WO2016037250A1

WO2016037250A1 - Jambu extract purification method, thus obtained purified extract, anaesthetic composition and bioadhesive containing purified jambu extract; and uses thereof

Info

Publication number: WO2016037250A1
Application number: PCT/BR2015/000094
Authority: WO
Inventors: Rodney Alexandre Ferreira RODRIGUES; Verônica Santana DE FREITAS; João Ernesto DE CARVALHO; Francisco Carlos Groppo
Original assignee: Universidade Estadual De Campinas - Unicamp
Priority date: 2014-09-11
Filing date: 2015-06-25
Publication date: 2016-03-17
Also published as: BR102014022486A2; BR102014022486B1

Abstract

The present invention relates to a method for purifying jambu (Spilanthes acmella (L.)L ) extract, the thus obtained purified jambu extract, an anaesthetic composition and a bioadhesive containing purified jambu extract. Another object of the invention is the use of the composition and of the bioadhesive as a topical anaesthetic agent against pain and/or inflammation. Also described is the use of the purified jambu extract for producing anaesthetic compositions.

Description

JAMBU EXTRACT PURIFICATION PROCESS, PURIFIED EXTRACT AS OBTAINED, ANESTHETIC COMPOSITION AND BIOADESIVE CONTAINING PURIFIED JAMBU EXTRACT; AND USES

FIELD OF INVENTION

[1] The present invention describes a purification process of Jambu extract (Spilant es acmelia (L.) L), thus obtained purified Jambu extract, anesthetic and bioadhesive composition containing purified Jambu extract. It is a further object the - use of the composition and the bioadhesive as topical anesthetic -associada the ^'pain- .and / cm inflammation. Also, the use of purified Jambu extract to produce anesthetic compositions.

[2] The present invention is. applied in the field of dentistry.

BACKGROUND OF THE INVENTION

[3] In dental treatment, one of the main triggers of fear is discomfort caused by anesthetic injection, as this procedure is most often associated with pain. This is a relatively common problem that reaches up to 30%. of the world population (Gordon et 1., 2013).

[4] The search for and use of methods that cause less patient discomfort are extremely valid given that there is, to date, no topical drug formulation capable of completely eliminating the pain caused by needle puncture during anesthetic injection, especially in the palatal region (Hmud &^' Walsh, 2009; Ogle & Mahjoubi, 2012).

[5] In this context, there is one. enormous therapeutic potential in the ^plants' medicinal and its metabóiitos only 5% of this universe underwent a phytochemical and biological study.

[6] Spilanthes acmella (L.) L. is a plant originating in South America and very common throughout Southeast Asia. In Brazil it is popularly known as watercress, watercress (SP), water pepper (PE), little pepper, pepper, pará (RJ), jambu (RJ), watercress do- Brazil (BA), watercress, jambu-rana, malacca herb, jambu-assú, mast or abecedaria. Spilanthes acmella is popularly used to treat toothache and other ailments that affect the gums and throat and is also used to combat tuberculosis, anemia and as an appetite stimulant (Di-Stasi et al. 2002; Lorenzi & Matos, 2008; Dubey et al., 2013). The anesthetic activity of jambu was promising in studies published by Jambu. some authors (Chakraborty et al., 2010; Fosquiera et al., 2012). These results coupled with its low toxicity (Chackraborty et al., 2004; Cilia-Lopez et al., 2010; Sharma et al., 2011 and Nomura et al., 2013) and widespread popular use, make jambu a good candidate in the world. development of a topical anesthetic for oral use.

[7] BRPI0500886-7A describes a preparation process that results in a chlorophyll free jambu extract. The process comprises the discoloration of a hot alcoholic extract using a strong base, an electrolyte and a liquid-liquid partition step. The aforementioned invention takes several steps and consumes many hours beyond the process resulting in the generation of waste and handling requires greater care for user safety. In the present invention, the renewal of the Extractor solvent which was reflected was a higher efficiency due to the better distribution coefficient of the spilantol compound when compared to a single extraction.

[8] WO2010010394A2 relates to an extractive process using the fresh plant, one of which takes 48 hours. That is, it is a long process that requires several steps to obtain the final extract. Moreover, the use of larger proportions of water results in an extract with different phytochemical profile, since spilantol is very little water soluble. The proposed invention has the advantage that extraction occurs with the aid of mechanical agitation, significantly reducing the time required to obtain the extract. In addition, there is no need for a sterile environment for extraction, and the final product has an appropriate appearance and color for consumption after pretreatment with charcoal.

[9] US 3720762 describes a process for obtaining the extract by distillation as the obtained extract is used as a flavoring agent. The proposed invention has the advantage of a more specific extraction process for the anesthetic compound, namely dynamic maceration with the use of ether as a solvent.

[10] JPH0790294 deals with the purification of spilanthol present in a ready-made formulation and specific dentifrice compositions by applying essential oil and not ethanolic extract. A method for increasing the spilanthol concentration of a finished product by liquid / liquid extraction is disclosed in the document. The proposed invention has the advantage of employing a solvent of The inventors of JPH0790294 technology employ hexane, acetonitrile and pentane in the liquid / liquid extraction step.

[11] WO 2013110883A2 deals with a method of concentrating spilanthol present in extracts by the use of solvents such as ethers and their derivatives and also by the use of strong acids and bases. It is about. a long and costly process with a high possibility of toxic waste generation. The proposed invention presents a simpler and faster process besides employing lower toxicity reagents.

[12] GB1438205A deals with dentifrice-specific compositions using jambu essential oil rather than ethanolic extract. Advantageously, the invention comprises fewer steps and less toxic solvent. GB1438205A cites the use of organic solvent, ethyl ether, which is quite hazardous as it is highly flammable and toxic compared to ethanol. In addition, additional steps are required which makes the process more laborious.

[13] Accordingly, none of the patent documents dealing with the purification of jambu extract to date use activated charcoal for this purpose, so the object of protection of this application is the purification process of jambu extract, jambu extract. chlorophyll-free activated charcoal, the anesthetic composition and the bioadhesive containing the purified extract, and the use of the extract to produce anesthetic compositions.

BRIEF DESCRIPTION OF THE INVENTION [14] The present invention describes a purification process of Jambu extract (Spilanthes acmella (L.) L), purified Jambu extract thus obtained, anesthetic and bioadhesive composition containing purified Jambu extract. It is an additional object to use the composition and bioadhesive as a topical anesthetic associated with pain and / or inflammation. Also, the use of purified Jambu extract to produce anesthetic compositions.

[15] The Jambu extract purification process comprises the steps of placing the Jambu extract in contact with activated carbon under heating and stirring; filtration; and extract concentration under vacuum and lyophilization.

[16] The purified extract obtained by this process comprises at least 40% spilanthol, is chlorophyll free and lyophilized.

[17] The anesthetic composition comprising standardized purified Jambu extract contains at least 40% spilanthol and a pharmaceutically acceptable carrier.

[18] The bioadhesive comprises a polymer, a solvent, and purified jambu extract standardized with at least 40% spilanthol.

[19] Finally, the present invention further describes the use of purified extract to manufacture anesthetic compositions in addition to the use of the composition and bioadhesive used as anesthetic.

BRIEF DESCRIPTION OF DRAWINGS

[20] In order to obtain a full view of the objects of the present invention, it is necessary to read this document and analyze the accompanying drawings and the which references are made as follows.

[21] FIGURE 1 - Figure 1 is a graph depicting the permeation profile of spilanthol across the pig esophageal mucosa obtained from the Franz cell permeation experiment.

[22] FIGURE 2 - Figure 2 is a graph illustrating the percentage of anesthetized animals as a function of time (min) in the Tail Flick test.

DETAILED DESCRIPTION OF THE INVENTION

[23] The present invention describes a purification process of Jambu extract (Spilanthes acmella (L.) L), purified Jambu extract thus obtained, anesthetic and bioadhesive composition containing purified Jambu extract. It is an additional object to use the composition and bioadhesive as a topical anesthetic associated with pain and / or inflammation. Also, the use of purified Jambu extract to produce anesthetic compositions.

[24] The purification process of Jambu extract (Spilanthes acmella (L.) L) is initiated when the ^' Jambu ethanolic extract is brought into contact with activated charcoal with preferred 800-900 mg iodine number 12 / g, molasses relative efficiency 100-130%, number of methylene blue (g / 100g) ≥ 14, humidity ≤ 10%, ashes ≤ 10%, mesh size 325 (50-80), 400 (> 90)% , from 2 to 6%, preferably in the proportion of 4% (w / w), under moderate heating from 35 to 45 ° C, preferably 40 ° C and magnetic or mechanical stirring, preferably at 100-300 rpm, preferably for one hour thereafter. filtration is performed under vacuum in a porous plate funnel with diatomaceous earth precoat in the proportion of 8 to 12%, preferably 10%, the extract is concentrated under vacuum, preferably in a rotary evaporation system at a temperature of 38 to 42 ° C, preferably 40 ° C, and lyophilized.

[25] In the present patent application, the purified Jambu extract is the purified Jambu extract obtained according to the purification process described above.

[26] The anesthetic composition comprises purified spambantol standardized jambu extract and contain at least 40% spilanthol from the jambu aerial parts and a pharmaceutically acceptable carrier.

[27] The anesthetic composition may contain optional ingredients to provide some desirable feature not achieved with the aforementioned components such as emollient, humectant, chelating agent, thickening system, antioxidant, preservative, coloring and flavoring.

[28] The dental bioadhesive comprises a polymer, solvent and purified Jambu extract standardized with at least 40% spilanthol, in a ratio ranging from 10 to 20% purified extract, preferably 20%.

[29] The bioadhesive may further comprise optional components such as chelating agent such as ethylenediamine tetraacetic acid (EDTA) and its salts; pH adjusting agent such as triethanolamine or inorganic hydroxides; preservative such as parabens, organic acids, imidazolidinines, diazolidines, phenolic alcohols, methyl hydroxybenzoate, propyl hydroxybenzoate, chlorocresol, benzalkonium chloride and glycol ethers; emollients such as alcohols and fatty acids, esters, ethers, mono-, di- or triglycerides, natural or synthetic hydrocarbons or organic carbonates and combinations thereof; dyes; flavoring agents; flavoring agents; humectants and permeation enhancers such as glycols, preferably glycerine, propylene glycol, butylene glycol and combinations thereof, ethoxydiglycol, more preferably ethoxydiglycol, in a ratio ranging from 5 to 10%, preferably 5%, based on the total bioadhesive mass.

[30] The polymer may be one of the following polymers: chitosan, polyvinyl alcohol (PVA), cellulose and its derivatives, alginate, gum arabic, cashew gum, other gums, agarose, in the proportion of 1 to 30%, preferably chitosan, in the proportion of 1.0 to 1.5%.

[31] The chitosan dissolving solvent should have a pH of less than 6.0 and can be any aqueous solution of weak or strong organic or inorganic acids, preferably acetic acid solution in a concentration ranging from 0.5 to 2.0%. (v / v), preferably 1.0%.

[32] The ethanolic extraction process of Jambu aerial parts to obtain extract is known from the state of the art.

[33] After purification with active charcoal there is a change in the color of Jambu ethanolic extract, justified by the removal of chlorophyll.

Preferred mode of bioadhesive

[34] The bioadhesive is prepared by slowly dissolving chitosan in 1.0% (v / v) acetic acid solution with the aid of homogenizer and reserved. In another container add 10 g of charcoal-treated jambu extract activated, 5 g ethoxydiglycol, 100 mg methylparaben and complete with freshly prepared chitosan gel to 100 g. 20 g of this mixture is poured into 55.4 cm ² polyacetal plates. Drying should be done in an oven without forced air circulation at 40 ° C.

[35] The homogenizer and ultrasound are important for complete incorporation of the extract and formation of homogeneous solution. This solution is then poured onto polystyrene, polytetrafluoroethylene, nylon or polyacetal plates, preferably polytetrafluoroethylene or polyacetal, except glass plates. Drying should be done in a greenhouse without forced air circulation at 40 ° C. The ideal thickness of the bioadhesives should respect the ratio of 2 to 3g / cm2.

[36] After complete drying, the bioadhesives should be cut into 2.68 cm2 (15 mm diameter) circles using a hole punch or changed. Thickness measurements should be made with a digital caliper at four different points of the bioadhesives (n = 7) to determine the mean thickness and its standard deviation. Measurements of bioadhesive mass (n = 7) should also be made on analytical balance to obtain the mean and standard deviation.

[37] Local absorption and efficacy of this invention may be improved by using different adjuvants in varying amounts or other chemical compounds known to facilitate absorption of the active compounds of the present invention.

Example 1: In vitro permeation assay of spilanthol contained in bioadhesives through pig esophageal mucosal epithelium in Franz type vertical diffusion cell

[38] Pig esophagus should be purchased from a slaughterhouse duly certified by the São Paulo State Department of Agriculture and Supply shortly after the sacrifice of the animals and transported in phosphate buffer (pH 7.4). For the separation of the 'esophagus' epithelium, the methodology described by Diaz Del Consuelo was used. et al., (2005). The mucosa was carefully separated from the adjacent tissue with the aid of a scalpel. The epithelium was separated from the connective tissue (lamina propria) with a spatula after immersion of the mucosa in distilled water in a heated bath. at 60 ° C for two minutes and used immediately.

[39] The permeation experiment through the pig esophageal mucosal epithelium was performed in the Franz-type vertical diffusion cell with a permeation area of 0.6 cm ² . Formulations containing 10 and 20% crude extract were used as well as the formulation with 10% active charcoal treated raw extract cut into 15 mm diameter circles.

[40] The epithelium was positioned ■ with the connective side over the regenerated cellulose filter (pores of 0.45 μπι diameter) and with the epithelium facing the donor compartment. The function of the cellulose filter is to support this fabric due to its great brittleness. The epithelium, the filter and the bioadhesive were placed together on the Franz cell, between donor and receiver compartments and the receptor compartment .It was filled with about 4 ml of 30% solution of physiologically ^"of .metanol in order to Ensure sink conditions, ie, the permeate amount of the asset has not reached 10% of its solubility.

[41] Franz's cell was then placed in a bath heated to 37 ° C under constant magnetic stirring. In At pre-established time intervals (30, 60, 90, 120, 150, 180, 210, 240, 270 and 300 min) aliquots were taken from the receptor solution (300 μΐι) and analyzed by GC-MS. After the removal of each aliquot, a volume corresponding to the one withdrawn was replaced.

[42] Following GC-MS quantification, the spilanthol permeation parameters were calculated such as: time required for initial permeation or time lag; flow and permeability coefficient. The experiments were performed in sextuplicates.

[43] For each cell, a graph was constructed from the amount of spilanthol accumulated in the receptor compartment as a function of time (time intervals of each collection). The inclination of the linear portion of the graphs represents the penetration flow of spilanthol through the mucosa and its intersection with the abscissa axis allowed to determine the time lag value. Thus, data obtained from permeation experiments were expressed as cumulative amounts of permeated spilanthol over time over a 5 h interval and analyzed according to equation 1:

Equation 1: J = P X Cd

[44] Where: J is the flow of spilanthol through the mucosa, P is the permeability coefficient and Cd is the concentration of spilantol present in the bioadhesive used in the donor compartment.

[45] Data were expressed as mean ± standard deviation (n = 6).

[46] Figure 1 shows the permeation profile of spilanthol from the different bioadhesives evaluated. THE statistical analysis (ANOVA and Tukey-Kramer) of the areas under the individual curves showed that the formulation of 10% active charcoal-treated crude extract (4%) showed larger area than the others in a statistically significant way (p <0.05), but there were no statistically significant differences between ^'the other formulations.

[47] The permeation parameters (flow, permeability coefficient and time lag) are presented in Table 1. Table 1 - Permeability parameters of spilanthol applied in finite dose condition through pig esophageal mucosal epithelium.

Formulation Flow Time lag Coefficient of

(h) permeability

([spilanthol (pg.cm- in mg]) UO ^ cm.h- ¹ )

1.97

treated with active charcoal).

[48] Statistical analysis (ANOVA and Tukey-Kramer) of the results revealed that there were no statistically significant differences between the EB10% and EB20% formulations, but both showed lower values than the EBTC10% for both flow and time lag. The permeability coefficient was higher for the EBTC10% than for the other groups. EB20% presented lower coefficient than EB10%.

Example 2: In vivo evaluation of jambu bioadhesive antinociceptive activity

[49] Male Swiss mice (25-40g) maintained at 25 ± 2 ° C in 12 h light-dark cycles (light phase starting at 7 h) and kept in a vivarium with water and ad libitum feed were used for at least 7 days before the experiments. The experiment was conducted after approval by the UNICAMP Animal Ethics Committee under No. 2851-1 and in accordance with good animal experimentation practices advocated by the Guide for Veterinary Care of Laboratory Animals by Voipio et al. (2008). The animals were divided into groups of 5-6 animals and each animal was used only once in the experiment.

[50] Evaluation of antinociceptive activity was performed by the tail flick model as described by de Araújo et al. , (2010). Initially, the mice were kept in crystal acrylic containers, keeping the distal portion of the tail free (10 cm) and the time required for tail removal (latency) was considered as aversive response to heat generated by incandescent light bulb. The basal value (baseline) of each animal was recorded before the beginning of the experiment and only those with a baseline value of up to 4 seconds were considered fit for the experiment. To avoid injury due to thermal injury, a maximum time of 10 s for cut-off contact was established. After determining the aversive response, the animals' tail was again exposed to heat generated by a light bulb. (55 ° C) and the response time determined. This study consisted of five groups: group 1: bioadhesive with 10% crude jambu extract (10% EB); group 2: bioadhesive with 20% crude jambu extract (20% EB); group 3: bioadhesive with 10% raw jambu extract treated with 4% active charcoal (10% EBTC); group 4: positive control 150 mg EMLA® / animal; group 5: negative control, only bioadhesive.

[51] The bioadhesive was applied 2 cm from the base of the animal's tail with the aid of tape for two min and after its removal the nociceptive stimulus was applied in the same region. The first measurement was performed shortly after the bioadhesive removal and the following measurements were taken every 15 min until the animal returned to its basal nociceptive stimulus response value. After handling the animals in the tail flick test, they were euthanized by cervical dislocation and placed in a freezer in plastic bags for subsequent incineration.

[52] Figure 2 shows the percentage of anesthetized animals (maximum time 10 s) over time for the formulations studied.

[53] Statistical analysis of the data (Gehan-Breslow-Wilcoxon test) showed that there were statistically significant differences (p <0.0245) between curves. Comparison between curves using the same test revealed that the 10% EBTC formulation was superior to all others (p <0.05), and there were no statistically significant differences (p> 0.05) between them. The average (± standard deviation) of anesthesia time (only animals that responded at the maximum time) was 3.7 (± 4.97) min for EB10%, 12 (± 14.42) min for EB20% and 49.2 (± 27.64) min for EBTC10%. As expected, there was no effect ^{'antinociception} in animals treated with chitosan bioadhesive without adding jambu extract as the chitosan does not have this pharmacological properties. (negative control - data not displayed). ^■

Example 3: Monitoring of spil'antol by GC / MS

[54] Analytical monitoring of spilanthol was performed by gas chromatography using a mass detector gas chromatograph (CG-MS, Hewlett Packard 5890, Series II, Hewlett Packard 5970 ΕΊ 7Q eV mass-selective mass detector). of WCOT fused silica, HP5-MS, Agilent brand of dimensions 30m x 0.25mm ID, 0.25 ym stationary phase thickness. The conditions of analysis were: injection temperature: 220 ° C, detector temperature: 250 ° C, temperature program: 60-240 ° C, (3 ° C / min). with . sample divider in ratio 1:40, carrier gas He 0,7 bar, ImL / rnin. The extracts, 15 mg, were dissolved in 1 mL. methanol and kept in a freezer at -20 ° C until the time of analysis. espilantol identification was done with the aid of the bank data -de ■ ^■ National Institute of Standards Technology (iNIST®) present in analysis software MS ChemStation D Agilent® at least 90% agreement.

[55] In the obtained chromatograms it was possible to verify that the spilanthol peak (retention time: 42 min) was the major one in all the performed tests.

^' [56] The normalized percentage area of spilanthol obtained for the crude extract. was 55 ± 1% and for the extract after Treatment with 4% active charcoal was 64.6 ± 1.15%. Thus, the extract treated with 4% active charcoal was chosen among the different treatments, as there was a relative increase in spilanthol content and pigment loss, despite the decrease in mass when compared to the original crude extract.

Example 4: Accelerated stability study of jambu extract-containing bioadhesive

[57] In accordance with ANVISA RE No. 1 of 2005 (Brazil, 2005) which deals with the regulations for this type of study, the bioadhesives were packaged in impermeable aluminum-lined plastic containers with airtight zip closure and submitted to the greenhouse. dries at 40 ° C ± 1 ° C for 120 days. The samples were analyzed at times 0, 90 and 120 days for appearance and pH. The spilanthol content was measured at times 0 and 120 days. The tests were performed in triplicate.

[58] After cutting with the hole punch (15 mm diameter), the bioadhesives looked as shown in Annex 1 and were then used for thickness measurement.

[59] Measurements of thickness (mm) and mass (g) are expressed as mean and standard deviation and presented in Table 2.

[60] Table 2 - Physical parameters of the developed bioadhesives.

Percentage and type of extract Thickness Mass

(mm) (g)

(n-7) M d a ± esv o r

[61] Bioadhesives maintained their physical integrity throughout the study period; There was, however, darkening of the activated carbon-treated bioadhesive, as can be seen in Annex 2. The photographic representation is individual, but the tests were performed in triplicate, with agreement between them.

Example 5: Evaluation of pH and spilanthol content of bioadhesives during stability study:

[62] The bioadhesives were weighed on an analytical balance and deionized water was added at a rate of 1% m / m. The mixture was brought to ul.tr.assom for ^'two minutes. As the pH of the resulting solution was held at pHmet.ro previously calibrated at pH 4 and · ^'7 at room temperature (Brazilian Pharmacopoeia; 2010). For the determination of the spilanthol content, the bioadhesives were weighed in analytical balance, methanol was added in the proportion of 1% w / w and the mixture was ultrasonized for ten. minutes At the end, 1.5 mL aliquots were taken and the spilanthol content was determined by GC-MS under the same assay conditions as described above.

[63] ^' 0 ^' pH remained stable without significant changes. After 120 days of storage in a greenhouse at 40 ° C, there was no significant degradation of spilanthol in formulations tested as shown in Table 3.

[64] Table 3 - Evaluation of pH ^* and spilanthol content in jambu bioadhesives.

PH percentage Spilanthol and type of

Extract ^{m {(3} ^/ ^{^)}

____ Time in days Time in days ^'

0 90 0 120

120 10%, EB 5.29 ± 5.04 ± 5.4, 18.58 ± 17.13 ± 0.03. _,., 0> 24 1,92_. 0.87 ^' j

20%, EB 5.09 ± 5.05 ± 5.14 ± 36.76 ± 35.87 ±

0, 02 0, 04 0, 05 4, 92 4, 02

[65] (n = 3) Mean ± standard deviation. EB: crude extract; EBTC: Active extract treated with active charcoal (4%).

[66] Results were subjected to analysis of variance considering a significance level of 5%. The statistical tests used were ANOVA, Tukey Kramer and Gehan-Breslow-Wilcoxon. All analyzes were performed using the GraphPad Prism, Instat statistical package (GraphPad Software, Inc.).

Claims

CLAIMS:

1. Purification process of jambu extract characterized by the following steps:

- the ethanolic extract of Jambu is placed in contact with activated charcoal in the proportion of 2 to 6%, preferably 4% (w / w), under heating and stirring preferably for one hour;

- filtration;

- The extract is concentrated under vacuum and lyophilized.

Process according to Claim 1, characterized in that the activated carbon has preferred characteristics of Iodine number 800-900 mg I2 / g, molasses relative efficiency 100-130%, methylene blue number (g / 100g). ≥ 14, humidity ≤ 10%, ashes ≤ 10%, mesh size 325 (50-80), 400 (> 90)%.

Process for purifying jambu extract according to claim 1, characterized in that the heating is from 35 to 45 ° C, preferably 40 ° C.

Jambu extract purification process according to claim 1, characterized in that the stirring is magnetic or mechanical, preferably at 100-300 rpm.

Process for purifying jambu extract according to claim 1, characterized in that the filtration is carried out under vacuum in a porous plate funnel with diatomaceous earth precoat in the proportion of 8 to 12%, preferably 10%.

Jambu extract purification process according to claim 1, characterized in that the extract concentration is preferably carried out in a rotary evaporation system at a temperature of 38 to 42 ° C, preferably 40 ° C.

Purified extract characterized in that it is obtained according to claims 1 to 6, comprises at least 40% spilanthol, is chlorophyll free and lyophilized.

Anesthetic composition comprising purified extract of. standard ambu ^' with at least 40% espilant.pl as described in claim 7 and a pharmaceutically acceptable carrier.

Anesthetic composition according to Claim 8, characterized in that it may comprise optional ingredients such as emollient, humectant, chelating agent, thickening system, antioxidant, preservative, coloring and flavoring.

10. bioadhesive comprising a polymer, a solvent and extract standardized AMBU purificado.de at least 40% of espilantol ⁵ as described in claim 7.

Bioadhesive according to Claim 10, characterized in that the polymer can be selected from the group chitosan, polyvinyl alcohol (PVA), cellulose and its derivatives, alginate, gum arabic, cashew gum, other gums and agarose, in the proportion of 1 30%, preferably chitosan.

Bioadhesive according to Claim 10, characterized in that it preferably comprises 1.0 to 1.5% chitosan.

Bioadhesive according to Claim 10, characterized in that said solvent is in a concentration ranging from 0.5 to 2.0% (v / v), preferably 1.0%.

Bioadhesive according to Claim 10, characterized in that the purified Jambu extract is in a concentration of 10 to 20%, preferably 10%.

15. bioadhesive according to claim 10 characterized in that such optional components may comprise ^"as chelating agent such as ethylenediaminetetraacetic acid (EDTA) and its salts; pH adjusting agent ^as' triethanolamine or inorganic hydroxides; preservative agent such as ^' parabens, acids propyl, chlorocresol, benzazonium chloride and glycol ethers; emollients such as alcohols and fatty acids, esters, ethers, mono-, di- or triglycerides, natural or synthetic hydrocarbons or organic carbonates and combinations thereof; dyes; flavoring agents; flavorants; humectants and permeation enhancers such as glycols, preferably glycerine, propylene glycol, butylene glycol and combinations thereof, ethoxydiglycol, more preferably ethoxydiglycol, in a ratio ranging from 5 to 10%, preferably 5%, based on the total bioadhesive mass.

Bioadhesive according to Claim 10, characterized in that the adjuvants may preferably be ethoxydiglycol in a proportion of 5-10%, preferably 5%, based on the total mass of the bioadhesive.

Use of the purified extract as described in claim 7 characterized in that it is for manufacturing anesthetic compositions.

Use of the composition as described in claim 8 characterized in that it is used as an anesthetic.

Use of the bioadhesive as described in claim 10 for use as an anesthetic.