On-water Sonochemical Synthesis and - PowerPoint Presentation

Slide1

On-water Sonochemical Synthesis and

in silico

Validation of

Trypanocidal

/

Leishmanicidal

Benzopyrazines

Shaila Akter Shetu

, Omar Espino, Debasish Bandyopadhyay

Slide2

Introduction

Bandyopadhyay 

et al., Curr. Med. Chem. 28(3), 472-495, (2021)Bandyopadhyay et al., Curr. Med. Chem. 24(41), 4677-4713, (2017)Bandyopadhyay et al., Curr. Med. Chem. 24(41), 4714-4725, (2017)

Figure 1:

Neglected Tropical Diseases

Slide3

Introduction

Bandyopadhyay 

et al. Front. Chem. 9:775327 (2021) DOI: https://doi.org/10.3389/fchem.2021.775327

Slide4

Introduction

Figure 2:

Global scenario of Two neglected tropical diseases: Chagas disease (American trypanosomiasis), and Leishmaniasis.Bandyopadhyay et al. Front. Chem. 9:775327 (2021)

Slide5

Introduction

Figure 3:

Representative examples of naturally occurring bioactive benzopyrazines

Slide6

Methods

Green Synthesis of Benzopyrazine

Determination of Drug-likeness​Trypanocidal and Leishmanicidal (in vitro) Evaluations

In Silico Molecular Docking Studies

Slide7

Sonochemical Synthesis of Benzopyrazines

Figure 4:

Ultrasonic Processor UP200St. Supports automatic frequency tunning 26kHz. 50% amplitude was used for all the reactions. Sonotrode S26d2 probe was used.

Slide8

Ortho

-phenylenediamine and

dicarbonyl compounds were mixed (1:1 molar ratio) in a hard glass test tube1 mL water added to the reaction mixtureThe mixture was sonicated using ultrasonic probeThe reaction was monitored by thin layer chromatography (TLC)The product was extracted with 9 ml (3x3 mL) of ethyl acetate

Finally, crystalline product was isolated

Sonochemical Synthesis of Benzopyrazines

Slide9

Determination of Drug-likeness

Drug-likeness indicates the possibility of a molecule becoming a drug

A drug molecule should have a balance of various physicochemical properties like molecular weight, hydrogen bond donor, hydrogen bond acceptor, total polar surface area, rotatable bond, hydrophilicity, lipophilicity, hydrophobicity, lipophobicity, bioavailability, half-life, etc.The druggability assessment of all the eleven benzopyrazines was performed in compliance with Lipinski’s rule of five (RO5)

Slide10

Trypanocidal

 and 

Leishmanicidal (in vitro) EvaluationsThe promastigotes of L. mexicana (MHOM/MX/ISETGS) clinical strain and the epimastigotes of T. cruzi (MHOM/MX/1994/NINOA) were used for the leishmanicidal and trypanocidal  growth inhibition assaySchneider’s Drosophila medium, supplemented with 10% fatal bovine serum, penicillin (10

IU/mL), and streptomycin (100 µg/mL), was used to culture the parasites 

Two first line commercial drug

nifurtimox (antichagasic drug) and

miltefosine

(

leishmanicidal

drug), were used as positive controls. Only the parasite-containing culture was used as negative control

The IC

50

values (the concentration required to inhibit 50% of parasite growth were calculated  (in µg/mL) were determined by

probit

analysis

Slide11

In Silico

Molecular Docking Studies

T. cruzi Histidyl-tRNA synthetase (PDB ID: 4YPF) and T. cruzi trans-sialidase (PDB ID: 1S0J) are considered as two major biological drug targets for American trypanosomiasis Leishmanial rRNA A-site (PDB ID: 4K32) and Leishmania major N-myristoyl transferase (PDB ID: 6QDA) are considered as the major biomolecular drug targets for leishmaniasis Binding site visualization was made feasible by Schrödinger Maestro, AutoDock4,

PyMOL, and other software

Slide12

Results

Green synthesis of diversely substituted Benzopyrazines

Drug likeness of BenzopyrazinesTrypanocidal and Leishmanicidal evaluations (in vitro) of the Benzopyrazines (1–11) In Silico

molecular docking of the compound

Slide13

Green Synthesis of Diversely Substituted Benzopyrazines

Figure 5:

Ultrasound-assisted on-water green synthesis of diverse benzopyrazines

Slide14

Green Synthesis of Diversely Substituted Benzopyrazines

(Isolated yield & Atom economy)

Table 1. The yield, and atom economy in the synthesis of benzopyrazines (1–11).

Slide15

X-ray crystallographic structure (ORTEP) of Compound 2 (Figure 5)

Figure

6: X-ray crystallographic structure (ORTEP) of Compound 2 (Fig. 5).

Slide16

Drug-likeness of Benzopyrazines

Table 2.

Validation† of drug-likeness of the benzopyrazines (1–11).

†Molinspiration

property engine v2018.10;

a

miLogP

:

Moriguchi

octanol-water partition coefficient, is based on quantitative structure-

LogP

relationships, by using topological indexes;

b

Hydrogen

bond acceptor;

c

Hydrogen

bond donor;

d

Total

polar surface area;

e

Number

of rotatable bonds.

Slide17

Trypanocidal

 and 

Leishmanicidal Evaluations (in vitro) of the Benzopyrazines (1–11)Table 3. IC50 (µM ± SD) of the benzopyrazines (1–

11

) against

epimastigotes

from

T.

cruzi

and promastigote from

L.

mexicana

.

Slide18

In Silico

Molecular Docking of the Compound 1 (Figure 5)

Table 4. Molecular docking scores of the compound 1 and the standard controls with the biomolecular targets

Slide19

In Silico

Molecular Docking of the Compound 1 (Fig. 5)

Table 5. Interactions between compound 1 with the four biomolecular targets (PDB IDs: 4YPF, 1S0J, 4K32 and 6QDA)

Slide20

In Silico

Molecular Docking of the Compound 1

Figure 7: The binding mode of the interactions between 1

with a) (PDB ID: 4YPF), b) (PDB ID: 1S0J), c) (PDB ID: 4K32), d) (PDB ID: 6QDA) active sites

7a

7b

7c

7d

Slide21

In Silico

Molecular Docking of the Compound 1

Figure 8: Results of the validation of

1 inside the a)

(PDB ID: 4YPF), b) (PDB ID: 1S0J), c) (PDB ID: 4K32), d) (PDB ID: 6QDA) active sites

8a

8b

8c

8d

Slide22

Discussion

 An expeditious on-water green synthesis carried out

A series of eleven benzopyrazines has been synthesizedmost of the synthesized compounds have drug-likeness following Lipinski's "Rule of 5 (RO5)“Our benzopyrazine 1 showed comparable in vitro activity against both the protozoa (L. mexicana

and T. cruzi

)

Benzopyrazine

1

demonstrated better binding affinity towards all the four proteins than the standard control drug

The molecules should have good pharmacokinetics and pharmacodynamics properties.

Slide23

Conclusion

Bandyopadhyay 

et al., Curr. Med. Chem. 28(3), 472-495, (2021),DOI: https://doi.org/10.3389/fchem.2021.725892

Slide24

Conclusion

On-water strategy contributes significantly towards green methodology development as sonication does as a green energy source

Our newly developed method satisfies several aspects of green chemistryBenzopyrazine 1 may find its application in the future drug development process against two major neglected tropical diseases: Chagas’ disease and leishmaniasis

Slide25

Future Aspects

Chemical Modification of our compounds (in particular Compound 1 (Fig. 5), so that it can target the related protein more precisely

To modify one or more functional groups/moieties to the existing molecule so that the new molecule can bind the target protein(s) more effectively than the currently available commercial drugs.To develop new drug(s) with higher potency and reduced side effects (toxicity) using this greener pathway and molecular docking studies.

Slide26

Acknowledgement

Special thanks to Jonathan Rock and Daniel Garcia for helping us in this project

I acknowledge the contribution of Dr. Gildardo Rivera, our collaborator from Mexico.

Slide27

References

1. Rock Jonathan, Garcia Daniel, Espino Omar, Shetu Shaila A., Chan-

Bacab Manuel J., Moo-Puc Rosa, Patel Navin B., Rivera Gildardo, Bandyopadhyay Debasish. Benzopyrazine-Based Small Molecule Inhibitors As Trypanocidal and Leishmanicidal Agents: Green Synthesis, In Vitro, and In Silico Evaluations, Frontiers in Chemistry, 9, 2021, 7572. Rivera G, Patel NB, Bandyopadhyay D. Editorial: Discovery and Development of Drugs for Neglected Diseases: Chagas Disease, Human African Trypanosomiasis, and Leishmaniasis. Front Chem. 2021 Oct 7;9:775327. doi: 10.3389/fchem.2021.775327. PMID: 34692649; PMCID: PMC8530326.3. Maheshwari KK, Bandyopadhyay D. Heterocycles in the Treatment of Neglected Tropical Diseases. Curr Med Chem. 2021;28(3):472-495. doi: 10.2174/0929867327666200219141652. PMID: 32072886.4. Bandyopadhyay D, Chavez A, Banik BK. Microwave-induced Bismuth Salts-mediated Synthesis of Molecules of Medicinal Interests. Curr

Med Chem. 2017;24(41):4677-4713. doi: 10.2174/0929867324666170320121142. PMID: 28322155.

5. Bandyopadhyay D,

Samano

S, Villalobos-Rocha JC, Sanchez-Torres LE,

Nogueda

-Torres B, Rivera G,

Banik

BK. A Practical Green Synthesis and Biological Evaluation of Benzimidazoles Against Two Neglected Tropical Diseases: Chagas and Leishmaniasis.

Curr

Med Chem. 2017;24(41):4714-4725.

doi

: 10.2174/0929867325666171201101807. PMID: 23317160.