Abstract:
The present invention relates to the method for preparing the anhydrous crystalline form of isoniazid-derived hydrazone (8-hydroxyquinoline-2-carboxaldehyde isonicotinoyl hydrazone), to the thus produced polymorph and to the use thereof for the treatment of Alzheimer&#39;s disease and parkinsonism, inter alia, and to a pharmaceutical composition.

Description:
FIELD OF THE INVENTION 
       [0001]    The present invention regards the preparation method of the anhydrous crystalline form of an isoniazid-derived hydrazone (namely, 8-hydroxyquinoline-2-carboxaldehyde isonicotinoyl hydrazone, or INHHQ), the polymorph produced and its use, in any pharmaceutical composition, for the treatment of Alzheimer&#39;s and Parkinson&#39;s diseases, as well as other neurodegenerative disorders. 
       BACKGROUND OF THE INVENTION 
       [0002]    With the general increase in life expectancy recorded in recent times, especially in developed countries, the prevalence of certain neurodegenerative diseases has been observed. Alzheimer&#39;s disease (AD), first characterized in 1906 by the german neuropathologist Alois Alzheimer, is currently the most common one. It is a primarily age-related disease and the most common cause of dementia in older people. Dementia is characterized by progressive loss of memory and cognitive functions, among other diagnostic criteria, being most of them present in the Diagnostic and Statistical Manual of Mental Disorders and described on the National Institute of Neurological and Communicative Disorders Association. 
         [0003]    The disease has two general classifications: 1) late onset, which occurs with the highest incidence at about 60 years of age, and 2) early onset, occurring around 40 years. In the US and Great Britain, it represents about 50% of cases of dementia, being estimated that it is the fourth leading cause of deaths of elderly in these countries. Regarding the neuropathological aspect, patients show diffuse cortical atrophy, presence of senile plaques and neurofibrillary tangles, neurovascular degeneration and neuronal loss. 
         [0004]    These senile plaques are characterized mainly by the presence of fibrillar deposits of β-amyloid peptide (Aβ), consisting of approx. 40 amino-acid residues. It has been observed high concentration of physiological metal ions such as Zn 2+  and Cu 2+ , in these plaques, which is considered indicative that the interaction of Aβ with these biometais is at the heart of events that lead to aggregation and toxicity of this peptide. The ferric ions, in turn, have also been related to the aggregation of neurofibrillary tangles, in addition to contributing to the oxidative processes that occur in the nerve cells of the body. Something similar occurs in Parkinson&#39;s disease with the protein α-synuclein. 
         [0005]    The already approved drugs for the treatment of AD are intended to combat deficits associated with reduced cerebral function and fall into two classes: acetylcholinesterase inhibitors and inhibitors of NMDA (N-Methyl-D-Aspartate) receptors. Such drugs seem to act in enhancing the remain of the cognitive function, however, are not able to prevent the progression of the disease, being, therefore, important the development of new therapeutic agents that hold the advance of neurodegeneration as well as, as far as possible, promote their regression. In this sense, an interesting approach is to obtain compounds that inhibit, specifically, abnormal metal-protein interactions. This class of drugs is known as MPACs (metal-protein attenuating compounds) and relate to the allocation and distribution normalization of physiological metal ions. 
         [0006]    A classic example is the clioquinol (CQ or PBT1), a substance belonging to the group of 8-hydroxyquinolines, which, however, was abandoned due to certain unwanted side effects, such as subacute myelo-optic neuropathy. 
         [0000]    
       
                 
         
             
             
         
       
       
         
           
             Structure of clioquinol (5-chloro-7-iodo-8-hydroxyquinoline). 
           
         
       
     
         [0008]    8-hydroxyquinoline-2-carboxaldehyde isonicotinoyl hydrazone (or INHHQ) was first described in a series of publications from the year 2009, in which its interactions with some rare earths ions, namely, dysprosium(III), europium(III), holmium(III), neodymium(III), and ytterbium(III) were examined. Erbium(III) and terbium(III) complexes were reported in 2010, while the samarium(III) compound was described in 2011. In those works, its rare earths complexes, and not INHHQ itself, were proposed as potential anticancer drugs, since they bind to DNA through an intercalation mechanism, besides they possess antioxidant properties, scavenging hydroxyl and superoxide radicals. Although the method of preparation described by the authors of those papers, Liu &amp; Yang, is similar to the one employed by us, some experimental details are different. Mainly, the use of acid catalysis (drops of concentrated hydrochloric acid) allow us to reduce the reflux period to just an hour. On the other hand, and more important yet, the use of methanol PA ACS (99.8%) in the recrystallization step, instead of the methanol:water 80:20 used by the Chinese authors, lead to the preparation of the anhydrous crystalline form of the compound, which is completely original. Although this is not commented in the original articles, data suggest that the Chinese authors worked with an hydrated form of INHHQ, different from the one obtained by us, which can be clearly seen in the major differences between the melting points of the compounds, as well as between the vibrational (FTIR and Raman) spectra of both samples. Besides, application of INHHQ in the treatment of neurodegenerative diseases is completely new. 
         [0009]    The anhydrous crystalline form described herein is characterized by its melting point, elemental analysis, crystal structure (single-crystal diffraction), powder diffraction pattern, and vibrational spectra. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0010]    The present invention will hereinafter be illustrated with reference to the attached figures, which represent: 
           [0011]      FIG. 1 : ORTEP representation of the compound INHHQ, showing the identification scheme of the constituent atoms; 
           [0012]      FIG. 2 : Perspective views of INHHQ showing: (a) the hydrogen-bonding network, (b) the π-π and the O1-H12 . . . π stacking interactions and (c) the 3D crystalline packing along the crystallographic axis a; 
           [0013]      FIG. 3 : Simulated and experimental X-ray diffractograms for INHHQ; 
           [0014]      FIG. 4 :Most stable calculated structure in the gas-phase (Conf. 5). Level of theory: B3LYP/6-311+G (d,p); 
           [0015]      FIG. 5 : FTIR (above) and Raman (below) spectra of INHHQ; 
           [0016]      FIG. 6 : 2D  1 H× 15 N HSQC contour plots showing that the addition of INHHQ (4 eq.) to a mixture containing the β-amyloid peptide and Zn(II) (1 eq.) leads to a partial intensity recovery for all Aβ signals, which can be considered as an indicative that the hydrazone INHHQ weakens Zn(II)-Aβ interactions; 
           [0017]      FIG. 7 :  1 H NMR spectra showing that the addition of INHHQ (4 eq.) to a mixture containinga-synuclein and Cu(II)/Cu(I) (1 eq.) leads to the recovery of the original methionine signals&#39; position; it is worth noting that methionine residues are the interaction point of copper with a-synuclein, which constitutes an indicative that INHHQ weakens Cu(II) or Cu(I)-α-synuclein interactions. 
       
    
    
     BRIEF DESCRIPTION OF THE INVENTION 
       [0018]    In the search for new MPACs, more effective and safer for patients, a hydrazone derived from the mycobactericidal agent isoniazid was produced, characterized in its crystalline form and successfully tested in modulating in vitro the interaction between β-amyloid peptide (or α-synuclein)and the physiological metals copper and zinc, which can have great impact on the treatment of Alzheimer&#39;s and Parkinson&#39;s diseases, as well as in other related diseases. 
       DETAILED DESCRIPTION OF THE INVENTION 
       [0019]    The isonicotinoyl hydrazone of 8-hydroxyquinoline-2-carboxaldehyde (or INHHQ) was synthesized, completely characterized in its anhydrous crystalline form and successfully tested in modulating the in vitro interaction between β-amyloid peptide (or α-synuclein) and the physiological metals copper and zinc, which can have applications in the treatment of Alzheimer&#39;s and Parkinson&#39;s diseases, amongst other neurodegenerative diseases (such as Huntington&#39;s). 
         [0000]    
       
                 
         
             
             
         
       
     
         [0020]    Structure of INHHQ (8-hydroxyquinoline-2-carboxaldehyde isonicotionoyl hydrazone). 
         [0021]    Connected to the 8-hydroxyquinoline portion, characteristic of CQ, INHHQ also contains the mycobactericidal drug isoniazid (INH), resulting in a potentially interesting hydrazone capable of coordinating metal ions of biological importance through its various N/O-donor sites. Furthermore, linking two molecules that have, individually, specific activity, creating a single species, has been an attractive approach to rational drug development, since a combination of the two original activities can be expected of the hybrid molecule. Besides, isoniazid-derived hydrazones are well known as iron chelating agents. However, to the best of our knowledge, there are no studies in the literature involving the coordination of INHHQ to any transition metal. 
         [0022]    In the context of the treatment of AD and Parkinson&#39;s disease, the coordination capacity of the ligand to essential transition metals (such as copper, zinc and iron) must be better understood. As a first approach to this problem, we present the results of a spectroscopic vibrational (FTIR/Raman) and a single-crystal X-ray diffraction structural study on the 8-hydroxyquinoline-2-carboxaldehyde isonicotinoyl hydrazone, obtained through the process herein described and claimed, including its crystal structure, vibrational spectra and their full assignment by computational methods based on the Density Functional Theory (DFT). 
       Preparation Methodology of the Anhydrous Crystalline Form of 8-hydroxyquinoline-2-carboxaldehyde isonicotinoyl hydrazone 
       [0023]    To 25 mL of a 8-hydroxyquinoline-2-carboxaldehyde (1.04 g, 6.0 mmol) solution in ethanol, 25 mL of ethanolic solution of isoniazid (0.82 g, 6.0 mmol) were dropwise added. Two drops of concentrated hydrochloric acid were added as a catalyst for the reaction. After refluxing for 1 h, the mixture was cooled to room temperature and the yellow precipitate obtained was filtered and vacuum dried. Next, the product was recrystallized in hot methanol methanol PA ACS (99.8%). A pale yellow crystalline solid was obtained after cooling and dried at room temperature. After few days, single-crystals of INHHQ were isolated from the mother liquor. Total yield of the process: 0.98 g (56%). 
         [0024]    Characterization: m.p.: 246-249° C. 
         [0025]    Elemental analysis: calculated for C 16 H 12 O 2 N 4 : C, 65.7%; H, 4.1%; N, 19.2%—found: C, 66.3%; H, 4.1%; N, 19.4%. 
         [0026]    Main IR bands (KBr, cm −1 ): 3396 (v OH); 3183 (v NH); 1656 (v C═O+β NH); 1647 (v C═N azomethine group); 1556 (v C═N e v C═C from the quinolinic ring) and 1545 (v C═C e v C═N from the pyridinic ring). 
       X-Ray Diffraction Analysis 
       [0027]    The X-ray diffraction (XRD) was performed using an appropriate single-crystal of INHHQ. The sample was measured on a diffractometer Enraf-Nonius Kappa-CCD with Mo K α  (λ=0.71073 Å) radiation. The unit cell parameters were based on all reflections. Data were collected at room temperature (293 K) using the computer program Collect being the integration and scaling of reflections made with Denzo-Scalepach system of HKL programs. The crystal structure was solved by direct method with SHELXS-97 and the atoms, except hydrogens, were anisotropically refined by the least squares method on F2 using the SHELXL-97 program. All aromatic and hydroxyl group&#39;s hydrogen atoms were placed in the calculated positions (C—H: 0.98 Å, O—H: 0.82 Å). Shift factors were taken as U(H) isot =1.2/1.5 Uhost. The H atoms attached to the C7 carbon and N2 nitrogen were located in the Fourier difference map and refined freely. The computer programs ORTEP-3 and Mercury (version 2.3) were used to draw the structures. On the other hand, powder X-ray diffraction experiments were performed in a Bruker D8 Discover XRD equipment using copper radiation. Experimental conditions: tension of 40 kV and current of 40 mA. The Bragg-Brentano geometry was used. 
       Spectroscopic Analyses 
       [0028]    IR spectra were obtained on a Perkin-Elmer 2000 FT-IR spectrometer, using KBr sampling. On the other hand, the Raman spectra of the solid sample were measured on a Perkin-Elmer 400 Station equipment, using the 785 nm line for excitation. 
       DFTcalculations 
       [0029]    The first step was to conduct a search of the conformational space of 8-hydroxyquinoline-2-carboxaldehyde isonicotinoyl hydrazone using the semi-empirical PM3 method, as implemented in the SPARTAN&#39;02 software, with the following set of parameters: Max-Confs=100; window=10 kcal mol −1 ; model=PM3. A total of 12 different conformations were found. The lowest energy conformation was of 41.387 kcal mol −1 . From this distribution, 5 conformations were selected for analysis by DFT. These conformations will be called, from now on, from Conf. 1 to Conf. 5. 
         [0030]    After the selection of these five conformations, a complete geometry optimization, without limitation, using the default convergence criteria and calculation of harmonic vibrational frequencies, was performed in gas phase, for each, using the Gaussian program package 03 Gauss. The three parameters of the exchange-correlation potential proposed by Becke with local and non-local correlations provided by Lee, Yang, and Parr (B3LYP functional) were selected with the basis set of triple zeta valence 6-311+G (d,p). Thermal contributions to the Gibbs free energy and other state functions were calculated at 298.15 K and 1 atm. The vibrational frequencies were scaled by a factor of 0.9381 for a better comparison with the experimental data. 
       Molecular Structure 
     Crystallographic Analysis 
       [0031]    The main crystal parameters used, as well as data of information collected and structural refinement parameters are summarized in Table 1. INHHQ crystallizes in the orthorhombic system, space group Pbca. The asymmetric unit of INHHQ is shown in  FIG. 1 . The compound adopts an (E) configuration with respect to the C7=N3 bond of the hydrazonic group. The INHHQ molecule is almost planarin solid state (rms deviation=0.2701° for all atoms, except H) and shows an intramolecular H bond involving the phenolic hydroxyl and the quinolinic nitrogen group: the O1-H donor interacts with N4 [O1 . . . N4=2,689 Å] receptor. In this process, a pseudo five-membered ring is formed. The distances and bond angles (Table 2) are not significantly different from those observed in similar compounds. The crystal packing is maintained by intermolecular H bonds involving the carbonyl oxygen O2 (acceptor) of a molecule and the N2-H group of the following molecule [moderate N2 . . . O2 i =2,966 Å, symmetry codes: (i) −x+½, y−½, z], connecting the molecules of INHHQ in zigzag chains running parallel to the crystallographic b axis ( FIG. 2 a   ). The molecules in each chain are interconnected by cross-stacking π-π interactions involving the quinoline rings. The calculated centroid-centroid distance is equal to 3.8303(9) Å. The adjacent chains are interconnected by O1-H12 . . . π interactions ( FIG. 2 b   ), being the distance H12-centroid (N4-C8-C9-C10-C11-C12) of 3.5339(17) Å [symmetry code: −½+x, y, ½−z]. As a result of this last interaction, zigzag columns run parallel to the crystallographic axis a ( FIG. 2 c   ). 
         [0000]    
       
         
               
             
               
               
             
           
               
                 TABLE 1 
               
               
                   
               
               
                 Selected crystallographic data for INHHQ 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Empirical formula 
                 C 16 H 12 O 2 N 4   
               
               
                 Molecular weight 
                 292.30 
               
               
                 Collecting temperature 
                 293(2) K 
               
               
                 Wave length 
                 0.71073 Å 
               
               
                 Crystalline system 
                 Orthorhombic 
               
               
                 Space group 
                 Pbca 
               
               
                 Unit cell dimensions 
                 a = 17.0761(4) Å 
               
               
                   
                 b = 8.25480(10) Å 
               
               
                   
                 c = 19.3549(4) Å 
               
               
                 Cell volume 
                 2728.26(9) Å 3   
               
               
                 Z 
                 8 
               
               
                 ρ(calculated) 
                 1.423 g cm −3   
               
               
                 Absorption coefficient 
                 0.098 mm −1   
               
               
                 F(000) 
                 1216 
               
               
                 Crystal size utilized 
                 0.484 × 0.236 × 0.171 mm 3   
               
               
                 θ range used in collect 
                 2.94 a 27.48° 
               
               
                 Index ranges 
                 −22.19; −10.10; −25.23 
               
               
                 Collected reflections 
                 27142 
               
               
                 Unique 
                 3100 [R (int)  = 0.1167] 
               
               
                 Completeness to θ = 27.48° 
                 99.0% 
               
               
                 Absorption correction 
                 None 
               
               
                 Refinement method 
                 “Full-matrix least-squares on F 2”   
               
               
                 Computational programs  a   
                 COLLECT, HKL Denzo and Scalepack, 
               
               
                   
                 SHELXS-97, SHELXL-97 
               
               
                 Data/restraints/parameters 
                 3100/0/208 
               
               
                 Goodness-of-fit on F 2   
                 1.042 
               
               
                 Final Rindexes [I &gt; 2σ (I)] 
                 R 1  = 0.0517, wR 2  = 0.1188 
               
               
                 R index (all data) 
                 R 1  = 0.0968, wR 2  = 0.1427 
               
               
                 Largest diff. peak and hole 
                 0.198 and −0.234 e Å −3   
               
               
                   
               
               
                   a  Used for data collection, data processing, structure solution, and structure refinement, respectively. 
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                 Main distances and bond angles, experimental and theoretical (Conf. 5), for 
               
               
                 INHHQ. The crystallographic identification scheme of the atoms was followed 
               
             
          
           
               
                 Distances (Å) 
                 Experimental 
                 Calculated 
                 Distances (Å) 
                 Experimental 
                 Calculated 
               
               
                   
               
             
          
           
               
                 O2—C6 
                 1.2268(18) 
                 1.211 
                 N4—C12 
                 1.362(2) 
                 1.355 
               
               
                 N2—C6 
                 1.345(2) 
                 1.388 
                 N3—C7 
                 1.270(2) 
                 1.280 
               
               
                 N2—N3 
                 1.3862(18) 
                 1.353 
                 O1—C13 
                 1.358(2) 
                 1.351 
               
               
                 N4—C8 
                 1.319(2) 
                 1.325 
                 N1—C1 
                 1.333(2) 
                 1.335 
               
               
                   
               
               
                 Angles (°) 
                   
                   
                 Angles (°) 
               
               
                   
               
               
                 C6—N2—N3 
                 118.44(13) 
                 120.9 
                 N2—C6—C5 
                 115.04(13) 
                 114.0 
               
               
                 C8—N4—C12 
                 117.83(13) 
                 118.6 
                 C1—N1—C2 
                 116.20(15) 
                 117.2 
               
               
                 C7—N3—N2 
                 115.54(13) 
                 117.4 
                 O1—C13—C12 
                 118.65(15) 
                 118.8 
               
               
                   
               
               
                 Dihedral angles (°) 
                   
                   
                 Dihedral angles (°) 
               
               
                   
               
               
                 C8—C7—N3—N2 
                 −177.63 
                 179.7 
                 N3—N2—C6—C5 
                 −178.62 
                 −178.6 
               
               
                 C7—N3—N2—C6 
                 −163.79 
                 −175.4 
                 N2—C6—C5—C3 
                 144.93 
                 −151.6 
               
               
                   
               
             
          
         
       
     
         [0032]    Moreover, an analysis of the INHHQ diffraction peaks was performed using Bragg-Brentano geometry (Table 3). The ultimate goal is to characterize, unambiguously, the obtained polymorph. The crystal structure-based simulated diffractogram in the range of 5&lt;2θ&lt;70°, was also calculated.  FIG. 3  displays the superposition of the experimental (red) and simulated (blue) diffractograms. 
         [0000]    
       
         
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                 Information on the diffraction peaks of INHHQ for 2θranging  
               
               
                 from 10.5 to 70.5° 
               
             
          
           
               
                   
                   
                   
                   
                 Interplanar 
                   
                 Structure 
               
             
          
           
               
                 Plane (h, k, l) 
                 Multiplicity 
                 distance (Å) 
                 2 theta  
                 factor 
               
             
          
           
               
                 h 
                 k 
                 l 
                 m 
                 d 
                 (°) 
                 F 2   
               
               
                   
               
             
          
           
               
                 1 
                 0 
                 0 
                 2 
                 8.34877 
                 10.58783 
                 1128.338 
               
               
                 0 
                 1 
                 0 
                 2 
                 7.07920 
                 12.49361 
                 331.562 
               
               
                 0 
                 1 
                 1 
                 4 
                 5.94931 
                 14.87873 
                 4309.398 
               
               
                 1 
                 0 
                 −2 
                 2 
                 5.93863 
                 14.90564 
                 3564.964 
               
               
                 1 
                 −1 
                 −1 
                 4 
                 5.50433 
                 16.08920 
                 1058.563 
               
               
                 0 
                 0 
                 2 
                 2 
                 5.48852 
                 16.13588 
                 185.920 
               
               
                 1 
                 1 
                 0 
                 4 
                 5.39942 
                 16.40394 
                 3734.208 
               
               
                 1 
                 −1 
                 −2 
                 4 
                 4.54974 
                 19.49495 
                 3562.456 
               
               
                 1 
                 1 
                 1 
                 4 
                 4.37713 
                 20.27167 
                 81.922 
               
               
                 2 
                 0 
                 −2 
                 2 
                 4.37659 
                 20.27421 
                 2544.993 
               
               
                 0 
                 1 
                 2 
                 4 
                 4.33757 
                 20.45853 
                 131.263 
               
               
                 2 
                 0 
                 0 
                 2 
                 4.17439 
                 21.26742 
                 743.827 
               
               
                 2 
                 −1 
                 −1 
                 4 
                 3.87923 
                 22.90666 
                 5227.304 
               
               
                 1 
                 0 
                 2 
                 2 
                 3.87110 
                 22.95541 
                 2615.373 
               
               
                 2 
                 −1 
                 −2 
                 4 
                 3.72262 
                 23.88427 
                 1558.836 
               
               
                 2 
                 1 
                 0 
                 4 
                 3.59579 
                 24.73979 
                 177.345 
               
               
                 0 
                 2 
                 0 
                 2 
                 3.53960 
                 25.13890 
                 1609.181 
               
               
                 1 
                 −1 
                 −3 
                 4 
                 3.53090 
                 25.20189 
                 4821.653 
               
               
                 1 
                 1 
                 2 
                 4 
                 3.39646 
                 26.21681 
                 4682.200 
               
               
                 0 
                 2 
                 1 
                 4 
                 3.36879 
                 26.43604 
                 783.016 
               
               
                 1 
                 −2 
                 −1 
                 4 
                 3.28146 
                 27.15290 
                 1838.082 
               
               
                 1 
                 2 
                 0 
                 4 
                 3.25881 
                 27.34522 
                 1594.341 
               
               
                 2 
                 −1 
                 −3 
                 4 
                 3.25355 
                 27.39035 
                 1187.558 
               
               
                 0 
                 1 
                 3 
                 4 
                 3.25049 
                 27.41659 
                 1027.571 
               
               
                 3 
                 0 
                 −2 
                 2 
                 3.08971 
                 28.87348 
                 81.686 
               
               
                 2 
                 1 
                 1 
                 4 
                 3.08859 
                 28.88417 
                 2178.500 
               
               
                 1 
                 −2 
                 −2 
                 4 
                 3.04049 
                 29.35123 
                 126085.008 
               
               
                 1 
                 0 
                 −4 
                 2 
                 3.03231 
                 29.43221 
                 46210.547 
               
               
                 1 
                 2 
                 1 
                 4 
                 2.98732 
                 29.88574 
                 3959.848 
               
               
                 0 
                 2 
                 2 
                 4 
                 2.97465 
                 30.01599 
                 23814.010 
               
               
                 2 
                 0 
                 −4 
                 2 
                 2.96931 
                 30.07124 
                 20257.656 
               
               
                 3 
                 −1 
                 −2 
                 4 
                 2.83175 
                 31.56923 
                 1316.031 
               
               
                 2 
                 −2 
                 −1 
                 4 
                 2.81367 
                 31.77739 
                 2147.033 
               
               
                 3 
                 −1 
                 −1 
                 4 
                 2.78865 
                 32.07025 
                 962.328 
               
               
                 1 
                 −1 
                 −4 
                 4 
                 2.78737 
                 32.08534 
                 5017.017 
               
               
                 2 
                 0 
                 2 
                 2 
                 2.78466 
                 32.11737 
                 5954.872 
               
               
                 3 
                 0 
                 0 
                 2 
                 2.78292 
                 32.13796 
                 198029.766 
               
               
                 2 
                 −2 
                 −2 
                 4 
                 2.75217 
                 32.50702 
                 166846.703 
               
               
                 0 
                 0 
                 4 
                 2 
                 2.74426 
                 32.60332 
                 78569.977 
               
               
                 2 
                 −1 
                 −4 
                 4 
                 2.73820 
                 32.67747 
                 7683.784 
               
               
                 2 
                 2 
                 0 
                 4 
                 2.69971 
                 33.15667 
                 5524.309 
               
               
                 3 
                 −1 
                 −3 
                 4 
                 2.69759 
                 33.18355 
                 5725.885 
               
               
                 1 
                 1 
                 3 
                 4 
                 2.69411 
                 33.22766 
                 3144.983 
               
               
                 1 
                 −2 
                 −3 
                 4 
                 2.67192 
                 33.51166 
                 5080.103 
               
               
                 1 
                 2 
                 2 
                 4 
                 2.61222 
                 34.30092 
                 184684.922 
               
               
                 3 
                 0 
                 −4 
                 2 
                 2.61018 
                 34.32857 
                 75081.414 
               
               
                 2 
                 1 
                 2 
                 4 
                 2.59139 
                 34.58541 
                 10269.332 
               
               
                 3 
                 1 
                 0 
                 4 
                 2.58999 
                 34.60470 
                 3123.402 
               
               
                 0 
                 1 
                 4 
                 4 
                 2.55873 
                 35.04100 
                 3632.206 
               
               
                 2 
                 −2 
                 −3 
                 4 
                 2.54551 
                 35.22898 
                 128.840 
               
               
                 0 
                 2 
                 3 
                 4 
                 2.54404 
                 35.24991 
                 2518.650 
               
               
                 2 
                 2 
                 1 
                 4 
                 2.46414 
                 36.43237 
                 9364.604 
               
               
                 3 
                 −1 
                 −4 
                 4 
                 2.44902 
                 36.66531 
                 6171.995 
               
               
                 0 
                 3 
                 0 
                 2 
                 2.35973 
                 38.10499 
                 2742.504 
               
               
                 3 
                 −2 
                 −2 
                 4 
                 2.32767 
                 38.65065 
                 49774.492 
               
               
                 1 
                 0 
                 4 
                 2 
                 2.32176 
                 38.75283 
                 27728.582 
               
               
                 4 
                 0 
                 −2 
                 2 
                 2.31874 
                 38.80530 
                 8126.180 
               
               
                 3 
                 1 
                 1 
                 4 
                 2.31772 
                 38.82317 
                 7422.130 
               
               
                 0 
                 3 
                 1 
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                 3 
                 4 
                 1 
                 4 
                 1.43522 
                 64.91979 
                 856.294 
               
               
                 0 
                 2 
                 7 
                 4 
                 1.43374 
                 64.99512 
                 3941.112 
               
               
                 6 
                 −1 
                 −1 
                 4 
                 1.43271 
                 65.04772 
                 9958.013 
               
               
                 5 
                 0 
                 2 
                 2 
                 1.43178 
                 65.09530 
                 464.878 
               
               
                 2 
                 −4 
                 −5 
                 4 
                 1.43122 
                 65.12385 
                 1343.613 
               
               
                 1 
                 −1 
                 −8 
                 4 
                 1.43049 
                 65.16140 
                 2849.458 
               
               
                 6 
                 −1 
                 −6 
                 4 
                 1.42884 
                 65.24586 
                 1632.203 
               
               
                 2 
                 1 
                 6 
                 4 
                 1.42677 
                 65.35220 
                 113.830 
               
               
                 1 
                 −4 
                 −5 
                 4 
                 1.42405 
                 65.49281 
                 2715.529 
               
               
                 5 
                 2 
                 1 
                 4 
                 1.42169 
                 65.61510 
                 2686.276 
               
               
                 6 
                 −2 
                 −3 
                 4 
                 1.41663 
                 65.87928 
                 845.347 
               
               
                 5 
                 −3 
                 −1 
                 4 
                 1.41622 
                 65.90063 
                 1809.560 
               
               
                 6 
                 −2 
                 −4 
                 4 
                 1.41588 
                 65.91867 
                 654.705 
               
               
                 0 
                 5 
                 0 
                 2 
                 1.41584 
                 65.92052 
                 1197.549 
               
               
                 4 
                 2 
                 3 
                 4 
                 1.41537 
                 65.94542 
                 244.526 
               
               
                 3 
                 1 
                 5 
                 4 
                 1.41356 
                 66.04055 
                 4426.744 
               
               
                 1 
                 4 
                 4 
                 4 
                 1.40751 
                 66.36081 
                 21200.830 
               
               
                 4 
                 −4 
                 −2 
                 4 
                 1.40684 
                 66.39668 
                 396.245 
               
               
                 5 
                 −3 
                 −5 
                 4 
                 1.40597 
                 66.44291 
                 29.975 
               
               
                 5 
                 0 
                 −8 
                 2 
                 1.40574 
                 66.45520 
                 3142.987 
               
               
                 0 
                 5 
                 1 
                 4 
                 1.40421 
                 66.53712 
                 7893.484 
               
               
                 5 
                 1 
                 2 
                 4 
                 1.40336 
                 66.58237 
                 970.894 
               
               
                 1 
                 1 
                 7 
                 4 
                 1.40288 
                 66.60856 
                 170.244 
               
               
                 4 
                 −4 
                 −3 
                 4 
                 1.40251 
                 66.62790 
                 1310.150 
               
               
                 2 
                 4 
                 3 
                 4 
                 1.40178 
                 66.66737 
                 2531.638 
               
               
                 2 
                 −3 
                 −7 
                 4 
                 1.40123 
                 66.69715 
                 8505.064 
               
               
                 5 
                 −2 
                 −7 
                 4 
                 1.40118 
                 66.69978 
                 401.337 
               
               
                 3 
                 −2 
                 −8 
                 4 
                 1.40053 
                 66.73479 
                 5147.162 
               
               
                 1 
                 −5 
                 −1 
                 4 
                 1.39767 
                 66.88894 
                 1543.957 
               
               
                 3 
                 −4 
                 −5 
                 4 
                 1.39761 
                 66.89248 
                 1963.978 
               
               
                 1 
                 5 
                 0 
                 4 
                 1.39591 
                 66.98464 
                 1123.055 
               
               
                 3 
                 −3 
                 −7 
                 4 
                 1.39498 
                 67.03543 
                 1584.166 
               
               
                 6 
                 −2 
                 −2 
                 4 
                 1.39432 
                 67.07096 
                 6423.831 
               
               
                 2 
                 −2 
                 −8 
                 4 
                 1.39368 
                 67.10576 
                 2221.480 
               
               
                 4 
                 0 
                 4 
                 2 
                 1.39233 
                 67.17963 
                 9756.798 
               
               
                 6 
                 −2 
                 −5 
                 4 
                 1.39218 
                 67.18803 
                 123.361 
               
               
                 6 
                 0 
                 0 
                 2 
                 1.39146 
                 67.22715 
                 61458.719 
               
               
                 4 
                 −4 
                 −1 
                 4 
                 1.38844 
                 67.39312 
                 1208.544 
               
               
                 4 
                 3 
                 2 
                 4 
                 1.38797 
                 67.41877 
                 146.200 
               
               
                 5 
                 −1 
                 −8 
                 4 
                 1.37882 
                 67.92709 
                 1203.279 
               
               
                 0 
                 4 
                 5 
                 4 
                 1.37785 
                 67.98154 
                 710.651 
               
               
                 1 
                 −5 
                 −2 
                 4 
                 1.37724 
                 68.01559 
                 3062.095 
               
               
                 4 
                 −4 
                 −4 
                 4 
                 1.37608 
                 68.08057 
                 27359.787 
               
               
                 1 
                 5 
                 1 
                 4 
                 1.37219 
                 68.30019 
                 694.557 
               
               
                 0 
                 0 
                 8 
                 2 
                 1.37213 
                 68.30382 
                 15200.482 
               
               
                 0 
                 5 
                 2 
                 4 
                 1.37096 
                 68.37016 
                 828.054 
               
               
                 1 
                 −3 
                 −7 
                 4 
                 1.36918 
                 68.47118 
                 3214.390 
               
               
                 4 
                 −2 
                 −8 
                 4 
                 1.36910 
                 68.47589 
                 45.553 
               
               
                 4 
                 1 
                 4 
                 4 
                 1.36616 
                 68.64389 
                 519.685 
               
               
                 6 
                 1 
                 0 
                 4 
                 1.36534 
                 68.69093 
                 1607.419 
               
               
                 5 
                 3 
                 0 
                 4 
                 1.36303 
                 68.82355 
                 2112.606 
               
               
                 3 
                 4 
                 2 
                 4 
                 1.36190 
                 68.88850 
                 1753.492 
               
               
                 6 
                 −1 
                 −7 
                 4 
                 1.36065 
                 68.96085 
                 2649.621 
               
               
                 2 
                 3 
                 5 
                 4 
                 1.36033 
                 68.97921 
                 2510.422 
               
               
                 2 
                 −5 
                 −1 
                 4 
                 1.35414 
                 69.33989 
                 3657.629 
               
               
                 6 
                 −2 
                 −1 
                 4 
                 1.35205 
                 69.46230 
                 243.801 
               
               
                 4 
                 −3 
                 −7 
                 4 
                 1.35190 
                 69.47100 
                 6915.945 
               
               
                 1 
                 −2 
                 −8 
                 4 
                 1.35018 
                 69.57237 
                 1099.760 
               
               
                 4 
                 4 
                 0 
                 4 
                 1.34986 
                 69.59140 
                 705.399 
               
               
                 5 
                 −3 
                 −6 
                 4 
                 1.34938 
                 69.61938 
                 756.755 
               
               
                 6 
                 −2 
                 −6 
                 4 
                 1.34879 
                 69.65417 
                 2227.248 
               
               
                 1 
                 3 
                 6 
                 4 
                 1.34807 
                 69.69667 
                 318.911 
               
               
                 2 
                 −5 
                 −2 
                 4 
                 1.34710 
                 69.75419 
                 61.678 
               
               
                 0 
                 1 
                 8 
                 4 
                 1.34706 
                 69.75684 
                 1471.690 
               
               
                 2 
                 2 
                 6 
                 4 
                 1.34705 
                 69.75717 
                 576.139 
               
               
                 2 
                 5 
                 0 
                 4 
                 1.34082 
                 70.12915 
                 2349.962 
               
               
                 3 
                 3 
                 4 
                 4 
                 1.34035 
                 70.15689 
                 35.647 
               
               
                 1 
                 −5 
                 −3 
                 4 
                 1.33737 
                 70.33644 
                 2229.274 
               
               
                 2 
                 −4 
                 −6 
                 4 
                 1.33596 
                 70.42175 
                 3337.051 
               
               
                 3 
                 2 
                 5 
                 4 
                 1.33592 
                 70.42439 
                 156.499 
               
               
                   
               
             
          
         
       
     
       Gas-Phase DFT Calculations 
       [0033]    The INHHQ structure was optimized in gas phase, using the DFT methodology, level of theory B3LYP/6-311+G (d,p). 
         [0034]    As described above, a total of 5 conformations with the smallest PM3 energy differences were selected to perform a DFT optimization and frequency calculations. Cis-trans isomerism was contemplated in this treatment. Energy values found for each of these structures indicated that there are, in fact, only 3 different conformations. Conf.5 is the one with the lowest free energy (ΔG) when compared to the other conformations. However, the energy differences between the structures are not appreciable, especially between the Conf. 1/2 with respect to the Conf. 5 (only 0.06 kcal mol −1 ), indicating that, indeed, all five conformations are possible, particularly Conf. 5. This last one was chosen for further deeper studies. 
         [0035]    It is shown (Table 2) that there is excellent agreement between the structural parameters found in the calculations (Conf. 5) and the refined X-ray structure and that, in turn, there is no significant difference between  FIGS. 1 and 4 . Crystallography shows that the phenolic hydrogen points in the direction of the quinolinic nitrogen, since a connection involving these H atoms is formed, producing a 2.691 Å donor-acceptor distance, in perfect agreement with the X-ray data (O1 . . . N4=2.689 Å). 
       Vibrational Analyses 
       [0036]    The experimental FTIR and Raman spectra of INHHQ, in the solid state, are shown in  FIG. 5 . The observed and calculated frequencies, as well as an attempt to assign the main bands, are given in Table 4. 
         [0000]    
       
         
               
             
               
               
               
             
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                 Assignment of the FTIR and Raman spectra of INHHQ (scale factor: 0.9381) 
               
             
          
           
               
                   
                 Theoretical (cm −1 ) 
                   
               
               
                   
                 B3LYP/6-311 + G(d, p) 
               
             
          
           
               
                 Experimental (cm −1 ) 
                 Unscaled 
                 Scaled 
                   
                   
               
               
                 FTIRRaman 
                 (cm −1 ) 
                 (cm −1 ) 
                 IR Intensity 
                 Vibrational Assignment 
               
               
                   
               
             
          
           
               
                 3396 
                 m 
                 3400 
                 br 
                 3662 
                 3435 
                 104.3584 
                 v OH 
               
               
                 3208 
                 sh 
                 3205 
                 w 
                   
                   
                   
                 — 
               
             
          
           
               
                 3183 
                 w 
                 — 
                 3502 
                 3285 
                 5.5099 
                 v NH 
               
               
                 3148 
                 sh 
                 — 
                 3211/3205 
                 3012/3007 
                 1.8906/3.2991 
                 v CH(Quin + Py)ip 
               
             
          
           
               
                 3073 
                 sh 
                 3081 
                 w 
                 3198 
                 3000 
                 9.2441 
                 v CH(Quin)ip 
               
               
                 3059 
                 sh 
                 3055 
                 w 
                 3185/3178 
                 2988/2981 
                 18.7464/9.9528  
                 v CH(Quin + Py)op 
               
             
          
           
               
                 3042 
                 w 
                 — 
                   
                   
                   
                 — 
               
             
          
           
               
                 3028 
                 sh 
                 3030 
                 w 
                   
                   
                   
                 — 
               
             
          
           
               
                 3016 
                 w 
                 — 
                   
                   
                   
                 — 
               
             
          
           
               
                 — 
                 2996 
                 w 
                   
                   
                   
                 — 
               
             
          
           
               
                 2959 
                 sh 
                 — 
                   
                   
                   
                 — 
               
               
                 2923 
                 w 
                 — 
                   
                   
                   
                 — 
               
               
                 2852 
                 w 
                 — 
                 3048 
                 2859 
                 40.4835 
                 vCH(Azomethine) 
               
               
                 2835 
                 sh 
                 — 
                   
                   
                   
                 — 
               
             
          
           
               
                 1656 
                 s 
                 1660 
                 sh 
                 1767 
                 1658 
                 350.6350 
                 vC═O + βNH 
               
               
                 1647 
                 vs 
                 1646 
                 w 
                 1673 
                 1569 
                 20.7477 
                 v C═N(Azomethine) + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 v C═C(Quin) + βC—OH 
               
               
                 1604 
                 w 
                 1603 
                 vs 
                 1659 
                 1556 
                 2.3397 
                 v C═N(Azomethine) + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 v C═C(Quin) + βC—OH 
               
               
                 1595 
                 w 
                 1595 
                 s 
                 1634 
                 1533 
                 5.5690 
                 v C═C(Quin) 
               
               
                 1556 
                 m 
                 1555 
                 w 
                 1597 
                 1498 
                 18.4149 
                 v C═N(Quin) + vC═C(Quin) 
               
             
          
           
               
                 1545 
                 m 
                 — 
                 1630 
                 1529 
                 14.2312 
                 v C═C(Py) + vC═N(Py) 
               
             
          
           
               
                 1507 
                 m 
                 1507 
                 w 
                 1555 
                 1459 
                 371.3416 
                 βNH + βCH(Quin) 
               
               
                 1490 
                 w 
                 1488 
                 vw 
                   
                   
                   
                 — 
               
               
                 1465 
                 m 
                 1468 
                 w 
                 1541 
                 1446 
                 168.8474 
                 Ring stretch(Quin) + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 βC—OH + β NH 
               
               
                 1437 
                 m 
                 1435 
                 s 
                 1519 
                 1425 
                 6.8860 
                 βCH(Py) 
               
             
          
           
               
                 1407 
                 w 
                 — 
                   
                   
                   
                 — 
               
             
          
           
               
                 1394 
                 sh 
                 1396 
                 m 
                   
                   
                   
                 — 
               
               
                 1371 
                 w 
                 1371 
                 m 
                 1495 
                 1402 
                 176.2051 
                 β C—OH + β NH + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 β CH(Quin) 
               
               
                 1330 
                 m 
                 1329 
                 m 
                   
                   
                   
                 — 
               
               
                 1299 
                 s 
                 1304 
                 m 
                 1360 
                 1276 
                 34.4059 
                 β CH(Quin + Azomethine) + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 v C═N(Quin) + v C—OH 
               
               
                 1280 
                 m 
                 1279 
                 sh 
                 1347 
                 1264 
                 31.7632 
                 β NH + β 
               
               
                   
                   
                   
                   
                   
                   
                   
                 CH(Azomethine + Py) + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 β C—OH 
               
             
          
           
               
                 1270 
                 sh 
                 — 
                   
                   
                   
                 — 
               
             
          
           
               
                 1252 
                 m 
                 1252 
                 s 
                 1309 
                 1228 
                 15.0576 
                 β CH(Quin + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 Azomethyne) + β C—OH 
               
             
          
           
               
                 1232 
                 m 
                 — 
                 1284 
                 1205 
                 117.6910 
                 v C—OH + β CH(Quin + 
               
               
                   
                   
                   
                   
                   
                   
                 Azomethine) 
               
             
          
           
               
                 1217 
                 w 
                 1220 
                 w 
                 1278 
                 1199 
                 32.0853 
                 v C═N(Py) + vC═C(Py) + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 β NH 
               
               
                 1204 
                 w 
                 1204 
                 vw 
                 1267 
                 1189 
                 56.8637 
                 β CH(Azomethine + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 Quin) + β C—OH 
               
               
                 1170 
                 w 
                 1172 
                 sh 
                 1253 
                 1175 
                 303.5474 
                 βCH(Py) + βNH 
               
               
                 1156 
                 m 
                 1156 
                 m 
                 1172 
                 1099 
                 380.9402 
                 v N—N + βCH(Py + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 Quin) 
               
               
                 1136 
                 w 
                 1133 
                 w 
                 1112 
                 1043 
                 10.8679 
                 βCH(Py) 
               
             
          
           
               
                 1122 
                 sh 
                 — 
                   
                   
                   
                 — 
               
             
          
           
               
                 1105 
                 w 
                 1105 
                 vw 
                   
                   
                   
                 — 
               
               
                 1090 
                 w 
                 1093 
                 vw 
                 1109 
                 1040 
                 19.7634 
                 βCH(Py + Quin) 
               
               
                 1072 
                 w 
                 1076 
                 w 
                 1092 
                 1024 
                 9.3109 
                 βCH(Py) 
               
             
          
           
               
                 — 
                 1063 
                 vw 
                 1079 
                 1012 
                 11.9293 
                 βCH(Quin) + δ NNC 
               
             
          
           
               
                 1044 
                 vw 
                 1044 
                 vw 
                 1068 
                 1002 
                 1.6514 
                 βCH(Quin) 
               
             
          
           
               
                 1007 
                 vw 
                 — 
                   
                   
                   
                 — 
               
             
          
           
               
                 992 
                 vw 
                 992 
                 w 
                 1010 
                 947 
                 1.7715 
                 Ring breath(Py) 
               
             
          
           
               
                 981 
                 vw 
                 — 
                 1008 
                 946 
                 1.5921 
                 γCH(Py) 
               
             
          
           
               
                 950 
                 vw 
                 952 
                 w 
                 959 
                 900 
                 14.8080 
                 γCH(Azomethine) 
               
               
                 931 
                 w 
                 932 
                 w 
                 914 
                 857 
                 2.9777 
                 Ring-deformation(Quin + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 Py) 
               
               
                 895 
                 w 
                 897 
                 vw 
                   
                   
                   
                 — 
               
             
          
           
               
                 881 
                 vw 
                 — 
                   
                   
                   
                 — 
               
             
          
           
               
                 867 
                 w 
                 869 
                 w 
                 898 
                 842 
                 0.0665 
                 γCH(Quin) 
               
               
                 856 
                 m 
                 855 
                 sh 
                 880 
                 826 
                 11.8041 
                 Ring-deformation(Quin + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 Py) 
               
               
                 837 
                 m 
                 835 
                 vw 
                 892 
                 837 
                 2.8606 
                 γCH(Py) 
               
             
          
           
               
                 — 
                 812 
                 vw 
                 804 
                 754 
                 2.0048 
                 γ C═C—C(Quin) + 
               
               
                   
                   
                   
                   
                   
                   
                 γ C═N—C(Quin) 
               
             
          
           
               
                 792 
                 vw 
                 780 
                 w 
                 789 
                 740 
                 5.7350 
                 β C═C—C(Quin + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 Azomethine) 
               
               
                 766 
                 s 
                 768 
                 vw 
                 767 
                 720 
                 9.2858 
                 γCH(Py) 
               
               
                 720 
                 m 
                 720 
                 m 
                 734 
                 689 
                 18.7702 
                 Ring-deformation(Quin) 
               
               
                 696 
                 w 
                 696 
                 vw 
                 763 
                 716 
                 38.2602 
                 γ CH(Quin) 
               
             
          
           
               
                 677 
                 s 
                 — 
                 719 
                 674 
                 19.3631 
                 β C═N—C(Py) + 
               
               
                   
                   
                   
                   
                   
                   
                 β C═C—C(Quin) 
               
             
          
           
               
                 670 
                 sh 
                 668 
                 w 
                 698 
                 655 
                 2.0785 
                 γ C═C—C(Quin) + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 β C═N—C(Py) 
               
               
                 656 
                 w 
                 657 
                 w 
                 693 
                 650 
                 58.0279 
                 Ring-deformation(Py) 
               
             
          
           
               
                 644 
                 sh 
                 — 
                 681 
                 639 
                 1.5003 
                 Ring-deformation(Py) 
               
               
                 616 
                 vw 
                 — 
                 627 
                 588 
                 2.3638 
                 Ring-deformation(Quin) 
               
               
                 587 
                 m 
                 — 
                 604 
                 567 
                 90.9456 
                 γ OH 
               
               
                 575 
                 w 
                 — 
                   
                   
                   
                 — 
               
             
          
           
               
                 547 
                 sh 
                 553 
                 w 
                 588 
                 552 
                 10.5255 
                 β C═C—C(Quin) 
               
               
                 532 
                 sh 
                 533 
                 vw 
                 559 
                 524 
                 1.6705 
                 β C═C—C(Quin) + γ NH 
               
               
                 522 
                 w 
                 523 
                 vw 
                 551 
                 517 
                 14.1393 
                 γCH(Quin) + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 β C═C—C(Quin) + 
               
               
                   
                   
                   
                   
                   
                   
                   
                 β C—OH 
               
               
                 482 
                 w 
                 485 
                 w 
                 539/534 
                 506/501 
                 16.0281/30.1104 
                 γ NH + γ C═C—C(Quin) 
               
               
                   
               
               
                 Quin: quinoline ring; Py: pyridine ring; vs: very strong; s: strong; m: medium; w: weak; vw: very weak; br: broad; sh: shoulder; ip: in-phase; op: out-of-phase; v: stretching; β: in-plane bending; γ: out-of-plane bending. 
               
             
          
         
       
     
       Carbonylic C═O Stretching 
       [0037]    The v C═O absorption is usually one of the most representative in an infrared spectrum and is also likely its most intense spectral feature. It appears in a wavenumber region relatively free of other vibrations (1800-1600 cm −1 ). On the other hand, this mode gives only weak or very weak absorptions in Raman spectroscopy. In our study, as expected, v C═O vibration originates one of the strongest bands of the infrared spectrum, at 1656 cm −1 , which is in excellent agreement with the calculated value of 1658 cm −1  (DFT calculations show a coupling between v C═O and β NH vibrations). 
         [0038]    This mode was assigned at 1663 cm −1  by Liu and Yang. 
         [0000]    Azomethine C═N stretching 
         [0039]    The C═N stretchings of azomethine groups show absorptions close to that of carbonyl stretching. This fact can difficult an accurate assignment. For example, the C═N stretching bands of alkylated Schiff bases are usually found in the range 1674-1649 cm −1 , inside the common region of v C═O absorption. If conjugations of the C═N moiety with phenyl groups are present, the stretching frequency shifts to 1650-1600 cm −1 . In this work, two frequencies involving azomethine C═N vibrations were calculated (1569/1556 cm −1 ), both of them coupled to v C═C of the quinoline ring and, to a lesser extent, to the phenol v C—OH. These values are in good agreement with the experimental frequencies observed in the infrared, at 1647 (vs) and 1604 (w) cm −1 , and Raman spectra, at 1646 (w) and 1603 (vs) cm −1 , respectively. 
         [0040]    Liu and Yang, though, attributed this mode to a single band at 1613 cm −1  in the IR spectrum, which was not observed in our study. 
       OH and NH Stretching Vibrations 
       [0041]    OH and NH groups are very characteristic and their stretching vibrations are observed, in many cases, around 3500-3300 cm −1 . This absorption, however, is highly influenced by chemical environment, mainly when OH or NH groups are involved in hydrogen bonding. This can occur within the same molecule (intramolecular H bonding) or with adjacent molecules (intermolecular H bonding). The presence of intramolecular H bonding causes a thinning of the band and makes its position unaffected by concentration changes. In the IR spectrum of INHHQ, we observed a sharp band of medium intensity located at 3396 cm −1 , assigned to v OH. A similar absorption, at 3418 cm −1 , was reported by Krishnakumar and Ramasamy in the infrared spectrum of 8-hydroxyquinoline (8-HQ). On the other hand, intermolecular hydrogen bonding usually leads to a broadening of the band, as can be seen in the case of the v NH absorption of INHHQ, which was attributed to the weak IR band at 3183 cm −1 . In a previous study on the isonicotinoyl hydrazone of 2-hydroxy-3-methoxybenzaldehyde, published by us, v NH vibration was observed as a weak band at 3157 cm −1 . Here, we found serious discrepancies concerning the assignments made by Liu and Yang, since these authors attributed an absorption of higher frequency (reported by them at 3576 cm −1 ) to the NH stretching mode, whereas the lower frequency band at 3193 cm −1  was credited to the OH stretching movement. 
         [0042]    Phenol C—OH Vibrations 
         [0043]    In this work, the C—OH stretching mode was assigned to the medium intensity infrared band at 1232 cm −1 . This vibration is Raman inactive and had its frequency calculated at 1205 cm −1 . A coupled mode involving this movement was also predicted at 1276 cm −1  [experimental: 1299 (infrared) cm −1  and 1304 (Raman) cm −1 ]. Another important vibration concerning the phenol group is the in-plane bending, which typically appears in the region 1440-1260 cm −1 , attributed to the weak infrared band (medium in the Raman spectrum) at 1371 cm −1 . Coupled modes are observed in FTIR at 1465, 1280, 1252 and 1202 cm −1 . Theoretical and experimental frequencies show good agreement (Table 4). 
       Biological Activity Studies 
       [0044]    This hydrazonic compound has the ability, as proved by 1D and 2D NMR experiments, of compete with the β-amyloid peptide (or α-synuclein), key-targets, respectively, in Alzheimer&#39;s and Parkinson&#39;s diseases, by physiological ions such as Zn 2+  Cu 2+  and Cu + , which can cause their precipitation or oligomerization and contribute to worsen the oxidative stress condition observed in the brains of patients suffering from these neurodegenerative disorders. INHHQ does not interact directly with the β-amyloid peptide and α-synuclein, nonetheless inhibits the interactions between these targets and metals through a mechanism that probably involves metal ion sequestering. 
         [0045]    In silico pharmacological analyses (mandatory studies in the development of new therapeutic agents, whose aim is to predict pharmacokinetic properties of novel molecules with a potential pharmacological action) show that the Lipinski&#39;s parameters presented by INHHQ (Table 5), calculated via 1D-QSAR method, are in agreement to ideal values, suggesting that this compound constitutes an excellent candidate to new drug, with good oral absorption and cellular permeability. The model also indicates that INHHQ possesses structural features which can allow it crossing the blood-brain barrier (BBB), resulting in therapeutic actions inside the patients&#39; brains. 
         [0000]    
       
         
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 Lipinski parameters calculated for 
               
               
                 INHHQ, along with reference values 
               
             
          
           
               
                   
                   
                 Ref. Oral 
                 Ref. BBB 
               
               
                 Parameters 
                 INHHQ 
                 Biodisponibility 
                 Crossing 
               
               
                   
               
             
          
           
               
                 HBD 
                 2 
                 ≦5 
                 ≦3 
               
               
                 HBA 
                 6 
                 ≦10 
                 ≦7 
               
               
                 MW 
                 292 
                 ≦500 
                 ≦400 
               
               
                 log P 
                 2.34 
                 −1 a 5 
                 −1 a 5 
               
               
                 log D (pH = 7.4) 
                 2.27 
                 −1 a 5 
                 −1 a 5 
               
               
                 log S 
                 −3.36 
                 −4 a 2 
                 −4 a 2 
               
               
                 PSA 
                 66.077 Å 
                 ≦140 Å 
                 ≦90 Å 
               
               
                 Rotatable bonds 
                 4 
                 ≦10 
                 ≦10 
               
               
                   
               
             
          
         
       
     
         [0046]    Furthermore, both the INHHQ molecule itself and its potential metabolites were shown, in theoretical comparison with the toxic fragments of over 3000 comercially available drugs, completely non-toxic. 
         [0047]    In fact, the intraperitoneal injection of up to 300 mg kg −1  of the compound, using 10% DMSO/saline solution as vehicle of injection, in Wistar male rats (acute toxicity test) showed that INHHQ was apparently not toxic to the animals throughout the 72 h of the experiment: no animals died and there were no behavioral changes noted in the injected rats. Also, after the animals&#39; sacrifice, there were no macroscopic abnormalities observed during its organs dissection.