Use of africanized bee venom for the treatment of fibromyalgia

BR102025002040A2Pending Publication Date: 2026-08-11

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Application Number: BR102025002040
Authority / Receiving Office: BR · BR
Patent Type: Applications
Publication Date: 2026-08-11

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001. This Invention Patent (PI) refers to the use of venom extracted from bees of the species Apis mellifera Linnaeus, originating from the Northeast region of Brazil, as an active element in pharmaceutical compositions for the treatment of fibromyalgia. The field of the invention encompasses pharmacology and pharmaceutical biotechnology, with emphasis on the development of new treatments for chronic pain conditions and neurological disorders. In this invention, the potential therapeutic effect of Africanized bee venom on low-conductance calcium-activated potassium channels and on the dopaminergic, glutamatergic, and serotonergic systems was evaluated using the application of reserpine in mice.The proposed invention aims to fill a pharmaceutical and technological gap by using Africanized bee venom as an active ingredient in pharmaceutical compositions and improving the bioavailability of this product, promoting a novel and innovative therapeutic action for the treatment of fibromyalgia. FUNDAMENTALS OF THE TECHNIQUE 002. Pain is defined by the International Association for the Study of Pain (IASP) as an unpleasant sensory and emotional experience associated with, or similar to, actual or potential tissue damage. This multidimensional perception involves not only the physical component but also emotional and cognitive aspects, reflecting the complexity of the phenomenon. Over the years, advances in understanding the mechanisms of pain have revealed that its manifestation results from a complex interaction between biological, psychological, and social factors (RAJA, SN; CARR, DB; COHEN, M.; FINNERUP, NB; FLOR, H., GIBSON, S.; KEEFE, FJ; MOGIL, JS; RINGKAMP, M.; SHIKA, K. A; SONG, X.; STEVENS, B.; SULLIVAN, M. D.; TUTELMAN, PR; Petition 870250008375, dated 01 / 31 / 2025, p. 16 / 99 2 / 25 Association for the Study of Pain definition of pain: concepts, challenges, and compromises. PAIN, 161 (9):p 1976-1982, 2020). 003. Fibromyalgia (FM) is a complex condition whose pathogenesis is not yet fully understood, but involves significant changes in pain processing in the brain. (SARZI-PUTTINI, P.; GIORGI, V.; MAROTTO, D.; ATZENI, F. Fibromyalgia: an update on clinical characteristics, aetiopathogenesis and treatment. Nature Reviews Rheumatology, n. 16, p. 645, 2020; SIRACUSA, R.; DI PAOLA, R.; CUZZOCREA, S.; IMPELLIZZERI, D.; Fibromyalgia: Pathogenesis, Mechanisms, Diagnosis and Treatment Options Update.) 004. Studies indicate dysfunctions in monoaminergic neurotransmission, with an increase in excitatory neurotransmitters, such as glutamate and substance P, in addition to a reduction in serotonin and norepinephrine in antinociceptive pathways (MARQUES, PA; SANTO, ASE; BERSSANETI, AA; MATSUTANI, LA; YUAN, SLR The prevalence of fibromyalgia: an updated literature review. Brazilian Journal of Rheumatology, v. 57, p. The prevalence is higher in women, possibly due to hormonal factors, a higher incidence of anxiety and depression, and an altered response to pain (BERNARDI, L; BERTUCCELLI, K.; FORMAGGIO, E.; RUBEGA, M.; BOSCO, G.; TENCONI, E.; CATTELAN, M.; MASIERO, F.; FELICE, AD Beyond physiotherapy and pharmacological treatment for fibromyalgia syndrome: tailored tACS as a new therapeutic tool. 005. The pathophysiology of the disease and its multifactorial alterations are not yet fully understood; for this reason, definitively effective mechanisms are needed for the treatment of this disease (SLUKA, KA; CALUW, DJ Neurobiology of fibromyalgia and chronic widespread pain. Elsevier Ltd, v. 338, p. 114-129, 2016). Pharmacotherapy, in turn, aims to provide pain relief, focusing on Petition 870250008375, dated 01 / 31 / 2025, page 17 / 99 3 / 25 underlying mechanism, induction of sleep quality, control of associated psychiatric symptoms and improvement of physical functions (BERNARDI, L.; BERTUCCELLI, K.; FORMAGGIO, E.; RUBEGA, M.; BOSCO, G.; TENCONI, E.; CATTELAN, M.; MASIERO, F.; FELICE, AD Beyond physiotherapy and pharmacological treatment for fibromyalgia syndrome: tailored tACS as a new therapeutic tool. Among the most used medications are anticonvulsants, such as Pregabalin, approved by the Food and Drug Administration (FDA) and with good cost-benefit. However, the currently available medications have limited efficacy and may cause adverse effects, as reported by the FDA (SLUKA, KA; CALUW, DJ Neurobiology of fibromyalgia and chronic widespread pain. Elsevier Ltd, v. 338, p. 114-129, 2016; POLUHA, RL; GROSSMAN, E.Does pregabalin improve sleep disorders in fibromyalgia, British Journal of Pain, v. 2, p. 163-166, 2018). 006. Scientific research has focused on innovative approaches to combat fibromyalgia, such as the promising use of bee venom (BV), derived from Africanized bees (Apis mellifera Linnaeus), which consists of a complex mixture of enzymes, lipids, amino acids, carbohydrates, and peptides, such as apamin and melittin (DANTAS, CG; NUNES, TLGM; PAIXÃO, AO DA.; REIS, FP; JÚNIOR, W; DE L.; CARDOSO, JC; GRAMACHO, KP; GOMES, MZ Pharmacological evaluation of bee venom and melittin. Brazilian Journal of Pharmacognosy, v. 24, p. previous studies (CHEN, L.; DELTHEIL, T.; TURLE-LORENZO, N.; LIBERGE, M.; ROSIER, C.; WATABE, I.; SRENG, L.; AMALRIC, M.; MOURRE, C.SK channel blockade reverses cognitive and motor deficits induced by. Petition 870250008375, dated 01 / 31 / 2025, p. 18 / 99 4 / 25 nigrostriatal dopamine lesions in rats. The International Journal of Neuropsychopharmacology, v. 17, n. 8, p. 1295-1306, 2014; PUCCA, M. B.; CERNI, F. A.; OLIVEIRA, I. S.; JENKINS, T. P.; ARGEMÍ, L.; SORENSEN, C. V.; AHMADI, S.; BARBOSA, J. E.; LAUSTSEN, A. H. Bee updated: Current knowledge on bee venom and bee envenoming therapy. Frontiers in Immunology, v. 10, n. 2090, p.1664-3224, 2019; SARGIN, D.; OLIVER, D. K.; LAMBE, E. K. Chronic social isolation reduces 5-HT neuronal activity via upregulated SK3 calcium-activated potassium channels. eLife, v.5, 2016; CHEN, L.; DELTHEIL, T.; TURLE-LORENZO, N.; LIBERGE, M.; ROSIER, C.; WATABE, I.; SRENG, L.; AMALRIC, M.; MOURRE, C. SK channel blockade reverses cognitive and motor deficits induced by nigrostriatal dopamine lesions in rats. The International Journal of Neuropsychopharmacology, v. 17, n. 8, p. 1295-1306, 2014). 007. Melittin, which constitutes approximately 50% of the dry weight of bee venom, has been linked to the modulation of dopaminergic transmission. In combination with apamin, which acts as a blocker of calcium-activated potassium channels, these compounds may increase the activity of dopaminergic neurons, suggesting their therapeutic potential in dopamine-related conditions (DANTAS, CG; PAIXÃO, AO DA.; NUNES, TLGM; SILVA IJF; LIMA, BS DOS.; ARAÚJO, AAS; ALBUQUERQUE-JÚNIOR, RLC DE.; GRAMACHO, KP; PADILHA, FF; COSTA, LP DA.; SEVERINO, P.; CARDOSO, JC; SOUTO, EB; GOMES, MZ; Africanized Bee Venom (Apis mellifera Linnaeus): Neuroprotective Effects in a Parkinson's Disease Mouse Model Induced by 6-hydroxydopamine. Toxics, v. 10, p. 583, 2022; CAGLAYAN, BC; CALIK, BB; KABUL, EG; KARASU, U. Investigation of effectiveness of reformer pilates in individuals with fibromyalgia: A randomized controlled trial.Clinical Rheumatology (Engl Ed), v. 1, p. 1825, 2023. Furthermore, there is evidence that bee venom has a promising therapeutic effect in models of behavioral depression. Petition 870250008375, dated 01 / 31 / 2025, page 19 / 99 5 / 25 (DANTAS, CG; SOUTO, RG; CANDIDO, EAF USE OF AFRICANIZED BEE VENOM FOR THE TREATMENT OF DEPRESSION. 2024, Brazil. Patent: Innovation Privilege. Registration number: BR10202401762, title: USE OF AFRICANIZED BEE VENOM FOR THE TREATMENT OF DEPRESSION, Registration institution: INPI - National Institute of Industrial Property. Filing date: 08 / 27 / 2024). 008. Bee venom also contains other compounds, such as adolapin, which has anti-inflammatory, antinociceptive, and antipyretic properties, possibly due to the inhibition of prostaglandin and cyclooxygenase synthesis (CHERNIACK, EP; GOVORUSHKO, S. To bee or not to bee: The potential efficacy and safety of bee venom acupuncture in humans. Toxicon, v. 158 p. Although not a disease notably of inflammatory etiology, it is known that the release of inflammatory cytokines occurs to some degree in the pathophysiological evolution of fibromyalgia, which would allow for a potential therapeutic summation of the effects of adolapin with those of other enzymes present in the venom of Africanized bees (DANTAS, CG; NUNES, TLGM; PAIXÃO, AO DA.; REIS, FP; JÚNIOR, W; DE L; CARDOSO, JC; GRAMACHO, KP; GOMES, MU.Z. Evaluation). pharmacological analysis of bee venom and melittin. Brazilian Journal of Pharmacognosy, v. 24, p. 67-77, 2014; SIRACUSA, R.; DI PAOLA, R.; CUZZOCREA, S.; IMPELLIZZERI, D.; Fibromyalgia: Pathogenesis, Mechanisms, Diagnosis and Treatment Options Update. International Journal of Molecular Sciences, v. 22, n. 8, p. 3891, 2021). 009. In addition, VA has several pharmacological effects such as antimutagenic, anti-nociceptive, radioprotective (CORNARA, L.; BIAGI, M.; XIAO, J.; BURLANDO, B. Therapeutic properties of bioactive compounds from different honeybee products. Frontiers in Pharmacology, v. 8, n.412, 2017), antimicrobial, and antiviral (CARPENA, M.; NUNEZ-ESTEVEZ, B.; SORIA-LOPEZ, A.; & SIMAL-GANDARA, J. Bee venom: An updating). Petition 870250008375, dated 01 / 31 / 2025, p. 20 / 99 6 / 25 review of its bioactive molecules and its health applications. Nutrients, v. 12, n. 11, p. 3360, 2020), anti-hepatotóxico, citoprotetor e antioxidante (HASSAN, S. A.; ALAZRAGI, R. S.; SALEM, N. A. Potential therapeutic effect of bee venom on cisplatin-induced hepatotoxicity. Journal of Pharmaceutical Research, v. 33, p. 200-210, 2021). 0010. Experimental models with reserpine reproduce symptoms similar to those of fibromyalgia in rodents, such as hyperalgesia, allodynia, depressive behaviors, and gastrointestinal dysfunction, characteristics common in patients with FM (HAO, Y.; GE, H.; SUN, M.; GAO, Y. Selecting an appropriate animal model of depression. International Journal of Molecular Sciences, v. 20, n. 19, p. 4827, 2019; BRUM, EDS; FIALHO, MFP; FISCHER, SPM; HARTMAN, DD; GONÇALVES, DF; SCUSSEL, R.; MACHADO-DE-ÁVILA, RA; CORTE, CLD; SOARES, FAA; OLIVEIRA, SM; Relevance of Mitochondrial Dysfunction in the Reserpine-Induced Experimental Fibromyalgia Model. Molecular Neurobiology, v. 57, n. 10, p. 4202-4217, 2020). Reserpine acts by reducing monoamine levels in the central nervous system, regulating the accumulation of these monoamines in synaptic vesicles and their reuptake from synapses, leading to their depletion (CZARNY, P.; WIGNER, P.; GALECKI, P.; SLIWINSKI, T.The interplay between inflammation, oxidative stress, DNA damage, DNA repair, and mitochondrial dysfunction in depression. Progress in Neuropsychopharmacology and Biological Psychiatry, v. 80, part. C, p. 30-321, 2018), and depressive disorders (VAGHEF, L. Cerebrolysin attenuates ethanol-induced spatial memory impairments through inhibition of hippocampal oxidative stress and apoptotic cell death in rats. Alcohol, v. 79, p. 127-135, 2019). Therefore, this study aims to evaluate the possible therapeutic effect of bee venom (Apis mellifera L.) in an experimental model of reserpine-induced fibromyalgia, an innovative approach since there is no research investigating this impact to date. Petition 870250008375, dated 01 / 31 / 2025, p. 21 / 99 7 / 25 STATE OF THE ART 0011. To substantiate the innovative nature of the present invention, a prior art search was conducted regarding the use of Africanized honeybee venom (Apis mellifera Linnaeus) in the databases of INPI, USPTO, WIPO Patentscope, Espacenet (Latipat), Worldwide, and LENS.ORG. Searches for scientific publications related to the topic were also conducted in Google Scholar, SciELO, and ScienceDirect. The most relevant patents and scientific publications are presented below; however, it should be noted that none of the documents found violate the novelty requirement of the invention patent requested in this document, given that the inventive concept proposed here differs considerably from the cited documents. 0012. Patent code PI010566-5 A describes an invention based on bee venom as the active ingredient in an ointment. According to the document, the effects of bee venom may be related to the modulation of the immune system, and the profile of its effects may vary depending on the amount of venom used. It has been shown that some of the peptides present in the venom have an amphipathic structure, capable of inducing a large release of histamine. The pharmaceutical composition is formulated as an ointment for topical use. The effect of bee venom in treatment was demonstrated by topical application to induced edema of the paw and ear. The proposed product differs from the aforementioned patent with regard to the mechanism of induction of fibromyalgia, since the present proposal uses reserpine, an irreversible inhibitor of the vesicular monoamine transporter 2, which regulates the accumulation of monoamines in synaptic vesicles and their reuptake from synapses.Thus, reserpine inhibits the presynaptic reuptake and storage of monoamines, leading to monoamine depletion and reproducing symptoms similar to those of fibromyalgia in rodents, such as hyperalgesia, allodynia, and behavioral changes. Petition 870250008375, dated 01 / 31 / 2025, page 22 / 99 8 / 25 depressive symptoms and gastrointestinal dysfunction. In this sense, the origin of the bee venom used also differs between studies, since the inventors cite the use of bee venom for the treatment of rheumatoid arthritis and other joint dysfunctions, which have a different pathological profile from fibromyalgia. 0013. Patent document PI9505366-2 refers to a production process for an ointment with therapeutic action, especially indicated for patients with rheumatism, arthritis, and arthrosis, an inflammatory joint process that causes pain in the affected areas, in addition to other secondary applications. The study was conducted using a mixture of petrolatum in an approximate proportional range of 64 to 68% of the total volume of the finished product, along with lanolin, in an approximate proportional range of 20 to 23% of the total volume of the finished product, and a given volume of dried bee venom, which will proportionally represent 0.15 to 0.16% of the total volume of the finished product. The proposed process differs from the aforementioned patent with regard to the venom content applied in the production process, since the dose of venom tested was not specified. In the present study, the doses tested were between 0.5 mg.Kg-1 and 1.5 mg.Kg-1, at a frequency of once a day, for 10 days.Furthermore, the aforementioned prior art document does not mention fibromyalgia as a disease to be treated through the present invention. 0014. The scientific article BEE VENOM ACUPUNCTURE FOR MUSCULOSKELETAL PAIN: A REVIEW presents a review showing evidence of the effectiveness of using bee venom combined with its injection into acupuncture points for the treatment of musculoskeletal pain, such as arthritis and rheumatic diseases. The article reports the use of solutions containing bee venom along with acupuncture needles for the relief of musculoskeletal pain, where an improvement in symptoms was observed; however, the small sample size makes it difficult to affirm that the improvement was statistically significant. Petition 870250008375, dated 01 / 31 / 2025, page 23 / 99 9 / 25 significant, suggesting the need for further experimental studies to also understand the mechanisms by which this technique works. Furthermore, the study did not specify fibromyalgia as a pain condition addressed in the review, thus not delimiting the present invention. 0015. Thus, based on the disclosed documents, it is highlighted that the development of the "USE OF AFRICANIZED BEE VENOM FOR THE TREATMENT OF FIBROMYALGIA" proposed in this document has unprecedented and innovative potential, and therefore meets the criterion of novelty, since to date no scientific or technical work included in the state of the art possesses a similar technology for development, use and application to that which is disclosed and requested. Finally, it is important to emphasize that the technology proposed in this patent application also meets the other patentability criteria, such as inventive activity and industrial application, requirements necessary for the granting of the requested patent. DESCRIPTION OF THE FIGURES 0016. The present invention, together with its additional advantages, can be better understood by reference to the accompanying drawings and description, which are for illustrative purposes only and do not limit the subject matter of the present invention. 0017. The invention will now be explained in detail, and the following drawings are presented by way of illustration: Figure 1 highlights the average time (in seconds) of immobility in the forced swimming test. The columns represent the group means on days 6 and 11, and the bars represent the standard error of the mean. Different symbols indicate significant differences: * (p < 0.05), ** (p < 0.05), *** (p < 0.05), and **** (p < 0.05). Two-way ANOVA. Figure 2 shows the result of the horizontal exploration parameter (EH) in the open field test. The columns represent the averages of horizontal farms in the groups on days 0, 4, and 14, and the bars represent the average number of horizontal farms in the groups on days 0, 4, and 14. Petition 870250008375, dated 01 / 31 / 2025, page 24 / 99 10 / 25 represents the standard error of the mean. Different symbols indicate significant differences: * (p < 0.05), ** (p < 0.05), and *** (p < 0.05). Two-way ANOVA. Figure 3 highlights the result of the vertical exploration (VE) parameter in the open field test. The columns represent the means of horizontal explorations in the groups on days 0, 4, and 14, and the bars represent the standard error of the mean. Different symbols indicate significant differences: * (p < 0.05), ** (p < 0.05), and *** (p < 0.05). Two-way ANOVA. Figure 4 shows the results for the fecal bolus parameter in the open field test. The columns represent the means of horizontal explorations in the groups on days 0, 4, and 14, and the bars represent the standard error of the mean. Different symbols indicate significant differences: * (p < 0.05) and ** (p < 0.05). Two-way ANOVA. 0018. Figure 5 highlights the result of the self-grooming parameter in the open field test. The columns represent the means of horizontal explorations in the groups on days 0, 4, and 14, and the bars represent the standard error of the mean. Different symbols indicate significant differences: * (p < 0.05), ** (p < 0.05), and *** (p < 0.05). Two-way ANOVA. 0019. Figure 6 shows the result of the average time (in seconds) that the animals remained in the closed arms in the elevated plus maze test. The columns represent the group means on days 5 and 13, and the bars represent the standard error of the mean. Different symbols indicate significant differences: ** (p < 0.05) and **** (p < 0.05). Two-way ANOVA. Figure 7 shows the average time (in seconds) that the animals remained in the open arms during the elevated plus maze test. The columns represent the group means on days 5 and 13, and the bars represent the standard error of the mean. Different symbols Petition 870250008375, dated 01 / 31 / 2025, p. 25 / 99 11 / 25 indicate significant differences: * (p < 0.05), ** (p < 0.05), *** (p < 0.05) and **** (p < 0.05). Two-way ANOVA. Figure 8 explores the results of the mean latency time of the thermal nociceptive threshold (in seconds) in the tail movement test. The columns represent the group means on days 0, 3, 4, 9, and 14, and the bars represent the standard error of the mean. Different symbols indicate significant differences: * (p < 0.05), ** (p < 0.05), *** (p < 0.05), and **** (p < 0.05). Two-way ANOVA. SUMMARY OF THE INVENTION 0020. The present invention patent (PI) aims to describe in detail the USE OF AFRICANIZED BEE VENOM FOR THE TREATMENT OF FIBROMYALGIA. More precisely, the invention addresses the use of bee venom as an active element in pharmaceutical compositions, for the treatment of reserpine-induced fibromyalgia in an experimental model using mice. 0021. The product exhibits potent anti-inflammatory and neuroprotective activity and a broad spectrum of action on the nervous system, with potential therapeutic action on an experimental model of fibromyalgia, improving locomotor behavior, reducing hypersensitivity, depression, and emotional behavior in animals. 0022. The present innovation consists of the use of venom from Africanized bees of the genus Apis, species A. mellifera, which have venom with a different chemical profile compared to European bees. Additionally, the use of Africanized bee venom for the treatment of fibromyalgia is proposed. The present invention consists of the subcutaneous application of venom from Africanized bees of the genus Apis, species A. mellifera (aiming at the synergy of action of all compounds), which have venom with a different chemical profile compared to European bees. 0023. The proposed invention aims at creating innovative pharmaceutical compounds to treat fibromyalgia, taking advantage of properties of Petition 870250008375, dated 01 / 31 / 2025, page 26 / 99 12 / 25 Africanized bee venom, focusing on improving the product's bioavailability and blocking ion channels related to dopaminergic and serotonergic neuronal activity. Furthermore, the study of the effect of apamin, a compound in the venom, on calcium-activated potassium channels, directs the invention toward a potential therapeutic advancement in the field of neurological and neuromuscular treatments. DETAILED DESCRIPTION OF THE INVENTION 0024. In order to ensure that the developed product, which is the subject of this patent application, can be understood and evaluated, a detailed description of the invention entitled USE OF AFRICANIZED BEE VENOM FOR THE TREATMENT OF FIBROMYALGIA is provided below: 0025. The present invention consists of the use of Africanized bee venom as an active ingredient in pharmaceutical compositions for the treatment of fibromyalgia. The capacity of Africanized bee venom as an active ingredient in pharmaceutical compositions was evaluated through controlled biological assays. Obtaining bee venom 0026. Bee venom was obtained from an apiary in Rio Grande do Norte - RN, in the Northeast region of Brazil, using electrical extraction. The electrical extraction collection technique consists of placing a collector, composed of glass plates and a pulse generator, connected to a battery or other power source, at the hive entrance. When bees land on the plate, they receive a shock and react by stinging the electrical collector plate, depositing a charge of venom between the glass and the protective material of the equipment, where it dries and is then scraped off and sold (HIDER, 1988; DUSSART; BARTHOLOMÉ, 2007; ZHOU et al., 2010). The main advantage of this method of obtaining venom is that it does not cause the death of the animal and preserves the bee's stinger (FERREIRA-JUNIOR et al., 2010). Petition 870250008375, dated 01 / 31 / 2025, page 27 / 99 13 / 25 Application 0027. Dilute the bee venom concentrations VA 0.5 mg. Kg-1 or VA 1.0 mg. Kg-1 or VA 1.5 mg. Kg-1 in 40 μL of saline solution. After dilution, administer the solution subcutaneously. Treatment 0028. Treatment is carried out continuously, once every 2 days. Chemical characterization of the venom by means of High Performance Liquid Chromatography (HPLC). 0029. Bee venom (10 mg) was diluted in 10 mL of trifluoroacetic acid (0.1% in Milli-Q water). The solution (concentration: 1 mg.mL) was centrifuged at 12000 rpm for 10 minutes (4 °C) and all the supernatant (approximately 9 mL) was collected for HPLC analysis. Prior to HPLC injection, the sample was filtered through a membrane filter (0.45 μm - PTFE). HPLC analysis was performed using a high-performance liquid chromatography system consisting of a DGU-20A3 degasser, two LC-20AD pumps, a SIL-20A HT autosampler, a CTO-20A column oven, an SPDM20Avp photodiode array detector (DAD), and a CBM-20A system controller. Chromatographic separation was performed using a C18 analytical column (4.6 x 250 mm, particle size: 5 μm). 0030. The mobile phase consisted of a gradient of: A: trifluoroacetic acid (0.1% in Milli-Q water) and B: acetonitrile. The mobile phase flow rate was 0.8 mL.min-1 and the sample injection volume was 20 μL. The elution gradient started with 5% B for 7 minutes, 5-10% B for 7-10 minutes, 10-17% B for 10-15 minutes, 17-25% B for 15-20 minutes, 25-35% B for 20-33 minutes, 35-40% B for 33-39 minutes, 40-5% B for 39-45 minutes, returning to initial conditions and ending the analysis. The detector (HPLC - DAD) was adjusted to 280 nm for peak analysis. Petition 870250008375, dated 01 / 31 / 2025, p. 28 / 99 14 / 25 0031. The compounds were identified through co-injections of standards by comparing retention times. Standard samples of apamin, melittin, and phospholipase A2 were diluted with Milli-Q water to 200 μg.mL⁻¹ (stock solution). The samples were filtered through a membrane filter (0.45 μm - PTFE) before HPLC injections. Quantitative analysis was performed using calibration curves for each identified standard compound. The curves were prepared at seven different concentrations: 0.8; 4; 8; 12; 16; 20 and 50 μg.mL⁻¹. All samples were filtered through a membrane filter (0.45 μm - PTFE) before HPLC injection and were analyzed in triplicate. Biological assay Animals and ethical guidelines 0032. Female Sw / ss mice (25-30 g) were kept in a temperature-controlled environment, under a 12-hour light-dark cycle, with free access to water and food. All procedures were in accordance with the standards of the Brazilian College of Animal Experimentation (COBEA), and the experimental protocol was approved by the Ethics Committee on the Use of Animals of Tiradentes University, Aracaju - SE, Brazil (approval number 010723). Sample 0033. For the development of the experimental model of depression, the animals were divided into the following groups: Reserpine; Control; Pregabalin; VA 0.5 mg.Kg-1; VA 1.0 mg.Kg-1 and VA 1.5 mg.Kg-1. Induction protocol for the reserpine-induced fibromyalgia experimental model. 0034. All animals, except those in the negative control group, received subcutaneous (sc) injections of reserpine (RES) once daily for 3 consecutive days to induce depletion of biogenic amines. RES (0.25 mg.kg-1) was diluted in Tween 80 to 0.5% v / v in phosphate-buffered saline (PBS), and pregabalin in saline solution (0.9% NaCl), volume of 10 mL.kg-1. (PEDRON, C.; ANTUNES, F. Petition 870250008375, dated 01 / 31 / 2025, p. 29 / 99 15 / 25 TT; REBELO, IN; CAMPOS, MM; CORREA, AP; KLEIN, CP; OLIVEIRA, IB; CORDEIRO, MN; GOMEZ, MV; SOUZA, AH. Phoneutria nigriventer Tx3-3 peptide toxin reduces fibromyalgia symptoms in mice. Neuropeptides, v. 85, p. 102094, 2021; FERRARINI, EG; PAES, RS; BALDASSO, GM; ASSIS PM; GOUVÊA, MC; CICCO, P.; RAPOSO, NRB; CAPASSO, R.; MOREIRA, ELG; DUTRA, RC. Broad-spectrum cannabis oil ameliorates a reserpine-induced fibromyalgia model in mice. Biomedicine & Pharmacotherapy, v. 154, p. 113552, 2022; MARTINS, CP; PAES, RS; BALDASSO, GM; FERRARINI, EG; SCUSSEL, R.; ZACCARON, RP; MACHADO-DE-ÁVILA, RA; SILVEIRA, PCL; DUTRA, RC. Pramipexole, a dopamine D3 / D2 receptor agonist, attenuates a reserpine-induced fibromyalgia-like model in mice. Neural Regeneration Research, v. 17, n. 2, p. 450-458, 2022. Treatment with bee venom (BV), pregabalin, or saline solution 0035. Treatments were performed by subcutaneous (sc) administration of Africanized bee venom (Apis mellifera L.) or saline solution (sc), or pregabalin (orally). Subcutaneous injections were administered in the dorsal region of the animal, alternating between the dorsal midline, right paramedian, and left paramedian regions to avoid irritation at the injection site. Needles measuring 0.45 x 13 mm were used for this purpose. The venom treatment groups were divided into: VA 0.5 mg.Kg-1; VA 1.0 mg.Kg-1; and VA 1.5 mg.Kg-1. The concentrations used were diluted in 40 μL of saline solution, as per a previous study. Animals in the pregabalin group received 30 mg.kg-1 of the drug dissolved in water, orally (PEDRON, C.; ANTUNES, FTT; REBELO, IN; CAMPOS, MM; CORREA, AP; KLEIN, CP; OLIVEIRA, IB; CORDEIRO, MN; GOMEZ, MV; SOUZA, AH. Phoneutria nigriventer Tx3-3 peptide toxin reduces fibromyalgia symptoms in mice. Neuropeptides, v. 85, p.102094, 2021). Petition 870250008375, dated 01 / 31 / 2025, page 30 / 99 16 / 25 The treatments were performed daily, from day 4 to day 14, once every 2 days (GOMES, MZ; GRAMACHO, KÁTIA P.; COSTA, LP; PADILHA, FF; DANTAS, CG; CARDOSO, JULIANA C.; ALBUQUERQUE JUNIOR, RLC. Use of bee venom in the treatment of Parkinson's disease and schizophrenia. 2017, Brazil). Behavioral assessments for an experimental model of fibromyalgia 0036. Functional effects were assessed using the following behavioral tests: forced swimming test (KAUR et al., 2019; BRUM et al., 2020; SINGH et al., 2021; GARCIA MENDES et al., 2021; MARTINS et al., 2022), open field test (KAUR et al., 2019; BRUM et al., 2020; MARTINS et al., 2022), elevated plus maze (KAUR et al., 2019; MARTINS et al., 2022), tail movement test (MARTINS et al., 2022), and cold thermal sensitivity test (BRUM et al., 2020). 0037. All behavioral assessments were performed before the injection(s) on the respective treatment and / or induction day (KAUR, A.; SINGH, l.; SINGH, N.; BHATTI, ,. S.; BHATTI, R. Ameliorative effect of imperatorin in chemically induced fibromyalgia: Role of NMDA / NFkB mediated downstream signaling. Biochemical Pharmacology, v. 166, p. 56-69, 2019), and were conducted by a trained evaluator blinded to the treatment. Furthermore, all devices were cleaned with 10% ethanol before the behavioral tests to eliminate any traces of odor. Forced swimming test 0038. The forced swimming test was performed as described by PORSOLT, RD; BERTIN, A.; JALFRE, M.; Behavioral despair in mice: a primary screening test for antidepressants. Archives Internacionales de Pharmacodynamie et de Therapie, v. 229, n.2, p. 327-336, 1977, in order to evaluate a possible antidepressant effect. The test consists of two swimming sessions, separated by a period of 24 h. In the first session (pre-test), the animals were placed in a water tank. Petition 870250008375, dated 01 / 31 / 2025, p. 31 / 99 17 / 25 circular (30 cm high) for 15 min. In the second session (test), the animals underwent the same procedure for 5 min, and then swimming and immobility times were evaluated. The pre-test was performed on the 10th day and the test was performed on the 11th day (WANG et al., 2017; DHAMIJA et al., 2017; PARK et al., 2018; ZHAO et al., 2019; PARK et al., 2020). 0039. In the forced swimming test, on day 6, a significant increase in immobility was observed in the injury group compared to the following groups: healthy control (p<0.0001), pregabalin (p<0.0001) and VA 0.5 mg.Kg-1 (p<0.0001). The VA 1 mg.Kg-1 and VA 1.5 mg.Kg-1 groups significantly increased immobility time compared to the healthy control group (p=0.02; p=0.0005, respectively). The VA 1.5 mg.Kg-1 group also significantly increased this parameter compared to the pregabalin group (p=0.009). The group treated with bee venom at a dose of 0.5 mg.Kg-1 significantly reduced immobility in the test compared to the groups treated with the highest doses of bee venom tested (1 and 1.5 mg.Kg-1, p=0.01; p=0.0002, respectively). 0040. On the 11th day of the experiment, in this test, there was a significant increase in immobility in the animals of the lesion group compared to all other groups tested: healthy control (p<0.0001), pregabalin (p<0.0001), VA 0.5 mg.Kg-1 (p=0.01), VA 1 mg.Kg-1 (p<0.0001) and VA 1.5 mg.Kg-1 (p<0.0001). The healthy control group significantly reduced this behavior compared to the pregabalin (p=0.01), VA 0.5 mg.Kg-1 (p<0.0001) and VA 1 mg.Kg-1 (p=0.01) groups - Figure 1. 0041. It is thus observed that the behavior of the lesion group reinforces the negative impact of fibromyalgia on the emotional state and physical disposition of the animals, while the groups treated with both pregabalin and venom suggest improvement in physical and motivational state (BRUM et al., 2020). The lowest dose tested in VA (0.5 mg.Kg-1) demonstrated greater efficacy in reducing immobility on the 6th day of the experiment, but on the 11th day of the research, all doses of VA tested Petition 870250008375, dated 01 / 31 / 2025, page 32 / 99 18 / 25 were effective, which denotes an important action of VA on depressive behavior. Brain regions such as the amygdala, hippocampus, prefrontal cortex, and anterior cingulate cortex are involved in mood disorders (Yao et al., 2020). A previous study points to VA acting in these brain areas, which possibly justifies the result found (DALKIRAN et al., 2023). Open field test 0042. To assess the locomotor activity of the animals, the open field test was performed on days 4 and 14, lasting 5 minutes (WHIMBEY, AE; DENENBERG, VH. Two independent behavioral dimensions in open-field performance. Journal of Comparative and Physiological Psychology, v. 63, n. 3, p. 500-504, 1967; MARTINS, CP; PAES, RS; BALDASSO, GM; FERRARINI, EG; SCUSSEL, R.; ZACCARON, RP; MACHADO-DE-ÁVILA, RA; SILVEIRA, PCL; 17, n. The equipment consists of a wooden quadrilateral covered with white Formica, with an area of 4830.25 cm2, walls 34.5 cm2 high and a base subdivided into sixteen quadrants (HONGXING, Z.; NANCAI, Y.; GUOFU, H.; JIANBO, S.; YANXIA, W.; HANJU, H.; QIAN, L.; WEI, M.; YANDONG, Y.; HAO, H.Neuroprotective effects of extracts of purslane herbs against D-galactose-induced neurotoxicity. ChemicoBiological Interactions, v. 170, n. 3, p. 145-152, 2007; O'LEARY, TP; GUNN, RK; BROWN, RE What are we measuring when we test strain differences in anxiety in mice?. Behavioral Genetics, v. 43, p. 34-50, 2013). 0043. To assess the animal's locomotor activity, the following parameters were evaluated: horizontal exploration - EH - (number of quadrants crossed / traveled by the animal during locomotion) and vertical exploration - EV - (number of times the animal remained Petition 870250008375, dated 01 / 31 / 2025, p. 33 / 99 19 / 25 on the hind legs). The test was performed on days 0 and 10 (PARK et al., 2018; ZHAO et al., 2019; PARK et al., 2020). The assessments were recorded on video for later analysis. 0044. In the open field test, regarding the horizontal exploration locomotor parameter (EH), it was observed that on day 0, the healthy control group showed a significant increase compared to the lesion group (p=0.02), pregabalin group (p=0.01), VA 0.5 mg.Kg-1 group (p=0.01) and VA 1 mg.Kg-1 group (p=0.0007). The group treated with VA 1.5 mg.Kg-1 showed a significant increase in this parameter compared to the lesion group (p=0.03) and the VA 1 mg.Kg-1 group (p=0.02). 0045. On day 4, the healthy control group maintained a significant increase in EH compared to the lesion group (p=0.02) and the VA 1 mg.Kg-1 group (p=0.002). The group treated with VA 1.5 mg.Kg-1 also maintained a significant increase in this motor parameter compared to the lesion group (p=0.01) and the VA 1 mg.Kg-1 group (p=0.03). A significant increase was also observed in the group treated with pregabalin compared to the group treated with bee venom at a dose of 1 mg.Kg-1 (p=0.03). 0046. On the 14th day of the experiment, it was observed that the healthy control group maintained a significant increase in EH compared to the lesion group (p=0.02). The pregabalin, VA 1 mg.Kg-1 and VA 1.5 mg.Kg-1 groups showed a significant increase in this behavior compared to the lesion group (p=0.03; p=0.03 and p=0.01, respectively) (Figure 2). 0047. It was observed, therefore, that the lesion group maintained a reduction in EH (Emotional Hygiene) compared to the healthy control group, from the beginning to the end of the experiment (MIYAHARA et al., 2021; BRUM et al., 2020). Treatment with bee venom at a dose of 1.5 mg.Kg-1 generated a significant increase in the animals' ambulation on the 4th day of the experiment, and this effect remained until the 14th day. The increase in exploratory activity, both horizontal and vertical, in this group indicates a symptomatic improvement, with greater willingness of the animals to explore the environment, which would be limited by pain and emotional symptoms (BRUM et al., 2020). Petition 870250008375, dated 01 / 31 / 2025, page 34 / 99 20 / 25 0048. In the evaluation carried out on the 14th day of the experiment, the dose of 1 mg.Kg-1 also generated a significant increase in this motor parameter. Dantas et al. (2014) reported the action of Apis mellifera L. venom on dopaminergic pathways, with a strong interaction on D1 and D2 receptors. Previous studies, which point to a relationship between apamin and calcium-activated potassium (SK) channels, identified that VA acts by blocking type 3 SK channels - SK3 (MAURICE et al., 2015). Thus, the pharmacological blockade of these channels is capable of reversing motor deficits and promoting dopaminergic neuronal activity (CHEN et al., 2014). 0049. Regarding vertical exploration (VE) behavior, on day 0, the lesion and VA 0.5 mg.Kg-1 groups showed a significant reduction compared to the healthy control group (p=0.04; p=0.008, respectively). On the 4th day of the experiment, the lesion group maintained a significant reduction in this motor behavior compared to the healthy control group (p=0.01), and a significant increase in VE was also observed in the group treated with VA 1 mg.Kg-1 compared to the lesion group (p=0.03). At the end of treatment (at 14 days), a significant increase was observed in the healthy control group (p=0.0002), pregabalin (p=0.008), VA 1 mg.Kg-1 (p=0.001), and VA 1.5 mg.Kg-1 (p=0.0009) compared to the lesion group, as shown in Figure 3. 0050. Treatment with VA at doses of 1.0 and 1.5 mg.Kg-1 and with pregabalin restored IV function in the animals. It is important to mention that pregabalin acts by binding to the alpha2delta subunit of voltage-dependent calcium channels in the central nervous system and acts to improve pain, fatigue, and sleep disturbances (TZADOK; ABLIN, 2020). The response similar to treatment with a drug already used for the disease, especially at a dose of 1 mg.Kg-1, suggests that the symptomatic response is promising and comparable to current treatments, but with a greater likelihood of reduced side effects (POLUHA et al., 2018). 0051. Regarding the fecal bolus parameter, it was found that on day 0, there was no significant difference between the groups tested. Petition 870250008375, dated 01 / 31 / 2025, p. 35 / 99 21 / 25 However, on the 4th day of the experiment, there was a significant increase in this parameter in the group treated with pregabalin compared to the lesion group (p=0.01). On the 14th day of the experiment, a significant increase in this parameter was observed in the pregabalin (p=0.01), VA 1 mg.Kg-1 (p=0.02) and VA 1.5 mg.Kg-1 (p=0.002) groups compared to the lesion group - Figure 4. 0052. In the self-grooming parameter, on the 4th day of the experiment, a significant reduction in this parameter was observed in the VA 1 mg.Kg-1 group compared to the VA 0.5 mg.Kg-1 group (p=0.04). On the 14th day of treatment, there was a significant reduction in self-grooming in the lesion group compared to the healthy control group (p=0.004), VA 1 mg.Kg-1 group (p=0.02), and VA 1.5 mg.Kg-1 group (p=0.0009). The group treated with bee venom, at a dose of 0.5 mg.Kg-1, also showed a significant reduction in this behavior compared to the group treated with the dose of 1.5 mg.Kg-1 (p=0.03), as shown in Figure 5. The action of pregabalin, together with the action of VA (at doses of 1 and 1.5 mg.Kg-1) on fecal bolus and self-cleaning parameters on the 14th day of treatment, indicates a significant reduction in the animal's anxiety levels (WINSLOW et al., 2023). Elevated plus maze test 0053. Used to assess fear and anxiety, the apparatus has a base that elevates it 45 cm above the ground. It consists of two open arms (50 x 10 cm) and two closed arms of the same size, with 40 cm high walls and uncovered at the top. In this test, the animal is placed in the center of the elevated plus-maze with its face turned towards the open arm. The number of entries and the time the animal spent in the open and closed arms was analyzed (PELLOW, S.; CHOPIN, P.; FILE, SE; BRILEY, M. Validation of open:closed arm entries in an elevated plus-maze as a measure of anxiety in the rat. Journal of Neuroscience Methods, v. 14, n. 3, p. 149-167, 1985), for 5 minutes (DHAMIJA, I.; PARLE, M.; & KUMAR, S. Antidepressant and anxiolytic effects of Garcinia indica fruit rind via Petition 870250008375, dated 01 / 31 / 2025, p. 36 / 99 22 / 25 monoaminergic pathway. 3 Biotech, v. 7, n. 2, p. 131, 2017). The test was performed on the following days: 5 and 13. 0054. In the elevated plus maze test, the average time the animals remained in the closed arms of the apparatus was observed. On day 5, there was a significant increase in the lesion group compared to all groups tested: healthy control (p<0.0001), pregabalin (p=0.001), VA 0.5 mg.Kg-1 (p<0.0001), VA 1 mg.Kg-1 (p<0.0001) and VA 1.5 mg.Kg-1 (p<0.0001). On day 13, this behavior was repeated, with a significant increase in the lesion group compared to the following groups: healthy control (p<0.0001), pregabalin (p<0.0001), VA 0.5 mg.Kg-1 (p<0.0001), VA 1 mg.Kg-1 (p<0.0001) and VA 1.5 mg.Kg-1 (p<0.0001) - Figure 6. 0055. In this same test, when observing the average time the animals remained in the open arms of the apparatus, a significant increase was observed on day 5 in the healthy control group (p<0.0001), pregabalin (p=0.04), VA 0.5 mg.Kg-1 (p=0.02) and VA 1.5 mg.Kg-1 (p=0.0004) compared to the lesion group. On the 13th day of the experiment, all tested groups showed a significant increase in this behavior compared to the lesion group, namely: healthy control (p<0.0001), pregabalin (p<0.0001), VA 0.5 mg.Kg-1 (p<0.0001), VA 1 mg.Kg-1 (p<0.0001) and VA 1.5 mg.Kg-1 (p<0.0001) - Figure 7. Tail movement test 0056. Used to assess mechanical hypersensitivity, the test was performed on days 0, 3, 4, 9, and 14 (MARTINS, CP; PAES, RS; BALDASSO, GM; FERRARINI, EG; SCUSSEL, R.; ZACCARON, RP; MACHADO-DE-ÁVILA, RA; SILVEIRA, PCL; DUTRA, RC). Pramipexole, a dopamine D3 / D2 receptor-preferring agonist, attenuates a reserpine-induced fibromyalgia-like model in mice. A brief immersion of the animal's tail in hot water (48 ± 1°C) was performed to measure the latency of the thermal threshold (TREVISAN, G.; MALDANER, G.; VELLOSO, NA; SANT'ANNA, G. DA. S.; ILHA, V.; VELHO GEWEHR, C. DE. C.; RUBIN, Petition 870250008375, dated 01 / 31 / 2025, p. 37 / 99 23 / 25 MA; MOREL, AF; FERREIRA, J. Antinociceptive effects of 14-membered cyclopeptide alkaloids. Journal of Natural Products, v. 72, p. 608-612, 2009; DALENOGARE, DP; FERRO, PR; DE PRA, SDT; RIGO, FK; DE DAVID ANTONIAZZI, CT; DE ALMEIDA, AS; DAMIANI, AP; STRAPAZZON, G.; DE OLIVEIRA SARDINHA, TT; GALVANI, NC; BOLIGON, AA; DE ANDRADE, VM; DA SILVA BRUM, E.; OLIVEIRA, SM; TREVISAN, G. Antinociceptive activity of Copaifera officinalis Jacq. L. oil and kaurenoic acid in mice. Inflammopharmacology, v. 27, p. 829-844, 2019). For the animal that did not exhibit nociceptive behavior after 15 seconds, the stimulus was withdrawn to avoid tissue damage (MARTINS, CP; PAES, RS; BALDASSO, GM; FERRARINI, EG; SCUSSEL, R.; ZACCARON, RP; MACHADO-DE-ÁVILA, RA; SILVEIRA, PCL; DUTRA, RC Pramipexole, a dopamine D3 / D2 receptor-preferring agonist, attenuates a reserpine-induced fibromyalgia model in mice.450-458, 2022). 0057. In the tail movement test (Figure 8), on day 0, an increase in the mean latency of the thermal threshold was observed in the pregabalin (p=0.03), VA 1 mg.Kg-1 (p=0.001) and VA 1.5 mg.Kg-1 (p=0.007) groups compared to the healthy control group. There was also a significant increase in these groups compared to the lesion group: pregabalin (p=0.003), VA 1 mg.Kg-1 (p=0.001) and VA 1.5 mg.Kg-1 (p=0.007). 0058. On day 3, the significant increase in this parameter was maintained in the pregabalin (p<0.0001), VA 1 mg.Kg-1 (p=0.002) and VA 1.5 mg.Kg-1 (p=0.0001) groups in relation to the healthy control group. There was a significant increase in the following groups in relation to the lesion group: pregabalin (p<0.0001), VA 0.5 mg.Kg-1 (p=0.01), VA 1 mg.Kg-1 (p=0.0005) and VA 1.5 mg.Kg-1 (p<0.0001). A significant increase in the mean latency of the thermal threshold was observed in the pregabalin group in relation to the groups treated with bee venom at doses of 0.5 mg.Kg-1 (p=0.005) and 1 mg.Kg-1 (p=0.001). Petition 870250008375, dated 01 / 31 / 2025, page 38 / 99 24 / 25 0059. Reserpine administration induced a significant decline in the thermal nociceptive threshold on the 4th day of the experiment compared to the pregabalin (p<0.0001), VA 0.5 mg.Kg-1 (p=0.002), VA 1 mg.Kg-1 (p<0.0001) and VA 1.5 mg.Kg-1 (p<0.0001) groups. Animals in the control group also showed a significant reduction in this behavior compared to the pregabalin (p<0.0001), VA 0.5 mg.Kg-1 (p=0.002), VA 1 mg.Kg-1 (p<0.0001) and VA 1.5 mg.Kg-1 (p<0.0001) groups. Animals treated with pregabalin showed a significant increase in the thermal nociceptive threshold compared to the groups treated with VA 0.5 mg.Kg-1 (p=0.01), VA 1 mg.Kg-1 (p=0.02) and VA 1.5 mg.Kg-1 (p=0.004). 0060. On the last 14th day of the experiment, a significant reduction in the thermal threshold was observed in the healthy control group compared to the pregabalin group (p=0.0007), VA 0.5 mg.Kg-1 (p=0.003) and VA 1.5 mg.Kg-1 (p=0.02). The lesion group showed a significant reduction compared to the pregabalin group (p<0.0001), VA 0.5 mg.Kg-1 (p=0.0008), VA 1 mg.Kg-1 (p=0.02) and VA 1.5 mg.Kg-1 (p=0.004). Animals treated with the lowest dose of VA 0.5 mg.Kg-1 showed a significant increase compared to animals treated with VA 1 mg.Kg-1 (p=0.01) and VA 1.5 mg.Kg-1 (p=0.02). Cold Sensitivity Test (Acetone Test) 0061. Evaluated cold thermal allodynia in mice, a characteristic clinical sign of neuropathic pain. The test was performed on days 0, 4, and 13 and used the volatile property of acetone to induce a sensation of cold (10 °C). The mice were placed in suspended cages with a floor. Statistical Analysis 0062. For normality analysis, a two-way analysis of variance (ANOVA) was performed. P-values < 0.05 were considered significant. 0063. Experimental treatment with bee venom, especially at doses of 1 mg.Kg-1 and 1.5 mg.Kg-1, appears to be effective. Petition 870250008375, dated 01 / 31 / 2025, page 39 / 99 25 / 25 in reducing hyperalgesia (antinociceptive effect) compared to the lesion group, and its efficacy is comparable to or even superior to that of pregabalin (DALENOGARE et al., 2019). The 1.5 mg.Kg-1 dose showed the best results regarding the reduction of pain sensitivity. 0064. These data reinforce that the venom, in appropriate doses, can be an effective alternative for controlling the symptomatology of fibromyalgia, both related to pain aspects such as hyperalgesia and allodynia, and to emotional aspects such as anxiety, motivation, and volition, with effects comparable to those of pregabalin. Such responses are consistent with what is expected when evaluating the potential pharmacological effects of the active principles present in the experimental treatment (DANTAS et al., 2014). 0065. It is important to point out that no side effects were observed from the use of this substance throughout the experiment, nor were there any animal deaths during the research. Euthanasia and obtaining tissue samples 0066. The animals were euthanized on the 12th day of the experiment by means of an overdose of a combination of dissociative anesthetics (ketamine) and alpha-2 adrenoreceptor agonists (xylazine), administered intraperitoneally. Subsequently, brain tissue samples were removed by dissection and individually placed in containers with formalin solution for processing and analysis at the Laboratory of Morphology and Structural Pathology (LMPE) of the Technological Research Institute (ITP).

Claims

1. USE OF BEE VENOM FOR THE TREATMENT OF FIBROMYALGIA, characterized by using doses of VA 0.5 mg.Kg-1; VA 1.0 mg.Kg-1 and VA 1.5 mg.Kg-1 for the treatment of fibromyalgia.

2. USE OF BEE VENOM FOR THE TREATMENT OF FIBROMYALGIA, according to claim 1, characterized in that the bee venom is from the species Apis mellifera Linnaeus.

3. USE OF BEE VENOM, according to claim 1, characterized by diluting Africanized bee venom in 40 μL of saline solution; 4. USE OF BEE VENOM according to claim 1, characterized by acting on the pathophysiology of diseases, to treat the symptoms of fibromyalgia.

5. USE OF BEE VENOM, according to claim 1, characterized by the dosage of bee venom being VA 0.5 mg. Kg-1, applied subcutaneously, continuously, once every 2 days.

6. USE OF BEE VENOM, according to claim 1, characterized in that the dosage of bee venom is VA 1.0 mg. Kg-1 - applied subcutaneously, continuously, once every 2 days.

7. USE OF BEE VENOM, according to claim 1, characterized by the dosage of bee venom being VA 1.5 mg. Kg-1 - applied subcutaneously, continuously, once every 2 days.

8. USE OF BEE VENOM, according to claim 1, characterized by the dosage of bee venom being VA 1.5 mg. Kg-1 for the reduction of hyperalgesia, applied subcutaneously, continuously, once every 2 days.