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![www.wjpps.com Vol 3, Issue 7, 2014. 956
Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences
SYNTHESIS, CHARACTERIZATION AND EVALUATION OF
SOME CYANOPYRIDINE DERIVATIVES
T.Rajkumar *, S.Paramesh, B.Ramesh, G.Pushapalatha, K.Bharathi & C.Ramesh
Department of Pharmaceutical Chemistry, Creative Educational Society’s
College of Pharmacy, Kurnool. AP. 518218
ABSTRACT
A new series of Cyanopyridine derivatives were synthesized by
condensing chalcones with malononitrile. First Chalcones were
prepared by treating various acetophenone derivatives with different
aromatic aldehydes by Claisen-Schimidt Condensation and then
refluxed with malononitrile to obtain cyanopyridines .All the
compounds were screened for their antitubercular and anthelmintic
activities. The structures of newly synthesized compounds were
characterized on the basis of IR, 1
H NMR and Mass spectral data.
Key Words: Cyanopyridine; Chalcone; Antitubercular activity;
Anthelmintic activity.
INTRODUCTION
Cyanopyridines are one of the most versatile synthetic intermediates used in pharmaceutical
industries because of their significant biological and pharmacological activities. The
importance of cyanopyridines in organic synthesis has increased over the past few decades.
Many fused cyanopyridines have drawn wide spectrum biological activities.
They are associated with wide spectrum of biological activities such as antiviral,
anthelmintic, anti-inflammatory, anticonvulsant and anti tubercular activity etc [1-3]
. Hence, it
worthwhile to prepare some new cyanopyridine derivatives.
In view of these observations and with a view to further explore the pharmacological profile
of this class of compounds; the present study includes synthesis of novel 3-cyanopyridines
(pyridine-3-carbonitriles).
WWOORRLLDD JJOOUURRNNAALL OOFF PPHHAARRMMAACCYY AANNDD PPHHAARRMMAACCEEUUTTIICCAALL SSCCIIEENNCCEESS
SSJJIIFF IImmppaacctt FFaaccttoorr 22..778866
VVoolluummee 33,, IIssssuuee 77,, 995566--996688.. RReesseeaarrcchh AArrttiiccllee IISSSSNN 2278 – 4357
Article Received on
04 May 2014,
Revised on 25 May
2014,
Accepted on 26 June 2014
*Correspondence for Author
T.Rajkumar
Department of Pharmaceutical
Chemistry, Creative
Educational Society’s College
of Pharmacy, Kurnool. AP](https://image.slidesharecdn.com/a5faa087-4ad1-4556-8580-bc834ecb302b-150827175457-lva1-app6891/85/WJPPS-2014-1-320.jpg)
![www.wjpps.com Vol 3, Issue 7, 2014. 957
Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences
MATERIALS AND METHODS
All the chemicals and solvents used in this study were of analytical grade (S.D. FINE Chem.
Limited, Mumbai). Melting points of the compounds were determined using Open Capillary
Melting Point apparatus and were reported uncorrected. FTIR spectra were detected in KBr
pellets using Bruker FTIR- spectrophotometer. The 1
H NMR spectra were recorded in
DMSO-d6 by NMR 300MHZ spectrometers using tetra methyl silane as an internal standard.
Reaction progress was checked by TLC in a solvent-vapor-saturated chamber on aluminium
plates coated with Silica Gel GF254 followed by visualization under UV light (254 nm).
Scheme I: Synthesis of Chalcones [4]
To a 20 ml solution of 20% sodium hydroxide in distilled water, acetophenone (0.001 mol)
was added and dissolved. To this benzaldehyde (0.001 mol) was added in small amounts with
continuous stirring with the help of a mechanical stirrer or about 2 h. The mixture was left in
refrigerator for 24 h. Then the yellowish product thus formed is filtered, dried and
recrystallized from ethanol. The synthesized compounds were monitored by TLC8
.
Scheme II: Synthesis of Cyanopyridine derivatives[5-6]
Compound C1-6 (0.01mol) in alcohol (40 mL) malononitrile (0.01mol) and ammonium
acetate was refluxed for 8 h. The reaction mixture was then cooled, poured into crushed ice
and product separated out was filtered, washed with water, dried and recrystallized from ethyl
alcohol.
Fig.1: General synthetic scheme](https://image.slidesharecdn.com/a5faa087-4ad1-4556-8580-bc834ecb302b-150827175457-lva1-app6891/85/WJPPS-2014-2-320.jpg)
![www.wjpps.com Vol 3, Issue 7, 2014. 958
Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences
Table 1: Type of Chalcones & Cyanopyridine Substitutions
Anti tubercular activity
Anti-TB activity using Alomar Blue Dye [7-8]
The anti mycobacterial activity of compounds were assessed against M. tuberculosis using
microplate Alamar Blue assay (MABA). This methodology is non-toxic, uses a thermally
stable reagent and shows good correlation with propotional and BACTEC radiometric
method. Briefly, 200µl of sterile deionzed water was added to all outer perimeter wells of
sterile 96 wells plate to minimized evaporation of medium in the test wells during incubation.
The 96 wells plate received 100 µl of the Middle brook 7H9 broth and serial dilutions of
compounds were made directly on plate. The final drug concentrations tested were 100 to 0.2
µg/ml. Plates were covered and sealed with para film and incubated at 37ºC for five days.
After this time, 25µl of freshly prepared 1:1 mixture of Almar Blue reagent and 10% tween
80 was added to the plate and incubated for 24 hrs. A blue color in the well was interpreted as
no bacterial growth, and pink color was scored as growth. The MIC was defined as lowest
drug concentration which prevented the color change from blue to pink.
Anthelmintic activity
Animals
Adult earthworm (Pheretima posthumous) was used to evaluate anthelmintic activity in vitro.
Earthworm was collected Govt. medical College Kurnool, The average size of earthworm
was 6-8 cm, Earth worm was identified in Dept. of Zoology, Govt. medical College Kurnool.
Reference Drug
Albendazole tablets (Mankind) was used during the experimental protocol.
Experimental protocol [9]
The anthelmintic assay was carried out as per the method of Ajaiyeoba et al . The assay was
performed in vitro using adult earthworm (Pheretima posthuma) owing to its anatomical and
physiological resemblance with the intestinal roundworm parasites of human beings for
Product R1 R2 R3
CP 1 4-OCH3 4-Cl CN
CP 2 2-OH 4-Cl CN
CP 3 4-OCH3 3-OCH3, 4-OH CN
CP 4 2-OH 3-OCH3, 4-OH CN
CP 5 4-OCH3 3-Cl CN
CP 6 2-OH 3-Cl CN](https://image.slidesharecdn.com/a5faa087-4ad1-4556-8580-bc834ecb302b-150827175457-lva1-app6891/85/WJPPS-2014-3-320.jpg)
![www.wjpps.com Vol 3, Issue 7, 2014. 959
Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences
preliminary evaluation anthelmintic activity. Test samples of the synthesized compounds
were prepared at the concentrations, 0.001,0.002,0.005 &0.10 µg/ml in distilled water,
Normal saline and ethanol and six worms i.e. Pheretima posthuma, were placed in each nine
cm petri dish containing 25 ml of above test solution of synthesized compounds. Albendazole
(10 mg/ml) was used as reference standard and distilled water as control. All the test solution
and standard drug solution were prepared freshly before starting the experiments.
Observations were made for the time taken for paralysis was noted when no movement of
any sort could be observed except when the worms were shaken vigorously. Time for death
of worms were recorded after ascertaining that worms neither moved when shaken
vigorously nor when dipped in warm water (500
C). All the results were shown in Table No.8
and expressed as a mean± SEM of six worms in each group.
Statistical methods
The results are expressed as means ± standard error of mean (S.E.M.). The data were
analyzed by one-way ANOVA followed by Dunnett’s test for comparing the control and the
test groups, using trial version of GraphPad InStat v 3. Statistical significance was assumed at
the 0.05 and 0.01 levels.
RESULTS & DISCUSSION
The structure, melting points, percentage yields, elemental analysis physical data of
chalcones and cyanopyridines are given in Table No 2 & 3, FTIR & 1
H NMR [10-11]
are given
in Table No 4 & 5 respectively.
Table 2: Physicochemical characterization data for synthesized Chalcones compounds
Compound
Mol.
Formula
Mol.Structure
Mol.
weight
Yield
( % )
M.P TLC
C 1 C16H13O2Cl 272.4 63% 1960
C 0.63
C 2 C15H11O2Cl 258 56% 2300
C 0.78](https://image.slidesharecdn.com/a5faa087-4ad1-4556-8580-bc834ecb302b-150827175457-lva1-app6891/85/WJPPS-2014-4-320.jpg)
![www.wjpps.com Vol 3, Issue 7, 2014. 967
Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences
The existing designed analogs (C 1-6) can be further modified so as to include substituted
pyrazolines, pyrimidines, Oxazoles, hydrazones, benzodiazepines etc which can produce
further substituted derivatives. Such analogs can be synthesized, and evaluated for their
Anthelmintic, anti-microbial and anti tubercular activities and other activities.
It would be interesting to deduce the structure of potent compounds for the activity claimed,
they may be our lead molecules and there is a possibility to extend the lead molecule towards
drug design using various drug design software’s like maestro, glide, scigress, autodock etc.
ACKNOWLEDGEMENT
The authors are thankful to the Principal, CES college of Pharmacy, HOD Department of
Pharmaceutical Chemistry for their support. Also we thank Dr. Murugesan, SAIF, IIT
Chennai and Dr. Kishore Bhat, Department of Microbiology, Maratha Mandal College of
Dental sciences, Belgaum for providing spectral and anti tubercular data respectively.
REFERENCES
1. Parikh KS & Patel RP. Synthesis and Characterization of Some Cyanopyridine
Compounds in Therapeutic Interest. Int.J. ChemTech Res, 2009; 1(3);581-586.
2. Hassan M Faidallah et al. Synthesis of some new 2-oxo-1,4-disubstituted-1,2,5,6-
tetrahydro-benzo[h]quinoline-3-carbonitriles and their biological evaluation as cytotoxic
and antiviral agents. J. Chem. Sci, 2012; 124(3); 625–631.
3. Kachhadia et al. Synthesis of Isoxazoles and Cyanopyridines Bearing Benzo(b)thiophene
Nucleus as Potential Antitubercular and Antimicrobial Agents. J. Sci. I. R. Iran, 2004;
15(1); 47-51.
4. Venkatesan J et al. synthesis and biological evaluation of 4, 6-diaryl substituted-4, 5-
dihydro-2-amino pyrimidines. Indian J.Pharm.Sci. 2007; 69(4); 586-589.
5. Anjani solankee et al. Anti microbial evaluation of some novel isoxazoles,
cyanopyridines and pyrimidinthiones. Indian J Chem.2013, 52 B;671-676.
6. Anjani Solankee et al. Synthesis and studies of chalcones and its Cyanopyridine and
Acetyl Pyrazoline Derivatives. Rasayan J. Chem. 2008; 1(3); 591-595.
7. L Collins, tuberculosis and Mycobacterium avium compounds against Mycobacterium
460 system for high-throughput screening of Microplate alamar blue assay versus
BACTEC. Antimicrob. Agents Chemother.1997; 41(5); 1004-1009.
8. Hussain et al. Synthesis and biological evaluation of some novel 2-mercapto pyrimidines
Int J Pharm Pharm Sci,2013;5(2); 471-473](https://image.slidesharecdn.com/a5faa087-4ad1-4556-8580-bc834ecb302b-150827175457-lva1-app6891/85/WJPPS-2014-12-320.jpg)
Uploaded byrajkutty1983
WJPPS-2014
This document describes the synthesis and characterization of cyanopyridine derivatives and their evaluation for antitubercular and anthelmintic activity. Chalcones were first synthesized through Claisen-Schmidt condensation and then reacted with malononitrile to obtain cyanopyridines. The structures of the compounds were characterized using techniques such as IR, NMR and mass spectrometry. The cyanopyridines were then evaluated for antitubercular activity against M. tuberculosis using the microplate Alamar Blue assay and for anthelmintic activity against earthworms. Some of the compounds showed promising activity in both assays.
WJPPS-2014
- 1. www.wjpps.com Vol 3,Issue 7, 2014. 956 Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences SYNTHESIS, CHARACTERIZATION AND EVALUATION OF SOME CYANOPYRIDINE DERIVATIVES T.Rajkumar *, S.Paramesh, B.Ramesh, G.Pushapalatha, K.Bharathi & C.Ramesh Department of Pharmaceutical Chemistry, Creative Educational Society’s College of Pharmacy, Kurnool. AP. 518218 ABSTRACT A new series of Cyanopyridine derivatives were synthesized by condensing chalcones with malononitrile. First Chalcones were prepared by treating various acetophenone derivatives with different aromatic aldehydes by Claisen-Schimidt Condensation and then refluxed with malononitrile to obtain cyanopyridines .All the compounds were screened for their antitubercular and anthelmintic activities. The structures of newly synthesized compounds were characterized on the basis of IR, 1 H NMR and Mass spectral data. Key Words: Cyanopyridine; Chalcone; Antitubercular activity; Anthelmintic activity. INTRODUCTION Cyanopyridines are one of the most versatile synthetic intermediates used in pharmaceutical industries because of their significant biological and pharmacological activities. The importance of cyanopyridines in organic synthesis has increased over the past few decades. Many fused cyanopyridines have drawn wide spectrum biological activities. They are associated with wide spectrum of biological activities such as antiviral, anthelmintic, anti-inflammatory, anticonvulsant and anti tubercular activity etc [1-3] . Hence, it worthwhile to prepare some new cyanopyridine derivatives. In view of these observations and with a view to further explore the pharmacological profile of this class of compounds; the present study includes synthesis of novel 3-cyanopyridines (pyridine-3-carbonitriles). WWOORRLLDD JJOOUURRNNAALL OOFF PPHHAARRMMAACCYY AANNDD PPHHAARRMMAACCEEUUTTIICCAALL SSCCIIEENNCCEESS SSJJIIFF IImmppaacctt FFaaccttoorr 22..778866 VVoolluummee 33,, IIssssuuee 77,, 995566--996688.. RReesseeaarrcchh AArrttiiccllee IISSSSNN 2278 – 4357 Article Received on 04 May 2014, Revised on 25 May 2014, Accepted on 26 June 2014 *Correspondence for Author T.Rajkumar Department of Pharmaceutical Chemistry, Creative Educational Society’s College of Pharmacy, Kurnool. AP
- 2. www.wjpps.com Vol 3,Issue 7, 2014. 957 Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences MATERIALS AND METHODS All the chemicals and solvents used in this study were of analytical grade (S.D. FINE Chem. Limited, Mumbai). Melting points of the compounds were determined using Open Capillary Melting Point apparatus and were reported uncorrected. FTIR spectra were detected in KBr pellets using Bruker FTIR- spectrophotometer. The 1 H NMR spectra were recorded in DMSO-d6 by NMR 300MHZ spectrometers using tetra methyl silane as an internal standard. Reaction progress was checked by TLC in a solvent-vapor-saturated chamber on aluminium plates coated with Silica Gel GF254 followed by visualization under UV light (254 nm). Scheme I: Synthesis of Chalcones [4] To a 20 ml solution of 20% sodium hydroxide in distilled water, acetophenone (0.001 mol) was added and dissolved. To this benzaldehyde (0.001 mol) was added in small amounts with continuous stirring with the help of a mechanical stirrer or about 2 h. The mixture was left in refrigerator for 24 h. Then the yellowish product thus formed is filtered, dried and recrystallized from ethanol. The synthesized compounds were monitored by TLC8 . Scheme II: Synthesis of Cyanopyridine derivatives[5-6] Compound C1-6 (0.01mol) in alcohol (40 mL) malononitrile (0.01mol) and ammonium acetate was refluxed for 8 h. The reaction mixture was then cooled, poured into crushed ice and product separated out was filtered, washed with water, dried and recrystallized from ethyl alcohol. Fig.1: General synthetic scheme
- 3. www.wjpps.com Vol 3,Issue 7, 2014. 958 Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences Table 1: Type of Chalcones & Cyanopyridine Substitutions Anti tubercular activity Anti-TB activity using Alomar Blue Dye [7-8] The anti mycobacterial activity of compounds were assessed against M. tuberculosis using microplate Alamar Blue assay (MABA). This methodology is non-toxic, uses a thermally stable reagent and shows good correlation with propotional and BACTEC radiometric method. Briefly, 200µl of sterile deionzed water was added to all outer perimeter wells of sterile 96 wells plate to minimized evaporation of medium in the test wells during incubation. The 96 wells plate received 100 µl of the Middle brook 7H9 broth and serial dilutions of compounds were made directly on plate. The final drug concentrations tested were 100 to 0.2 µg/ml. Plates were covered and sealed with para film and incubated at 37ºC for five days. After this time, 25µl of freshly prepared 1:1 mixture of Almar Blue reagent and 10% tween 80 was added to the plate and incubated for 24 hrs. A blue color in the well was interpreted as no bacterial growth, and pink color was scored as growth. The MIC was defined as lowest drug concentration which prevented the color change from blue to pink. Anthelmintic activity Animals Adult earthworm (Pheretima posthumous) was used to evaluate anthelmintic activity in vitro. Earthworm was collected Govt. medical College Kurnool, The average size of earthworm was 6-8 cm, Earth worm was identified in Dept. of Zoology, Govt. medical College Kurnool. Reference Drug Albendazole tablets (Mankind) was used during the experimental protocol. Experimental protocol [9] The anthelmintic assay was carried out as per the method of Ajaiyeoba et al . The assay was performed in vitro using adult earthworm (Pheretima posthuma) owing to its anatomical and physiological resemblance with the intestinal roundworm parasites of human beings for Product R1 R2 R3 CP 1 4-OCH3 4-Cl CN CP 2 2-OH 4-Cl CN CP 3 4-OCH3 3-OCH3, 4-OH CN CP 4 2-OH 3-OCH3, 4-OH CN CP 5 4-OCH3 3-Cl CN CP 6 2-OH 3-Cl CN
- 4. www.wjpps.com Vol 3,Issue 7, 2014. 959 Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences preliminary evaluation anthelmintic activity. Test samples of the synthesized compounds were prepared at the concentrations, 0.001,0.002,0.005 &0.10 µg/ml in distilled water, Normal saline and ethanol and six worms i.e. Pheretima posthuma, were placed in each nine cm petri dish containing 25 ml of above test solution of synthesized compounds. Albendazole (10 mg/ml) was used as reference standard and distilled water as control. All the test solution and standard drug solution were prepared freshly before starting the experiments. Observations were made for the time taken for paralysis was noted when no movement of any sort could be observed except when the worms were shaken vigorously. Time for death of worms were recorded after ascertaining that worms neither moved when shaken vigorously nor when dipped in warm water (500 C). All the results were shown in Table No.8 and expressed as a mean± SEM of six worms in each group. Statistical methods The results are expressed as means ± standard error of mean (S.E.M.). The data were analyzed by one-way ANOVA followed by Dunnett’s test for comparing the control and the test groups, using trial version of GraphPad InStat v 3. Statistical significance was assumed at the 0.05 and 0.01 levels. RESULTS & DISCUSSION The structure, melting points, percentage yields, elemental analysis physical data of chalcones and cyanopyridines are given in Table No 2 & 3, FTIR & 1 H NMR [10-11] are given in Table No 4 & 5 respectively. Table 2: Physicochemical characterization data for synthesized Chalcones compounds Compound Mol. Formula Mol.Structure Mol. weight Yield ( % ) M.P TLC C 1 C16H13O2Cl 272.4 63% 1960 C 0.63 C 2 C15H11O2Cl 258 56% 2300 C 0.78
- 5. www.wjpps.com Vol 3,Issue 7, 2014. 960 Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences C 3 C17H16O4 284 87.2% 2400 C 0.55 C 4 C16H14O4 270 64.5% 2630 C 0.84 C 5 C16H13O2Cl 272.4 55% 1960 C 0.75 C 6 C15H11O2Cl 258 70% 2300 C 0.56 Table 3: Physicochemical characterization data for synthesized Cyanopyridines Compound Mol. Formula Mol. Structure Mol. weight Yield ( % ) M.P TLC CP1 C19H13ClN2O 320 56% 1960 C 0.63 CP2 C18H11ClN2O 306 65% 2300 C 0.56
- 6. www.wjpps.com Vol 3,Issue 7, 2014. 961 Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences CP3 C20H16N2O3 332 76% 2260 C 0.65 CP4 C19H14N2O3 318 75% 2360 C 0.78 CP5 C19H11ClN2O 320 65% 2560 C 0.56 CP6 C18H11ClN2O 306 77% 2360 C 0.65 Table No 4: FT-IR spectral data of Chalcones & Cyanopyridines CHALCONES CYANOPYRIDINES COMPOUND WAVE NUMBER (cm-1 ) FUNCTIONAL GROUP COMPOUND WAVE NUMBER (cm-1 ) FUNCTIONA L GROUP C 1 1600 1658 2836 1182 756.68 C=C (Stetching) C-O Group stretching CH-Aromatic stretching CO-CH2 stretching Aromatic bending Cl CP 1 1572.88 2211.35 834.40 1649.26 2930.65 C=N C-N Cl C-O Group stretching CH-Aromatic stretching
- 7. www.wjpps.com Vol 3,Issue 7, 2014. 962 Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences Table No 5: 1 HNMR spectral data of Chalcones & Cyanopyridines C 2 1453.42 3203 2975.17 687.64 CO Group stretching OH Group Aromatic bending Cl CP 2 1553.88 2197.08 760.33 1228.99 2925.11 C=N C-N Cl OH stretching CH-Aromatic stretching C 3 1580.86 1228.16 1656.63 3262.01 C=C (Stetching) OH bending C-O Group stretching CH-Aromatic stretching CP 3 1567.33 2227.63 728.34 1611.34 2976.26 C=N C-N Cl C-O Group stretching CH-Aromatic stretching C 4 1228.16 1656.90 3263.14 1581.12 OH Bending C-O stretching CH-Aromatic stretching CH-CH stretching CP 4 1567.34 2227.81 728.65 1188.49 2976.25 C=N C-N Cl OH stretching CH-Aromatic stretching C 5 2975.28 1664.93 1170.96 778.91 CH-Aromatic stretching C-O stretching CO-CH2 stretching Cl CP 5 1569.82 2215.57 836.49 1252.18 2931.47 C=N C-N Cl C-O Group stretching CH-Aromatic stretching C 6 1584 755.91 2958 1207 CO Group stretching Cl CH-Aromatic stretching CH-Aromatic stretching CP 6 1553.00 2195.59 759.45 1228.58 2903.09 C=N C-N Cl OH Group stretching CH-Aromatic stretching CHALCONES CYANOPYRIDINES COMPOUND 1 HNMR(CDC13,ppm) COMPOUND 1 HNMR(CDC13,ppm) C1 7.602-7.752(1H;d;CH-Ar) 7.285-7.418 (1H;d;CO-H=) 6.875-8.308(8H;m,Ar-H) CP1 7.285(2H;d;CH-Ar) 3.877 (3H;S;OCH3) 7.048-8.285(8H;m,Ar-H) C2 7.612-7.802(1H;d;CH-Ar) 7.261-7.425 (1H;d;CO-H=) 6.192-8.310(8H;m,Ar-H) 3.872(3H;s;OCH3) CP2 7.285(2H;d;CH-Ar) 4.28-5.0 (1H;OH) 7.038-8.176(8H;m,Ar-H) C3 7.648-7.697(1H;d;CH-Ar) 7.285-7.492 (1H;d;CO-H=) 7.048-8.285(8H;m,Ar-H) 3.76(3H;s;OCH3) CP3 7.270(1H;d;CH-Ar) 3.866 (3H;S;OCH3) 6.891-8.293(8H;m,Ar-H) 4.52 (1H;OH) C4 7.573-7.828(1H;d;CH-Ar) CP4 7.3(1H;d;CH-Ar)
- 8. www.wjpps.com Vol 3,Issue 7, 2014. 963 Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences Table No 6: Elemental Analysis Compound ELEMENTAL ANALYSIS C % H % N % CP 1 65.42 13.54 4.66 CP 2 63.95 12.60 5.12 CP 3 56.51 14.81 4.56 CP 4 63.01 15.33 4.90 CP 5 62.23 14.56 4.84 CP 6 67.48 13.49 4.40 Table No 7: Anti Tubercular activity of Cyanopyridine derivatives Compounds 100 µg/ml 50 µg/ml 25 µg/ml 12.5 µg/ml 6.25 µg/ml 3.125 µg/ml 1.6 µg/ml 0.8 µg/ml 1 CP1 S S R R R R R R 2 CP2 S S S S S R R R 3 CP3 S R R R R R R R 4 CP4 S S R R R R R R 5 CP 5 S S R R R R R R 6 CP 6 S S R R R R R R S – Sensitive R – Resistant Here are the standard values for the Anti-tb test which was performed. Pyrazinamide- 3.125µg/ml ; Streptomycin- 6.25µg/ml Here are the standard values for the Anti-TB test which was performed. Pyrazinamide- 3.125µg/ml Streptomycin- 6.25µg/ml 7.202-7.503 (1H;d;CO-H=) 8.143-8.406 (8H;m,Ar-H) 3.872(3H;s;OCH3) 3.956 (3H;S;OCH3) 6.637-8.101(8H;m,Ar-H) 4.5(1H;OH) C5 7.602-7.752(1H;d;CH-Ar) 7.285-7.418 (1H;d;CO-H=) 6.875-8.308(8H;m,Ar-H) CP5 8.035(2H;d;CH-Ar) 3.879 (3H;S;OCH3) 6.552-8.627(8H;m,Ar-H) C6 7.612-7.802(1H;d;CH-Ar) 7.261-7.425 (1H;d;CO-H=) 6.192-8.310(8H;m,Ar-H) 3.872(3H;s;OCH3) CP6 7.285(2H;d;CH-Ar) 4.35-5.0 (1H;OH) 6.552-8.627(8H;m,Ar-H)
- 9. www.wjpps.com Vol 3,Issue 7, 2014. 964 Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences Table No 8: Anthelmintic Effect of Synthesized cyanopyridines on Pheretima posthuma S.No Treatment Concentration Time taken for paralysis ( Mean ± SEM) in Sec Time taken for death ( Mean ± SEM) 1. Control 0.001 In ethanol 0 0 0.002 In ethanol 0 0 0.005 In ethanol 0 0 0.01 In ethanol 0 0 2. Albendazole 0.001 In ethanol 13.51±1.29 61±2.34 0.002 In ethanol 12.05±0.82 60±0.13 0.005 In ethanol 10.73±0.55 59±12.92 0.01 In ethanol 9.6±0.49 55.3±4.89 3. CP 1 0.001 In ethanol 13.19±0.94 60.4±0.15 0.002 In ethanol 10.43±0.7 59.2±0.036 0.005 In ethanol 11.23±0.32 58.05±0.036 0.01 In ethanol 9.48±0.45 56.2±3.92 4. CP 2 0.001 In ethanol 17.63±0.86 62±16.6 0.002 In ethanol 14.76±0.76 60.7±12.3 0.005 In ethanol 11.9±1.01 59.2±16.4 0.01 In ethanol 10.33±1.12 54.7±0.06 5. CP 3 0.001 In ethanol 12.17±0.94 64.2±0.008 0.002 In ethanol 10.5±0.82 61.02±0.10 0.005 In ethanol 10.2±0.49 60.4±0.008 0.01 In ethanol 9.52±0.29 56.6±0.04 6 CP 4 0.001 In ethanol 10.9±0.34 53.3±0.02 0.002 In ethanol 9.2±0.29 50.8±0.03 0.005 In ethanol 9.55±1.03 48±0.05 0.01 In ethanol 9±0.87 45.34±0.07 7 CP 5 0.001 In ethanol 12.85±1.103 60.01±0.01 0.002 In ethanol 10±1.080 56.02±0.02 0.005 In ethanol 10.5±0.64 59.03±0.03 0.01 In ethanol 9±1.080 60.1±0.04 8 CP 6 0.001 In ethanol 9±0.40 50.08±0.03 0.002 In ethanol 7.75±0.47 53.02±0.03 0.005 In ethanol 8.25±0.85 50.3±0.05 0.01 In ethanol 7.75±0.25 50.9±0.03
- 10. www.wjpps.com Vol 3,Issue 7, 2014. 965 Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences Fig No 2: Anti-TB Alamar blue assay Standard Fig No 3: Anti-TB Alamar blue assay for cyanopyridines Fig.No 4:Anthelmentic activity of compounds Fig No 5: Anthelmentic activity of (in case of Paralysis) Albendazole (in case of Paralysis)
- 11. www.wjpps.com Vol 3,Issue 7, 2014. 966 Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences Fig.No 6:Anthelmentic activity of compounds Fig No 7: Anthelmentic activity of (in case of death) Albendazole (in case of death) CONCLUSION The synthesized compounds were characterized by TLC, melting point, IR spectroscopy, 1 HNMR Spectroscopy. The results obtained from this study confirmed that the product has formed. Henceforth viewing these characteristic properties more compounds can be synthesized and subjected to pharmacological evaluation. These Cyanopyridine derivatives have biological activities like Anthelmintic, and anti tubercular may be a pave for synthesis and characterization of some new derivatives. The result of the Anthelmintic activity was given in table 8 and figure 4 to 7. From this table it was found that CP 4, CP 5 & CP 6 showed more activity than all other derivates. The standard is Albendazole. The Results of anthelmintic activity were presented as mean ±SEM (standard error of mean). The Tubercular activity of synthesized Cyanopyridine CP 1 to CP 6 were screened against M.Tuberculosis H37RV strain in the Middle Brook 7H9 (MB7H9 Broth) by using streptomycin& Pyrazinamide as standard drug. Results are given in Table No. 7 and Figures are shown in 2 & 3,. The results of Anti tubercular activity revealed that compounds CP2 exhibited activity at 6.25µg|ml the remaining compounds were not shown activity up to 50µg of CP 2. It is clear from the present study that the presence of OH & Cl group at para positions of both aromatic rings may be necessary for showing anti tubercular activity.
- 12. www.wjpps.com Vol 3,Issue 7, 2014. 967 Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences The existing designed analogs (C 1-6) can be further modified so as to include substituted pyrazolines, pyrimidines, Oxazoles, hydrazones, benzodiazepines etc which can produce further substituted derivatives. Such analogs can be synthesized, and evaluated for their Anthelmintic, anti-microbial and anti tubercular activities and other activities. It would be interesting to deduce the structure of potent compounds for the activity claimed, they may be our lead molecules and there is a possibility to extend the lead molecule towards drug design using various drug design software’s like maestro, glide, scigress, autodock etc. ACKNOWLEDGEMENT The authors are thankful to the Principal, CES college of Pharmacy, HOD Department of Pharmaceutical Chemistry for their support. Also we thank Dr. Murugesan, SAIF, IIT Chennai and Dr. Kishore Bhat, Department of Microbiology, Maratha Mandal College of Dental sciences, Belgaum for providing spectral and anti tubercular data respectively. REFERENCES 1. Parikh KS & Patel RP. Synthesis and Characterization of Some Cyanopyridine Compounds in Therapeutic Interest. Int.J. ChemTech Res, 2009; 1(3);581-586. 2. Hassan M Faidallah et al. Synthesis of some new 2-oxo-1,4-disubstituted-1,2,5,6- tetrahydro-benzo[h]quinoline-3-carbonitriles and their biological evaluation as cytotoxic and antiviral agents. J. Chem. Sci, 2012; 124(3); 625–631. 3. Kachhadia et al. Synthesis of Isoxazoles and Cyanopyridines Bearing Benzo(b)thiophene Nucleus as Potential Antitubercular and Antimicrobial Agents. J. Sci. I. R. Iran, 2004; 15(1); 47-51. 4. Venkatesan J et al. synthesis and biological evaluation of 4, 6-diaryl substituted-4, 5- dihydro-2-amino pyrimidines. Indian J.Pharm.Sci. 2007; 69(4); 586-589. 5. Anjani solankee et al. Anti microbial evaluation of some novel isoxazoles, cyanopyridines and pyrimidinthiones. Indian J Chem.2013, 52 B;671-676. 6. Anjani Solankee et al. Synthesis and studies of chalcones and its Cyanopyridine and Acetyl Pyrazoline Derivatives. Rasayan J. Chem. 2008; 1(3); 591-595. 7. L Collins, tuberculosis and Mycobacterium avium compounds against Mycobacterium 460 system for high-throughput screening of Microplate alamar blue assay versus BACTEC. Antimicrob. Agents Chemother.1997; 41(5); 1004-1009. 8. Hussain et al. Synthesis and biological evaluation of some novel 2-mercapto pyrimidines Int J Pharm Pharm Sci,2013;5(2); 471-473
- 13. www.wjpps.com Vol 3,Issue 7, 2014. 968 Rajkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences 9. Biswajit Chandra Das et al. Anthelmintic and anti-microbial activity of some novel chalcone derivatives J. Chem. Pharm. Res, 2010; 2(1);113-120. 10. P.S. Kalsi, Spectroscopy of Organic Compounds, (Sixth Edition)Published by New Age International (P) Limited, 2012 11. Silverstein RM, The Spectrometric Identification of Organic Compounds (7th Edition) Published by John Wiley & Sons, 2005.