synthesis and study of new indazole …phdthesis.uaic.ro/phdthesis/visu (cheptea), corina, synthesis...
TRANSCRIPT
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UNIVERSITY “AL. I. CUZA” IAŞI
FACULTY OF CHEMISTRY
DEPARTMENT CHEMISTRY
Corina VISU
(married CHEPTEA)
SYNTHESIS AND STUDY OF NEW INDAZOLE DERIVATIVES
WITH POTENTIAL PHARMACOLOGICAL ACTIVITY
RESUME OF THE DOCTORATE THESIS
Scientific adviser:
Prof. univ. em. dr. Valeriu ȘUNEL
IAȘI -2012-
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CONTENTS
INTRODUCTION………………………………………………………….................................................................... 5
PART I Theoretical considerations I.1. General considerations on indazole combinations......................................................................................................................................................................
8 I.1.1 Syntheses of indazoles............................................................................................................................................... 8 I.1.1.1. From o-hydrazine-cinamic acid............................................................................................................................. 8 I.1.1.2. Reaction of o- and nitro-o-toluidine with nitrous acid.................................................................................................................................................................
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I.1.1.3. From acidic chloride compounds and esthers of furan-2-carboxylic acid......................................................................................................................................................
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I.1.1.4. Syntheses from aldehyde....................................................................................................................................... 10 I.1.1.4.1. Cyclization between o-nitro-benzaldehyde and phenyl-hydrazine.........................................................................................................................................................
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I.1.1.4.2. Cyclization of 2-fluoro-5-nitro-benzaldehyde with hydrazine....................................................................................................................................................................
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I.1.1.4.3. Reaction of salicyl-aldehyde with hydrazine...................................................................................................... 11 I.1.1.4.4.From pentafluorobenzaldehyde and hydrazine.................................................................................................... 11 I.1.1.5. Reaction between aromatic acids and thionyl chloride..................................................................................................................................................................
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I.1.1.6. From Schiff bases.................................................................................................................................................. 12 I.1.1.7. From oxymes......................................................................................................................................................... 13 I.1.1.8. From 2-chlorobenzonitril and hydrazine............................................................................................................... 13 I.1.1.9. Reaction of 2-chloro-5-nitrobenzoic acid with hydrazine....................................................................................................................................................................
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I.1.1.10. From imides of the dicarboxylic acids................................................................................................................. 15 I.1.1.11. From azides throughCurtius transposition .......................................................................................................... 15 I.1.1.12. From cyclohexanone and ethyl formiate............................................................................................................. 16 I.1.2. Characteristics of indazoles...................................................................................................................................... 17 I.1.2.1. Indazoles tautomery............................................................................................................................................... 17 I.1.2.2. Indazoles reactions................................................................................................................................................. 19 I.1.2.2.1. Salt formation with acids and bases.................................................................................................................... 19 I.1.2.2.2. Reduction............................................................................................................................................................ 20 I.1.2.2.3. Oxidation............................................................................................................................................................ 20 I.1.2.2.4. Reactions at imino- group................................................................................................................................... 21 I.1.2.2.4.1. Reaction with alkyl halogenated compounds.................................................................................................. 21 I.1.2.2.4.2. Reaction with acidic chloride and anhydride compounds............................................................................... 22 I.1.2.2.4.3. Reaction with propylenoxide........................................................................................................................... 23 I.1.2.2.4.4. Reaction with acrylonitril................................................................................................................................ 23 I.1.2.2.4.5. Reaction with epichlorhydrine......................................................................................................................... 24 I.1.2.2.4.6. Reaction with phenyl-isothiocyanate............................................................................................................... 24 I.1.2.2.4.7. Reaction with ethyl-chloroacetate................................................................................................................... 25 I.1.2.2.4.8. Mannich reaction............................................................................................................................................. 25 I.1.2.2.4.9. Reaction with aromatic halogenated derivatives............................................................................................. 26 I.2.2.5. Reactions within the benzene ring......................................................................................................................... 26 I.2.2.5.1. Nucleofile substitution........................................................................................................................................ 26 I.2.2.5.2. Electrofile substitution........................................................................................................................................ 27 I.2.2.5.2.1. Nitrogenation reaction..................................................................................................................................... 27 I.2.2.5.2.2. Halogenation reaction...................................................................................................................................... 27 I.2.2.5.2.3. Sulphonation reaction...................................................................................................................................... 28 I.2. General considerations on 4-substituted 1-acyl-thiosemicarbazides ...........................................................................................................................
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I.2.1. Methods of synthesis................................................................................................................................................ 29 I.2.1.1. Carboxylic acid hydrazides addition to isothiocyanates...............................................................................................................................................................
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I.2.1.2. N,N’-diacyl-thioureas reaction with hydrazides…………………………………………………………….…... 32 I.2.1.3. From 4-substituted thiosemicarbazide and acidic chloride compounds.................................................................................................................................................
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I.2.2. Structure and reactivity of 4-substituted 1-acyl-thiosemicarbazides............................................................................................................................
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I.3. General considerations on 1,3,4-thiadiazoles.............................................................................................................. 35
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I.3.1. Methods of synthesis................................................................................................................................................ 36 I.3.2. Characteristics.......................................................................................................................................................... 39 I.4. General considerations on thiazolidines...................................................................................................................... 40 I.4.1. Methods of synthesis................................................................................................................................................ 40 I.4.2. Characteristics.......................................................................................................................................................... 44 I.5. General considerations on 1,3,4-oxadiazoles.............................................................................................................. 47 I.5.1. Methods of preparation............................................................................................................................................ 47 I.5.2.Characteristics........................................................................................................................................................... 50 I.6. General considerations on 1,2,4-triazoles………………………………………………………………...…………. 52 I.6.1. Methods of synthesis……………………………………………………………………………...………………. 52 I.6.2. Characteristics………………………………………………………………………………….………………….. 57
PART II Personal research II.1. Synthesis of new compounds with indazole structure................................................................................................
61 II.1.1. Synthesis of the ethylic ester of 5-nitroindazole-1-yl-acetic acid......................................................................................................................................... II.1.2. Synthesis of amide type derivatives of 5-nitroindazole-1-yl–acetic acid........................................................................................................................................
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II.1.3. Synthesis of the 5-nitroindazole-1-yl-acetic acid hydrazide........................................................................................................................
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II.1.4. Synthesis of N-acyl-hydrazones (hydrazidones) of 5-nitroindazole-1-il-acet-hydrazides..................................................................................................................................
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II.1.5. Synthesis of 1-(5’-nitroindazole-1’-yl-acetyl)- 4-aryl-thiosemicarbazides……………………………………..………………………………………………..………..
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II.1.6. Synthesis of 2 substituted 5-aryl-amino-1,3,4-thiadiazolecompounds........................................................................................................................
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II.1.7. Synthesis of2,3-disubstituted 1,3-thiazolidine compounds.................................................................................... 88 II.1.7.1. Characterization through HyperChem calculus of the 3-[(5’-nitroindazole-1’-yl-acetamidyl)]-2-aryl-4-oxo-1,3-thiazolidines ...........................................................................
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II.1.7.2. Factorial experiment for optimized synthesis of the 3-[(5’-nitroindazole-1’-yl-acetamidyl)]-2-aryl-4-oxo-1,3- thiazolidines.......................................................................................................................................................................
99 II.1.8. Synthesis of 2-substituted 5-aryl-amino-1,3,4-oxadiazoles.........................................................................................................................................
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II.1.9. Synthesis of3,4-disubstituted 5-mercapto-1,2,4-triazoles.................................................................................................................................................
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II.1.10. Grafting of the N-mustard group on the molecule of 5-nitroindazole derivatives................................................................................................................................................
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II.2. Study of the biological properties of the synthesized compounds.....................................................................................................................................................
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II.2.1. Toxicity evaluation of the synthesized substances................................................................................................. 129 II.2.2. Testing of the cytostatic activity on animals with experimental tumours.................................................................................................................................................
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II.2.2.1. Anti-cancer activity of the 2,3-disubstituted 4-oxo-1,3-thiazolidines......................................................................................................................................................
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II.2.2.2. Anti-cancer activity of the N-mustards with 5-nitroindazole support..............................................................................................................................................
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II.2.3. Testing of the depressant activity on the vegetal cell ofthe3-[(5’-nitroindazole-1’-yl-acetamidyl)]- 2-aryl-4-oxo-1,3-thiazolidines...........................................................................................................................................
135 II.2.4. Testing of the anti-microbial activity of 1,3,4-thiadiazoles and 1,2,4-triazoles..............................................................................................................................................................
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II.2.5. Testing of anti-pyretic actions of 2-[(5’-nitroindazole- 1’-yl-methyl)]-5-(p-tolyl-amino)-1,3,4-oxadiazole……………………………………………………………..………..
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II.3. Experimental procedures............................................................................................................................................ 143 PART III General Conclusions........................................................................................................................................................
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BIBLIOGRAPHY............................................................................................................................................................ 185
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INTRODUCTION
Indazole and its derivatives make up an important category of heterocyclic compounds, one that continues to raise the interest of chemical engineers, researchers, and other specialists in the field.
The appearance of a new branch of chemistry, chemistry of the pharmacological drugs, lead to the develeopment of new substances, particularly substances with biological activity.
Contemporary sicentific researchin the field of pharmacology focuses on the synthesis of new substances which could become extremely useful through their pharmacological and biological actions.
Indazole structure has been noted to induce a wide range of biological effects that are enhanced, by substitution/addition within its molecule, of different functional groups – as it has been illustrated in the literature.
Amongst the different biological effects, most commonly mentioned in the literature, are: anti-viral, anti-arrhythmic, anti-inflammatory, anti-pyretic, anti-malaria, anti-tumoural and anti-microbial activities.
Our research focused on the insertion/substitution, within the 5-nitroindazole molecule, of different chemically reactive groups and by this, to render the „supporting” molecule, certain pharmacological properties. Thus, we obtained the ester and the respective hydrazide for amidic compounds, hydrazines, acyl-thyocarbazides, as well as cyclization products, thiadiazoles, thiazolidines, oxadiazoles, triazoles. We also obtained alkylating products, derivatives of 5-nitroindazole, that have in their molecule the di-(beta-chloroethyl)-amine group.
The compounds obtained have been fully characterized, their structure being confirmed by means of elemental and spectral chemical analysis (FT-IR, 1H-RMN, 13C-RMN, SM). We also established the mechanisms of the reactions for some of the methods of synthesis used.
We also made physical measurements for some of the indazole derivatives with a view to optimize their method of synthesis – factorial experiment, molecular geometry, length of atomic bonds, distribution of atomic charges, by means of the programme HyperChem-5.
Taking into account the structural components of the synthesized compounds, we determined their toxicity, and, for some of these compounds, we compared their toxicity levels against their anti-tumoural, anti-microbial, and anti-pyrexial activities. Our experiments underline once more, the importance of the existing relationship between the chemical structure and the biological properties of the synthesized compounds (structure-function relationship).
The doctoral thesis entitled “Synthesisand study of new indazole derivatives with potential pharmacological actions“ has the following structure:
Part I has a monographic character and presents, based on information from literature, the methods of synthesis for indazoles, as well as their characteristics. Separate chapters have general considerations regarding synthesis and reactivity of compounds belonging to thiosemicarbazides, thiadiazoles, thiazolidines, oxadiazoles, and triazoles – some of these substances being newly synthesized derivatives.
Part II deals with personal research, particularly in the field of obtaining new derivatives of 5-nitroindazole - both synthesis and the physico-chemical characterization of these new derivatives. It also describes the results of the biological experiments done with some of these compounds. We describe the
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experiments performed, their methodology, as well as the systematic nomenclature of all the synthesized compounds.
Part III of the paper contains the general conclusions that result from the research undertaken. Bibliography, ending the paper, illustrates the literature, the scientific papers, and documents
concerning the field researched, up to 2012. The thesis is accompanied by a list of documents and an annex, again, illustrating scientific papers
and literature in extenso, both published and that actually based on the research done for this paper. The resume contains, in a synthesized manner, the results of the personal research, general
conclusions, and, part of the bibliography. The labelling of the compounds, nomenclature, tables, schematics, and mathematical formulas
used, as well as the bibliographical items used in this resume, are all identical to the ones used in the thesis.
PART II
Personal contributions (excerpts)
II.1. Synthesis of new compounds with indazole structure
Indazole and its derivatives make up an important class of heterocyclic compounds that can be found in the composition of many substances with extremely different pharmacological actions: anti-histamine [44], anti-viral [184], anti-microbial [185], cytostatic [49], anti-inflammatory [43], analgesic [195], anti-psychotic [194], anti-arrhythmic [196], anti-HIV [197], anti-malaria [198], anti-fungal [199]. Some of these drugs also have functions as neuronal inhibitors [12, 204], as well as antagonists of glucocorticoid receptors [205].
Such considerations justify all the efforts made in this field of obtaining new compoundsd with indazole structure that have potential biological activity.
The research was meant to enlarge the range of these new biologically active compounds. The aim was to insert in the molecule of the 5-nitro-indazole, groups of chemically active
compounds, leading to biologically active substances. These substances were characterized from a physic-chemical point of view and their activity was tested, while, at the same time, their toxicity was compared and analysed against a control substance of known toxicity.
II. 1. 1. Synthesis of the ethylic ester of 5-nitroindazole-1-yl-acetic acid
In order to obtain derivatives of 5-nitroindazole with biological activity, it was necessary to obtain first the 5-nitroindazole-yl acetic acid ester I by means of condensation of 5-nitroindazole with ethyl monochloroacetate, in the presence of sodium ethoxide, refluxing in pure ethyl alcohol [206].
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The advantage of this method consists in avoiding the separation of the sodium salt of 5-nitroindazole in solid phase.
The separated compound, dried and recrystallized from ethyl alcohol, has a reaction productivity of 85% and a melting point of 155-156°C.
The structure of the ester I was investigated by means of elemental and spectral chemical analysis (FT-IR, 1H-RMN, 13C-RMN, mass spectroscopy).
Fig. 2. The1H-RMN spectre of the ethylic ester of the 5-nitroindazole-1-yl-acetic acid I
II. 1. 2. Synthesis of amide type derivatives of 5-nitroindazole-1-yl-acetic acid
Interest in this type of compounds is justified by the importance awarded to the amidic structures
within the drugs, lending them significant pharmacological actions, such as: antihelmintic [207], antihistaminic [209], antimicrobial [213], tuberculostatic [214], anti-inflammatory [83,216], antiepileptic, as well as anticonvulsant [217], analgesic [218], and cytostatic [219-221].
In our situation, we obtained this type of compounds by starting from the ethyl ester of 5-nitroindazole-1-yl-acetic acid I. This was subjected to a reaction of condensation with a number of amines (monoethanolamine, diethanolamine, diethylaminoethylamine, isopropylamine, alylamine) leading to the corresponding series of amides II-VI [222]:
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Purification of these compounds was done from boiling ethyl alcohol. The resulting final products
were obtained with an efficacy of 71-87%. The structure of compounds II–VI suggested by chemical analysis, was also confirmed by means
of spectral analysis (FT-IR, 1H-RMN, 13C-RMN, mass spectroscopy).
Fig. 6. The1H-RMN spectre of the di-(β-hydroxyethyl)-amide of
5-nitroindazole-1-yl-acetic acid II
II. 1. 3. Synthesis of 5-nitroindazole-1-yl-acetic acid hydrazide
Taking into account the fact that numerous hydrazide derivatives have a wide range of practical applications [227], we thought it useful to synthesize the hydrazide of the 5-nitroindazole-1-yl-acetic acid, and this constituted a basic intermediary in the further synthesis of the great majority of the 5-nitroindazole derivatives.
During our studies we obtained the substance VII by treating ethyl ester of the 5-nitroindazole-1-yl-acetic acid I with hydrazine hydrated 98% in methanol, at boiling temperature, with the end result of a substance that through recrystallization from ethyl alcohol, melts at 174-175°C. The yield for the end product is about 80% [206].
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With an yield of even greater value, of about 92%, and at even higher melting temperature, of about 175-176°C, we obtained this hydrazide by using an excess of hydrated hydrazine at room temperature in an ethanolic solution [206, 236].
The structure of 5-nitroindazolyl-acet-hydrazide VII was confirmed by both spectral and elemental analysis (FT-IR, 1H-RMN, 13C-RMN, mass spectroscopy).
Fig. 10. The1H-RMN spectre of 5-nitroindazole-1-yl-acetic acid hydrazideVII
II.1.4. Synthesis of some N-acyl-hydrazones (hydrazidones) of the 5- nitroindazole-1-yl-acet-hydrazide
The pharmacological significance of hydrazones was demonstrated for diverse biological
activities, such as: antibacterial [239], anticonvulsant [241], antifungal [243], anti-inflammatory [244], antimalaria [246], anti-tumoral [249], as well as anti-HIV [250].
Taking into account all of the above, we decided to condense the 5-nitroindazole-1-yl-acetic acid hydrazide VII with different aromatic aldehydes with the goal of obtaining a new series of N-acyl hydrazones, that would contain in their structure the mentioned hydrazone.
In our particular case, hydrazide VII was condensed with benzoic aldehyde, o- and p-nitrobenzoic, o-hydroxybenzoic aldehyde, leading to four new hydrazones with the structures VIII-XI [206].
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The structure of hydrazones VIII-XI, as suspected through synthesis, was confirmed through spectral and elemental analysis(FT-IR, 1H-RMN, 13C-RMN, mass spectroscopy).
Fig.14. The1H-RMN spectre of N1-(5-nitroindazole-1-yl-acetil)-N2-benzaldehyde-hydrazone VIII
II.1.5. Synthesis of some 1-(5’-nitroindazole-1’-yl-acetil)- 4-aryl-thiosemicarbazides
There is awhole lot of information in the literature supporting the biological importance of such
compounds [ 84, 104, 214, 261]. Based on this information, we decided to achieve the addition of different aromatic isocyanate
compounds to the 5-nitroindazole-1-yl-acetic acid hydrazide. Synthesis of 1-(5’-nitroindazole-1’-yl-acetil)-4-aryl-thiosemicarbazides XII–XVII was achieved
according to the methods described in literature for some other types of thiosemicarbazides [82, 214, 263]: in methyl alcohol solution, through heating to boiling of a mixture of a 5-nitroindazole-1-yl-acetic
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acid hydrazide VII with isothiocyanates: phenyl, p-tolyl, p-methoxyphenyl, p-bromphenyl, p-chlorphenyl and p-iodophenyl [264].
NN
O2N
CH2 CO NH NH2
S=C=N R+N
NO2N
CH2 CO NH NHXII - XVIIVII
C NH R
S
The thiosemicarbazides XII–XVII were purified through recrystallization from methanol, and the yield for the final products was79-82%.
The structure of the synthesized substances was confirmed through elemental analysis, as well as by spectral analysis IR, 1H-RMN, 13C-RMN, mass spectroscopy.
Fig. 18. The1H-RMN spectre of the 1-(5’-nitroindazole-1’-yl-acetil)-4-(p-methoxyphenyl)
thiosemicarbazide XIV
II.1.6. Synthesis of 2-substituted 5-aryl-amino-1,3,4-thiadiazoles
Data from the literature underline a wide range of pharmacological actions for derivatives of 1,3,4-thiadiazole, depending on the functional groups grafted on the heterocycle, such as: anti-bacterial [127, 268], antifungal [117], tuberculostatic [142, 272], analgesic [115, 274], anti-inflammatory [83,115, 275],antidepressive [268],anticonvulsant [241], antitumoural [279].
Based on such assumptions, we extended the field of our research, synthesizing new derivatives of 5-aryl-amino-1,3,4-thiadiazole that contain in position 2 remains of 5-nitroindazole.
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To this end, we cyclized, in an acidic environment, a series of 1-acyl-4-aryl-thiosemicarbazides
compounds XII-XVII derived from the hydrazide of the 5-nitroindazole-1-yl-acetic acid, following a protocol mentioned in the literature for some other types of thiadiazoles [82, 83, 86].
The cyclization reaction lead to the elimination of a molecule of water under the dehydrating action of the sulphuric acid, resulting in the 2-substituted 5-aryl-amino-1,3,4-thiadiazoles compounds XVIII-XXIII, with fixed melting points and yield of 73-77% [264].
In order to obtain reliable information based on physical and chemical research methods about the structure of the thiadiazoles XVIII-XXIII, we employed both elemental and spectral analysis IR, 1H-RMN, 13C-RMN, mass spectroscopy.
Fig. 22. The1H-RMN spectre of the 2-(5’-nitroindazole-1’-yl-methyl)-5-(phenyl-amino)-1,3,4-
thiadiazoles XVIII
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II.1.7. Synthesis of 2,3-disubstituted 1,3-thiazolidines
The thiazolidinic nucleus and its derivatives are well known as some of the most active pharmacological agents with actions such as antimicrobial [150, 291], cytotoxic [294], anti-inflammatory [295], hypoglicemic [296], antifungal [147], anticancer [297], antituberculosis [298], and antiviral [299]. Synthesis of organic substances that contain in their molecule 4-oxo-thiazolidine and 5-nitro-indazole, appears interesting and seems to be justified by the results of research, particularly in this highly specialized domain.
Having at our disposal the series of N1-(5-nitroindazole-1-yl-acetil)-N2-aryl-hydrazones VIII-XI, some with a substituent at the benzene ring, we synthesized new thiazolidinesby means of the hydrazones’ reaction with thyoglycolic acid in the presence of anhydrous zinc chloride as a dehydrating agent.
Practically, the hydrazones VIII-XI and thyoglycolic acid are heated in anhydrous dioxane together with anhydrous zinc chloride, for 6 hours, thus, allowing for intramolecular cyclization within thiazolidines XXIV-XXVII, while water is eliminated [300].
The end products were purified through recrystallization from ethyl alcohol, the efficacy of the reactions varying between 61 and 74%.
The chemical structure of the substances was established by elemental and spectral analysis (FT-IR, 1H-RMN, 13C-RMN, mass spectroscopy).
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Fig. 26. The 1H-RMN spectre of 3-[(5’-nitroindazole-1’-yl-acetamidyl)]-2-phenyl-4-oxo-
1,3-thiazolidine XXIV
In order to fully characterize 2,3-disubstituted 4-oxo-1,3-thiazolidines XXIV-XXVII, we made use of the HyperChem-5 programme, and this gave us informations regarding the molecular geometry, the length of atomic bonds, as well as the distribution of atomic charges within the molecules [301].
II. 1. 8. Synthesis of 2-substituted 5-aryl-amino-1,3,4-oxadiazoles
Justification for synthesizing such substances is based on numerous observations in the literature that underline the fact that oxadiazol derivatives have numerous pharmacological applications. Scientific evidence show antimalaria [312], anti-inflammatory, antipyretic [312, 316], antihypertensive [317], local anesthetic [318], antispastic [319], analgesic [321], antitumoral [322], insecticidal [323], antibacterial [322,326], antifungal [328], hypoglycemic [329] effects.
All these data that suggest the pharmacologic potential of the oxadiazolic heterocycle, gave direction to our research towards synthesizing new 1,3,4-oxadiazoles of 5-nitroindazole.
Synthesis consisted of heating 4-substituted 1-acyl-thiosemicarbazides with α-halogenated ester in ethanolic solution and in the presence of anhydrous sodium acetate.This method is a substantial improvement when compared to the synthesis methods described in the literature [159, 160].
Practically, 1-(5’-nitroindazole-1’-yl-acetyl)-4-aryl-thiosemicarbazides XII-XVII and ethyl chloroacetateare heated in ethanolic solution and ahydrous sodium acetate for 11 hours, thus allowing cyclization of oxadiazoles XXVIII-XXXIII, simultaneously with elimination of ethyl mercaptoacetate, as well as, acetic acid and sodium chloride [332].
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XXVIII R = - C6H5; XXIX R = - C6H4 - CH3(p); XXX R = - C6H4 - OCH3(p);
XXXI R = - C6H4 - Br(p); XXXII R = - C6H4 - Cl(p); XXXIII R = - C6H4 - I(p).
The derivatives 1,3,4-oxadiazoles XXVIII-XXXIII were purified by means of boiling ethylic alcohol, the end result being white-cream crystals, with melting points within a narrow range. The yield of the end product was 62-72%.
The structure of these derivatives was established by means of elemental and spectral analysis (FT-IR, 1H-RMN, 13C-RMN, mass spectroscopy).
Fig. 34. The1H-RMN spectre of 2-[(5’-nitroindazole-1’-yl-methyl)]-5-phenyl-amino-1,3,4-
oxadiazole XXVII
II. 1. 9. Synthesis of 3,4-disubstituted 5-mercapto-1,2,4-triazoles
It was noted that some of the triazoles have antifungal [333], antimicrobial[105, 115, 141, 335], anticonvulsant [336], anti-inflammatory [83, 115, 337], hypnotic and sedative [338], as well as analgesic [115, 167, 339], cardioprotective [340], and antitumoural [141] actions.
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The newly synthesized derivatives of 1,2,4-triazole, have a particular characteristic, and that is, are substituted in position 4 with a phenyl group. They may also have substitution in the para position. It would be extremely interesting to follow up whether these compounds with their triazolic nucleus have any influence on the biological activity of the respective molecule.
Transformation of 1,4- disubstituted acyl-thiosemicarbazides XII-XVII, in the corresponding 1,2,4-triazoles XXXIV-XXXIX [264], was achieved through one of the methods published in literature for other types of triazoles [82, 83, 335], in the presence of hydroxyl ions [264].
The synthesized mercapto-triazoles after recrystallization from ethylic alcohol at boiling, give a
yield of 56-78%. The structure of the triazole derivatives has been established by elemental and spectral analysis
(FT-IR, 1H-RMN, 13C-RMN, mass spectroscopy).
Fig. 38. The1H-RMN spectre of 3-(5’-nitroindazole-1’-yl-methyl)-4-(p-metoxyphenyl)-5-
mercapto-1,2,4-triazol XXXVI
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II.1.10. Grafting of the N-mustard group on the molecule of
5-nitroindazole derivatives
Cancer, both from an etiologic and therapeutic point of view, represents one of the most important problems of clincal medicine today, prompting an avid search for a solution within the boundaries of chemotherapy, plant chemistry as well as biology.
The literature offers methods of synthesis for some N-mustardcompounds where, varying the support for the cytotoxic functional group, one can obtain alkylating substances with an advantageous chemotherapeutic index, as well as reduced/lesser toxicity [219, 220, 348, 354].
In our research our aim was focused on grafting the di-(β-chloroethyl)-amine group, in various modalities, on pyrazole nucleus from 5-nitroindazole [264].
To this end, dissolution at high temperature, of 5-nitroindazole in an alcoholic solution containing sodium ethoxide lead to the sodium salt of 5-nitroindazole XL, which on turn, in combination with sodium monochloroacetate lead to the sodium salt of 5-nitroindazole-1-yl-acetic acid XLI.
Through the reaction of sodium salts XL and XLI with tris-(β-chloroethyl)-amine, with heated anhydrous dioxane, new organic substances have been obtained: their alkylating group binds the indazole substrate through an ethyl bridge N-[di-(β-chloroethyl)-aminoethyl]-5-nitroindazole XLII. Alternatively, the above mentioned bridge can also present as an ester: N-[di-(β-chloroethyl)-aminoethyl-carboxy-methyl]-5-nitroindazole XLIII.
Attachement of the alkylating group to the indazole via either the ester or ethyl bridge could be a
useful characteristic, since this could ease the splitting of the molecule in the human body, freeing the alkylating portion to act directly on the tumoral cell.
By reaction of the sodium salt XL with chloride monochloroacetic acid, we obtained N-chloroacetyl-5-nitroindazole XLIV, and this, by reacting with di-(β-chloroethyl)-amine, in anhydrous dioxane, leads to N-[di-(β-chloroethyl)-aminoacetyl]-5-nitroindazole XLV, with an efficacy of 65%.
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The composition and structure of the compounds XL-XLV was established by spectral and
chemical analysis (FT-IR, 1H-RMN, 13C-RMN).
Fig. 43. The1H-RMN spectre of N-[di-(β-chloroethyl)-aminoetil]-5-nitroindazole XLII
II. 2. Study of biological properties of some of the synthesized compounds
Experimental testing on animals folosite was done according to Ordinance no. 37/2002 from
30/01/2002 published in Monitorul Oficial, Part I, no. 95 on the 02/02/2002.
II.2.1. Evaluation of toxicity for the synthesized substances
The DL50 was determined using Spearman-Karber method (363). The results can be seen in table 11.
Table 11. DL50 values for substances I - XLV
Compound
DL50(mg/kg body weight) 24 hours 48 hours 7 days Average
1 2 3 4 5 I 9850 9850 9800 9853 II 8450 8470 8430 8450 III 8870 8900 8840 8870
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IV 7740 7780 7700 7740 V 7380 7400 7360 7380 VI 7250 7270 7230 7250 VII 8260 8260 8240 8253 VIII 1280 1295 1265 1280 IX 1050 1060 1040 1050 X 1025 1035 1015 1025 XI 1520 1550 1490 1520 XII 8350 8350 8320 8340 XIII 9120 9120 9100 9113 XIV 9550 9550 9500 9533 XV 9100 9100 9080 9093 XVI 9000 9000 8800 8933 XVII 9190 9190 9150 9176 XVIII 8850 8850 8820 8840 XIX 9480 9480 9460 9473 XX 9515 9515 9500 9510 XXI 9390 9390 9370 9383 XXII 9270 9270 9240 9260 XXIII 9340 9340 9310 9330 XXIV 1310 1310 1295 1305 XXV 1120 1120 1105 1115 XXVI 1100 1100 1090 1096 XXVII 1585 1585 1540 1570 XXVIII 8970 8970 8920 8953 XXIX 9685 9685 9625 9665 XXX 9715 9715 9425 9618 XXXI 9520 9520 9480 9506 XXXII 9220 9220 9180 9207 XXXIII 9315 9315 9279 9303 XXXIV 8950 8950 8920 8940 XXXV 9570 9570 9550 9563 XXXVI 9700 9700 9650 9683 XXXVII 9490 9490 9460 9480 XXXVIII 9200 9200 9170 9190 XXXIX 9295 9295 9275 9288
XL 1675 1675 1640 1663 XLI 1775 1775 1730 1760 XLII 1580 1580 1550 1570 XLIII 1620 1620 1600 1613 XLIV 1590 1590 1565 1581 XLV 1555 1555 1535 1548 Analysis of the 45 synthesized compounds has shown that from a toxicological point of view, they
lack significant general toxicity, as well as hematological toxicity and the DL50 values so established, recommend them for further biological testing.
II.2.2. Testing of the cytostatic activity on animals with experimental tumours
Anti-tumoral activity of some of the newly synthesized derivatives was studied against three
experimental tumours: carcinosarcoma Walker 256, Jensen sarcoma, and Guérin TB sarcoma.
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II.2.2.1. Anti-cancer activity of 2,3-disubstituted 4-oxo-1,3-4-oxo-1,3-thiazolidines
For the purpose of studying the cytostatic action we used Wistar rats weighing 100-130 g (±15g) with carcinosarcoma Walker 256 and Jensen sarcoma, all, sourced from the biobase of the Oncologic Institute “ Prof. Dr. Ion Chiricuță” Cluj-Napoca.
The solid tumour transplantation was done subcutaneously, from donors with 21-day-old tumours.The administration of substances in concentrations of 400, 200 and 40 mg/kg body weight was done by single injections on the 14th day after transplantation. The percentage inhibition produced by the investigated substances was worked out using the formula indicated in the literature [366], 14 days after administration for both tumours.
I% = ீିீீ
.100
GC, GT - the weight of the tumours of the animals, control (C) and respectively, treated (T).
Table 12. %inhibition by 5-nitroindazole and 2,3-disubstituted1,3-thiazolidines XXIV-XXVII. Compound %inhibition
Walker carcinosarcoma Jensen sarcoma
(mg/kg body weight)
400 200 40 400 200 40
5-nitroindazole 35 26 19 31 22 17
XXIV 45 40 34 41 36 31
XXV 44 39 35 42 37 33
XXVI 50 42 34 41 36 32
XXVII 54 44 40 50 45 40
It is noticeable that the values for percentage inhibition for carcinosarcoma Walter 256 are
superior to those corresponding the Jensen sarcoma, the substance XXVII being the most active one.
II. 2. 2. 2. Anti-cancer activity of N-mustards 5-nitroindazole based
For the study of the compounds XLII, XLIII, XLV [359], fragments of Guérin TB carcinoma
were implanted under the skin of 15 male rats weighing 100-150g. Administration of investigated substances started when the volume of the tumor reached 5 - 6 cm3. The substance was diluted with normal saline and administered by means of gastric tube to 10 of the animals, the other 5 being used as control. Daily dose was 20 mg/animal. The rats were treated for 10-15 days with close follow-up on the haematopoietic system – peripheral blood smear and cell count variation.
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Inhibition of tumoral growth was assessed by means of weighing the tumor 24 hours after the last administration of the drug and was worked up using the same formula [366] as in the case of carcinosarcoma Walker 256 and Jensen sarcoma.
Table 13 shows tumoral growth inhibition for the new compounds XLII, XLIII, XLV.
Table 13. Anti-tumoral activity of N-mustard compounds XLII, XLIII, XLV. Compound Route ofadministration Experimental animals %inhibition
Guérin TB carcinoma XLII p.o. rats 60 XLIII p.o. rats 70 XLV p.o. rats 56
Endoxane 84 Substances tested have a remarkable anti-tumoral activity, the most selective being N-mustard
XLIII – it produces an inhibition of the Guérin carcinoma similar to that produced by endoxane (reference cytostatic). The reason for this is that besides the alkylating effect, these compounds have also an antimetabolite effect.
II. 2. 3. Testing of the depressant activity on the vegetal cell of the 3-[(5’-nitroindazole-1’-yl-acetamidyl)]-2-aryl-4-oxo-1,3-thiazolidines
The experiment was done within the framework of a biotest that brought the investigated
substances in a suspension of 1% methylcelulose together with two drugs (IOB-82 and Methotrexate) used for chemotherapy.
The vegetal tissue used for the test [300] is the roots of the Lepidium sativum L. species, and the control is represented by a solution of 1% methylcelulose.
The depressant activity of the investigated substances XXIV-XXVII was assessed by the value of the coefficient of inhibition of growth of the roots (Table 14).
I% = ಾ ష ು ಾ
. 100
where: LM – medium length of the roots of the control lot; LP– medium length of the roots of the lot that was acted upon with the investigated compounds.
Table 14. % inhibition achieved by the 2,3-disubstituted 4-oxo-1,3-thiazolidines XXIV-XXVII on the
growth of the Lepidium sativum L.roots. Compound mg/mL
3,3 1,6 0,8 0,4 0,2 XXIV 72 67 61 65 60 XXV 70 68 62 57 52 XXVI 74 71 68 63 61 XXVII 76 72 70 68 66 IOB-82 88 86 83 80 77
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Methotrexate 73 70 66 61 60
The four compounds in the series of 2,3-disubstituted thiazolidines have values of inhibition of over 50% for all concentrations, while substances XXVI and XXVII have values greater than methotrexate and smaller than the cytostatic IOB-82.
II. 2. 4. Testing of antimicrobial activity of the 1,3,4-thiadiazoles and 1,2,4-triazoles
Research in this domain was initiated on the basis of the existing relationship between chemical structure and antimicrobial activity.
Testing was done according to the international norms and regulations NCCLS [371] against the following strains: Staphylococcus aureus ATCC-25923, Bacillus subtilis ATCC-6638, Bacillus aureus ATCC-10876, Escherichia coli ATCC-25922 and Salmonela entiritidis P1131, from the Microbiology Laboratory, Department of Epidemiology, from the Institute of Public Health Iasi.
The results of the antimicrobial effects of the analysed substances were expressed as the diameter of the area of inhibition of bacterial growth – diffusometric method Kiry-Bauer [371] (Table 15) using as reference substance Kanamycin.
Table 15. Antimicrobial activity of 1,3,4-thiadiazoles XVIII-XXIII and 1,2,4-triazoles XXXIV-XXXIX.
Compound Diameter of area of inhibition (mm) S. aureus B. subtilis B. cereus E. coli S. enteritidis
XVIII 16-17 4-5 6-7 11-12 5-6 XIX 18-19 5-6 8-10 12-13 8-9 XX 19-20 6-7 6-7 14-15 7-8 XXI 20-21 8-9 9-10 15-16 9-10 XXII 19-20 6-7 6-7 13-14 7-8 XXIII 23-24 6-7 7-8 16-17 8-9
XXXIV 7-8 18-19 19-20 6-7 5-6 XXXV 6-7 20-21 19-20 7-8 6-7 XXXVI 8-9 19-20 18-19 7-8 5-6 XXXVII 6-7 23-24 22-23 9-10 9-10 XXXVIII 5-6 21-22 20-21 10-11 7-8 XXXIX 9-10 25-26 23-24 6-7 8-9
Kanamycin 31-32 27-28 30-31 29-30 28-29 From the tested compounds, the most active are the heterocycle derivatives that contain in their
molecule remains of p-bromophenyl XXI, XXXVII, and p-iodophenyl XXIII, XXXIX, respectively. The antimicrobial activity is appreciable, but inferior to the reference substance, kanamycin.
II.2.5. Testing of anti-pyretic activity of 2-[(5’-nitroindazole-1’- yl-methyl)]-5-(p-tolyl-amino)-1,3,4-oxadiazole
Literature shows that oxadiazole and its derivatives have a privileged structure in the sense that
allowance is made for a wide range of biological activities amogst which, one of particular interest, is the anti-inflammatory/anti-pyretic action [373].
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We looked at the anti-pyretic activity of one of the synthesized compounds, the one with the lowest toxicity in the series 2-[(5’-nitroindazole-1’-yl-methyl)-5(p-tolyl-amino)]-1,3,4-oxadiazole XXIX [332].
Anti-pyretic activity was assessed by the pyrexia test in the rat (intraperitoneal injection of the specified substance) and comparing it to indomethacin, phenylbutasone and acetil-salicylic acid. The results are shown in Table 17.
Table 17. Antipyretic activity of 1,3,4-oxadiazole XXIX
Compound Dose mg/Kg body
weight p.o.
Temperature, °C Temperature, °C after treatment, excluding the controls
Initially One hour after administration of the substance
1 hour 2 hours 3 hours 4 hours
Control - 36,72±0,14 37,80±0,07 38,40±0,11 38,65±0,05 38,95±0,04 39,20±0,12 Indomethacin 100 36,73±0,15 37,90±0,08 36,93±0,11 36,50±0,14 36,44±0,11 36,40±0,11
Phenylbutasone 100 36,73±0,15 37,82±0,07 37,71±0,11 37,40±0,08 37,36±0,12 37,46±0,13 Acetil-salicylic
Acid 100 36,71±0,09 37,75±0,12 37,08±0,17 37,06±0,22 36,93±0,16 36,70±0,24
Derivative of 1,3,4-oxadiazole,
XXIX
100 36,68±0,06 37,70±0,11 37,11±0,17 37,08±0,15 36,95±0,11 36,77±0,08
The 2,5-disubstituted 1,3,4-oxadiazole XXIX manifested a remarkable antipyretic activity, close
to the acetil-salicylic acid one, probably due to the action of inhibition of the synthesis of prostaglandins, characteristic to the oxadiazoles [376].
PART III
General Conclusions
Research undertaken for this paper refers to the synthesis and the study of the physico-chemical, and biological properties of new 5-nitroindazole derivatives. The results obtained allow for the following general conclusions:
1. The ethylic ester of the 5-nitroindazole-1-yl-acetic acid was obtained through the reaction of 5-nitroindazole with ethyl monochloroacetate, in the presence of sodium ethoxide, refluxing on ethylic alcohol. The advantage of the method we employed consists in the elimination of the separation phase of the sodium salt of 5-nitroindazole in solid form.
2. Having in mind the pharmacological significance of these substances, we synthesized new amidic type derivatives of 5-nitroindazole-1-yl-acetic acid and analysed their potential biological activities in that respect.
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3. We obtained the 5-nitroindazole-1-yl-acetic acid hydrazide by the reaction of ethylic ester of the 5-nitroindazole-1-yl-acetic acid with hydrazine hydrate in alcoholic solution at boiling temperature.
We synthesized this hydrazine with greater efficiency and of better purity just working at room temperature and using an excess of hydrazine hydrate. This substance represented the starting point for the synthesis of new compounds with an indazole nucleus in their structure.
4. In order to establish the influence of substituent groups (–NO2 and –OH) on the level of toxicity of the synthesized compounds, we obtained new hydrazones from condensation of 5-nitroindazolyl-acet-hydrazide with aromatic aldehydes.
5. We analysed the behaviour of the hydrazones obtained by by subjecting them to the cyclization reaction of 2,3-disubstituted 1,3-thiazolidines with thyoglycolic acid in the presence of zinc chloride as a dehydrating agent. We noticed that the substituent groups in the benzene ring (–NO2 in ortho or para position, and –OH in ortho position) in the structure of thiazolidines markedly influence their pharmacologic actions.
6. Applying a factorial experiment, we managed to optimize the synthesis reaction of the 2,3-disubstituted 1,3-thiazolidines, derived from acetamidyl-5-nitroindazole.
7. For full characterization of 2,3-disubstituted 4-oxo-1,3-thiazolidines, we used the HyperChem-5.0 programme that gave us important information regarding the geometry of the molecule, the of the atomic bonds, as well as the distribution of atomic charges within the molecules.
8. By the addition of 5-nitroindazole-1-yl-acetic acid hydrazide to the aromatic isothiocianates, we obtained the corresponding series of new acyl-thiosemicarbazides, which contain in their molecule both the thiosemicarbaside group and the remains of 5-nitroindazole.
9. From the synthesized acyl-thiosemicarbasides we analysed the reactivity of the thisemicarbaside group during cyclization under diverse experimental conditions.
a). By cyclization in an acid environment, we synthesized new thiadiazole derivatives that contain in their structure 5-nitroindazole. We also analysed their biological activity.
b). Cyclization of acyl-thiosemicarbasides with ethyl ester of monochloroacetic acid, in the presence of sodium acetate, lead to synthesis of 2,5-disubstituted 1,3,4-oxadiazoles, that had their toxicity, as well as their anti-pyrectic activity determined.
c). Through cyclization in an alkaline environment, of the acyl-thiosemicarbasides, we obtained 3,4-disubstituted mercapto-triazoles, and they have also been thoroughly tested from a biological point of view.
10. We synthesized new N-mustard derivatives by grafting di-(β-chloroethyl)-aminic group via an ethyl, ester or acetyl bridge from the5-nitroindazole molecule.
We hoped to achieve a break down of the molecule in the body, thus allowing the free alkylating fragment to attack the tumour cells. For the N-mustards synthesized, we established their DL50, while at the same time, we determined their cytostatic actions on experimental tumours.
11. We clarified the reaction mechanisms for closure of the thiadiazolic, thiazolidinic, oxadiazolic and cycles triazolic, function of the experimental circumstances.
12. The structure of the synthesized compounds was confirmed by means of quantitative determinations of the elements, as well as by spectral analysis (FT-IR, 1H-RMN, 13C-RMN, mass spectroscopy).
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13. Toxicological studies of the newly synthesized compounds have shown that they almost completely lack both general and haematopoietic toxicity and the values found experimentally for their DL50, recommends them for further biological testing.
14. Biological testing was done for 20 of the synthesized compounds. The experiments looked particularly at their cytostatic, mitodepressive, antimicrobial, and antipyretic actions.
a). A number of 7 compounds from the thiazolidines (XXIV-XXVII) and N-mustards (XLII, XLIII; XLV) have been tested for their cytostatic actions against experimental carcinosarcoma Walker 256, Jensen sarcoma and Guérin TB carcinoma. The results of the research show that these combinations behave as antitumoral agents and that they do have lesser toxicity and better selectivity with respect to the neoplastic cells. The most active are thiazolidine XXVII against carcinosarcoma Walker 256 and N-mustard XLIII against Guérin TB carcinoma.
b). Testing of the depressant action on the vegetal cell was done for four compounds (XXIV-XXVII) belonging to the 2,3-disubstituted 1,3-thiazolidines. The analysed compounds have an inhibiting effect on the growth of the roots of Lepidium sativum L. species greater than 50%, while compounds XXVI and XXVII have inhibiting values superior to the ones of methotrexate and inferior (lower) than the ones of the cytostatic IOB-82.
c). From a microbiological point of view, we tested the antimicrobial activity against six microbial strains, for a number of 12 substances (1,3,4-thiadiazoles XVIII-XXIII and 1,2,4-triazoles XXXIV-XXXIX). The most sensitive strains proved to be Staphylococcus aureus, Bacillus subtilis, and Bacillus cereus.
The substances having in their structure remains of p-bromophenyl XXI, XXXVII, and p-iodophenyl, respectively, XXIII, XXXIX, proved to have a remarkable antimicrobial activity, inferior though, to kanamycin (reference substance).
d). We assessed, on experimental animals, the antipyretic actions of 1,3,4-oxadiazole XXIX, using the pyrexia test and compared the results against indomethacin, phenylbutasone, and acetil-salicylic acid (reference drugs). The research showed that the substance tested has antipyretic activity close to that of the acetil-salicylic acid, an effect probably due to the inhibition of the biosynthesis of prostaglandines by the oxadiazolic heterocyle within the structure of compound XXIX.
15. With the research undertaken, we synthesized 45 intermediary and final compounds that are completely new, never before described in the specialised literature.
16. The original results presented in this paper have been published or accepted for publication as follows: 1 paper in on overseas journal (accepted for publication, graded ISI and SRI), 6 papers (5 published, 1 accepted for publication) in international journals (6 graded ISI, 5 graded SRI), 1 paper in a local journal (not graded ISI), 3 papers in overseas proceedings (graded ISI), 3 posters at national scientific meetings/congresses, first author of a book for tests and practical exercises in organic chemistry for under/postgraduate study (accreditation CNCSIS).
All of these are found with details in the publications list attached. 17. Other research done in this field has been published in 2 other papers that can be found in the
research contract CEEX No.9/2005-2008 (valued at 225.000 RON) titled: Advanced polymere biomaterials – structure and characteristics, applications in the food and pharmacy industry (scientific coordinator prof. dr. Valeriu Șunel) with beneficiary Academy of Medical Sciences-București, of whom, I am also an active research member.
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List of scientific works completed during the preparation of the Ph.D.
Articles published in journals abroad and the country: 8
1. Cheptea, C., Șunel, V., Desbrieres, J., Popa, M. Synthesis and antimicrobial activity of new derivatives of 1,3,4-thiadiazoles and 1,2,4-triazoles with 5-nitroindazole as support
J. Heterocycl. Chem., 2012, (ISI 1,22; influence RS 0,401)- accepted for publication, decision ID JHET-12-0102.R1 2. Cheptea, C., Șunel, V., Holban, M., Desbrieres, J., Popa, M., Lionte, C. Enhanced antipyretic activity of new 2,5-disubstituted 1,3,4-oxadiazoles encapsulated in alginate/gelatin particulated systems
Cell. Chem. Technol., 46, 2012, p 19-25, 2012 (ISI 0,55; influence RS 0,295) 3. Cheptea, C., Dulcescu, M. M., Dorohoi, D. O., Șunel, V., Desbrieres, J., Design, synthesis and molecular modelling of new thiazolidines 2,3-disubstituted with antitumoral activity
Dig. J. Nanomater. Bios., 7, 2012, p. 287-297 (ISI 1,2) 4. Cheptea, C., Dascălu, C. F., Șunel, V., Dorohoi, D. O. New derivatives based 1,3-thiazolidine-2,3-disubstituted with support of 5-nitroindazole-1’-acetamidyl with mitodepresive activity. Reaction optimization in factorial experiment
Rev. Chim., București, 63, 2012, p. 319-323. (ISI 0, 599; influence RS 0,125) 5. Cheptea, C. , Șunel, V., Stan, C., Dorohoi, D. O. New derivatives of 5-nitroindazole with potential antitumor activity
Rev. Roum. Chim., București, 57 (2), 2012, p. 229-234. (ISI 0,418; influence RS 0,152) 6. Cheptea, C., Holban, M., Peptu, C., Lionte, C., Șunel, V., Popa, M., Desbrieres, J. Synthesis and antimicrobial activity of new amidic derivatives of 5-nitroindazol-1-yl-acetic acid encapsulated into alginate/pectin particles
Cell. Chem. Technol., 2012 (ISI 0,55; influence RS 0,295) – accepted for publication, decision from may 2012 7. Murariu, C., Murariu, A., Harnagea, M., Ciovică, S., Cheptea, C., Șunel, V. A polymer-drug system based on regenerated cellulose used in textile industry
Cellulose Chem. Technol., 44, 2010, p 223-230 (ISI 0,320; influence RS 0,295) 8. Cheptea, C., Șunel, V., Profire, L., Popa, M., Lionte, C. New hydrazones of 5-nitroindazol-1-yl-acethydrazide with pharmacological potential
Bul. Inst. Polit., Iași, S.IIc, 55 (59), 2009, p 85-90.
Papers published as proceeding's: 3 1. C. Cheptea, V. Șunel, M. Crețu, J. Desbrieres, M. Holban, N. Bibire Synthesis of new 5-aryl-amino-2-substituted-1,3,4-oxadiazoles with antipyretic activity 15th Panhellenic Pharmaceutical Congres, Athens, Greece, May 13-15, 2011. J. Eur. Drug Metab. Ph., 36, 12, (2011) (ISI 0,479) 2. C. Cheptea, V. Șunel, L. Profire, C. Lionte, D. Stoican
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On the synthesis and toxicity of some new hydrazones of 5-nitroindazole-1-yl -acethydrazide 14th Panhellenic Pharmaceutical Congres, Athens, Greece,May 9-11, 2009, J. Eur. Drug Metab. Ph., 34, 13 (2009). (ISI 0.479 ) 3. C. Cheptea, V. Șunel, N. Dulea, M. Popa, L. Profire, C. Lionte New derivatives of 5-nitroindazole with potential antitumoral activity. 13th Panhellenic Pharmaceutical Congres, Athens, Greece,May 12-14, 2007, J. Eur. Drug Metab. Ph., 32, 8, (2007). (ISI 0,479)
Papers presented at scientific meetings: 3
1. C. Cheptea, V, Șunel, M. Zagnat New antitumoral compounds based on nitroindazole derivatives Conferința Națională ”Zilele Facultăţii de Bioinginerie Medicală“ Iaşi, 19-22 mai, 2011(poster 34) 2. C. Cheptea, V. Șunel, C. Lionte, M. Popa, J. Desbrieres Sinteza unor noi derivați ai 5-nitroindazolului cu potențială activitate antitumorală Sesiunea de Comunicări Ştiinţifice ”Zilele Universităţii“ Iaşi, 12-13 noiembrie, 2010 (poster P-21) 3. C. Cheptea, C. Maftei, V. Șunel, M. Popa Noi alchilanţi antitumorali cu suport de derivaţi ai 5-nitroindazolului. Sesiunea de Comunicări Ştiinţifice ”Zilele Universităţii“ Iaşi, 27-28 octombrie, 2006 (poster P-19).
Books published in accredited central publishing CNCSIS: 1
1. C. Cheptea, J. Cozma, M. Moise, V. Şunel. Probleme și exerciții de chimie organică (ISBN: 978-973-702-638-5; 206 pagini) Ed. Tehnopress, Iaşi, 2009
Research Contracts: 1
1. Member of research team of grant CEEX 9/3.10.2005-2008 with topic of Advanced polymeric biomaterials with structure and directed properties, with applications in pharmaceutical and food industry . Client: Academy of Medical Sciences, Bucharest Project manager: prof. Ph. Dr. Valeriu Şunel Value: 225.000 RON
Works outside the field of research: 2 1. Stoican, D., Sunel, V., Popa, M., Profire L., Cheptea, C. Synthesis of some di-chloroethyl amidic derivatives of the benzothiazolyl-2-mercaptoacetic acid and testing of their cytostatic activity 14th Panhellenic Pharmaceutical Congres, Athens, Greece,May 9-11, 2009, J. Eur. Drug Metab. Ph., 34, 7 (2009). (ISI 0.479 ). 2. Șunel, V., Lionte, C., Băsu, C., Cheptea, C. New antitumour alkylating compounds with N-[m-(arylthiocarbamoyl)-aminobenzoyl]-asparagic acids as supports Chem. Indian Journal, 2, 2005, p. 1-6.