SYNTHESIS OF PYRIMIDINE SUBSTITUTED PYRAZOLE
DERIVATIVES AND EVALUATION OF THEIR
PHARMACOLOGICAL ACTIVITY
M.Pharm Dissertation Protocol Submitted to
Rajiv Gandhi University of Health Sciences, Karnataka
Bangalore 560041
By
Mr. PATEL JIGNESHKUMAR JAYANTIBHAI, B.Pharm
Under the Guidance of
Dr. C. SREEDHAR, M.Pharm. Ph.D
Professor and Head
Department of Pharmaceutical Chemistry
Department of Pharmaceutical Chemistry,
Acharya & B.M. Reddy College of Pharmacy,
Soldevanahalli, Chikkabanavara Post,
Bangalore -560090
2009-2010
1
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
KARNATAKA, BANGALORE.
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1
Name of the candidate and
address
Mr. PATEL JIGNESHKUMAR
JAYANTIBHAI
#27, Nikunj society, Radhanpur road,
Mahesana-384002, Gujarat.
2
Name of the institution
ACHARYA & B.M. REDDY COLLEGE OF
PHARMACY.
89/90, Soldevanahalli, Chikkabanavara post,
Hesaraghatta main road,
Bangalore - 560090
3
Course of study and subject
4
Date of the admission
5
Title of the topic:
MASTER OF PHARMACY
(PHARMACEUTICAL CHEMISTRY)
29th May 2009
Synthesis of pyrimidine substituted pyrazole derivatives and evaluation of
their pharmacological activity
2
6.0
Brief resume of the intended work
6.1 Need for study:
The inflammatory response is accompanied by the clinical signs of
erythema, edema, hyperalgia and pain (algesia). Inflammation may lead to
osteoarthritis and rheumatoid arthritis.
The existing non steroidal anti-inflammatory drugs have side effects like
gastric or intestinal ulceration and bleeding that sometimes can be accompanied by
anemia from the resultant blood loss. Other side effects of these drugs that result
from blockade of the synthesis of endogenous prostaglandins and thromboxane-A2
include disturbances in platelet functions and changes in renal function.1
Pyrazole is a 5 membered heterocyclic aromatic organic compound
containing three carbon atoms and two nitrogen atoms in adjacent positions.
Pyrazoles are produced synthetically through the reaction of α,β-unsaturated
aldehydes with hydrazine and subsequent dehydrogenation. In medicine, pyrazoles
are used for their analgesic, anti-inflammatory, antipyretic, antiarrhythmic,
tranquilizing,
muscle
relaxing,
psychoanaleptic
and
anticonvulsant,
monoamineoxidase inhibiting, antidiabetic and antibacterial activities. Structurally
related compounds are pyrazoline and pyrazolidine.2
3
Pyrimidine is a six membered heterocyclic aromatic organic compound
containing two nitrogen atoms at positions 1 and 3. It is isomeric with two other
forms of diazine. A pyrimidine has many properties in common with pyridine, as the
number of nitrogen atoms in the ring increases the ring π electrons become less
energetic and electrophilic aromatic substitution gets more difficult while
nucleophilic aromatic substitution gets easier. Reduction in resonance stabilization of
pyrimidines may lead to addition and ring cleavage reactions rather than
substitutions. Pyrimidines can also be prepared within the laboratory by organic
synthesis. One method is the classic Biginelli-reaction. Many other methods rely on
condensation of carbonyls with amines for instance the synthesis of 2-thio-6methyluracil from thiourea and ethyl acetoacetate or the synthesis of 4methylpyrimidine with 4,4-dimethoxy-2-butanone and formamide . A novel method
is by reaction of certain amides with carbonitriles under electrophilic activation of the
amide with 2-chloro-pyridine and trifluoromethanesulfonic anhydride. Pyrimidine are
used for their CNS depressant, neuroleptic, antimicrobial, tuberculostic and antiinflammatory activity. So, we have planned to synthesize pyrimidine substituted
pyrazole derivatives and evaluation of their anti-inflammatory and antimicrobial
activity.2
4
6.2 Review of literature:
1. Flora
FB
et
al
synthesized
compounds
utilizing
in
vivo
acute
carrageenan-induced paw edema standard method in rats exhibited that many of
the tested compounds reveal considerable anti-inflammatory properties. PGE2
inhibitory properties of the highly promising synthesized anti-inflammatory active
agents were determined by PGE2 assay kit technique.3
2. Adnan AB et al synthesized compounds were examined for their in vivo antiinflammatory activity in two different bioassays namely; cotton pellet-induced
granuloma and carrageenan-induced paw edema in rats. The in vitro inhibitory
activity of the most active compounds towards human COX-1 and COX-2 enzymes
was also estimated.4
3. Holla BS et al synthesized 5-substituted 6-methyl-1-[8-(trifluromethyl)quinolin-4yl]-1,5-dihydro-4h-pyrazolo[3,4-d]pyrimidin-4-ones,
respectively
structure
of
product have been determined by chemical reaction and spectral studies. All the
compounds of the series have been screened for their antibacterial and antifungal
activity.5
4. Tarik EA et al synthesized derivatives of dipyrazolo[1,5-a:3,4-d]pyrimidine
Structures of the products have been determined by elemental analysis and spectral
studies. All compounds have been screened for their antibacterial and antifungal
activities.6
5. Adnan AB et al synthesized pyrazolyl benzenesulfonamide derivatives celecoxib
was shown to be a potent and gastrointestinal safe anti-inflammatory analgesic agent.
It is considered a typical model of pyrazole containing compound, pyrazofurin which
demonstrated abroad spectrum antimicrobial activity consequently, Several pyrazole
derivatives that exhibited antimicrobial activity were reported
cowarker.7
5
by tanitane and
F 3C
N
N
HO
HO
NH2
HO
S
O
O
NH2
O
O
HO
Celecoxib
N
N
H
purazofurin
6. Sherif AFR et al synthesis of new1-(4-chlorophenyl)-4-hydroxy-1H-pyrazole-3carboxylicacidhydrazide analogs and some derived ring systems and evaluated for
their antitumor activity.8
7. Ram S et al synthesized of pyrimidine and pyrazole based acyclo-C-nucleosides as
carbohydrieds was obtimizd and developed. The structure of celebrex contain
pyrazole shows that anti-inflammatory activity.9
8. Franco C et al synthesized 1-propanyol-3,5-diphenyl-4,5-dihydro-(1H)-pyrazole
derivative and assayed as inhibitors of MOA-A and MOA-B isoforms. Most of the
tested compound showed inhibitory activity with micromolar value and MAO-A
selectivity.10
6
9. Anand KT et al synthesized a series of pyrazolines were prepared both by
conventional and microwave methods. Their N-phenyl and acetyl derivatives were
also prepared. The yields of these compounds by microwave method were found to
be 70-85% for simple pyrazolines and 65-80% for N-phenyl substituted
pyrazolines.11
10. Michonl V et al synthesized some unsymmetrical Ri-exocyclic and N-endocyclic
derivatives from benzoylation of 3,5-aminopyrazol and evaluate their anticonvulsant
activity towards the MES and scMET tests, only the N-exe-pyrazole benzamides
showed good protection with respect to these tests.12
11. Aymn ER et al used 5-amino-1-substituted-1H-pyrazole-4-carbonitrile derivative as
a precursor for preparation of some novel substituted pyrazole and pyrazolo[3,4d]pyrimidine derivatives, and also prepared sugar hydrazone derivative and their
bannelated C-nucleosides and evaluated for antiviral activity.13
12. Laszlo V et al investigated the reaction of chalcones and guanidine in the presence of
an oxidizing agent, by addition of a 4,6-diaryl-pyrimidine-2-ylamine or 2-amino-5,5disubstituted-3,5-dihydro-imidazol-4-one.14
13. Jacobus PDV et al synthesized substituted dicyanopyridines and aminopyrimidines,
yielding two series of cyano-substituted diphenylaminopyrimidine, the new
compounds were generally selective for adenosine A1 receptor, although affinity for
the adenosine A2A receptor was also found.15
14.Yong X et al synthesized a series of novel 1-arylmethyl-3-aryl-1H-pyrazole-5
carbohydrazide hydrazone derivatives were synthesized and the effects of all the
compounds on a 549 cell growth were investigated.16
15. Neslihan D et al synthesized 5-oxo-[1,2,4]triazole ring and evaluate for
antimicrobial and antifungal activity,
some compound shows an antimicrobial
activity against one or more microorganism, but no antifungal activity has been
observed against yeast like fungi.17
7
16. Mehdi A et al synthesized 2,4,6-Triarylpyrimidines via a simple, one-pot, fourcomponent reaction between aryl methyl ketones, benzaldehydes, aromaticnitriles,
and hydroxylamine under microwave irradiation and solvent-free condition give
excellent yields.18
17. Rahaman SA et al synthesized novel pyrimidines by the condensation of chalcones
of 40-piperazine acetophenone with guanidine hydrochloride, also screened for
antihistaminic activity, some compound shows a significant anti-histaminic activity
when compared to the reference anti-histaminic drug mepiramine.19
18. Anu A et al a series of 2,4,6-trisubstituted pyrimidine derivatives was synthesized
and evaluated for their in vivo pregnancy interceptive activity in hamsters. Out of the
17 compounds synthesized three compounds showed 100% activity at a dose of
10mg/kg.20
19. Anjani S et al base catalysed condensation of ketone 5 with different aldehydes
give chalcones, these chalcones on cyclization with hydrazine hydrate in the presence
of glacial acetic acid, guanidine nitrate in the presence of alkali and malononitrile in
the presence of ammonium acetate give the corresponding acetyl pyrazolines.21
6.3 Objectives of the study:
1. To synthesize some newer derivatives of pyrimidine substituted pyrazole
2. To characterize the synthesized compounds by different analytical techniques such as
IR, NMR and Mass spectral data.
3. To screen the synthesized compounds for their anti-inflammatory and antimicrobial
activities.
8
7.0
Materials and methods:
7.1 Sources of data
Databases like Chemical abstracts, Biological abstracts, Medline, and Journal of
Chemistry section B, Indian Journal of Heterocyclic Chemistry, European Journal of
Medicinal Chemistry, Bioorganic and Medicinal Chemistry Letters, Acta
crystallographica, Helinet of RUGHS etc.
7.2 Method of collection of Data:
A) Synthesis of the compounds:
Chemicals and other reagents required for synthesis will be procured from standard
company sources. Compounds will be synthesized by using standard techniques. The
reactions will be monitored by TLC and purification of the compound will be done
by recrystallization method.
B) Characterization of the compounds:
The synthesized compounds will be characterized by preliminary laboratory
techniques such as melting point, boiling point etc. Compounds synthesized will be
confirmed by FTIR, Mass Spectroscopy and NMR spectral data. The Mass and NMR
spectral data of the synthesized compound will be collected by sending the
compounds to research center at IISc, Bangalore.
C) 1) Screening of anti-inflammatory activity:4,5
In vivo anti-inflammatory activity study:
Method Used: Carrageenan-induced paw edema model.
Animals Used: Albino Wistar rats.
Number of animals used: 72 numbers.
9
Carrageenan-induced paw edema model:
A 1% w/v suspension of carrageenan is prepared freshly in normal saline and
injected into subplantar region of left hind paw (usually 0.1mL in rats and 0.0250.05mL in mice). In control group animals, only vehicle is injected. Test drug is
usually administered orally or intraperitoneally, according to body weight
immediately or half an hour or one hour before (depending on the expected peak
effect) carrageenan challenge. A mark is made on the ankle joint of each rodent. Paw
volume up to the ankle joint is measured in drug treated and untreated groups before
and 3 hours after carrageenan challenge using a plethysmograph filled with mercury.
2) Screening of antimicrobial activity:6,7
Screening of antibacterial activity by disc diffusion method:
Antimicrobial studies will be carried out on both Gram positive and Gram
negative organisms like Staphylococcus aureus, Pseudomonas aeruginosa,
Escherichia coli and Bacillus subtilis etc using sterile media like Mueller-Hinton
Agar etc by Disc Diffusion Method. Zone of inhibition of the compounds synthesized
will be noted and compared with that of standard drugs like Amoxicillin,
Ciprofloxacin etc. The entire work will be done using horizontal Laminar Flow hood.
10
7.3 Does the study require any investigation or interventions to be
conducted on patients or other humans or animals?
YES
7.4 Has ethical clearance been obtained from your institution in case of
7.3?
APPLIED FOR CLEARANCE
11
8.0
REFERENCES:
1. Jackson RL, Jason DM. Goodman and Gilman′s the pharmacological basis of
therapeutics. 10th ed. New York: McGraw-Hill Medical publication division; 2001.
2. “http://en.wikipedia.org/wiki”
3. Flora FB, Adel SG. Facile synthesis of bis(4,5-dihydro-1H-pyrazole-1-carboxamides)
and their thio-analogues of potential PGE2 inhibitory properties. Eur J Med Chem
2009;44:2172-177.
4. Adnan AB, Hesham TVF, Sherif AFR, Azz MB. Design and synthesis of some
substituted
1H-pyrazolyl-thiazolo[4,5-d]pyrimidines
as
anti-
inflammatory,
antimicrobial agent. Eur J Med Chem 2003;38(1):27-36.
5. Holla BS, Manjathuru M, sitambaram KM, Padiyath MA, Nalilu SS. Synthesis of
some novel pyrazolo[3,4-d]pyrimidine derivatives as potential antimicrobial agent.
Bioorg Med Chem 2006;14:2040-47.
6. Tarik EA. Synthesis of some novel pyrazolo[3,4-b]pyrimidine and pyrazolo[3,4d]pyrimidine derivatives bearing 5,6-diphenyl-1,2,4-triazine moiety as potential
antimicrobial agents. Eur J Med Chem 2009;44:4385-92.
7. Adnan AB, Hayam MAA, Yasser SA, Alaa EAB, Azza B. Synthesis and biological
evaluation of somethiazolyl and thiadiazolyl derivatives of 1H-pyrazole as antiinflammatory antimicrobial agents. Eur J Med Chem 2008;43:456-63.
8. Sherif AFR, Manal AS, Maha AED. Synthesis of new 1-(4-chlorophenyl)-4-hydroxy1H-pyrazole-3- carboxylicacid hydrazide analogs and some derivedring systems. A
novel class of potential antitumor and anti-HCVagents. Eur J Med Chem
2003;38:959-74.
12
9. Ram S, Moon-JK, Seung BP. An improved synthesis of pyrimidine-and pyrazolebased acyclo-C-nucleosidesascarbohybrids. Tetrahedron Lett 2008;49:5080-83.
10. Franco C, Rossella F, Adriana B, Fedele M, Paola C, Daniela S. Synthesis, molecular
modeling studies and selective inhibitory. Activity against MAO of N1-propanoyl3,5-diphenyl-4,5-dihydro-(1H)-pyrazole derivatives. Eur J Med Chem 2008;43:226267.
11. Anand KT, Shrishaailappa B, Prashantha BRK, Santosh KHD, Ravi S, Durai AKT.
Microwave assisted synthesis of pyrazoline derivatives and their antiangiogenic and
antioxidant activities. Indian J H Chem 2008;16:333-6.
12. Michon V, Hervk CP, Tombret F, Gillardin JM, Lepage F, Berthon L. Preparation,
structural analysis and anticonvulsant activity of 3-and saminopyrazole N-benzoyl
derivatives. Eur J Med Chem 1995;30:147-55.
13. Aymn ER, Mohamed IH, Randa EA, Nahed F, Farouk MEA. Synthesis and antiHSV-1 evaluation of some pyrazoles and fused pyrazolopyrimidines. Eur J Med
Chem 2009;44:3285-92.
14. Laszlo V, Tamas N, Istvan K, Jordi BB, Gyorgy D, Laszlo UFD. Solutionphaseparallel synthesis of 4,6-diaryl-pyrimidine-Yl amines and 2-amino-5,5disubstituted-3,5-dihydro-imidazol-4-ones
via
a
rearrangement.
Tetrahedron
2003;59:655-62.
15. Jacobus PDV, Lisa CWC, Jacobien KFDK, Thea MK, Regina SL, Margot WB. et al.
A new generation of adenosine receptor antagonists From di-to trisubstituted amino
pyrimidines. Bioorg Med Chem 2008;16:2741-52.
13
16. YongXia, Chuan DF, Bao XZ, Jing Z, Dong SS, Jun YM. Synthesis and structure
activity relationships of novel 1-arylmethyl-3-aryl-1H-pyrazole-5-carbohydrazide
hydrazone derivatives as potential agents against A549 lung cancer cells. Eur J Med
Chem 2008;43:2347-53.
17. Neslihan D, Sengül AK, Ahmet D, Kemal S. Synthesis and antimicrobial activities of
some new 1-(5-phenylamino-[1,3,4]thiadiazol-2yl)methyl-5-oxo-[1,2,4]triazoleand1(4-phenyl-5-thioxo-[1,2,4]triazol and 1(4-phenyl-5-thioxo-[1,2,4]triazol-3-yl)methyl5-oxo-[1,2,4]triazole derivatives. Eur J Med Chem 2004;39:793-804.
18. Mehdi A, Niusha M, Mohammad M, Hamid RB. Microwave-assisted simple, onepot, four-component synthesis of 2,4,6-triaryl pyrimidines under solvent-free
conditions. Tetrahedron Lett 2006;47:9365-68.
19. Rahaman SA, Rajendra PY, Phani K, Bharath K. Synthesis and anti-histaminic
activity of some novel pyrimidines. Journal of the Saudi Pharmaceutical Society
2009;16:255-8.
20. Anu A, Brijesh K, Purshottam KM, Prem MSC. 2,4,6-Trisubstituted pyrimidine
derivatives as pregnancy interceptive agents. Bioorg Med Chem 2005;13:1893-9.
21. Anjani S, Kishor K, Ana C, Marina S, Irini D, Athina G. Synthesis of some new striazine based chalcones and their derivatives. As potent antimicrobial agents. Eur J
Med Chem 2009;xxx:01-09.
14
9.0
Signature of the Candidate
10.0
Remarks of the Guide
11.0
Name and Designation of
11.1Guide
Dr. C. Sreedhar
Professor & Head
Department of Pharmaceutical Chemistry
Acharya & B.M. Reddy College of Pharmacy,
Bangalore-90
11.2 Signature
11.3 Co-Guide
NIL
11.4 Signature
11.5 Head of the Department
Dr. C. Sreedhar
Professor & Head
Department of Pharmaceutical Chemistry,
Acharya & B.M. Reddy College of Pharmacy,
Bangalore-90
11.6 Signature
12.0
12.1Remarks of Principal
12.2 Name of the Principal
Dr. Divakar Goli
Principal,
Acharya & B.M. Reddy College of Pharmacy,
Bangalore-90
12.3 Signature
15