic acid binding

1goy, 1rpf, 1w4q, 3dgy, 1s9g, 1rca, 1i0v,1rga, 2rok, 1rds, 3lpz, 1iyb

3.112 0.343

18 Cell division protein Involved in protein kinase activity 1gih, 1di8, 3.06 0.364