BR102019021448A2 - PHARMACEUTICAL PREPARATION CONTAINING MORINA - Google Patents

Abstract

pharmaceutical preparation containing morine. the present invention patent application deals with an inclusion complex containing the flavonoid morina in hydroxypropyl-ß-cyclodextrin, as well as antihyperalgesic and anti-inflammatory activity of the complexed flavonoid. the invention has potential for application in the clinical treatment of painful and inflammatory conditions.

Description

PHARMACEUTICAL PREPARATION CONTAINING MORINA

(001) The present patent application deals with a pharmaceutical preparation containing morine complexed with hydroxypropyl-β-cyclodextrin, as well as anti-hyperalgesic and anti-inflammatory activity of the complex.

(002) The work was developed seeking to provide technological innovation focused on the new use of the pharmaceutical preparation containing a flavonoid, the morine, from the complexation in hydroxypropyl-β-cyclodextrin, which has physico-chemical properties capable of forming inclusion complexes that will improve the solubility, chemical stability, as well as the anti-hyperalgesic and anti-inflammatory effect of morina.

(003) Inflammatory conditions and pain are an important global health problem, affecting millions of people worldwide. Thus, a significant part of the world population is affected by some type of pain arising from inflammatory reactions, causing a decrease in the quality of life (DE SANTANA et al., 2015; SALA et al., 2018). The most effective clinical resource for the treatment of inflammation and pain of inflammatory origin is pharmacological therapy. Among the drugs used for this purpose, the class of non-steroidal anti-inflammatory drugs (NSAIDs) stands out. However, about 40 to 60% of patients do not respond to this pharmacotherapy, in addition to the fact that prolonged or high doses of these drugs produce undesired effects, such as: gastric lesions, nephrotoxicity, nausea and vomiting (DWIVEDI et al ., 2015; LICATA, 2016). Therefore, it is necessary to search for new anti-inflammatory drugs that have efficacy and less adverse effects for the development of drugs for the treatment of inflammatory diseases.

(004) Flavonoids are a class of secondary metabolites that occur naturally in plants, which are found in various foods and drinks, thus being widely distributed in the human diet. In addition, they are molecules that can be obtained easily in a synthetic way, with low cost, high yield and purity. In the past two decades to the present day, flavonoids have attracted numerous studies by researchers and the pharmaceutical industry, as these compounds have several pharmacological activities, with an emphasis on anti-inflammatory and antioxidant activities. Thus, flavonoids appear as an important pharmacological alternative, as they have the potential for innovation, play an important role in the prevention and treatment of pain / inflammation, in addition to the fact that they are linked to a possible reduction of adverse effects in the short and long term , when compared with clinically accepted drugs (WANG et al., 2013; DUTRA et al., 2016; WANG; LI; BI, 2018).

(005) Morina (2 ', 3,4', 5,7-pentahydroxyflavone) is a flavonoid widely found in plants of the Moraceae and Myrtaceae family (NASO et al., 2013). It has antioxidant (LI et al., 2016), anti-inflammatory (FANG et al., 2003; HEEBA; MAHMOUD, 2014), anti-cancer (NASO et al., 2013; ZHANG et al., 2018), anti-diabetic (VANITHA et al., 2014), antihyperuricemic (ZHANG et al., 2016), among others.

(006) Given these activities, morine can be considered a promising substance for the development of pharmaceutical products. However, like the other flavonoids, morina has low water solubility, limiting its absorption in oral administration and, consequently, its bioavailability (JULLIAN et al., 2008; HEEBA; MAHMOUD, 2014; WANG; LI; BI, 2018). Thus, it becomes necessary to carry out the incorporation of morine in drug delivery systems, such as cyclodextrins (CDs), which are considered a viable alternative to improve the physico-chemical and pharmacological properties of bioactive compounds (PÉREZ-ABRIL et al., 2017).

(007) CDs are oligosaccharides obtained from enzymatic degradation of starch, having a cyclic structure formed by D-glucopyranose units linked through α (1-4) bonds. These molecules have trunk-conical conformation, with a hydrophilic external surface, due to the orientation of the hydroxy groups towards the outside of the cavity, which accredit them as efficient carriers of hydrophobic molecules, since they are capable of being solubilized in an aqueous medium and at the same time. At the same time, incorporate hydrophobic substances into your cavity. DCs are excipients used during the development of pharmaceutical formulations in order to increase the chemical stability and solubility of drugs, resulting in increased bioavailability and pharmacological effect. In addition, it is possible to decrease the dose of drug administered, enabling the reduction of its adverse effects (DEL VALLE, 2004; MURA, 2015; JANSOOK; OGAWA; LOFTSSON, 2018).

(008) Natural CDs are called alpha (α-), beta (β-) and gamma (γ-) CDs and have 6, 7 and 8 units of D-glucopyranose, respectively. The aqueous solubility of natural CDs is low due to the binding strength of the CD molecules in the crystalline state. Β-CD, for example, by forming intermolecular hydrogen bonds, has little interaction with the surrounding water molecules and, therefore, is the natural DC of less aqueous solubility. Despite this, it is the most used by the pharmaceutical industry because it has the lowest cost and high yield. The low water solubility of natural CDs, especially β-CD, increases their tendency to crystallize, limiting their use in parenteral solutions. In addition, β-CD has nephrotoxicity when administered intravenously, as it is not metabolized and accumulates in the form of insoluble crystalline complexes in the kidneys. Thus, it became necessary to make modifications to the natural CDs, with the objective of improving their solubility, reducing toxicity and inserting other functionalities (UEKAMA; HIRAYAMA; IRIE, 1998; JANSOOK; OGAWA; LOFTSSON, 2018).

(009) Among the modified CDs, hydroxypropyl-β-cyclodextrin (HP-β-CD) stands out, obtained from the reaction of β-CD with propylene oxide in alkaline solution. In this reaction, one or more hydroxyl groups on the β-CD bind to the hydroxypropyl radicals, thus reducing its crystallinity, becoming amorphous, resulting in better aqueous solubility and lower toxicological profiles. These characteristics make HP-β-CD one of the most used modified CDs today, especially in the development of formulations for oral or parenteral application, due to its high solubility in water, generating more soluble inclusion complexes, increasing the bioavailability of drugs (LOFTSSON ; DUCHÊNE, 2007; PÉREZ-ABRIL et al., 2017; JANSOOK; OGAWA; LOFTSSON, 2018).

(010) There are numerous patents with complexes for the inclusion of bioactive compounds with CDs. Among them, the patent BR 10 2016 001855 2 A2, which deals with the application of essential oil extracted from the leaves of Citrus sinensis (L.) complexed with β -CD with potential application in the treatment of Alzheimer's disease. The patent BR 10 2015 018668 1 A2 also related to the application of an inclusion complex, which refers to the inclusion of citronellar, a monoterpene, in β-CD and its action in the treatment of pain, that is, the analgesic effect . The patent BR 10 2015 005044 5 A2 that deals with a pharmaceutical preparation containing essential oil of Hyptis pectinata complexed in β-CD with analgesic and anti-inflammatory activity. The patent BR 10 2013 030024 1 A2 which is related to the complexation of carvacrol in β-CD and the antinociceptive activity of this monoterpene against cancer pain. The patent BR 10 2013 024308 6 A2, which refers to the antihypertensive activity of the inclusion complex between the cyclic monoterpene β-Pinene and β-CD. However, in a search carried out in a database of national and international patents, no inventions were found with the purpose of evaluating the anti-hyperalgesic and anti-inflammatory effect of morina complexed in HP-β-CD, as well as, of free morine.

(011) In this context, the objective of the present invention is to obtain the complex of inclusion of morine with HP-β-CD, by the lyophilization method and to demonstrate its anti-hyperalgesic and anti-inflammatory effect. The invention provides a pharmaceutical preparation that can be used in the treatment of painful and inflammatory conditions.

(012) The description presented below serves as a detailed example of the invention, not delimiting the scope of the invention. The inclusion complex was prepared in a 1: 1 molar ratio of morine and HP-β-CD, based on the molecular weights of these substances, by the lyophilization method. For comparison of results, the physical mixture of morina and HP-β-CD was prepared in a 1: 1 molar ratio, by manually stirring both substances with the aid of a mortar and pistil for 10 minutes until a homogeneous mixture was obtained. Once the samples were obtained, the stage of physical-chemical and morphological characterization was initiated through the techniques of differential scanning calorimetry (DSC), X-ray diffraction (XRD), scanning electron microscopy (SEM) and nuclear magnetic resonance (NMR) .

(013) DSC analyzes of morina, HP-β-CD, physical mixture and the inclusion complex morina / HP-β-CD were carried out in a temperature range of 30 -300 ° C using a DSC-60 Shimadzu equipment ®, under dynamic nitrogen atmosphere (100 mL / min) and heating rate of 10 ° C / min, in aluminum capsules containing approximately 2 mg of the sample. The DSC-60 was previously calibrated with standard metallic indium (purity ≥ 99%) before the analyzes, which were carried out in triplicate (n = 3) (LIU et al., 2013a; CARVALHO et al., 2017).

(014) The diffraction patterns of morina, ΗΡ-β-CD, physical mixture and the inclusion complex morina / HP-β-CD were obtained with monochromatic radiation Cu Κα (wavelength = 1.54056 Å), using a Bruker® X-ray diffractometer model D8 Advance Da Vinci. The powder samples were placed in a rectangular aluminum apparatus before exposure to the X-ray beam. The scanning regions of the 2θ diffraction angle were 5 - 50 ° and the radiation was detected with a proportional LYNXEYE detector, with aperture 3.3 ° and 0.2 ° divergence cracks. The measurements were carried out in triplicate (n = 3) with a voltage of 40 kV, an electric current of 40 mA, a step of 0.02 ° with a counting time of 0.5 seconds / step, at room temperature (YAO et al., 2014).

(015) The SEM technique was used to evaluate the morphology of morina, HP-β-CD, physical mixture and the inclusion complex morina / HP-β-CD. The electronic photomicrographs were obtained using a scanning electron microscope JEOL®, model JSM - 6610. To obtain the images, the samples were placed on double-sided tapes, placed on aluminum metallic stubs and coated with metallic gold under vacuum to become conductive. The samples were analyzed with an acceleration potential of 15 kV in 500x increase (LIU et al., 2013).

(016) The 1H NMR spectra were obtained on a Bruker® NMR spectrometer (DRX 500), operating at 400 MHz hydrogen frequency. The HP-β-CD and the morina / HP-β-CD inclusion complex were diluted in deuterated water (D2O), which presents a chemical displacement of 4.8 ppm (QIU et al., 2014).

(017) Complexation efficiency (EC), that is, the amount of morina complexed on HP-β-CD was determined by HPLC using a previously validated analytical method. Initially, 10 mg of the morina / HP-β-CD inclusion complex was dissolved in 10 ml of acetonitrile and the obtained solution was kept under magnetic stirring for 24 hours, at a speed of 400 rpm, at room temperature, to allow the morina complexed is present in solution. Then, the solution was centrifuged (5000 rpm - 30 minutes) to remove the HP-β-CD from the solution, leaving only the active compound (ABARCA et al., 2016; CARVALHO et al., 2017). The supernatant was collected, filtered through membrane filters (PTFE - 0.45 μm), subjected to sonicating ultrasound for 10 minutes and analyzed by HPLC. The samples were prepared and analyzed in triplicate (n = 3). The EC was calculated according to the following equation: (%) = mass of the recovered morina in the complex / mass of the morina added in the complex x 100. (WEI et al., 2017).

(018) The morine DSC curve showed an endothermic peak at 296.54 ° C, corresponding to the melting point of its crystalline form. The DSC curve of the HP-β-CD showed the natural amorphous state of this CD, presenting itself as a flat line with a thermal event around 95 ° C, which is related to the loss of water. Regarding physical mixing, the DSC curve showed the presence of the characteristic event of HP-β-CD reported as water loss and also the crystalline peak of morina, which is indicative of its melting point. Thus, the DSC curve of the physical mixture was basically an overlap between the morina and HP-β-CD curves, which suggests a weak interaction between these molecules and thus, there was no formation of the inclusion complex. However, in the DSC curve of the morina / HP-β-CD complex, the endothermic peak of the morine has completely disappeared, which suggests that the morina was incorporated into the ΗΡ-β-CD cavity, forming the inclusion complex. In addition, there was a reduction in the event related to water loss, which is also indicative of complexation. As the amount of water in the DC cavity decreases, it means that the water molecules are leaving, due to the entry of the drug, giving rise to the inclusion complex.

(019) The diffraction pattern of the morina obtained by the XRD technique, presented several sharp peaks, characteristic of its crystalline natural state, distributed in several diffraction angles: 8.4 °; 10.2 °; 12.8 °; 14.3 °; 16.8 °; 18.6 ° and small peaks in the 24 - 31 ° range. On the other hand, ΗΡ-β-CD showed an amorphous diffraction pattern, that is, without the presence of crystalline peaks, only with two wide diffraction halos, characteristic of this molecule around 10 and 20 °. The diffraction pattern of the physical mixture exhibited an amorphous characteristic of the ΗΡ-β-CD with some crystalline peaks characteristic of the morina clearly distinguished, that is, the XRD pattern of the physical mixture was an overlap of the morina and ΗΡ-β-CD patterns , demonstrating that no interaction was formed between these molecules during manual agitation and both remained with their original physical characteristics. However, the morina / HP-β-CD inclusion complex showed a diffraction pattern with broad amorphous halos, similar to ΗΡ-β-CD and the crystalline peaks of the morina disappeared completely. This result indicates that the morine was encapsulated in the ΗΡ-β-CD cavity with consequent loss of its crystallinity, suggesting the formation of a new compound in the form of an inclusion complex.

(020) The SEM photomicrographs showed that the morina has an acicular shape, that is, a shape similar to small crystalline needles with different sizes, while the ΗΡ-β-CD presented its typical structure, that is, spherical, amorphous particles with cavities. In the physical mixture, the small needle-shaped crystals characteristic of morina and the spherical particles (or fragments of these particles) of the ΗΡ-β-CD are present concurrently, demonstrating that there were no changes in the shape of these substances, due to the absence or weak interaction between compounds during manual stirring. On the other hand, the morina / HP-β-CD inclusion complex appeared as irregular particles of varying sizes, with uniform internal appearance in the form of small blocks. In the complex, the original morphology of both components has completely disappeared, making it impossible to differentiate between morine and HP-β-CD. This drastic change in particle morphology demonstrates a strong interaction between the morine and the HP-β-CD suggesting the formation of the inclusion complex.

(021) To obtain the 1H NMR spectra of HP-β-CD and the morina / HP-β-CD inclusion complex, the samples were diluted in deuterated water (D2O). In the HP-β-CD spectrum, the displacement of H-1, H-2, H-3, H-4 and H-5 was 4.994; 3,535; 3,935; 3,473 and 3,774 ppm, respectively, which are values equivalent to those found in the literature. In the spectrum of the inclusion complex, there were some changes in the chemical shifts of HP-β-CD in relation to its free form. The values of the difference in chemical displacements between the protons of HP-β-CD in the presence and absence of morine were determined by the equation: Δδ = δ morina / HP-β-CD - δ ΗΡ-β-CD. Protons H-1, H-2 and H-4 showed a small displacement difference with values of -0.009; -0.012 and -0.015 ppm, respectively. On the other hand, H-3 and H-5 showed a relatively high displacement, with values of -0.033 and -0.052 ppm, respectively. It is known that H-2 and H-4 are located on the external surface of HP-β-CD, while H-3 and H-5 are located in the internal cavity. Thus, the major change in the chemical displacements of H-3 and H-5 was probably due to the presence of the guest molecule (morine) in the HP-β-CD cavity, evidencing the formation of the inclusion complex. In addition, as H-3 is close to the wide side of the cavity and H-5 is close to the narrow side, it is suggested that the morine be incorporated into the HP-β-CD through the narrow side of the cavity.

(022) The EC was 98.3 ± 0.36% and this value close to 100%, demonstrates that practically every morina was incorporated into the HP-β-CD cavity, thus, the method of obtaining the inclusion complex morina / HP-β-CD by lyophilization is adequate and efficient, obtaining a high yield and great interaction between the two molecules. In addition, the high EC value can be attributed to the chemical structure and physicochemical properties of HP-β-CD, which has high solubility and an internal and external cavity diameter suitable for the incorporation of morine.

(023) After the physical-chemical, morphological and CE characterization showed the formation of the morina inclusion complex with the HP-β-CD, this pharmaceutical preparation was submitted to a dissolution study in comparison with the free morine to prove the increased solubility. As a dissolution medium, an aqueous solution of Tween 80 (0.1%) was used and the test was carried out in a time interval of 0 - 240 minutes. Free morine demonstrated a limited dissolution rate, since only 28.01 ± 0.5% was dissolved in 240 minutes. The low percentage of dissolution of free morine can be associated with the low solubility of this molecule, which was not encapsulated in any release system capable of increasing its solubility. Thus, its incorporation in release systems such as CDs is a viable alternative to improve its physical and chemical properties. On the other hand, the morina / HP-β-CD inclusion complex exhibited a faster dissolution than the free morina in the initial phase, releasing 70.12 ± 0.9% morina in 15 minutes, reaching a maximum dissolution of 97 , 84 ± 0.3% in 240 minutes. The high value of the dissolved morine quantity is related to the increase in solubility and reduction of the crystallinity of the morine due to the complexation with HP-β-CD. Therefore, it is expected that the increase in the solubility and dissolution of morina will occur in in vivo models, improving the bioavailability and, consequently, the pharmacological effects of this substance.

(024) After in vitro characterization and dissolution studies, the anti-hyperalgesic and anti-inflammatory activities of the morina / HP-β-CD inclusion complex were evaluated in vivo. For that, the tests of carrageenan-induced hyperalgesia and carrageenan-induced pleurisy were performed, with subsequent quantification of leukocytes and tumor necrosis factor (TNF-α).

(025) The studies were carried out on male Swiss mice, weighing between 25 - 30 grams and aged 2 - 3 months. The animals were obtained from the sectoral vivarium of the physiology department of the Federal University of Sergipe (UFS), kept in a dark light cycle from 12: 00h / 12: 00h (light: 6 to 18 hours; dark: 18 to 6 hours) , at a temperature of 25 ± 0.5 ° C and exhaust air, with free access to water and feed up to 60 minutes before the experiments. In the present study, the ethical principles established by Law No. 11,794 / 2008, which establishes the experimental procedures for the scientific use of animals, and by Law No. 8,366 / 2017, which establishes rules for the protection, defense and preservation of animals in the state of São Paulo, were respected. Sergipe. The experimental protocols for this research were approved by the UFS Animal Research Ethics Committee under approval number 18/2018 (CEPA / UFS # 18/2018).

(026) Mechanical hyperalgesia was evaluated on a digital analgesometer according to the methodology described by Cunha et al., 2004. 30 minutes before the test, the animals were placed in acrylic boxes (12 x 10 x 17 cm) with floors grid. The researcher was trained to apply the tip perpendicularly to the central area of the hind paw with a gradual increase in pressure. The “end point” was characterized by the removal of the paw followed by clear “flinching” movements. After this response, the pressure intensity was automatically registered. The intensity of the stimulus was obtained by the average of three measurements performed with minimum intervals of 3 minutes. The animals were evaluated before and after treatment.The animals were divided into four groups (n = 6) and treated with vehicle (0.9% saline, oral), morina inclusion complex / HP-β-CD (100 mg / kg, orally), free morine (100 mg / kg, orally) or indomethacin (10 mg / kg, orally) .1 hour after treatment, 20 μL of carrageenan (300 μg / paw) was injected subcutaneously in the subplantar region of the hind leg. Mechanical hyperalgesia was evaluated at 1, 2, 3 and 4 hours after the injection of the hyperalgesic agent.

(027) In the carrageenan-induced pleurisy assay, the animals were divided into three groups (n = 6) and treated with vehicle (0.9% saline, oral), morina inclusion complex / HP-β-CD (100 mg / kg, orally) or free morine (100 mg / kg, orally), 1 hour before the pleural injection of carrageenan. Pleurisy was induced in mice through the intrapleural administration of 100 μL of 1% (w / v) carrageenan suspension in saline (OLIVEIRA, A. M. et al., 2012). 4 hours after pleurisy induction, the animals were euthanized and the pleural inflammatory exudate was collected by pleural lavage with 1 mL of phosphate buffer (PBS) containing ethylene diaminetetraacetic acid (EDTA; 10 mM). The exudate volume was centrifuged (1500 rpm - 10 min) and the supernatant was collected to determine the levels of cytokines in the pleural fluid. The cells were resuspended in 1000 μL of PBS and an aliquot of 10 μL was diluted with Turk's solution (1:20). Total leukocytes were counted in a Neubauer chamber, examining four external quadrants, using a light microscope (VINEGAR; TRUAX; SELPH, 1973).

(028) TNF-α levels in the pleural cavity were assessed 4 hours after the injection of carrageenan. TNF-α was quantified in the cell-free supernatant by the ELISA technique, according to the protocols established by the manufacturer (BD-Bioscience Pharmingen, San Diego, CA).

(029) The results were analyzed statistically according to the mean ± standard deviation of the analyzes. The data obtained were evaluated by one-way analysis of variance (ANOVA) followed by the Tukey test. In all cases, the differences were considered significant if p <0.05.

(030) In carrageenan-induced hyperalgesia, the morina / HP-β-CD inclusion complex and free morine demonstrated anti-hyperalgesic effect in 1 (p <0.001) 2 (p <0.001) 3 (p <0.001) and 4 (p <0.001) hours after induction, when compared to the control (vehicle). The present research revealed that the morina / HP-β-CD complex was able to inhibit the mechanical hyperalgesia induced by carrageenan demonstrating anti-hyperalgesic effect in the model of inflammatory pain induced by carrageenan.

(031) Regarding carrageenan-induced pleurisy, the results showed that the administration of carrageenan in the pleural space of mice induces an inflammatory process with an increase in the total leukocyte count and dysregulation in the release of TNF-α in the pleural fluid. The treatment with the inclusion complex morine / HP-β-CD and with free morine caused a significant decrease in the total leukocyte count and a significant reduction in the levels of TNF-α in the pleural fluid when compared to the control group (vehicle), demonstrating the anti-inflammatory potential of the morina / HP-β-CD inclusion complex and free morina.

(032) In the in vivo tests performed, the doses of free morine and the complex were nominal doses, that is, although both doses were 100 mg / kg, they were not equivalent doses. Therefore, the amount of morine present in the complex is less than the amount of free morine. To prepare the free morine solution, 18 mg (n = 6) of morine were weighed and diluted in the vehicle before administration to mice. For the complex group it also weighed 18 mg (n = 6), however, 18 mg of the complex is equivalent to 3.17 mg of morine. Therefore, the dose of the free morine group was approximately 6 times greater than the dose of morine present in the complex group.

(033) This characteristic in the experiment is remarkable, because DCs have the ability to increase or maintain the pharmacological effect of drugs, even in a lower dose. In addition, lower doses decrease the adverse effects of drugs, which is one of the advantages of complexing molecules with CDs (JANSOOK; OGAWA; LOFTSSON, 2018). Thus, the results obtained in our study demonstrate that the free morine and the Μο / ΗΡ-β-CD complex have an anti-inflammatory effect, however, the complexed morina on the HP-β-CD has greater therapeutic potential, since the complex it allows a decrease in the dose of the drug and still, allowing a more significant anti-inflammatory effect.

(034) The present research presents an innovation with morina, since the complexation of this flavonoid in HP-β-CD, obtaining the morina / HP-β-CD inclusion complex, increases the pharmacological potential. The improvement in the pharmacological effect is due to improvements in the properties of morina after complexing with HP-β-CD, such as increased solubility, dissolution and oral bioavailability, in addition to the fact that it is possible to use smaller doses, decreasing the possibility of adverse effects. Therefore, the morina / HP-β-CD complex has great potential to act as an anti-hyperalgesic and anti-inflammatory agent.

(035) The results of this study clearly demonstrate the formation of the morina inclusion complex with HP-β-CD, which was evidenced by the techniques of DSC, XRD, SEM and NMR. The EC demonstrated that practically every morina is incorporated in the HP-β-CD by the lyophilization method. The in vitro dissolution study proved the increase in the solubility of morina after complexation in HP-β-CD. In addition, the morina / HP-β-CD inclusion complex has anti-hyperalgesic and anti-inflammatory activity, in lower doses than free morine.

• a) Formas farmacêuticas sólidas para uso oral: comprimidos (revestidos ou não revestidos), comprimidos mastigáveis, comprimidos solúveis (não efervescentes), comprimidos efervescentes, drágeas, cápsulas moles ou duras;
• b) Formas farmacêuticas líquidas para uso oral: xaropes, emulsões ou suspensões;
• c) Formas farmacêuticas para uso tópico: cremes, pomadas, gel e emulsões;
• d) Forma farmacêuticas para aplicação parenteral.

(036) Thus, the morina inclusion complex with HP-β-CD can be used in different pharmaceutical forms for human use, such as:

• a) Solid dosage forms for oral use: tablets (coated or uncoated), chewable tablets, soluble tablets (non-effervescent), effervescent tablets, pills, soft or hard capsules;
• b) Liquid pharmaceutical forms for oral use: syrups, emulsions or suspensions;
• c) Pharmaceutical forms for topical use: creams, ointments, gels and emulsions;
• d) Pharmaceutical forms for parenteral application.

Claims (8)

PHARMACEUTICAL PREPARATION, characterized by containing flavonoids, the flavonoid being morine. PHARMACEUTICAL PREPARATION according to claim 1, characterized in that the flavonoid is complexed in cyclodextrin, preferably hydroxypropyl-β-cyclodextrin. INCLUSION COMPLEX CONTAINING FLAVONOID, characterized by greater solubility than free flavonoid. INCLUSION COMPLEX CONTAINING FLAVONOID, according to claim 3, characterized by having a better anti-hyperalgesic effect when compared to free flavonoid. INCLUSION COMPLEX CONTAINING FLAVONOID, according to claim 3, characterized by having a better anti-inflammatory effect when compared to free flavonoid. INCLUSION COMPLEX CONTAINING FLAVONOID according to claim 3, characterized by having greater bioavailability than pure flavonoid. USE OF FLAVONOID MORINA, characterized by being in pharmaceutical preparations, preferably for oral use. USE OF THE FLAVONOID MORINA, characterized for being to treat pain and inflammation.