Synthesis, characterization and antimicrobial study of novel substituted Curcumin Derivatives

 

Goyal Rupendra1*, Jain Suman2, Agarwal D D3

1Assistant Professor, SOS in Pharmaceutical Sciences, Jiwaji University, Gwalior (M.P.)

2Director, SOS in Pharmaceutical Sciences, Jiwaji University, Gwalior (M.P.)

3Professor, SOS in Chemistry, Jiwaji University, Gwalior (M.P.)

*Corresponding Author E-mail: goyalrk09@gmail.com

 

ABSTRACT:

Chemically, curcumin is a diaryl heptanoid, belonging to the group of curcuminoids, which are natural phenols responsible for turmeric's yellow color. It is a Keto–enol tautomer, existing in enolic form in organic solvents and in keto form in water. The one pot three component of hydrazide substituted benzaldehyde was used for the synthesis of curcumin derivatives. In this work the synthesis of curcumin derivatives was done using an environmental friendly procedure via Mg-Al-CO3 hydrotalcites which have been used as efficient catalysts. These catalysts are inexpensive and non-toxic powders. Data of Spectra and other analytical technique supported identification of compound curcumin derivatives. The present work offers many merits such as the reaction conditions were extremely simple, there was operational simplicity, reaction time was short, easy to work up and purification was also easy for the products via simple means of re-crystallization. The antibacterial activity of three compounds B2, B4 and D3 was found to be near to the significant with standard compounds amoxicillin.

 

KEYWORDS: Curcumin, Antimicrobial activity, Characterization, Hydrotalcites.

 

 


INTRODUCTION:

Curcumin incorporates a seven carbon linker and three major functional groups: an α,β-unsaturated β-diketone moiety and an aromatic O-methoxy-phenolic group. The aromatic ring systems, which are phenols, are connected by two α,β-unsaturated carbonyl groups1. Curcumin derivatives have been well-documented due to their natural antioxidant, antimicrobial and anti-inflammatory activities. Curcuminoids have also gained widespread recognition due to their wide range of other activities which include anti-infective, anti-mutagenic, anticancer, anti-coagulant, antiarthrititc, and wound healing potential2. Co-administration of curcumin with metformin3, phenytoin and sodium valproate4, 2,4-D induced toxicity5 produced improved in efficacy of these drugs. Promising antimicrobial activity of curcumin stabilized Z nanoparticles6, PGV-17, 4 Piperidone8, PGV-6, HGV-6 and GVT -6 analogues9 of curcumin.

 

 

 

Despite of having a wide range of activities, the inherent physicochemical characteristics (poor water solubility, low bioavailability, chemical instability, photodegradation, rapid metabolism and short half-life) of curcumin derivatives limit their pharmaceutical significance. Aiming to overcome these pharmaceutical issues and improving therapeutic efficacy of curcuminoids, newer strategies have been attempted in recent years10.

 

They further represent as an efficient and effective tool for generating libraries of small molecule compounds and also exhibit indispensability for the studies of SAR i.e. structure activity relationship. Some of the uncommon scaffolds are generated via MCRs, their utility in biological realm is explored by the key of exhibiting the ability to functionalize or modify them11. These MCRs are most preferred because of their effectiveness and efficiency in synthesizing highly functionalized organic molecules. The availability of their starting materials is also easy which are further combined in one pot process including synthesizing the product and keeping diversity and complexity leading to time, cost as well as reduction of waste effluents12,13.

MATERIALS AND METHODS:

Material and instrument used:

All chemicals were obtained from Merck, High media and SD fine chemicals ltd. All solvents were redistilled and dried before use. Reactions were routinely monitored by thin layer chromatography and spots were visualized by exposure to iodine vapour or UV light. All the synthesized compounds were purified by column chromatography followed by recrystallization. Melting points were determined by using open capillary method and are uncorrected. Fourier Transform Infra Red spectra (FTIR) were recorded on Shimadzu FTIR-8400S spectrophotometer using potassium bromide pellets and sodium chloride cell. Nuclear Magnetic Resonance spectra (1H-NMR and 13C-NMR) were recorded on JEOL-300 MHz spectrophotometer in CDCl3 using TMS as an internal standard. Chemical shifts (δ) are expressed in parts per million (ppm). Mass spectra were recorded on HEWLETT 180017, PACKARD GCD System mass spectrophotometer using electron ionization detector.

 


 

Synthetic scheme I:

Fig 1: Synthetic scheme of aldehyde derivatives of curcumin moiety and R-substituted aromatic aldehyde derivatives

 

Synthetic schemes-II:

 

Fig 2: Synthetic scheme of hydrazine derivatives of curcumin moiety and R-Substituted hydrazine derivatives


Table 1: Targeted derivatives

Comp code

R1

R2

R3

R4

R5

Substrate name

B1

-H

-H

-H

-H

-H

Benzaldehyde

B2

-OH

-H

-H

-H

-H

o-Salicylaldehyde

B3

-H

-H

-OH

-H

-H

p- Salicylaldehyde

B4

-H

-H

-OCH3

-H

-H

4-methoxybenzaldehyde

B5

-H

-OCH3

-OH

-H

-H

4-hydroxy-3-methoxybenzaldehyde

B6

-H

-H

-N(CH3)2

-H

-H

4-(dimethylamino) benzaldehyde

D1

-H

 

Hydrazine

D2

-C6H4

Phenyl hydrazine

D3

- C6H3(NO2)2

2,4-Dinitro phenyl hydrazine

D4

-CONH2

Semicarbazide

D5

-CSNH2

Thiosemicarbazide

 


General Synthetic Procedure for Compounds B1-B6:

A solution of substituted benzaldehyde (0.01mol) in ethanol along with curcumin (0.01mol) and Isoniazide (0.01mol) in the presence of hydrotalcite was refluxed for 4 hours upto 80oC. The excess solvent was distilled off under reduced pressure. The cooled residual mass was washed with distilled water14. It was filtered and dried. The crude product was recrystallised from methanol to yield compound B1 to B6 reaction was monitored by TLC.

 

General Synthetic Procedure for Compounds D1-D5:

A solution of substituted hydrazine (0.01mol) in ethanol and benzaldehyde (0.01mol) with curcumin (0.01mol) in the presence of hydrotalcite was refluxed for 4 hours up to 80oC. The excess solvent was distilled off under reduced pressure. The cooled residual mass was washed with distilled water. It was filtered and dried14,15. The crude product was recrystallised from methanol to yield compound D1 to D5 reaction. The reaction was monitored by TLC.

 

RESULT AND DISCUSSION:

Effect of physicochemical parameter on synthesized derivatives:

Effect of catalyst:

The reaction using different hydrotalcites as catalysts were studied and the product yield was analyzed. The product yield is definitely affected by the type of catalysts16. The Highest yield of product (89.64%) was obtained with Mg-Al-CO3hydrotalcites. It was followed by Ca-Al-CO3 giving 81% yield and lowest yield of the product (32.51%) was obtained with Al-Mn-Cu-CO3. No product formation were obtained in the absence of catalysts. The result shows the hydrotalcites having basic metals give highest yield.

 

Table 2 Effect of various catalysts in the product yield

S.N.

Catalysts

Yield (%)

1

Ca-Al- CO3

81.05

2

Zn-Al- CO3

78.13

3

Mg-Al-CO3

89.64

4

Ni-Al- CO3

74.26

5

Ni-Fe- CO3

71.88

6

Ni-Cu-Al- CO3

67.07

7

Ni-Ca-La- CO3

60.01

8

Ni-Ca-Al- CO3

48.26

9

Ni-Zn-Al-Fe- CO3

41.38

10

Al-Mn-Cu- CO3

32.51

11

No catalyst

0

 

 

Figure.3: Effect of various catalysts on % yield

 

Effect of temperature:

The yield at various temperature ranges from room temperature to 100oC was studied. The reaction was highly sensitive to temperature. When temperature is raised to 50oC the yield was 67%. Increase in temperature further increased the yield. Higher yield (89.64%) was obtained at 800C17. when temperature was further raised the yield decreases hence 80oC was considered as optimum temperature for future studies.

 

Figure 4: Effect of temperature on % yield

 

Effect of catalysts concentration:

The effect of concentration of optimized catalyst on the formation ofcurcumin derivatives was studied. Concentration of catalyst range from 50 to 150 mg and observed the % yield of product18. The highest yield (89.64%) was observed at 100 mg weight of hydrotalcites.

 

Figure 5: Effect of Concentration of catalyst on % yield

 

Recyclability of catalyst:

Catalyst can be used again with small reduction in the weight of catalysts and without any loss in their ability. The stability of catalyst in recyclability upon 2ndrecycleproducts yield was 82%. This can be either some loss of catalyst during workup17,19. The 3rd recycle shows yield 69%. This shows slight loss of yield. So hydrotalcites found stable as recyclable catalyst.

 

Figure 6:  Recyclability of catalyst

 

So the final optimized condition for the synthesis of curcumin derivatives as benzaldehyde (0.01mol) in ethanol, curcumin (0.01mol) and hydrazine (0.01mol) in the presence of hydrotalcite 100 mg was refluxed for 4 hours upto 80oC.

 

Table 3: Physiochemical data of synthesized compound

Compound

% yield

*Rf value

Mol. Formula

Mol. Mass g/mol

B1

89.64

0.65

C34H37N3O6

583

B2

72.64

0.71

C34H37N3O7

599

B3

73.27

0.82

C34H37N3O7

599

B4

78.62

0.58

C35H39N3O7

613

B5

67.48

0.52

C35H39N3O8

629

B6

57.13

0.70

C36H42N4O6

626

D1

71.36

0.63

C27H30N2O5

462

D2

78.58

0.50

C34H38N2O5

554

D3

56.12

0.49

C34H36N4O9

644

D4

64.36

0.81

C29H35N3O6

521

D5

46.02

0.55

C29H35N3O5S

537

 

Characterization of synthesized compounds:

CMP-B1:(5-(3-hydroxy-4-methoxyphenethyl)-4-(1-hydroxy-2- (3,methoxy 4 hydroxyyphenyl) ethyl)-3- phenyl-1H-pyrazol-2 (3H)-yl)(pyridin-4-yl)methanone.

 

CMP-B2:(5-(3-hydroxy-4-methoxyphenethyl)-4-(1-hydroxy-2-(3, methoxy 4 hydroxyyphenyl) ethyl)-3-(1-hydroxyphenyl-1H-pyrazol-2(3H)-yl)(pyridin-4-yl)methanone.

 

CMP-B3:(5-(3-hydroxy-4-methoxyphenethyl)-4-(1-hydroxy-2-(3, methoxy 4 hydroxyyphenyl) ethyl)-3-(3-hydroxyphenyl-1H-pyrazol-2(3H)-yl)(pyridin-3-yl)methanone.

 

CMP-B4:(5-(3-hydroxy-4-methoxyphenethyl)-4-(1-hydroxy-2-(3, methoxy 4 hydroxyyphenyl) ethyl)-3-(3-Methoxyyphenyl-1H-pyrazol-2(3H)-yl)(pyridin-3-yl)methanone.

 

CMP-B5:(5-(3-hydroxy-4-methoxyphenethyl)-4-(1-hydroxy-2-(3, methoxy 4 hydroxyyphenyl) ethyl)-3-(2-hydroxy3-Methoxyyphenyl-1H-pyrazol-2(3H)-yl)(pyridin-3-yl)methanone.

 

CMP-B6: (5-(3-hydroxy-4-methoxyphenethyl)-4-(1-hydroxy-2-(3, methoxy 4 hydroxyyphenyl) ethyl)-3-(3-N,N-dimethylphenyl-1H-pyrazol-2(3H)-yl)(pyridin-3-yl)methanone.

 

CMP-D1:(5-(3-hydroxy-4-methoxyphenethyl)-4-(1-hydroxy-2-(3, methoxy 4 hydroxyyphenyl) ethyl)-3-(3-phenyl-1H-pyrazol-2(3H)-yl

 

CMP-D2:5-(2-(4-(1-hydroxy-3-(3-methoxy 4 hydroxyphenyl) propyl)-1,5-diphenylpyrazolidin-3-yl)ethyl)-2-methoxyphenol.

 

CMP-D3:5-(2-(4-(1-hydroxy-3-(3-methoxy 4 hydroxyphenyl) propyl)-1,5-diphenylpyrazolidin-3-yl)ethyl)-2-phenyl2,4-dinitro.

 

CMP-D4:5-(2-(4-(1-hydroxy-3-(3-methoxy 4 hydroxyphenyl) propyl)-1,5-diphenylpyrazolidin-3-yl)ethyl)-2-amide.

 

CMP-D5:5-(2-(4-(1-hydroxy-3-(3-methoxy 4 hydroxyphenyl) propyl)-1,5-diphenylpyrazolidin-3-yl)ethyl)-2-thiamide structures were confirmed by FTIR, Mass, 13C and 1H NMR Techniques.

 

Antimicrobial activity of synthesized compounds:

All the compounds synthesize in the present investigation were screen for their anti-bacterial activity. Antibacterial activities were tested on nutrient medium against, Staphylococcus aureus and Escherchia coli which are representative types of gram positive and gram negative organisms respectively. The antibacterial activities of the compounds were assessed by disc-diffusion method.

 

Preparation of test solutions:

10 mg of the compound was dissolved in 10 ml of DMF. From this 1 ml of solution was taken and diluted up to 10 ml with DMF. Now the concentration of the test solution was 100 mg/ml. From the stock solution 1ml of solution was taken and diluted with 1ml of DMF now the concentration was 50mg/ml20.

 

Preparation of Standard Antibiotic Solution:

Amoxicillin were use as standard antibiotics for comparison and solutions were prepare by using sterile water, as they were water-soluble. The solutions are dilute by using sterile water so that the concentrations of the solutions were 100 mg/ml and 50 mg/ml20.

 

Preparation of Discs:

Discs of 6-7 mm in diameter were punch from NO. 1 Whatman filter paper with sterile cork borer of same size. These discs were sterilized by keeping in oven at 1400C for 60 minutes. Then standard and test solutions were added to each disc and discs were air-dried.

 

Method of Testing:

The sterilize media was cooled to 45oC with gentle shaking to bring about uniform cooling and then inoculate with 18-24 hrs old culture under aseptic conditions, mixwell by gentle shaking. This was poured in to sterile Petri dishes (properly labeled) and allows the medium to set. After solidification all the Petri dishes were transfer to laminar flow unit. Then the discs which were previously prepared were carefully kept on the solidified media by using sterilized forceps. These Petri dishes were kept as it is for one-hour diffusion at room temperature and then for incubation at 370C for 24 hours in an incubator21. The extent diameter of inhibition after 24 hours was measured as the zone of inhibition in millimeters.

 

Screening of anti-bacterial activity:

Table no. 4: Zone of inhibition of synthesized compounds

Sr. No

Compound code

Concentration μg/ml

E.coli

S.

Aureus

1

B1

50

8

8

2

 

100

9

10

3

B2

50

13

12

4

 

100

14

14

5

B3

50

9

10

6

 

100

10

11

7

B4

50

13

13

8

 

100

14

14

9

B5

50

11

10

10

 

100

12

11

11

B6

50

11

9

12

 

100

13

11

13

D1

50

9

10

14

 

100

11

12

15

D2

50

10

11

16

 

100

12

14

17

D3

50

12

13

18

 

100

14

14

19

D4

50

10

9

20

 

100

12

10

21

D5

50

10

11

22

 

100

12

13

23

Amoxicilline

50

14

13

24

 

100

15

14

Range of Zone of inhibition of synthesized compounds: * 6-8 mm poor activity, 9-11 mm moderate activity, 12-15 above good.

 

CONCLUSION:

All the compounds B1-D5 were evaluated for antimicrobial activity against gram positive bacteria, Staphylococcus aureus, gram negative bacteria Escherichia coli, and fungi Candida albicans. The compound B2 presence of electron withdrawing group –OH, and compound B4 having methoxy group –OCH3 at p-position and compound D3 possess the nitro (-NO2) groups of is important for the antibacterial activity and all three compounds B2, B4 and D3 near to significant with standard compounds amoxiciliine, other synthesized compounds having less active than amoxicilline was used as standard drugs. A series of new curcumin derivatives (B1-D5) were synthesized in good yield and were characterized by different spectral studies.

 

ACKNOWLEDGMENT:

Authors are highly acknowledge to Central Instrument Facility lab of Jiwaji University, Gwalior for providing help to characterize compounds and also acknowledge to Faculty and Research Scholars of Department of Pharmacy.

 

CONFLICT OF INTEREST:

All authors declare that they have no conflict of interest

 

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Received on 26.06.2021           Modified on 23.01.2022

Accepted on 29.05.2022         © RJPT All right reserved

Research J. Pharm. and Tech. 2022; 15(7):3091-3095.

DOI: 10.52711/0974-360X.2022.00517