O

N
N

7a
O O

1
 Design, Synthesis, Antimicrobial evaluation of Novel
Coumarin Derivatives and Process Optimization for
the Synthesis of Desidustat

Under the Presented

Guidance
Prof. ofNanduri
Srinivas : By : A. WARAKE
RUTURAJ
Dept. of Process chemistry O O Dept. of Process chemistry
PTPC/2020/517

2
 Flow of presentation

Introduction

Literature Review

Work Plan
Chapter 1
Work Done

Future Plan of Work

Discussion

References

Introduction

Literature Review
Chapter 2
Work Plan

References
3
 Chapter 1: Design, synthesis and antimicrobial evaluation of coumarin derivatives

Introduction

• Antimicrobial Agent:
An agent which kills microorganisms or prevent their
growth.

• Antimicrobial resistance:
The ability of the microbes to resist the lethal effects of
earlier used antimicrobial agents.

• Emergence of multidrug resistance shown by certain

strains of gram-negative bacteria such as Pseudomonas,
Klebsiella, Enterobacter, Salmonella species and gram- Fig 1. Mechanism of drug resistance
positive organisms such as Staphylococcus,
Enterococcus and Streptococcus species.

1. Annu. Rev. Genet. 43, 167–195 (2009)

2. Clinics 66, 1699–1705 (2011)
 Introduction

Inhibition of DNA Gyrase

ATP Synthase

Inhibition of Interference
membrane with cell wall
function synthesis

ANTIBACTERIAL
TARGETS

Inhibition of a Inhibition of
metabolic protein
pathway synthesis
Interference
with nucleic
acid synthesis
5
3. Cold Spring Harb. Perspect. Med. 6, 1–7 (2016)
4. Trends Microbiol. 5, 102–109 (1997)
5. Appl. Microbiol. Biotechnol. 92, 479–497 (2011)
 Literature Review

Coumarins Pharmacological Role

OH
OH
H
O N
O
OH O
O
O O O O
H
N OH Cl
O
O O
2
1 Anti-Microbial
Anti-Coagulant

O O OH

O O

HO O O
O
O O O 4
3 Anti-Tuberculosis
N O Anti-Depressant Anti-Cancer
Anti-HIV Anti-Oxidant
Anti-Hyperlipedimic

• Due to potential applications in bacterial infections, extensive studies have been carried out on the design and synthesis of
coumarin derivatives with improved antibacterial activity.
• Aminocoumarins such as novobiocin, clorobiocin, and coumeramycin-A are the known inhibitors of GyrB ATPase.
6
6. Curr. Med. Chem.2008, 15 (26), 2664–2679
7. Braz. J. Med. Biol. Res.2017, 50 (11)
8. Curr. Pharm. Des.2004, 10 (30), 3797–3811
 Literature Review
• Heide and colleagues reported various novobiocin analogues with antibacterial activity. Compound 5 had excellent activity
(0.06 mg/mL) against MRSA and compound 6 has an S. aureus MIC of 1 mg/mL and is equally potent against a
novobiocin-resistant strain.

O O
OH N
H
O O O O O N N
O
O O
N O
H OH
OH
HO 5 6
MIC values MIC values
MRSA=0.06 mg/mL Novobiocin-resistant strain = 1 mg/mL

• Compound 7 (MIC=0.25 mg/L) showed considerable inhibitory activity than ciprofloxacin (MIC=0.5 mg/L) against
Escherichia coli and 8 (MIC=0.05 mg/L) exhibited excellent antibacterial activity than ciprofloxacin (MIC=0.25 mg/L)
against Salmonella.

HOOC EtOOC
O N O N
Cl H
NH N NH N
H O O (Et)2N O O

7 Cl Cl
8
Escherichia coli MIC values = 0.25 mg/L Salmonella MIC values= 0.05 mg/L

7
9. Chemistry & biology 2007, 14, 955-967
10. Med. Chem. Lett. 2007, 17, 4708-4714
11. Med. Chem. Res. 2015, 24, 1626-1634
 Literature Review
• Biphenyl rings attached chromene-2-one derivatives 9 and 10 with good antibacterial activity against multiple
ciprofloxacin resistant MRSA strains with MIC values of 1.56 and 0.78 µg/ml against S. aureus and 0.65 µg/ml against B.
anthracis respectively.
CH3 O CH3 O
OH OH
O O O O O O

Cl

9 10

MIC values
MIC values
S.aureus = 1.56 µg/ml
S.aureus = 1.56 µg/ml
B.anthracis = 0.78 µg/ml
B.anthracis = 0.65 µg/ml

• The 5,7-dihydroxy coumarin derivative 11 showed superior inhibitory profile towards most of the tested bacterial strains
with MIC values ranging between 0.16-5 mg/mL against the gram-negative and 0.02-0.63 mg/mL against the gram-positive
bacterial strains. Derivative 12 showed potent antibacterial activity against S. aureus and B. cereus and L. monocytogenes
strains with MIC values in the range of 11-44 µg/ml.

OH

OH

O O OH

O O OH 12
11
S. aureus and B. cereus MIC values = 11-44 µg/mL
Gram-positive strain MIC values = 0.02-0.63 mg/mL For Gram-negative strains MIC values = >350µg/mL)
Gram-negative strain MIC values = between 0.16-5 mg/mL 8

12. J. Med. Chem.2012, 55 (24), 10896–10908

13. Chemico-Biological Interactions. 2015, 231: 10-17
 Work Plan

Scheme: 1
OH Cl
POCl3, TEA

Reflux
O O O O
O
2 O
1 O
HO
N O R
NH N
Cl N R
OH OH N N

O
HCl IN DIOXANE (4N) O HATU, DIPEA O
(Boc)2O O O

N N
THF, 0°C O O DCM, rt DMF, rt
K2CO3, DMF,rt O O
O O
N N 6 7
H 5
7a R= Me,
O O 7b R= Cl
3 TEA, ethanol, reflux
S R
C
N S
4 R
N N
H
N

O O
8a R= H,
8 8b R= Cl

9
 Work Plan

Scheme: 2
O O

O
S
R
N N
R H H
SCN
5
5a R= OCH3
TEA, ACN,
O O 5b R=CH3
O O Reflux, 80°C
O O
OH K2CO3 NH4Cl, Fe
DMF, rt O O
O2N EtOH/H2O,
Cl 80°C
1 2 O2N H2N O O
HATU, DIPEA.
3 4 DMF, rt

O
O
R
HO
N
H
O 6a R= Br
R 6b R= OH
6

10
 Work done

Scheme 1:
Step 1: Synthesis of 4-Chloro coumarin (2):

OH Cl
POCl3

TEA, reflux
O O O O
1 2

Step 2: Synthesis of tert-butyl 4-(4-hydroxyphenyl) piperazine-1-carboxylate (4):

OH OH

(boc)2O

N N
THF, 0°C

N N
H
O O
3

4
11
 Work done

Step 3: Synthesis of tert-butyl 4-(4-((2-oxo-2H-chromen-4-yl) oxy)phenyl)piperazine-1-carboxylate (5):

O

N O
Cl N
OH

O
O O

N
K2CO3, DMF, rt O O

N 5
O O

Step 4: Synthesis of 4-(4-(piperazin-1-yl) phenoxy)-2H-chromen-2-one (6):

O

N O
NH
N
N
O HCl IN DIOXANE (4N)
O

DCM, rt
O O
O O
12
5 6
 Work done

Step 5a: Synthesis of 4 substituted (4-(4-benzoylpiperazin-1-yl) phenoxy)-2H-chromen-2-one:

O
O
NH N
HO R
N R N

O O
HATU, DIPEA
DMF, rt
O O O O

6 7 7a R= Me,
7b R= Cl

Step 5b: Synthesis of 4-(4-((2-oxo-2H-chromen-4-yl) oxy) phenyl)-N-phenylpiperazine-1-carbothioamide:

S
NH R
N N N
S R H
C N
N
O
TEA, ethanol, reflux O

O O
O O
6 8a R= H,
8 8b R= Cl
13
 Work done
Scheme 2:
Step 1: Synthesis of 4-(4-nitrophenoxy)-2H-chromen-2-one (3):
O O
O O
NO2 K2CO3

DMF, rt O
HO
Cl
1 2 O2N
3

Step 2: Synthesis of 4-(4-aminophenoxy)-2H-chromen-2-one (4):

O O O O

NH4Cl, Fe
O O
EtOH/H2O Reflux

O2N H2N
3 4

Step 3: Synthesis of 4 substituted 1-(4-((2-oxo-2H-chromen-4-yl) oxy)phenyl)-3-phenylthiourea:

O O NCS O O
R

O O
TEA, ACN, S
R
Reflux, 800C
H2N N N
H H 5a R= OCH3 14
4 5 5b R=CH3
 Work done

Step 4: Synthesis of 4 substituted N-(4-((2-oxo-2H-chromen-4-yl) oxy)phenyl)benzamide:

O O
O O R
HO

O
O
O O
HATU, DIPEA.
DMF, rt N
H2N R H
6a R= Br
4 6 6b R= OH

15
FUTURE PLAN OF WORK

 To synthesis more number of analogues.

 To submit the synthesized analogues for biological evaluation.

 To perform the molecular docking studies to understand mode of inhibition.

Conclusion

 8 derivatives were synthesized and characterized by 1H NMR and13C NMR spectral studies.

16
 REFERENCES
1. Andersson, D. I. & Hughes, D. Gene amplification and adaptive evolution in bacteria. Annu. Rev. Genet. 43, 167–195 (2009).
2. Neuenschwander, L. C. et al. Plasma levels of procalcitonin and eight additional inflammatory molecules in febrile neutropenic patients. Clinics
66, 1699–1705 (2011).
3. Silver, L. L. Appropriate targets for antibacterial drugs. Cold Spring Harb. Perspect. Med. 6, 1–7 (2016).
4. Maxwell, A. DNA gyrase as a drug target. Trends Microbiol. 5, 102–109 (1997).
5. Collin, F., Karkare, S. & Maxwell, A. Exploiting bacterial DNA gyrase as a drug target: Current state and perspectives. Appl. Microbiol.
Biotechnol. 92, 479–497 (2011).
6. Hiasa, H. DNA topoisomerases as targets for antibacterial agents. Methods Mol. Biol. 1703, 47–62 (2018).
7. Heide, L. New aminocoumarin antibiotics as gyrase inhibitors. Int. J. Med. Microbiol. 304, 31–36 (2014).
8. Hooper, D. C. & Jacoby, G. A. Mechanisms of Action and Resistance. 1–21 (2016).
9. Sahoo, C. R. et al. Coumarin derivatives as promising antibacterial agent(s). Arab. J. Chem. 14, 102922 (2021).
10. Veselinović, J. B. et al. Antibacterial potential of selected 4-phenyl hydroxycoumarins: Integrated in vitro and molecular docking studies. Med.
Chem. Res. 24, 1626–1634 (2015).
11. Pisano, M. B. et al. Antibacterial Activity and Molecular Docking Studies of a Selected Series of Hydroxy-3-arylcoumarins. Molecules 24, 1–14
(2019).
12. Liu, H. et al. Novel coumarin-pyrazole carboxamide derivatives as potential topoisomerase II inhibitors: Design, synthesis and antibacterial
activity. Eur. J. Med. Chem. 157, 81–87 (2018).
13. Liu, H. et al. Novel coumarin-thiazolyl ester derivatives as potential DNA gyrase Inhibitors: Design, synthesis, and antibacterial activity.
Bioorg. Chem. 100, 103907 (2020). 17
 Chapter 2: Process optimization for the synthesis of Desidustat
INTRODUCTION

Desidustat (also known as ZYAN1) is a drug for the treatment

of anemia of chronic kidney disease.

Brand name: Oxemia O
N O
O
Discovered and Developed: Zydus Life Sciences H
N
OH
MOA: a HIF prolyl-hydroxylase inhibitor OH O

Uses: Desidustat helps by reducing hepcidin, reducing inflammation, Desidustat

and better iron mobilisation.

A clinical trial to evaluate the efficacy and safety of desidustat tablet for the management of Covid-19 patients is
ongoing in Mexico.

18
1. Clinical Pharmacokinetics, 57(1), pp.87-102
2. American Journal of Nephrology, 49(6), pp.470-478
3. Drug research, 66(02), pp.107-112
 Literature Review

O O
K2CO3 Hydrazine hydrate
+
HO N Br O N O
450C - 550C DCM, 20-350C, NH2
O 3 hrs O 18 hrs 3
2-Hydroxy- O
isoindoline-1,3-dione 1 2
O O
K2CO3, 20-350C,
OH O O
I 18 hrs
H2SO4 O
O NH
O +
I Ethanol, 80-850C, O O
OEt
20 hrs
6 4
5

Tolouene, Co(I), K2CO3, ethanol,

Glycine reflux at 78-850C, 24 hrs OEt
O O O
O
Boc H EtO OH
Scheme: 1 N
O Methanolic HCl
N
O POCl3, N
O
O O O
DCM, 20-300C, ACN, Pyridine,
OEt 3 hrs OEt 0-100C, 45 mins 9
OEt
7 8

Methanol, NaOMe, 10-200C, 30 mins

1N HCl

OH O OH O
OH O
EtOOC NH2
O N COOEt OEt
N H Glycine ethylester HCl
H MeOH, HCl
OH N O N O
N O NaOH/H2O
O THF, DIPEA, 65-70 C, 0 O
O 30-400C, 2 hrs
18 hrs
19

4. Patent Application Publication Ii KfiH Ir. 1, 11 10

12
(2000).
 Literature Review

Scheme: 2

Step 1
O
i) S Cl
O
OH I
TEA, Ethanol
ii) Sodium iodide, reflux 2
1 O O
O OH O
O OH NaOMe
O OO O O
step 2 Cs2CO3, DMF ACN, Pyridine
O N O MeOH N O
O O rt POCl3
sodium O O O
N+ acetate trihydrate N
O- O- H
O 7
stannous chloride OH 5
N O
O dihydrate H 6 H Cl DIPEA, Dioxane
H2N
3 4 O

OH O OH O
OH NaOH O
N N
H H
O O
N O TMF, rt N O
O O

9 8

20

5. W. A. C - A l A l A l. vol. 1 (2017).
 Work Plan

O O

+
HO N Br O N O
NH2
O O
O

O O

O O
I
O
NH
O +
O O
OEt

OEt
O
O
Boc H
N N
O N
O O
O O
O
OEt OEt OEt

OH O OH O
OH O
O N COOEt OEt
N H
H
OH N O N O
N O
O O
O

21
 REFERENCES

1. Kansagra, K.A., Parmar, D., Jani, R.H., Srinivas, N.R., Lickliter, J., Patel, H.V., Parikh, D.P., Heading, H., Patel, H.B., Gupta, R.J. and
Shah, C.Y., 2018. Phase I clinical study of ZYAN1, a novel prolyl-hydroxylase (PHD) inhibitor to evaluate the safety, tolerability, and
pharmacokinetics following oral administration in healthy volunteers. Clinical Pharmacokinetics, 57(1), pp.87-102.
2. Parmar, D.V., Kansagra, K.A., Patel, J.C., Joshi, S.N., Sharma, N.S., Shelat, A.D., Patel, N.B., Nakrani, V.B., Shaikh, F.A., Patel, H.V. and
ZYAN1 Trial Investigators, 2019. Outcomes of desidustat treatment in people with anemia and chronic kidney disease: a phase 2
study. American Journal of Nephrology, 49(6), pp.470-478.
3. Jain, M.R., Joharapurkar, A.A., Pandya, V., Patel, V., Joshi, J., Kshirsagar, S., Patel, K., Patel, P.R. and Desai, R.C., 2016. Pharmacological
characterization of ZYAN1, a novel prolyl hydroxylase inhibitor for the treatment of anemia. Drug research, 66(02), pp.107-112.
4. Chrysafis, C., Said, A., Drukarev, A., Islam, A. & Pearlman, W. A. C - A l A l A l. vol. 1 (2017).
5. Sharma, R. et al. ( U ) Patent Application Publication Ii KfiH Ir. 1, (2000).

22