viva 01032011
TRANSCRIPT
![Page 1: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/1.jpg)
111
Synthesis & Antibacterial Evaluation of Juglone
Derivatives MSc candidate: Daniel Tan Teoh Chuan (P-
KM0010/06(R))Main supervisor: Assoc. Prof. Dr. Hasnah Osman
Co-Supervisor: Prof. Azlina Harun @ Kamaruddin
![Page 2: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/2.jpg)
222
Introduction- Juglone (1)
A naturally-occuring naphthoquinone.1
Mainly found in the black walnut tree (Juglans Nigra).1
IUPAC name: 5-Hydroxy-1,4-naphthalenedione.
Molecular formula: C10H6O3 (MW: 174.15 g mol-1).
Dark orange-brown solid. Sparingly soluble in water. Applications: natural dye and
folkloric treatment for skin infections.
O
OOH
(1)
1
2
3
45
6
7
89
10
1. Thomson, R. H. (1971). Naturally Occurring Quinones. London, Academic Press.
ww
w.academ
yfloor.com/index.php%
3...pter%3D
0
http://www.morethanalive.com/Black-Walnut-hull-powder
![Page 3: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/3.jpg)
3
Introduction- Reported Synthesis of Juglone (1)Year Authors Oxidant [O] Yield
(%)
1887 Bernthsen & Semper
Chromic acid -
1984 Wakamatsu et. al.
O2 + salcomine catalyst (3)
71
1990 Barret & Daudon
Hypervalent iodine oxidants [eg. (4)]
91
2006 Oelgemöller et. al.
O2 + sunlight + rose bengal sensitiser
79
OH
OH
[O]
O
OOH(1)(2)
N N
O OCo
(3)
IOCOCF3
OCOCF3(4)
F3CF2CF2CF2CF2CF2C
![Page 4: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/4.jpg)
4
Introduction- The Use of Juglone (1) & 2-Bromo Acetyl Juglone (7) as Organic Reactants Diels-Alder Reactions.2,3
OMe
O
OOH
O
OOH(1)
(i) B(OAc)3 (1equivalent), CH2Cl2, 20oC
(5) rac-(ochromycinone)
+i
O O
75%
2. Guingant, A. & Barreto, M. M. (1987). Tetrahedron Letters 28 (27), 3107.
3. Krohn, K. et. al. (2004). Tetrahedron Asymmetry 15, 713.
O
OOAc
Br+
(7) (8)
O
OOH
O
OOH
O
(+)-(19)
(+)-(ochromycinone)
i
ii
iii(i) Toluene, 80-100oC, 14 h;(ii) K2CO3, MeOH; (iii) h, O2
44%
31%
![Page 5: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/5.jpg)
5
Introduction- The Use of Juglone (1) as an Organic Reactant Direct amination.4
O
OOH
O
OOH
NH3 (g)CH3OH, rt, 24 h
NH2
(1) (9)20%
4. Arnone, A. et. al. (2007). Synthetic Communications 37, 2569.
![Page 6: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/6.jpg)
6
Introduction- Synthetic Juglone Derivatives from Non-juglone Reactants
OAc
OAc
O
OAc O
BrNBS (4.0 eq), AcOH
60oC, 45 min,
(10) (7)>90%
i. S8, H2S2O7ii. H2O
(11)
NH2
OOH
O
(12)
NO2
NO2
5. Heinzman, S. W. & Grunwell, J. R. (1980). Tetrahedron Letters 21 (45), 4305. 6. Fariña, F. et. al. (1985). Synthesis 1985 (8), 781.
2-Bromo acetyl juglone (7) 8-Aminojuglone (12)
![Page 7: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/7.jpg)
77
Introduction- Reported Biological Activities of Juglone Allelopathic activity against potato, tomato and alfalfa.1, 7
Antimicrobial activity against pathogenic fungus8 and bacteria.9
Antitumour activity against intestinal10 and melanoma11 tumours. Antihypertensive activity.12, 13
Larvicidal & molluscicdal activities.14
1. Thomson, R. H. (1971). Naturally Occurring Quinones. London, Academic Press.7. Rietveld, W. J. (1983). Journal of Chemical Ecology 9 (2), 295.8. Clark, A. M. et. al. (1990). Phytotherapy Research 4 (1), 11.9. Cai, L. N. et. al. (2000). Journal of Agricultural and Food Chemistry 48 (3), 909.10. Sugie, S. et. al. (1998). Cancer Letters 127 (1-2), 177.11. Aithal, B. K. et. al. (2009). Cell Biology International 33 (10), 1039.12. Neuhauscarlisle, K. et. al. (1997). Phytomedicine 4 (1), 67.13. Bhosale, S. H. et. al. (1999). Indian Journal of Pharmacology 31 (3), 222.14. Ribeiro, K. A. L. et. al. (2009). Acta Tropica 111, 44.
![Page 8: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/8.jpg)
888
Research Objectives
1. To synthesise juglone derivatives from juglone. 2. To synthesise juglone derivatives from non-juglone
starting materials.3. To characterise all synthetic compounds.4. To evaluate the antibacterial activities of all synthetic
compounds.
![Page 9: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/9.jpg)
9
Research Methodology- Synthesis from Juglone Reactant The Diels-Alder synthesis of ochromycinone precursor (19).
O
OOH
O
OOH
(1)
(i) Dry CHCl3, Ar, rt, 3 h (ii) DBU (2 eq.), air, rt, 13 h
(50)
+i
OSiMe3
ii
(19)
![Page 10: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/10.jpg)
10
Research Methodology- Synthetic Preparation of Siloxy diene (50)
O EtOC HOH2C OHC
i ii iii iv
(i) NaH, (EtO)2POCH2COOEt, dry THF, rt, 5 h; (ii) LiAlH4, dry toluene, ref lux (120oC), 3 h; (iii)NMO, TPAP, 4 Å molecular sieves, dry CH2Cl2, 3 h; (iv) Dry ZnCl2 (liq), dry Et3N, dry TMSCl,dry toluene, Ar
OSiMe3(15) (47) (48) (49) (50)
O
15. Krohn, K. et. al. (1995). Liebigs Annalen 1995 (8), 1529.16. Hasnah, O. (2005). Synthetic Approaches to the Angucycline Antibiotics, Dunedin, University of Otago, PhD.
![Page 11: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/11.jpg)
11
Research Methodology- Synthesis from Juglone Reactant Direct nitration
17. Anuradha, V. et. al. (2006). Tetrahedron Letters 47, 4933.
OH
O
OOH
O
O
Ni(NO3)2.6H2O, p -TsOH
acetone, ref lux, >24 h
(1) (51)
NO2
![Page 12: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/12.jpg)
12
Research Methodology- Synthesis from Non-juglone Reactants Friedel-Crafts Acylation
OH
+ O
O
O
O
OOH
X R
(26) Cl Me
(27) Br Me
(65) Br H
(66) Cl H
(67) Cl Et
(26, 27, 65, 66, 67) (68) (30, 31, 52, 53, 54)
XR
XR
i
(i) AlCl3 (liq), NaCl (liq), 180-185oC, 2 mins. (ii) H3O
ii
X R
(30) Cl Me
(31) Br Me
(52) Br H
(53) Cl H
(54) Cl Et
18. Mahapatra, et. al. (2007). Bioorganic & Medicinal Chemistry 15, 7638.19. Musgrave, O. C. & Skoyles, D. (2001). Journal of the Chemical Society, Perkin Transaction 1, 1318.
![Page 13: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/13.jpg)
13
Research Methodology- Separation and Purification of Synthetic Compounds
Silica gel column chromatography. Single-solvent recrystallisation.
![Page 14: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/14.jpg)
14
Research Methodology- Characterisation of Synthetic Compounds
Melting point. High Resolution Electron Spray Ionisation Mass
Spectrometry (HRESIMS). Gas Chromtography-Mass Spectrometry (GC-MS). Ultraviolet spectroscopy (UV). Infrared spectroscopy (IR). Nuclear magnetic resonance spectroscopy (NMR). X-ray crystallography.
![Page 15: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/15.jpg)
15
Research Methodology- Antibacterial Assay Broth microdilution method. Conducted in triplicates. 96-Well tissue culture plate. Sample: DMSO: water (1:9). Initial concentration: 250 g ml-1. Reference: juglone. Negative control: antibiotic. Positive control: Müller-Hinton
broth, MHB. Colour indicator: MTT [3-(4,5-
Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] www.opticsplanet.net/bd-falcon-u...lid.html
![Page 16: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/16.jpg)
16
Results & Discussion- Preparation of (±)-(E)-Trimethyl(2-(5-methylcyclohex-1-enyl)vinyloxy) (50)
O EtOC HOH2C OHC
i ii iii iv
(i) NaH, (EtO)2POCH2COOEt, dry THF, rt, 5 h; (ii) LiAlH4, dry toluene, ref lux (120oC), 3 h; (iii)NMO, TPAP, 4 Å molecular sieves, dry CH2Cl2, 3 h; (iv) Dry ZnCl2 (liq), dry Et3N, dry TMSCl,dry toluene, Ar
OSiMe3
(15) (47) (48) (49) (50)
O
79% 56% 71% 75%
Overall yield 24%
![Page 17: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/17.jpg)
17
Results & Discussion-Synthesis of (±)-8-Hydroxy-3-methyl-1,2,3,4-tetrahydrotetraphene-7,12-dione (19)
Non-Lewis acid promoted. Mild reaction condition. DBU base: elimination &
further aromatisation.
O
OOH
O
OOH
(1)
(i) Dry CHCl3, Ar, rt, 3 h (ii) DBU (2 eq.), air, rt, 13 h
(50)
+i
OSiMe3
ii
(19)
47%
![Page 18: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/18.jpg)
18
Results & Discussion- Synthesis of Ochromycinone Precursor (19)
O
OOH
(1) (50)
+
OSiMe3
O
OOH
H
HO
O
OOH
(55)
(60)
H
B
O
OOH
H H
B = DBU
+ BH
SiMe3
+ SiMe3O
H
tautomerism
O O
OH
OOH
OO O
OOH
O
OH
H
B
O
OOH
(19)
+ BH +HO2
20. Kim, K. et. al. (1992). Journal of Organic Chemistry 57 (21), 5557.
![Page 19: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/19.jpg)
19
Results & Discussion- Synthesis of 5-Hydroxy-8-nitro-1,4-naphthalenedione (51)
OH
O
OOH
O
O
Ni(NO3)2.6H2O, p -TsOH
acetone, ref lux, >24 h
(1) (51)
NO2
11%
The para-nitro substituted juglone is the sole isomeric product. Contrary to Anuradha et. al., 2006.
O
O
H
H
H
H
O
N
H
O O(51)
Hydrogen bond
![Page 20: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/20.jpg)
202020
Results & Discussion- Syntheses of Halogen Substituted Juglone Derivatives
Compound Melting Point (oC) Yield (%)
158-160 (lit. 160-161) 8
148-150 (lit. 154) 10
206-209 24
202-204 (lit. 201-202) 10
108-111 14
O
OOH(53)
Cl
O
OOH
Br
(31)
O
OOH
Cl
(54)
O
OOH(30)
Cl
O
OOH(52)
Br
![Page 21: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/21.jpg)
2121
Results & Discussion- Syntheses of Halogen Substituted Juglone Derivatives
OH
+ O
O
O
O
OOH(26) (68) (30)
Cl Cl
i
(i) AlCl3 (liq), NaCl (liq), 180-185oC, 2 mins. (ii) H3O
ii+
O
OOH
OH
OH(69)
8% 1%
18. Musgrave, O. C. & Skoyles, D. (2001). Journal of the Chemical Society, Perkin Transaction 1, 1318.
(69): Helminthosporin, 1,5,8-trihydroxy-3-methyl-9,10-anthracenedione
![Page 22: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/22.jpg)
22
Results & Discussion- Syntheses of Halogen Substituted Juglone Derivatives
OH
+ O
O
O
O
OOH(26) (68) (30)
Cl Cl OH
OHOH
Cl
[H]
O
O
O
(68)
OH
OHOH
O
O
O
OOH
OH
OH
ClCltautomerism
O
OOH
OH
OH(69)
1. [H], H
2. -HCl
18. Musgrave, O. C. & Skoyles, D. (2001). Journal of the Chemical Society, Perkin Transaction 1, 1318.
![Page 23: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/23.jpg)
232323
Results & Discussion- Antibacterial AssayCompound MIC (mM)
B. cereus S. aureus E. Coli K. Pneumoniae
0.36 0.72 0.36 0.36
0.21 0.43 0.43 0.43
0.29 0.14 0.57 0.57
0.56 1.1 0.56 0.56
O
OOH(1)
O
OOH(19)
O
OOH
NO2
(51)
O
OOH
Cl
(30)
![Page 24: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/24.jpg)
242424
Results & Discussion- Antibacterial AssayCompound MIC (mM)
B. cereus S. aureus E. Coli K. Pneumoniae
0.47 0.94 0.47 0.47
0.50 0.99 0.50 0.50
0.23 0.46 0.23 0.23
0.53 1.1 0.53 0.53
O
OOH(31)
Br
O
OOH(69)
OH
OH
O
OOH
Br
(52)
O
OOH
Cl
(54)
![Page 25: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/25.jpg)
252525
Results & Discussion- Antibacterial Assay
Compound MIC (mM)
B. cereus S. aureus E. Coli K. Pneumoniae
0.60 0.60 0.60 0.60
Gentamicin - - 0.027 0.014
Tetracycline 0.016 <0.004 - -
O
OOH(53)
Cl
![Page 26: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/26.jpg)
26
Conclusion
The non-Lewis acid promoted, mild Diels-Alder reaction between juglone (1) & siloxydiene (50) afforded the ochromycinone precursor (19) in 47% yield.
8-Nitrojuglone (51) was obtained as the sole para isomeric product in 11% yield from the direct nickel(II) nitrate nitration of juglone (1).
The Friedel-Crafts acylation gave five 8-halojuglone derivatives (30, 31, 52, 53 & 54) in 8-24% yields. Two of which (52 & 54) are new derivatives.
A helminthosporin anthraquinone, (69) was obtained as the minor product of one of the Friedel-Crafts acylation reactions.
![Page 27: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/27.jpg)
27
Conclusion
8-Nitrojuglone (51) exhibited the most significant activity against S. aureus (MIC= 0.14 mM).
Helminthosporin (69) displayed high activities against the Gram-negative bacteria tested.
All of the five 8-halojuglone derivatives showed lower antibacterial activities as compared to juglone (1).
![Page 28: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/28.jpg)
2828
Acknowledgements
Assoc. Prof. Dr. Hasnah Osman (Main supervisor). Prof. Azlina Harun @ Kamaruddin (Co-supervisor). Madam Suriyati Mohamad. Ms. Anis Safirah. USM School of Chemical Sciences. USM School of Biological Sciences. USM Fellowship Scheme. PURGS grant: 1001/PKIMIA/832017. RU grant: 1001/PKIMIA/811016.
![Page 29: Viva 01032011](https://reader035.vdocuments.us/reader035/viewer/2022062522/58a9574b1a28ab77408b5c77/html5/thumbnails/29.jpg)
292929