WO2018068120A1 - Use of a compound, synthetic intermediate, pharmaceutical composition, and neuromodulatory therapeutic method - Google Patents

Use of a compound, synthetic intermediate, pharmaceutical composition, and neuromodulatory therapeutic method Download PDF

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Publication number
WO2018068120A1
WO2018068120A1 PCT/BR2017/050314 BR2017050314W WO2018068120A1 WO 2018068120 A1 WO2018068120 A1 WO 2018068120A1 BR 2017050314 W BR2017050314 W BR 2017050314W WO 2018068120 A1 WO2018068120 A1 WO 2018068120A1
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WIPO (PCT)
Prior art keywords
compound
hydrogen
pharmaceutical composition
nfkf
preparation
Prior art date
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PCT/BR2017/050314
Other languages
French (fr)
Portuguese (pt)
Inventor
Ricardo Amaral Remer
Andrea Sterman Heimann
Original Assignee
Proteimax Biotecnologia Ltda
Remer Consultores Assessoria Empresarial Ltda
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Priority claimed from BR102016023848-0A external-priority patent/BR102016023848A2/en
Priority claimed from BR102017010169-0A external-priority patent/BR102017010169A2/en
Application filed by Proteimax Biotecnologia Ltda, Remer Consultores Assessoria Empresarial Ltda filed Critical Proteimax Biotecnologia Ltda
Priority to IL273868A priority Critical patent/IL273868B2/en
Publication of WO2018068120A1 publication Critical patent/WO2018068120A1/en
Priority to PCT/BR2018/050156 priority patent/WO2018209415A1/en
Priority to JP2019564034A priority patent/JP7241032B2/en
Priority to CN201880047283.1A priority patent/CN110891963B/en
Priority to US16/612,220 priority patent/US20220098237A1/en
Priority to BR112019002655A priority patent/BR112019002655A2/en
Priority to EP18802073.9A priority patent/EP3626727A4/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/08—Antiepileptics; Anticonvulsants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/12—Drugs for disorders of the metabolism for electrolyte homeostasis
    • A61P3/14—Drugs for disorders of the metabolism for electrolyte homeostasis for calcium homeostasis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/14—Drugs for disorders of the endocrine system of the thyroid hormones, e.g. T3, T4

Definitions

  • the present invention is in the fields of pharmacy, medicine, chemistry and biotechnology. More specifically, the present invention describes a compound and its use for preparing a binder of diagnostic and / or therapeutic interest, a synthetic intermediate in the preparation of compounds of pharmaceutical interest, the use of a compound for preparing a cannabinoid receptor modulating medicament and / or muscarinic, a pharmaceutical composition containing said compound and a therapeutic method.
  • the pharmaceutical composition of the invention comprises a peptidic compound and is particularly useful for the modulation of metabolic function or neuromodulation and may be administered orally.
  • oral administration of the composition of the invention demonstrated significant and surprising results in the curative or prophylactic treatment of seizures.
  • Test results indicate that the compound of the invention interacts with and / or modulates the activity of cannabinoid (CB) receptors, especially CB1 and / or CB2, the pharmaceutical composition of the invention being a promising alternative to the administration of cannabinoid compounds currently known as anticonvulsants, such as cannabidiol.
  • CBD cannabinoid
  • the present invention describes a compound and its use for the preparation of pharmaceutical compositions useful for a varied set of medical conditions.
  • the pharmaceutical composition of the invention is useful under medical conditions related to the central nervous system.
  • the composition of the invention comprises a peptide active
  • oral administration has provided brain effects in animals.
  • in vivo tests with the composition of the invention demonstrated surprising results as to its neuromodulatory activity.
  • the pharmaceutical composition of the invention when administered previously to animals, provided surprising, potent and long protection against damage resulting from the subsequent administration of substances known to be harmful to neurons. Accordingly, neuromodulation or neuroprotection provided by the compound of the invention is particularly useful as a therapeutic alternative for various medical conditions, including those related to neuronal excitability disorders.
  • in vivo tests with the compound of the invention have shown surprising results regarding its anticonvulsant activity.
  • the compound of the invention when used as an anticonvulsant, additionally provides the advantage of being a good candidate to substitute the cannabinoid compounds known for their acting as anticonvulsants, such as Canabidiol.
  • Cannabidiol despite its proven effects as an anticonvulsant, has been facing regulatory problems due to its origin, the Cannabis sativa plant.
  • the present invention provides an additional therapeutic approach for patients suffering from seizures and having difficulty in obtaining drugs, being based on a peptide and not using Cannabis sativa derivatives.
  • the results showed that the compound of the invention, when used as an anticonvulsant, provides other surprising technical advantages in use, including greater therapeutic effect, oral use, lower dosage, less occurrence of side effects such as prostration and nasal bleeding, among others advantages.
  • Pilocarpine (commonly called "Pilo") is an alkaloid extracted from the leaves of the jaborandi plant (Pilocarpus jaborand ⁇ ), a plant used for centuries by the Tupi-Guarani Indians who inhabit Brazil and take advantage of their properties to produce sweat and saliva .
  • Pilocarpine is a non-specific muscarinic agonist, slowly degraded and has no effects on the nicotinic receptors and was introduced in clinical practice by the Brazilian physician Sifrônio Coutinho, in 1874, through extracts of the jaborand ⁇ leaf to obtain diaphoretic effect (sweat production) and silagogue (saliva production).
  • pilocarpine at high concentrations induces the occurrence of convulsions, being used as an experimental model for both.
  • Pilocarpine-induced seizures lead to neurotoxicity at the cellular level and may be related to increased cerebral oxidative stress and changes in the concentration of certain amino acids (Santos et al, 201 1).
  • pilocarpine causes cholinergic alterations capable of inducing status epilepticus (SE) associated with convulsive stereotyped movements.
  • SE status epilepticus
  • Pilo is able to induce epilepticus status both administered directly in the brain and intraperitoneally.
  • Acetylcholine through its muscarinic receptor (mAChRs) plays an important role in cognitive functions, such as learning and memory.
  • MAChRs are receptors that form G protein-receptor complexes on the cell membranes of certain neurons and other cells. They play several roles, including acting as the ultimate end receptor stimulated by acetylcholine released from postganglionic fibers in the parasympathetic nervous system.
  • Muscarinic receptors are so called because they are more sensitive to muscarin than to nicotine. Its counterparts are nicotinic acetylcholine receptors (nAChRs), channels of receptor ions that are also important in the autonomic nervous system. Many drugs and other substances (eg, pilocarpine and scopolamine) manipulate these two distinct receptors acting as selective agonists or antagonists.
  • nAChRs nicotinic acetylcholine receptors
  • Many drugs and other substances eg, pilocarpine and scopolamine
  • the mAChRs are among the most well characterized among the transmembrane receptors (7TM), being widely expressed in the central nervous system (CNS).
  • Seven mAChR subtypes were cloned (M1, M2, M3, M4 and M5) and are generally divided into two distinct classes based on signal transduction.
  • mAChR M1, M3 and M5 are subtypes that signal through Gq / 11 proteins and activate phospholipase-C and mobilize intracellular calcium.
  • M2 and M4 mAChR predominate through Gi / o proteins inhibiting adenylate cyclase and reducing the intracellular concentration of cAMP.
  • the predominant mAChR in the CNS is the M1 subtype, which is located in the cortex, hippocampus, striatum and thalamus, where it is found post-synaptic.
  • the M2 mAChRs are located predominantly in the brainstem and thalamus, but also in the cortex, hippocampus and striatum, where they control the release of acetylcholine.
  • the MAChRs of M3 and M5 are expressed at much lower levels than Mh or Mh MAChRs in the CNS, but Mh MAChRs are found in the cortex and hippocampus, while M5 mAChRs have a very discrete location in the substantia nigra.
  • MAChR M4 are found in many regions of the brain, including the cortex and hippocampus, but are more prominent in the striatum, where they are thought to play a role in the control of dopamine release and modulate locomotor activity.
  • test results presented in the present patent application indicate that the composition of the invention interacts with cannabinoid and / or muscarinic receptor receptors.
  • the results show that the compound of the invention modulates the action of neurons and is also neuroprotective and anticonvulsive.
  • CB cannabinoid
  • the cannabinoid system which comprises the CB1 and CB2 receptors and their endogenous ligands, acts on various metabolic functions, including control of food intake, energy and / or lipid metabolism, regulation of intestinal motility, immune system, in the balance of the calcium cycle, among others.
  • Cannabinoid receptors are widely expressed in the brain, including the cortex, hippocampus, amygdala, pituitary, and hypothalamus.
  • CB receptors, particularly CB1 have already been identified in numerous peripheral organs and tissues, including the thyroid gland, adrenal gland, reproductive organs, adipose tissue, liver, muscle, and gastrointestinal tract.
  • WO 2014/008567 discloses compounds and compositions useful for treating metabolic disorders comprising obesity, diabetes, systemic arterial hypertension (or disease, condition related and / or associated comorbidities); prevention of overweight; regulation of appetite; induction of satiety; prevention of weight gain after successful weight loss; increased energy consumption; aesthetic weight reduction; or bulimia.
  • No report or suggestion of use of the compound of the invention as neuromodulator, neuroprotective or as anticonvulsant is made in said document, because this approach was not imagined by the inventors nor was it obvious from the state of the art.
  • WO 201 01/01187 discloses the use of hemopressin for the treatment of obesity in a subject and further discloses that hemopressin is a compound that binds effectively to the CB1 receptor.
  • the present invention differs from said document, among other reasons, to provide a composition for other therapeutic uses, in addition to revealing other compounds with surprising activity.
  • the test results performed by the inventors have been surprising, especially in regard to neuromodulation, neuroprotection and inhibition of seizures, facts not described or suggested in said document.
  • WO 2013/021 196 discloses the use of hemopressin as an agent that interferes with oligodentine differentiation and is useful as an anti-demyelinating agent.
  • the present invention differs from said document, among other reasons, in that it provides other compounds, other uses and provides benefits when compared to hemopressin, including ease of preparation, improved stability and activity, among other advantages.
  • the test results performed by the inventors have been surprising, especially in regard to neuromodulation, neuroprotection and / or inhibition of the occurrence of seizures with the compounds of the invention, which are neither described nor suggested in said document.
  • MAIOROV V. N .
  • CRIPPEN G. M. Significance of root-mean-square deviation in comparing three-dimensional structures of globular proteins. Journal of Molecular Biology, v. 235, p. 625-634, 1994.
  • MORGAN C.A .
  • HURLEY T. D. Characterization of two distinct structural classes of selective aldehyde dehydrogenase 1 A1 inhibitors. Journal of Medicinal Chemistry, v. 58, p. 1964-1975, 2015.
  • PERTWEE, R. G. The diverse CB1 and CB2 receptor pharmacology of three plant cannabinoids: A9-tetrahydrocannabinol, cannabidiol and ⁇ 9-tetrahydrocannabivarin. British Journal of Pharmacology, v. 153, p. 199-215, 2008.
  • RAMACHANDRAN G. N .
  • RAMAKRISHNAN C
  • SASISEKHARAN V. Stereochemistry of polypeptide chain configurations. Journal of Molecular Biology, v. 7, p. 95-99, 1963.
  • the present invention addresses a number of known problems of the prior art, for example, to provide: a stable peptidic compound, a synthetic intermediate in the preparation of compounds of pharmaceutical interest, use of the compound to prepare a binder of interest and / or neuromodulatory modulatory pharmaceutical composition containing said compound; the use of a medicament for modulating the cannabinoid and / or muscarinic system; the use of the peptidic compound for the preparation of a medicament for the curative or prophylactic treatment of seizures; a molecular entity that provides these and / or other technical effects without the drawbacks arising from the use of the cannabinoid substances such as prostration and nasal bleeding, among others; a pharmaceutical composition useful as a therapeutic alternative for neuromodulation and / or for preventing or treating seizures; a therapeutic method.
  • AA is an amino acid selected from the group consisting of F, W, L, I, V, P, G; AA 2 is hydrogen or an amino acid selected from the group consisting of F, W, L, I, V, P, G;
  • R 1 is hydrogen, the amino acid V, or dipeptide PV
  • R 2 is absent when AA 2 is hydrogen or is hydrogen, the amino acid L, or the dipeptide LS when R 1 is hydrogen,
  • AA is F, W, or L.
  • Ri and R2 are both hydrogen.
  • Embodiments of the peptidic compound of the invention useful in the various objects thereof include one or more peptides selected from the group consisting of consists of: NFK, NWK, NLK, NWK, NLKF, NWKF, NWKF, NWKWL, NWKW, NLKW, NLKW, NWKL, NWKLL, NWKLL, NWKLL, NLKLL, , VNFKF, VNWKW, VNLKL, VNFKFL, VNWKWL, VNLKLL, PVNFKF, PVNWKW, PVNLKL, NFKFLS, NWKWLS, NLKLLS, as well as modified, cyclic, amidated, methylated, PEGylated forms thereof; their salts or combinations thereof.
  • the peptidic compound of the invention is selected from the group comprising tripeptide, NFK, NFKF tetrapeptide, NFKL tetrapeptide, or combinations thereof.
  • the compound of the invention provides improved stability and / or ease of handling, being particularly useful in pharmaceutical and drug preparations.
  • the compound of the invention also provides oral administration to a mammalian animal.
  • the compound of the invention provides advantages in the administration, bioavailability and / or therapeutic action in an animal when compared to other peptides, such as hemopressin and known variants, usually of larger size.
  • Another object of the present invention is a synthetic intermediate in the preparation of compounds of pharmaceutical interest which comprise the compound described above and may include chemical modifications, substitutions, inclusion of other functional groups.
  • Another object of the present invention is the use of the compound described above for the preparation of other compounds of pharmaceutical interest, said compounds comprising the compound described above and chemical modifications, substitutions, other functional groups.
  • Administration of the compound of the invention to a mammal provides neuromodulatory, neuroprotective and / or anticonvulsant activity; enables oral administration in animals; does not have or entails the drawbacks arising from the production, storage, transport and use of cannabinoid substances, in addition to providing additional advantages in the preparation of therapeutic products for mammals in their administration and / or effects.
  • administering provides important and surprising technical advantages, including superior anticonvulsant activity over hemopressin, the use of which as an anticonvulsant is the subject of the co-pending co-pending application of the same inventors.
  • Another object of the present invention is a therapeutic method for metabolic modulation and / or neuromodulation comprising administering to an animal the peptide described above.
  • Another object of the invention is a neuromodulatory, neuroprotective pharmaceutical composition and / or for the curative or prophylactic treatment of seizures in a mammal, said composition comprising a pharmaceutically acceptable carrier; and, as active principle, the compound described above.
  • the pharmaceutical composition is in the form of a tablet, tablet, gel, oral liquid or syrup, capsule, suppository, injectable solution or inhalable or adhesive forms, and may optionally comprise other active principles.
  • Figure 1 shows the results of comparative stability tests of the compound of the invention vs Hp (PVNFKFLSH).
  • Statistical significance data are also shown, the asterisks indicating: ( * ) P ⁇ 0.05 vs Control ( *** ) P ⁇ 0.005 vs Control; ( **** ) P ⁇ 0.0001 vs Control.
  • Figure 2 shows the results of neuroprotection tests with the compound of the invention NFKF in the pilocarpine model, the survival / death profile of the animals being indicated by administration of the peptide of the invention.
  • A) the survival profile of the animals to which the control was administered (saline only) is shown;
  • B) the profile is shown survival rate of the animals administered cannabidiol 30mg / kg;
  • C) the survival profile of the animals to which the compound of the invention was administered NFKF 50C g / kg;
  • all profiles are shown in a single graph. It is interesting to note that in the group treated with the neuromodulator of the invention NFKF 500 ⁇ g / kg only two animals died.
  • Figure 3 shows the test results of the compound of the invention NFKF in the pilocarpine model, indicating the time for the first salivation to occur with the administration of cannabidiol (30mg / kg), or the compound of the invention NFKF (500 ⁇ g / kg).
  • cannabidiol 30mg / kg
  • compound of the invention NFKF 500 ⁇ g / kg
  • Figure 4 shows the test results of the compound of the invention NFKF as an anticonvulsant in the pilocarpine model, indicating the time for the first seizure to occur with the administration of cannabidiol (30mg / kg) or the peptide of the invention NFKF ( 500 ⁇ g / kg).
  • the asterisks indicate the statistical significance: ( ** ) P ⁇ 0.02 vs Control; ( *** ) P ⁇ 0.002 vs Control.
  • Figure 5 shows the test results of the compound of the invention NFKF compared to the results of hemopressin tests, both in the pilocarpine model.
  • the times for the first salivation occur with administration of the following treatment doses: control (saline); hemopressin (Hp or PVNFKFLSH, 0.551334 ⁇ / kg); hemopressin (0.91889 ⁇ / kg); the compound of the invention NFKF (0.540882 ⁇ / kg); NFKF (0.901469 ⁇ / kg); the compound PEP-19 (DIIADDEPLT, 0.9081 17 ⁇ l / kg).
  • the asterisks indicate the statistical significance: ( * ) P ⁇ 0.05 vs Control; ( ** ) P ⁇ 0.01 vs Control; the + sign indicates P ⁇ 0.05 vs Hp 0.91889 ⁇ / kg.
  • Figure 6 shows the test results of the compound of the invention NFKF as an anticonvulsant compared to the results of tests of hemopressin as an anticonvulsant, both in the pilocarpine model.
  • control saline
  • hemopressin Hp or PVNFKFLSH, 0.551334 ⁇ / kg
  • hemopressin 0.0.91889 ⁇ / kg
  • the compound of the invention NFKF 0.540882 ⁇ l-iol / kg
  • NFKF (0.901469 ⁇ - ⁇ / kg)
  • the compound PEP-19 DIIADDEPLT, 0.9081 17 ⁇ l / kg.
  • the asterisks indicate the statistical significance: ( * ) P ⁇ 0.05 vs Control; ( ** ) P ⁇ 0.01 vs Control; the + sign indicates P ⁇ 0.05 vs Hp 0.91889 ⁇ - ⁇ / kg.
  • Figure 7 shows an overview of the three-dimensional structure of a GPCR, in this case, the cannabinoid receptor of subtype 1.
  • Figure 8 shows the overlap of the AM6538 structure of the crystallographic structure (PDB 5TGZ), and the result obtained after validation by the Goldscore function (when the figure appears colored, the crystal structure is purple and the result of the Goldscore function in blue / cyan, although for the purposes of this application such colors are irrelevant).
  • Figure 9 shows the main interactions observed for AM6538 at the CB1 receptor binding site.
  • Figure 10 shows the major interactions observed for rimonabant at the CB1 receptor binding site.
  • Figure 11 shows the major interactions observed for cannabidiol at the CB1 receptor binding site.
  • Figure 12 shows the major interactions observed for the peptide of the invention NFKF at the binding site of the CB1 receptor.
  • Figure 13 shows the results of the process of obtaining CB2 receptor structure.
  • A) the three-dimensional structure of the obtained CB2 receptor is shown; in B) the Ramachandran Graph for the obtained human CB2 model is shown.
  • Figure 14 shows the major interactions between the CB2 receptor and the AM6538 linker.
  • Figure 15 shows the major interactions between the CB2 receptor and rimonabant.
  • Figure 16 shows the major interactions between the CB2 receptor and cannabidiol.
  • Figure 17 shows the major interactions between the CB2 receptor and the NFKF tetrapeptide.
  • the compound of the invention has demonstrated several surprising uses and results in in vitro and in vivo tests.
  • the compound of the invention is useful for the preparation of other compounds of pharmaceutical interest.
  • the compound of the invention is useful for the preparation of a pharmaceutical composition.
  • the pharmaceutical composition of the invention is cannabinoid and / or muscarinic receptor modulator, being particularly useful in modulating metabolic functions and / or neuromodulation of a mammalian animal.
  • AAi is an amino acid selected from the group consisting of F, W, L, I, V, P, G;
  • AA 2 is hydrogen or an amino acid selected from the group consisting of F, W, L, I, V, P, G;
  • R 1 is hydrogen, the amino acid V, or dipeptide PV
  • R 2 is absent when AA 2 is hydrogen or is hydrogen, the amino acid L, or the dipeptide LS when R 1 is hydrogen, and / or modified, cyclic forms thereof, amidated, methylated, PEGylated versions, salts thereof; and / or combinations thereof for the preparation of a composition of pharmaceutical interest.
  • compound of pharmaceutical interest means any molecular entity comprising the compound described as inventive concept common to the present application, including also molecular entities obtained by chemical derivatization thereof, with the inclusion of other functional groups, linear or branched side chains, alteration of hydrophilicity or hydrophobicity, among others, provided that they comprise as nucleus the entity R1-N-AA1-K-AA2-R2 as defined above, with the exception of entities natural and already known.
  • composition is to be understood as any and all compositions containing an active principle, for prophylactic, palliative and / or curative purposes, acting in a manner to maintain and / or restore the homeostasis, and may be administered orally, topically, parenterally, enterally and / or intrathecally.
  • a "pharmaceutically acceptable formulation” is meant a formulation containing pharmaceutically acceptable excipients and carriers well known to those skilled in the art, such as the development of convenient doses and treatments for use in particular compositions which can be described in a series of treatment regimens, including oral, parenteral, intravenous, intranasal, intravitreal and intramuscular, intracerebral, intracerebroventricular and intraocular and their administration and / or formulation.
  • modified peptide is to be understood as a non-naturally occurring, artificially modified or synthesized peptide, including cyclized, amidated, methylated, PEGylated, or salt forms thereof, as well as a peptide comprising one or more unnatural amino acid, such as d-amino acid forms.
  • the peptidic compound may be pegylated using techniques known to those skilled in the art, such as, for example, pegylation with reagents containing the succinimidyl group, which preferentially react with primary amines present in the N-terminal region of the peptide.
  • cyclic, cyclized or" circular peptide is to be understood as a peptide which has a covalent bond between the two ends of a linear peptide molecule by any method known in the art, particularly by the activity of enzymes.Cyclic peptide can be used instead of the linear peptide because it is more difficult to be degraded, since its ends or zones of attack by hydrolyzing enzymes are not as exposed as in a linear peptide.
  • agonist is to be understood as a drug, drug, hormone, neurotransmitter or other signaling molecule which forms a complex with a receptor site, thereby triggering an active response of a cell.
  • inverse agonist or antagonist as agent (s) (for example, drugs, drugs, hormones or enzymes) which binds to agonist receptors and produces pharmacological effects opposite to agonists, in such a way that the action of one partially or totally inhibits the effect of the other.
  • agent for example, drugs, drugs, hormones or enzymes
  • a compound is an inverse agonist when it acts in the presence of an agonist, but reducing its activity; an antagonist is a compound that will totally block the activity of the agonist.
  • the concept of "equivalent dose in humans” is the dose at which, in humans, the same magnitude of effects observed in animals at a given dose is expected, as advocated in Guidance for Industry Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers "published by the US Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and Research (CDER), July 2005 Pharmacology and Toxicology.
  • the conversion of the dose observed in animals (mg / kg) to Equivalent Dose in Humans (mg / kg) implies dividing the result obtained in rats by 6.2 and the result obtained in mice by 12.3. These values apply to a human with 60 kg of standard weight.
  • DEH Equivalent Human Dose
  • the compound of the invention is useful for modulating the cannabinoid system, either by modulating the CB1 receptor, the CB2 receptor, both concomitantly, by modulating the binding or action of other substances interacting in the cannabinoid system, by modulating proteases or peptidases which lead to the generation or degradation of active peptides in the cannabinoid system, or combinations thereof.
  • the term "modulating muscarinic receptor function" should be understood as an interaction leading to neuronal changes, including muscarinic acetylcholine receptor (mAChRs), which plays an important role in cognitive functions, such as such as learning and memory, control of dopamine release, modulation of locomotor activity, its modulation being also useful in the control of Alzheimer's disease and / or control of addiction or addiction to drugs of abuse. It is understood that the change is positive when an antagonist or inverse agonist effect occurs at muscarinic receptors and that the change is negative when an agonist effect occurs at muscarinic receptors.
  • the tests presented in the present patent application suggest that the compound of the invention interacts with and / or modulates muscarinic receptors.
  • this term is understood to include modulation of: energy and / or lipid metabolism; arterial hypertension, regulation of intestinal motility; Imune system; balance of the calcium cycle, conditions associated with the thyroid gland, peripheral organs and tissues, including reproductive organs, adipose tissue, liver, muscles and gastrointestinal tract, being useful in the treatment of obesity, diabetes, diseases or immune / inflammatory disorders, osteopenia, osteoporosis , cancer.
  • neuromodulator or “neuromodulator” is understood as the function of modulating the function neuronal / neurological, including modulation of brain activity, cortex, hippocampus, amygdala, pituitary, hypothalamus; adrenal gland.
  • Neuromodulation includes the beneficial modulation of neuroprotection against agents or conditions leading to pathophysiological processes. Neuroprotective agents or compounds are preferably used prior to (or during) the prodromal stage of disease, which often begins many years before the onset of symptoms.
  • a neuromodulator is potentially useful in the curative or prophylactic treatment of a variety of neurological conditions or diseases, including essential tremor, migraine, neuropathic pain, psychiatric disorders such as anxiety, schizophrenia or bipolar disorder, Alzheimer's, Parkinson's, autism, and is also potentially useful in modifying the pathophysiological processes involved in the occurrence of seizures and / or epilepsy, as well as in other clinical conditions related to disorders of neuronal excitability or neuronal lesions due to ischemia, hypoxia or other harmful conditions.
  • neurological conditions or diseases including essential tremor, migraine, neuropathic pain, psychiatric disorders such as anxiety, schizophrenia or bipolar disorder, Alzheimer's, Parkinson's, autism, and is also potentially useful in modifying the pathophysiological processes involved in the occurrence of seizures and / or epilepsy, as well as in other clinical conditions related to disorders of neuronal excitability or neuronal lesions due to ischemia, hypoxia or other harmful conditions.
  • the surprising pharmaceutical action of the invention may be linked to action on CB1 and / or CB2 and / or muscarinic or possibly linked to uptake modulation adenosine, GGPR55, PPAR ⁇ receptors, intracellular calcium level, or combinations thereof.
  • any therapeutic indication related to these targets may benefit from the present invention.
  • said peptide is selected from the group consisting of: NFK, NWK, NLK, NFKF, NWKF, NLKF, NFKW, NWKW, NLKW, NFKL, NWKL, NLKL, VNFK, VNWK, VNLK, NFKFL , NWKFL, NLKLL, NWKLL, NWKLL, NWKLL, NWKLL, NWKLL, NWKLL, NWKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL
  • a pharmaceutical composition modulating metabolic functions in a mammal comprising a pharmaceutically acceptable carrier; and, as active principle, the compound described above.
  • a curative or prophylactic pneumodulatory pharmaceutical composition in a mammal comprising a pharmaceutically acceptable carrier; and, as active principle, the compound described above.
  • a pharmaceutical composition for the curative or prophylactic treatment of seizures in a mammal comprising a pharmaceutically acceptable carrier; and, as active principle, the compound described above.
  • Neuromodulatory pharmaceutical composition comprising the NFK tripeptide, the NFKF tetrapeptide, the NFKL tetrapeptide, or combinations thereof.
  • a therapeutic modulator of metabolic functions of an animal comprising administering the peptide described above.
  • the peptide of the invention is preferably modified so that its molecular weight is greater, minimizing or preventing the passage thereof through the blood-brain barrier.
  • Therapeutic method for neuromodulation, neuroprotection and / or curative or prophylactic treatment of seizures comprising administering, to an animal, the compound described above.
  • the compound of the invention has been shown to be much more stable than hemopressin, which in addition causes the technical problems of fiber formation or fibrils, as it does with its known larger variants.
  • the neuromodulator / neuroprotective effect is evidenced by the absence of symptoms, brain damage, and deaths associated with the administration of substances known to be harmful, such as pilocarpine.
  • the invention provides the use of said compound for the preparation of a neuromodulatory, neuroprotective medicament and / or for the curative or prophylactic treatment of seizures in a mammal.
  • Administration, to an animal, of the compound of the invention provides neuromodulatory, neuroprotective and / or anticonvulsant activity; makes oral administration feasible; does not have or entails the drawbacks arising from the production, storage, transport and use of cannabinoid substances, in addition to providing additional advantages in the preparation of therapeutic products for mammals in their administration and / or effects.
  • administering the compound of the invention to an animal provides important and surprising technical advantages, including superior anticonvulsant activity with respect to hemopressin, the use of which as an anticonvulsant is the subject of the co-pending co-pending application of the same inventors.
  • the present invention describes the use of a compound for the preparation of pharmaceutical compositions useful for a varied set of medical conditions, including those related to the central nervous system.
  • the active compound of the invention is peptidic or predominantly peptidic, oral administration has provided brain effects in animals.
  • in vivo tests with the composition of the invention demonstrated surprising results regarding its neuroprotective activity.
  • the composition of the invention when administered previously to animals, provided surprising, potent and long protection against damage resulting from the subsequent administration of substances known to be harmful to neurons. Accordingly, the neuroprotection provided by the compound of the invention is particularly useful as a therapeutic alternative for various medical conditions, including those related to disorders of neuronal excitability, such as seizures.
  • in vivo tests with the compound of the invention demonstrated surprising results regarding its anticonvulsant activity.
  • the compound of the invention when used as an anticonvulsant, additionally provides the advantage of being a good candidate to substitute the cannabinoid compounds known for their acting as anticonvulsants, such as Canabidiol.
  • Cannabidiol despite its proven effects as an anticonvulsant, has been facing regulatory problems due to its origin, the Cannabis sativa plant.
  • the present invention provides an additional therapeutic approach for patients suffering from seizures and having difficulty in obtaining drugs, being based on a peptide, ie, does not use derivatives of Cannabis sativa.
  • the results showed that the compound of invention, when used as an anticonvulsant, provides other surprising technical advantages in use, including greater therapeutic effect, oral use, lower dosage, less occurrence of side effects such as prostration and nasal bleeding, among other technical advantages.
  • composition of the invention provides surprising technical advantages in use, including greater therapeutic effect, viability of oral use, lack of use of carrier oil (which in many cases causes side effects), lower dosage and lower occurrence of side effects such as prostration and nasal bleeding, among others.
  • the pharmaceutical composition of the invention is also useful for the treatment of diseases associated with modulation of the activity of the cannabinoid system, cannabinoid (CB) and / or muscarinic receptors - with various technical advantages and without known undesirable effects of the available congeners in the state of the art.
  • the compound as described above is useful for the preparation of a medicament modulator of metabolic functions.
  • the compound as described above is useful for the preparation of a neuromodulator, neuroprotective medicament and / or a medicament for the treatment of seizures.
  • the use of the compound of the invention in the preparation of a neuromodulatory, neuroprotective and / or anticonvulsant drug provides for the delivery of a medicament orally administrable to a mammal.
  • the results of the tests revealed significant brain action, suggesting that administration of the compound of the invention provides that the active element crosses the blood brain barrier.
  • the results show / support the use of the compound of the invention regardless of whether the compound of the invention is the active which acts directly on the target, ie, does not degrade during oral ingestion, or the compound is a precursor which, upon modification chemistry, acts on the target - in this case, being characterized as a pro-drug.
  • the present application discloses a pharmaceutical composition
  • a pharmaceutical composition comprising the compound described above.
  • Said pharmaceutical composition also comprises a pharmaceutically acceptable carrier, optionally also comprising other pharmaceutically acceptable actives and / or salts thereof.
  • the compound of the invention is one or an active component of the pharmaceutical composition of the invention, which is administered in the form of a tablet, gel, capsule, oral liquid or syrup, a suppository, an injectable solution or other suitable forms of administration for pharmaceutical and medical purposes.
  • the neuromodulatory / neuroprotective / anticonvulsive effects of the composition of the invention have been evaluated in vivo by oral administration to animals.
  • the pharmaceutical composition of the invention was administered to mammals (Mus musculus or mouse) with an oral dose of treatment with different embodiments of the compound of the invention, as compared to other compounds or the saline control.
  • the test compounds were administered orally 10 minutes prior to (intraperitoneal) administration of pilocarpine.
  • Pilocarpine hydrochloride 320 mg / kg, Merck
  • dissolved in 0.9% sterile saline was administered intraperitoneally for induction of SE (status epileticus) (Turski et al., 1983).
  • SE status epileticus
  • the neurotoxic effects begin about 15-25 minutes after the injection of Pilo, with the occurrence of motor and limbic seizures, the animals evolving to a state of continuous (clonic) seizures that characterize SE Sanabria and Cavalheiro, 2000).
  • Example 1 Stability tests under extreme conditions
  • Hp hemopressin
  • PVNFKFLSH hemopressin
  • the compound R-N-AA -K-AA 2- F 2 is the NFKF tetrapeptide, which has been synthesized by chemical synthesis.
  • Said peptide was used in the preparation of a liquid pharmaceutical composition for oral use comprising between 2,7x10 "4 Molar said peptide and a pharmaceutically acceptable carrier.
  • said carrier solution is saline, the pharmaceutical composition being a solution for oral use.
  • Said composition was used for oral in vivo administration to mammals according to examples 3-6 below.
  • the pharmaceutical composition is in the form of a tablet, gel, oral liquid or syrup, capsule, suppository, injectable solution or inhalable or adhesive forms, optionally comprising other active principles.
  • Example 3 Neuromodulatory pharmaceutical composition comprising compound NFKF - in vivo test results
  • the neuromodulatory effect of the composition of the invention prepared according to example 2 was evaluated by prior administration of the inventive composition and subsequent administration of pilocarpine to animals.
  • Other test compounds were also evaluated as described below.
  • Administration of pilocarpine leads to severe brain injury, neurotoxicity and usually culminates in the death of the animals.
  • This substance was used in the experiments described below but its harmful neuronal / encephalic effects were inhibited by the prior administration of the composition of the present invention: the vast majority of the animals subjected to these experiments had no symptoms related to brain lesions and survived without apparent damage, in contrast to groups of animals treated with other known substances.
  • FIG. 2 shows the results of neuroprotection tests with the compound of the invention NFKF in the pilocarpine model, the survival / death profile of the animals being indicated by administration of the peptide of the invention.
  • A) the survival profile of the animals to which the control was administered (saline only) is shown;
  • B) the survival profile of the animals administered cannabidiol 30mg / kg is shown;
  • C) the survival profile of the animals to which the peptide of the invention is administered NFKF 500 ⁇ g / kg;
  • all profiles are shown in a single graph.
  • the anticonvulsive effect of the composition of the invention prepared according to example 2 was evaluated by prior administration of the inventive composition and subsequent administration of pilocarpine to animals.
  • Figure 3 presents the results of the pilocarpine model, indicating the time for the first salivation to occur with the administration of cannabidiol (30mg / kg) of the compound of the invention R-N-AA-K-AA 2 -F 2, in which embodiment it is the NFKF tetrapeptide (500 ⁇ g / kg).
  • the data between control and other test compounds do not present statistical significance among themselves under the conditions tested.
  • Figure 4 presents the results of tests with the pilocarpine model, indicating the time for the occurrence of the first convulsion with the administration of cannabidiol (30mg / kg) and the peptide of the invention NFKF (50C gg / kg).
  • the asterisks indicate: ( ** ) P ⁇ 0.02 vs Control; ( *** ) P ⁇ 0.002 vs Control.
  • Example 5 Comparative pharmaceutical composition comprising the compound NFKF with the pharmaceutical composition comprising Hp - in vivo test results
  • Figure 5 shows the test results of the compound of the invention NFKF as compared to the results of hemopressin tests, both in the pilocarpine model.
  • control saline
  • hemopressin Hp or PVNFKFLSH, 0.551334 ⁇ / kg
  • hemopressin 0.0.91889 ⁇ -iol / kg
  • the peptide of the invention NFKF 0.540882 ⁇ - ⁇ / kg
  • NFKF (0.901469 ⁇ / kg
  • PEP-19 DIIADDEPLT, 0.9081 17 ⁇ / kg.
  • the asterisks indicate the statistical significance: ( * ) P ⁇ 0.05 vs Control; ( ** ) P ⁇ 0.01 vs Control; the + sign indicates P ⁇ 0.05 vs Hp 0.91889 ⁇ / kg.
  • salivation induced by administration of pilocarpine is indicative of changes in muscarinic receptors. Consequently, the substantial change in the time profile for the occurrence of the first salivation observed with prior administration of the compound of the invention suggests modulation, either directly or indirectly, of muscarinic receptors.
  • Example 6 Comparative anticonvulsant pharmaceutical composition comprising the compound NFKF with the anticonvulsant pharmaceutical composition comprising Hp - in vivo test results
  • the anticonvulsive effects of the composition of the invention were compared to the pharmaceutical composition containing Hemopressin (Hp or PVNFKFLSH), the use of which as an anticonvulsant is co-pending co-pending application by the same inventors.
  • Hp or PVNFKFLSH Hemopressin
  • Figure 6 shows the test results of the compound of the invention NFKF as an anticonvulsant compared to the results of tests of hemopressin as an anticonvulsant, both in the pilocarpine model.
  • the percentages of time (relative to the control) for the first seizure occur with administration of the following treatment doses: hemopressin (Hp or PVNFKFLSH, 0.551334 ⁇ / kg); hemopressin (0.91889 ⁇ / kg); the peptide of the invention NFKF (0.540882 ⁇ / kg); NFKF (0.901469 ⁇ / kg); PEP-19 (DIIADDEPLT, 0.9081 17 ⁇ / kg) and control (saline).
  • the asterisks indicate: ( * ) P ⁇ 0.05 vs Control; ( ** ) P ⁇ 0.01 vs Control; the + sign indicates P ⁇ 0.05 vs Hp 0.91889 ⁇ / kg.
  • results of Figure 6 clearly show that oral administration of the compound of the present invention provides substantially higher seizure-modulating activity substantially greater than that observed with oral administration of Hp.
  • the same effect is observed with half the dose of the compound of the invention as compared to Hp.
  • the results of the tests performed in examples 3 to 6 above show significant and significant in vivo results, in dosage ranges of the order of 500 to 1000 ⁇ g of compound of the invention per kg of animal. Considering the tests in mice and the conversion to the Human Equivalent Dose mentioned above, effects of the same magnitude are expected in humans in the dosage range of 40 to 80 ⁇ g of compound of the invention per kg of human and, Safety ranges, concentrations between 4 to 800 ⁇ g / kg for administration to humans are considered in the present invention.
  • Example 7 In silico liqation tests / interaction of compound NFKF to CB1 receptor
  • the cannabinoid receptor 1 corresponds to GPCR (Expressed Receptors to Protein G) most expressed in the human brain and is found at high levels in the Central Nervous System in general ( Figure 7). It is activated by endocannabinoids and has been indicated as a promising therapeutic target for the treatment of various diseases such as pain and inflammation, multiple sclerosis and neurodegenerative diseases (agonist effect) and obesity, liver fibrosis and nicotine dependence (antagonistic effect) et al., 2016).
  • Figure 7 shows an overview of the three-dimensional structure of a GPCR, in this case, the cannabinoid receptor of subtype 1.
  • Table 1 RMSD values obtained from the validation of the scoring functions of the GOLD v. 5.5 for the redocking of AM6538 in the CB1 receptor.
  • Figure 8 shows visually the overlap between the crystallographic structure complex (PDB 5TGZ) and the resulting complex from the AM6538 redocking experiment using the validated methodology.
  • the resulting docking complex of the NFKF tetrapeptide in CB1 obtained a score of 100.66, substantially greater than the score achieved by the reference ligands. This fact can be explained by additional hydrogen bonds observed between the tetrapeptide and residues Ser-123, Thr-197, Ser-167 and Ser-383. In addition, the hydrophobic interactions observed for AM6538 are also present in the NFKF interaction mode ( Figure 12).
  • Example 8 In silico liqation tests / interaction of compound NFKF to CB2 receptor
  • the construction of the 3D model of CB2 was done from a search in the UniProt database of the amino acid sequence of this receptor.
  • the criterion used for sequence selection was the species (Homo sapiens).
  • the sequence selected for carrying out the molecular modeling studies was that of code P34972.
  • the Template Identification tool of the SwissModel server was used to identify and select the protein template.
  • the sequence identified by the server as having the highest structural identity to the human CB2 sequence was that of PDT 5TGZ code (HUA et al., 2016) belonging to the species Homo sapiens, corresponding to the CB1 receptor.
  • the target and template sequences were then aligned using ClustalW2 software linked to the UniProt database to compare the sequences to establish the percentage of identity between them.
  • the 3D homology model of human CB2 was constructed using the Automated Mode tool available on the Swiss-model server page and, for the validation of the generated model, were analyzed the overall structural quality and the stereochemical quality of the model through the value presented for the GMQE parameter and the analysis of the Ramachandran graph.
  • the stereochemical quality of the model was analyzed using the Ramachandran graph (Figure 13B).
  • the Ramachandran graph corresponds to a mathematical model, which relates the dihedral angles ⁇ (angle C-N-Ca-C) on the x-axis and ⁇ (angle N-Ca-C-N) on the y-axis.
  • This graph is divided into regions capable of representing the probability of a combination between the angles ⁇ and ⁇ (RAMACHANDRAN; RAMAKRISHNAN; SASISEKHARAN, 1963). These regions are classified as: favorable, permitted, generously permitted and prohibited.
  • the Ramachandran graph constructed for the human CB2 model showed the presence of approximately 96% of the amino acid residues in the most favorable regions and more than 4% of the residues in acceptable regions. No amino acid residue was found in forbidden regions, indicating the stereochemical validation of the model created.
  • the docking studies on CB2 of the NFKF tetrapeptide, AM6538 antagonist, the cannabidiol endogenous ligand and the rimonabant antagonist were performed.
  • Table 2 summarizes the comparison between the scores obtained by the docking studies on both receptors (CB1 and CB2) in examples 7 and 8 above:
  • Table 2 Comparison between the scores obtained in the docking studies for CB1 and CB2 receptors.
  • the results of the in silico experiments of examples 7 and 8 indicate that the NFKF tetrapeptide is a CB1 receptor linker.
  • CB2 receptors less favorable interactions of this compound were evidenced in CB2 receptors, indicating a possible selectivity profile.
  • the results of the experiments of examples 7 and 8, especially those shown in Figures 12 and 17, show that NFK amino acids participate much more strongly in the binding of CB1 and CB2 than the amino acid F in the C-terminal position, indicating that this tripeptide is a potential candidate for ligand / modulator of such receptors.
  • binding techniques were used to measure the affinity of the NFKF tetrapeptide (powder in 100% purity and prepared as stock solution 10mM in DMSO) by the cannabinoid receptor CB1 at the concentrations of 1 and 10 ⁇ .
  • Results are expressed as the percentage of specific control binding according to the formula:
  • IC50 values concentration that causes half of maximum inhibition of specific control binding
  • Hill (nH) coefficients were determined by nonlinear regression of the competition curves generated with the mean values of the replicates using the Hill equation .
  • results showing inhibition or stimulation greater than 50% are considered to represent significant effect of the test compound. Results showing inhibition or stimulation between 25% and 50% are indicative of low to moderate effect of the test compound. Results showing inhibition or stimulation of less than 25% can be considered as minor. Results showing inhibition greater than or equal to 50% are indicative non-specific or allosteric effects of the test compound.
  • Example 10 In vitro in vitro binding of the compound NFKF to the CB2 receptor
  • the in vitro affinity profile of the compound of the invention R-N-AA-K-AA 2 -R 2 , in which is the NFKF tetrapeptide, is evaluated with the cannabinoid receptor CB2.
  • binding techniques were used to measure the affinity of the NFKF tetrapeptide (powder in 100% purity and prepared as stock solution 10mM in DMSO) by the cannabinoid receptor CB2 at concentrations of 1 and 10 ⁇ .
  • in vitro test results demonstrate that the compound of the invention can be used as a binder of diagnostic interest, for example radioactive labeling or with chromophores for subsequent identification of the sites of binding in cells and / or tissues.
  • the inventive concept herein disclosed and exemplified in one or more ways was treated as an industrial secret and was not previously disclosed until the filing of this patent application.
  • This industrial secret is immaterial asset of the depositor.
  • the possible future publication of the patent application does not in itself constitute authorization for use by third parties, serving only as: (i) scientific knowledge to third parties of the existence of said industrial secret at the time of filing; (ii) unequivocal indication of its holder; and (iii) stimulating the development of new improvements based on the concept to avoid reinvestment in the development of the same asset already held by the depositor.

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Abstract

The present invention relates to the fields of chemistry, pharmacy, biotechnology and medicine. More particularly, the present invention describes: a compound; the use of said compound; a synthetic intermediate in the preparation of compounds of pharmaceutical interest; the use of the compound to prepare a binder of diagnostic and/or therapeutic interest; the use of the compound to prepare a metabolic modulator drug or a neuromodulator drug; a pharmaceutical composition containing the claimed peptide compound; and a therapeutic method. The claimed compound binds to and/or modulates the activity of cannabinoid (CB) receptors, particularly CB1 and/or CB2, and/or muscarinic receptors, and has proven to be extremely useful for modulating the respective systems. The claimed composition is useful for modulating metabolic function and/or for neuromodulation. In one embodiment, the claimed compound provides excellent neuroprotective and anticonvulsant results when administered orally to a mammal, and, in addition to other advantages and technical effects, does not entail the drawbacks arising from the use of cannabinoid substances such as cannabidiol, and provides a stronger therapeutic effect in a smaller dosage and with fewer side effects.

Description

Relatório Descritivo de Patente de Invenção  Patent Specification Report
Uso de Composto, Intermediário de Síntese, Composição Farmacêutica e  Use of Compound, Synthesis Intermediate, Pharmaceutical Composition and
Método terapêutico neuromodulador  Neuromodulatory therapeutic method
Campo da Invenção Field of the Invention
[0001] A presente invenção situa-se nos campos da farmácia, medicina, química e biotecnologia. Mais especificamente, a presente invenção descreve um composto e seu uso para preparar um ligante de interesse diagnóstico e/ou terapêutico, um intermediário de síntese na preparação de compostos de interesse farmacêutico, o uso de um composto para preparar um medicamento modulador de receptores canabinoides e/ou muscarínicos, uma composição farmacêutica contendo o referido composto e um método terapêutico. A composição farmacêutica da invenção compreende um composto peptídico e é particularmente útil para a modulação de função metabólica ou neuromodulação e pode ser administrada por via oral. Em uma concretização, a administração oral da composição da invenção demonstrou importantes e surpreendentes resultados no tratamento curativo ou profilático de convulsões. Resultados de testes indicam que o composto da invenção interage com e/ou modula a atividade de receptores canabinoides (CB), especialmente CB1 e/ou CB2, a composição farmacêutica da invenção sendo uma promissora alternativa à administração de compostos canabinoides atualmente conhecidos como anticonvulsivantes, como, por exemplo, o canabidiol.  The present invention is in the fields of pharmacy, medicine, chemistry and biotechnology. More specifically, the present invention describes a compound and its use for preparing a binder of diagnostic and / or therapeutic interest, a synthetic intermediate in the preparation of compounds of pharmaceutical interest, the use of a compound for preparing a cannabinoid receptor modulating medicament and / or muscarinic, a pharmaceutical composition containing said compound and a therapeutic method. The pharmaceutical composition of the invention comprises a peptidic compound and is particularly useful for the modulation of metabolic function or neuromodulation and may be administered orally. In one embodiment, oral administration of the composition of the invention demonstrated significant and surprising results in the curative or prophylactic treatment of seizures. Test results indicate that the compound of the invention interacts with and / or modulates the activity of cannabinoid (CB) receptors, especially CB1 and / or CB2, the pharmaceutical composition of the invention being a promising alternative to the administration of cannabinoid compounds currently known as anticonvulsants, such as cannabidiol.
Antecedentes da Invenção Background of the Invention
[0002] A presente invenção descreve um composto e seu uso para a preparação de composições farmacêuticas úteis para um variado conjunto de condições médicas. Em uma concretização, a composição farmacêutica da invenção é útil em condições médicas relacionadas ao sistema nervoso central. Surpreendentemente, embora a composição da invenção compreenda um ativo peptídico, a administração oral proporcionou efeitos cerebrais em animais. [0003] Em uma concretização, testes in vivo com a composição da invenção demonstraram resultados surpreendentes quanto à sua atividade neuromoduladora. A composição farmacêutica da invenção, quando administrada previamente a animais, proporcionou surpreendente, potente e longa proteção contra danos decorrentes da subsequente administração de substâncias sabidamente danosas aos neurónios. Assim sendo, a neuromodulação ou neuroproteção proporcionada pelo composto da invenção é particularmente útil como alternativa terapêutica para diversas condições médicas, incluindo aquelas relacionadas a distúrbios de excitabilidade neuronal. The present invention describes a compound and its use for the preparation of pharmaceutical compositions useful for a varied set of medical conditions. In one embodiment, the pharmaceutical composition of the invention is useful under medical conditions related to the central nervous system. Surprisingly, although the composition of the invention comprises a peptide active, oral administration has provided brain effects in animals. In one embodiment, in vivo tests with the composition of the invention demonstrated surprising results as to its neuromodulatory activity. The pharmaceutical composition of the invention, when administered previously to animals, provided surprising, potent and long protection against damage resulting from the subsequent administration of substances known to be harmful to neurons. Accordingly, neuromodulation or neuroprotection provided by the compound of the invention is particularly useful as a therapeutic alternative for various medical conditions, including those related to neuronal excitability disorders.
[0004] Em várias concretizações, testes in vivo com o composto da invenção demonstraram resultados surpreendentes quanto à sua atividade anticonvulsivante. O composto da invenção, quando usado como anticonvulsivante, adicionalmente proporciona a vantagem de ser um bom candidato a substituto dos compostos canabinoides conhecidos por sua atuação como anticonvulsivantes, como é o caso o Canabidiol. O Canabidiol, apesar de seus efeitos comprovados como anticonvulsivante, tem enfrentando problemas regulatorios devido à sua origem, a planta Cannabis sativa. A presente invenção proporciona uma adicional abordagem terapêutica para os pacientes que sofrem de convulsões e que têm dificuldade de obter medicamentos, sendo baseada em um peptídeo e não usa derivados da Cannabis sativa. Além disso, os resultados mostraram que o composto da invenção, quando usado como anticonvulsivante, proporciona outras surpreendentes vantagens técnicas no uso, incluindo maior efeito terapêutico, uso oral, menor dosagem, menor ocorrência de efeitos colaterais como a prostração e sangramento nasal, dentre outras vantagens técnicas.  In various embodiments, in vivo tests with the compound of the invention have shown surprising results regarding its anticonvulsant activity. The compound of the invention, when used as an anticonvulsant, additionally provides the advantage of being a good candidate to substitute the cannabinoid compounds known for their acting as anticonvulsants, such as Canabidiol. Cannabidiol, despite its proven effects as an anticonvulsant, has been facing regulatory problems due to its origin, the Cannabis sativa plant. The present invention provides an additional therapeutic approach for patients suffering from seizures and having difficulty in obtaining drugs, being based on a peptide and not using Cannabis sativa derivatives. In addition, the results showed that the compound of the invention, when used as an anticonvulsant, provides other surprising technical advantages in use, including greater therapeutic effect, oral use, lower dosage, less occurrence of side effects such as prostration and nasal bleeding, among others advantages.
[0005] A pilocarpina (comumente chamada "Pilo") é um alcalóide extraído das folhas da planta jaborandi {Pilocarpus jaborandí), planta usada há séculos pelos índios Tupi-Guarani que habitam o Brasil e tiram proveito de suas propriedades de produzir suor e saliva. A pilocarpina é um agonista muscarínico não específico, lentamente degradado e sem efeitos sobre os receptores nicotínicos e foi introduzida na prática clínica pelo médico brasileiro Sifrônio Coutinho, em 1874, através de extratos da folha do jaborandí para obter efeito diaforético (produção de suor) e silagogo (produção de saliva). Pilocarpine (commonly called "Pilo") is an alkaloid extracted from the leaves of the jaborandi plant (Pilocarpus jaborandí), a plant used for centuries by the Tupi-Guarani Indians who inhabit Brazil and take advantage of their properties to produce sweat and saliva . Pilocarpine is a non-specific muscarinic agonist, slowly degraded and has no effects on the nicotinic receptors and was introduced in clinical practice by the Brazilian physician Sifrônio Coutinho, in 1874, through extracts of the jaborandí leaf to obtain diaphoretic effect (sweat production) and silagogue (saliva production).
[0006] A despeito de suas propriedades farmacêuticas, a pilocarpina em elevadas concentrações induz a ocorrência de convulsões, sendo usada como um modelo experimental para tanto. As convulsões induzidas pela pilocarpina levam à neurotoxicidade em nível celular e podem ser relacionadas ao aumento do estresse oxidativo cerebral e das alterações na concentração de certos aminoácidos (Santos et al, 201 1 ). Despite its pharmaceutical properties, pilocarpine at high concentrations induces the occurrence of convulsions, being used as an experimental model for both. Pilocarpine-induced seizures lead to neurotoxicity at the cellular level and may be related to increased cerebral oxidative stress and changes in the concentration of certain amino acids (Santos et al, 201 1).
[0007] A administração de pilocarpina neste modelo experimental leva a lesões encefálicas severas, neurotoxicidade e normalmente culmina na morte dos animais. Entretanto, antes deste desfecho, a pilocarpina acarreta alterações colinérgicas capazes de induzir o status epilepticus (SE) associado a movimentos estereotipados convulsivos. A Pilo é capaz de induzir status epilepticus tanto administrada diretamente no encéfalo como por via intraperitoneal. Esta substância foi usada em alguns experimentos descritos neste pedido de patente, seus efeitos neuronais/encefálicos danosos tendo sido inibidos pela composição da presente invenção: a vasta maioria dos animais submetidos a estes experimentos não apresentou sintomas relacionados às lesões cerebrais e sobreviveu sem danos aparentes, em contraste aos grupos de animais tratados com outras substâncias conhecidas.  Administration of pilocarpine in this experimental model leads to severe brain injury, neurotoxicity and usually culminates in the death of animals. However, prior to this outcome, pilocarpine causes cholinergic alterations capable of inducing status epilepticus (SE) associated with convulsive stereotyped movements. Pilo is able to induce epilepticus status both administered directly in the brain and intraperitoneally. This substance was used in some experiments described in this patent application, its harmful neuronal / encephalic effects having been inhibited by the composition of the present invention: the vast majority of animals subjected to these experiments had no symptoms related to brain lesions and survived without apparent damage, in contrast to groups of animals treated with other known substances.
[0008] Grande parte do entendimento sobre os mecanismos das epilepsias vem de estudos em modelos experimentais animais, principalmente em ratos e camundongos. Neste contexto, a administração de pilocarpina em roedores mimetiza a epilepsia (ELT) de humanos e recebe geralmente o nome de "modelo da pilocarpina". Esse modelo foi desenvolvido em 1983 por Turski e colaboradores, e é hoje um dos mais utilizados modelos de epilepsia, tendo em vista que suas características histológicas, bioquímicas, farmacológicas, eletrofisiológicas e comportamentais (Turski et al., 1983) reproduzem similarmente as encontradas em humanos portadores de ELT. [0009] O modelo da pilocarpina também é útil para evidenciar alterações nos receptores muscarínicos, como é o caso da salivação. A acetilcolina, através de seu receptor muscarínico (mAChRs) desempenha um papel importante nas funções cognitivas, tais como a aprendizagem e memória. Os mAChRs são receptores que formam complexos proteína-receptor G nas membranas celulares de certos neurónios e outras células. Eles desempenham vários papéis, incluindo atuar como o principal receptor final estimulado pela acetilcolina liberada a partir de fibras pós-ganglionares no sistema nervoso parassimpático. [0008] Much of the understanding of the mechanisms of epilepsy comes from studies in experimental animal models, especially in rats and mice. In this context, the administration of pilocarpine in rodents mimics epilepsy (ELT) in humans and is generally referred to as the "pilocarpine model". This model was developed in 1983 by Turski et al., And is now one of the most widely used models of epilepsy, considering that its histological, biochemical, pharmacological, electrophysiological and behavioral characteristics (Turski et al., 1983) similarly reproduce those found in human carriers of ELT. The pilocarpine model is also useful for evidence of changes in muscarinic receptors, such as salivation. Acetylcholine, through its muscarinic receptor (mAChRs) plays an important role in cognitive functions, such as learning and memory. MAChRs are receptors that form G protein-receptor complexes on the cell membranes of certain neurons and other cells. They play several roles, including acting as the ultimate end receptor stimulated by acetylcholine released from postganglionic fibers in the parasympathetic nervous system.
[0010] Os receptores muscarínicos são assim chamados porque são mais sensíveis à muscarina do que à nicotina. Suas contrapartes são receptores nicotínicos de acetilcolina (nAChRs), canais de íons receptores que também são importantes no sistema nervoso autónomo. Muitas drogas e outras substâncias (por exemplo, pilocarpina e escopolamina) manipulam estes dois receptores distintos atuando como agonistas ou antagonistas seletivos.  Muscarinic receptors are so called because they are more sensitive to muscarin than to nicotine. Its counterparts are nicotinic acetylcholine receptors (nAChRs), channels of receptor ions that are also important in the autonomic nervous system. Many drugs and other substances (eg, pilocarpine and scopolamine) manipulate these two distinct receptors acting as selective agonists or antagonists.
[0011] Os mAChRs estão entre os mais bem caracterizados dentre os receptores transmembranares (7TM), sendo amplamente expressos no sistema nervoso central (SNC). Cinco subtipos mAChR foram clonados (M1 , M2, M3, M4 e M5) e geralmente são divididos em duas classes distintas com base na transdução de sinal. mAChR M1 , M3 e M5 são subtipos que sinalizam através de proteínas Gq/1 1 e ativam a fosfolipase-C e mobilizam cálcio intracelular. Os mAChR M2 e M4, no entanto, predominam através de proteínas Gi/o inibindo a adenilato ciclase e reduzindo a concentração intracelular de AMPc. O mAChR predominante no SNC é o subtipo M1 , que está localizado no córtex, hipocampo, estriado e tálamo, onde é encontrado pós-sináptico. Os mAChR M2 estão localizados predominantemente no tronco encefálico e no tálamo, embora também no córtex, hipocampo e estriado, onde controlam a liberação de acetilcolina. Os MAChR de M3 e M5 são expressos em níveis muito mais baixos do que MAChRs M1 ou M2 no SNC, mas M3 MAChRs são encontrados no córtex e hipocampo, enquanto que M5 mAChRs têm uma localização muito discreta na substância negra. Os MAChR M4 são encontrados em muitas regiões do cérebro, incluindo o córtex e o hipocampo, mas são mais proeminentes no estriado, onde se pensa que eles desempenham um papel no controle da liberação de dopamina e modulem a atividade locomotora. The mAChRs are among the most well characterized among the transmembrane receptors (7TM), being widely expressed in the central nervous system (CNS). Five mAChR subtypes were cloned (M1, M2, M3, M4 and M5) and are generally divided into two distinct classes based on signal transduction. mAChR M1, M3 and M5 are subtypes that signal through Gq / 11 proteins and activate phospholipase-C and mobilize intracellular calcium. M2 and M4 mAChR, however, predominate through Gi / o proteins inhibiting adenylate cyclase and reducing the intracellular concentration of cAMP. The predominant mAChR in the CNS is the M1 subtype, which is located in the cortex, hippocampus, striatum and thalamus, where it is found post-synaptic. The M2 mAChRs are located predominantly in the brainstem and thalamus, but also in the cortex, hippocampus and striatum, where they control the release of acetylcholine. The MAChRs of M3 and M5 are expressed at much lower levels than Mh or Mh MAChRs in the CNS, but Mh MAChRs are found in the cortex and hippocampus, while M5 mAChRs have a very discrete location in the substantia nigra. MAChR M4 are found in many regions of the brain, including the cortex and hippocampus, but are more prominent in the striatum, where they are thought to play a role in the control of dopamine release and modulate locomotor activity.
[0012] Dado o amplo e variado perfil de expressão dos mAChRs no SNC, não é surpreendente que todos os subtipos tenham sido avaliados como potenciais alvos de fármacos. Alguns desses alvos são bem aceitos na literatura, como o envolvimento do M1 na doença de Alzheimer, enquanto outros são relativamente novos, como o M5 que tem sido correlacionado à dependência e ao vício de drogas de abuso. Given the wide and varied expression profile of mAChRs in the CNS, it is not surprising that all subtypes have been evaluated as potential drug targets. Some of these targets are well accepted in the literature, such as the involvement of M1 in Alzheimer's disease, while others are relatively new, such as M5 that has been correlated to addiction and addiction to drugs of abuse.
[0013] Os resultados dos testes apresentados no presente pedido de patente indicam que a composição da invenção interage com receptores canabinoides e/ou receptores muscarínicos. Os resultados mostram que o composto da invenção modula a ação de neurónios, sendo também neuroprotetor e anticonvulsivante.  The test results presented in the present patent application indicate that the composition of the invention interacts with cannabinoid and / or muscarinic receptor receptors. The results show that the compound of the invention modulates the action of neurons and is also neuroprotective and anticonvulsive.
[0014] Dados experimentais indicam que o composto da invenção interage com e/ou modula a atividade de receptores canabinoides (CB). O sistema canabinóide, que compreende os receptores CB1 e CB2 e seus ligantes endógenos, atua em diversas funções metabólicas, incluindo o controle de ingestão de alimentos, o metabolismo de energia e/ou de lipídeos, na regulação da motilidade intestinal, no sistema imune, no equilíbrio do ciclo do cálcio, entre outros. Os receptores canabinoides são amplamente expressos no cérebro, incluindo córtex, hipocampo, amígdala, pituitário e hipotálamo. Os receptores CB, particularmente CB1 , já foram identificados em inúmeros órgãos periféricos e tecidos, incluindo glândula tiroidal, glândula adrenal, órgãos reprodutivos, tecido adiposo, fígado, músculos e trato gastrointestinal.  Experimental data indicate that the compound of the invention interacts with and / or modulates the activity of cannabinoid (CB) receptors. The cannabinoid system, which comprises the CB1 and CB2 receptors and their endogenous ligands, acts on various metabolic functions, including control of food intake, energy and / or lipid metabolism, regulation of intestinal motility, immune system, in the balance of the calcium cycle, among others. Cannabinoid receptors are widely expressed in the brain, including the cortex, hippocampus, amygdala, pituitary, and hypothalamus. CB receptors, particularly CB1, have already been identified in numerous peripheral organs and tissues, including the thyroid gland, adrenal gland, reproductive organs, adipose tissue, liver, muscle, and gastrointestinal tract.
[0015] Diversos compostos já foram detectados na técnica possuindo atividade de modulação desses receptores. Entre estes, alguns têm como alvo o desenvolvimento de fármacos para redução de peso e afinamento da cintura, como o rimonabanto. Entretanto, esse composto foi subsequentemente associado ao aumento da ocorrência de doenças psiquiátricas em humanos e foi removido do mercado mundial. [0016] As buscas de antecedentes revelaram documentos apenas parcialmente relevantes para a presente invenção. Tais documentos serão descritos abaixo, sendo os mesmos colocados aqui unicamente com o objetivo de servir de base ao estado da técnica atual, uma vez que nenhum deles antecipa ou sequer sugere qualquer dos objetos da presente invenção. Several compounds have already been detected in the art having modulation activity of these receptors. Among these, some target the development of drugs for weight reduction and thinning of the waist, such as rimonabant. However, this compound was subsequently associated with increased occurrence of psychiatric diseases in humans and was removed from the world market. Background searches have disclosed only partially relevant documents for the present invention. Such documents will be described below, and are set forth herein solely for the purpose of serving as a basis for the state of the art, since none of them anticipate or even suggest any of the objects of the present invention.
[0017] O documento US 2007/213302 apresenta compostos de interação com o receptor CB1 , consistindo de pirazóis e seus sais farmaceuticamente aceitáveis que atuam como antagonistas ou agonistas inversos do receptor CB1 . A presente invenção difere do referido documento, dentre outros motivos, por apresentar um composto que compreende um peptídeo, que não compreende pirazóis ou seus sais, e cujos resultados mostram-se surpreendentes, especialmente em relação à neuromodulação, neuroproteção e à inibição da ocorrência de convulsões, fatos não descritos e nem sugeridos no referido documento. US 2007/213302 discloses CB1 receptor interaction compounds, consisting of pyrazoles and their pharmaceutically acceptable salts which act as antagonists or inverse agonists of the CB1 receptor. The present invention differs from said document, among other reasons, by having a compound comprising a peptide, which does not comprise pyrazoles or their salts, and the results of which are surprising, especially in relation to neuromodulation, neuroprotection and inhibition of the occurrence of convulsions, facts not described or suggested in said document.
[0018] Alguns estudos apresentados por um dos presentes inventores em "Novel Natural Peptide Substrates for Endopeptidase 24.15 Neurolysin, and Angiotensin-converting Enzyme" (Vanessa Rioli, Fabio C. Gozzo, Andrea S. Heimann, Alessandra Linardi, José E. Krieger, Cláudio S. Shida, Paulo C. Almeida, Stephen Hyslop, Marcos N. Eberlin, Emer S. Ferro; 7 de Março de 2003) demonstram uma técnica eficaz de "triagem" de novos peptídeos. A presente invenção difere desse documento, dentre outras razões, por apresentar uma nova abordagem terapêutica para a neuromodulação, neuroproteção e/ou para as convulsões, cujos resultados mostraram-se surpreendentes.  Some studies presented by one of the present inventors in Novel Novel Peptide Substrates for Endopeptidase 24.15 Neurolysin and Angiotensin Converting Enzyme (Vanessa Rioli, Fabio C. Gozzo, Andrea S. Heimann, Alessandra Linardi, Joseph E. Krieger, Cláudio S. Shida, Paulo C. Almeida, Stephen Hyslop, Marcos N. Eberlin, Emer S. Ferro, March 7, 2003) demonstrate an effective technique of "screening" of new peptides. The present invention differs from this document, among other reasons, because it presents a new therapeutic approach for neuromodulation, neuroprotection and / or seizures, the results of which have been surprising.
[0019] O documento WO 2014/008567, de um dos presentes inventores, revela compostos e composições úteis para o tratamento desordens metabólicas compreendendo obesidade, diabetes, hipertensão arterial sistémica (ou doença, estado relacionado e/ou comorbidades associadas); prevenção de sobrepeso; regulação do apetite; indução de saciedade; prevenção de ganho de peso após perda de peso com sucesso; aumento do consumo de energia; redução de peso estética; ou bulimia. Nenhum relato ou sugestão de uso do composto da invenção como neuromodulador, neuroprotetor ou como anticonvulsivante é feito no referido documento, até porque esta abordagem não havia sido imaginada pelos inventores nem era óbvia do estado da técnica. WO 2014/008567, one of the present inventors, discloses compounds and compositions useful for treating metabolic disorders comprising obesity, diabetes, systemic arterial hypertension (or disease, condition related and / or associated comorbidities); prevention of overweight; regulation of appetite; induction of satiety; prevention of weight gain after successful weight loss; increased energy consumption; aesthetic weight reduction; or bulimia. No report or suggestion of use of the compound of the invention as neuromodulator, neuroprotective or as anticonvulsant is made in said document, because this approach was not imagined by the inventors nor was it obvious from the state of the art.
[0020] O documento WO 201 1 /01 1847 revela o uso da hemopressina para o tratamento da obesidade em um indivíduo e adicionalmente revela que hemopressina é um composto que se liga eficazmente ao receptor CB1 . A presente invenção difere do referido documento, dentre outros motivos, por proporcionar uma composição para outros usos terapêuticos, além de revelar outros compostos com atividade surpreendente. Além disso, os resultados de testes realizados pelos inventores mostraram-se surpreendentes, especialmente em relação à neuromodulação, neuroproteção e à inibição da ocorrência de convulsões, fatos não descritos e nem sugeridos no referido documento. WO 201 01/01187 discloses the use of hemopressin for the treatment of obesity in a subject and further discloses that hemopressin is a compound that binds effectively to the CB1 receptor. The present invention differs from said document, among other reasons, to provide a composition for other therapeutic uses, in addition to revealing other compounds with surprising activity. In addition, the test results performed by the inventors have been surprising, especially in regard to neuromodulation, neuroprotection and inhibition of seizures, facts not described or suggested in said document.
[0021] O documento WO 2013/021 196 revela o uso da hemopressina como agente que interfere na diferenciação de oligodentritos, sendo útil como agente anti-desmielinizante. A presente invenção difere do referido documento, dentre outros motivos, por proporcionar revelar outros compostos, outros usos e por proporcionar benefícios quando em comparação à hemopressina, incluindo mais facilidade de preparo, maior estabilidade e atividade, entre outras vantagens. Além disso, os resultados de testes realizados pelos inventores mostraram-se surpreendentes, especialmente em relação à neuromodulação, neuroproteção e/ou à inibição da ocorrência de convulsões com os compostos da invenção, que não são descritos nem sugeridos no referido documento.  WO 2013/021 196 discloses the use of hemopressin as an agent that interferes with oligodentine differentiation and is useful as an anti-demyelinating agent. The present invention differs from said document, among other reasons, in that it provides other compounds, other uses and provides benefits when compared to hemopressin, including ease of preparation, improved stability and activity, among other advantages. Furthermore, the test results performed by the inventors have been surprising, especially in regard to neuromodulation, neuroprotection and / or inhibition of the occurrence of seizures with the compounds of the invention, which are neither described nor suggested in said document.
[0022] Outras referências de literatura científica que circunscrevem a invenção, porém sem antecipá-la ou sugeri-la, incluem os seguintes documentos. Other references of scientific literature which circumscribe the invention, but without anticipating or suggesting it, include the following documents.
[0023] ASPRONI, B. et al. Novel pyrrolocycloalkylpyrazole analogues as CB1 ligands. Chemical Biology and Drug Design, p. 1 -13, 2017. [0023] ASPRONI, B. et al. Novel pyrrolocycloalkylpyrazole analogues as CB1 ligands. Chemical Biology and Drug Design, p. 1 -13, 2017.
[0024] HILDEBRANDT, A. K. et al. Efficient computation of root mean square deviations under rigid transformations. Journal of Computational Chemistry, v. 35, p. 765-771 , 2014. [0025] HUA T. et al. Crystal structure of the human cannabinoid receptor CB1 . Celi, v. 167, p. 750-762, 2016. HILDEBRANDT, AK et al. Efficient computation of root mean square deviations under rigid transformations. Journal of Computational Chemistry, v. 35, p. 765-771, 2014. [0025] HUA T. et al. Crystal structure of the human cannabinoid CB1 receptor. Celi, v. 167, p. 750-762, 2016.
[0026] MAIOROV, V. N.; CRIPPEN, G. M. Significance of root-mean-square deviation in comparing three-dimensional structures of globular proteins. Journal of Molecular Biology, v. 235, p. 625-634, 1994.  [0026] MAIOROV, V. N .; CRIPPEN, G. M. Significance of root-mean-square deviation in comparing three-dimensional structures of globular proteins. Journal of Molecular Biology, v. 235, p. 625-634, 1994.
[0027] MORGAN, C. A.; HURLEY, T. D. Characterization of two distinct structural classes of selective aldehyde dehydrogenase 1 A1 inhibitors. Journal of Medicinal Chemistry, v. 58, p. 1964-1975, 2015.  [0027] MORGAN, C.A .; HURLEY, T. D. Characterization of two distinct structural classes of selective aldehyde dehydrogenase 1 A1 inhibitors. Journal of Medicinal Chemistry, v. 58, p. 1964-1975, 2015.
[0028] PERTWEE, R. G. The diverse CB1 and CB2 receptor pharmacology of three plant cannabinoids: A9-tetrahydrocannabinol, cannabidiol and Δ9 - tetrahydrocannabivarin. British Journal of Pharmacology, v. 153, p. 199-215, 2008.  [0028] PERTWEE, R. G. The diverse CB1 and CB2 receptor pharmacology of three plant cannabinoids: A9-tetrahydrocannabinol, cannabidiol and Δ9-tetrahydrocannabivarin. British Journal of Pharmacology, v. 153, p. 199-215, 2008.
[0029] RAMACHANDRAN, G. N.; RAMAKRISHNAN, C; SASISEKHARAN, V. Stereochemistry of polypeptide chain configurations. Journal of Molecular Biology, v. 7, p. 95-99, 1963.  [0029] RAMACHANDRAN, G. N .; RAMAKRISHNAN, C; SASISEKHARAN, V. Stereochemistry of polypeptide chain configurations. Journal of Molecular Biology, v. 7, p. 95-99, 1963.
[0030] SANTOS, P. S. et al. Efeitos do ácido lipóico nas concentrações de glutamato e taurina no hipocampo de ratos após convulsões induzidas por pilocarpina. Arq. Neuro-Psiquiatr. [online]. 201 1 , vol.69, n.2b, pp.360-364. .  [0030] SANTOS, P. S. et al. Effects of lipoic acid on glutamate and taurine concentrations in rat hippocampus after pilocarpine-induced seizures. Arq. Neuro-Psiquiatr. [online]. 201 1, vol.69, n.2b, pp.360-364. .
[0031] MUNRO et al. Nature 365:61 -65, 1993. [0031] MUNRO et al. Nature 365: 61-65, 1993.
[0032] RINALDI-CARMONA M. et al. J. Pharmacol. Exp. Ther. 278:871 -878, 1996.  [0032] RINALDI-CARMONA M. et al. J. Pharmacol. Exp. Ther. 278: 871-878, 1996. [Medline]
[0033] Langmead CJ1 , Watson J, Reavill C. Muscarinic acetylcholine receptors as CNS drug targets. Pharmacol Ther. 2008 Feb;1 17(2):232-43. Epub 2007 Dec 20.  [0033] Langmead CJ1, Watson J, Reavill C. Muscarinic acetylcholine receptors as CNS drug targets. Pharmacol Ther. 2008 Feb; 17 (2): 232-43. Epub 2007 Dec 20.
[0034] Do que se depreende da literatura pesquisada, não foram encontrados documentos antecipando ou sugerindo os ensinamentos da presente invenção, muito menos seus surpreendentes efeitos técnicos, de forma que a solução aqui proposta, aos olhos dos inventores, possui novidade e atividade inventiva frente ao estado da técnica. Sumário da Invenção From the literature, no documents were found anticipating or suggesting the teachings of the present invention, let alone their surprising technical effects, so that the solution proposed here, in the eyes of the inventors, has novelty and inventive activity to the state of the art. Summary of the Invention
[0035] A presente invenção vem resolver uma série de problemas conhecidos do estado da técnica como, por exemplo, proporcionar: um composto peptídico estável, um intermediário de síntese na preparação de compostos de interesse farmacêutico, uso do composto para preparar um ligante de interesse diagnóstico e/ou terapêutico, o uso para preparar um medicamento modulador do sistema canabinoide e/ou muscarínico, uma composição farmacêutica moduladora de funções metabólicas e/ou neuromoduladora contendo o referido composto; o uso do composto peptídico para a preparação de um medicamento para o tratamento curativo ou profilático de convulsões; uma entidade molecular que proporciona estes e/ou outros efeitos técnicos sem os inconvenientes decorrentes do uso das substâncias canabinoides como a prostração e sangramento nasal, dentre outros; uma composição farmacêutica útil como alternativa terapêutica para neuromodulação e/ou para prevenir ou tratar convulsões; um método terapêutico.  The present invention addresses a number of known problems of the prior art, for example, to provide: a stable peptidic compound, a synthetic intermediate in the preparation of compounds of pharmaceutical interest, use of the compound to prepare a binder of interest and / or neuromodulatory modulatory pharmaceutical composition containing said compound; the use of a medicament for modulating the cannabinoid and / or muscarinic system; the use of the peptidic compound for the preparation of a medicament for the curative or prophylactic treatment of seizures; a molecular entity that provides these and / or other technical effects without the drawbacks arising from the use of the cannabinoid substances such as prostration and nasal bleeding, among others; a pharmaceutical composition useful as a therapeutic alternative for neuromodulation and / or for preventing or treating seizures; a therapeutic method.
[0036] É, portanto, um dos objetos da presente invenção um composto peptídico com até 6 aminoácidos de fórmula:  It is therefore one of the objects of the present invention a peptidic compound having up to 6 amino acids of the formula:
R1-N-AA1-K-AA2-R2 R 1 -N-AA 1 -K-AA 2 -R 2
onde: at where:
AA é um aminoácido selecionado do grupo que consiste de F, W, L, I, V, P, G; AA2 é hidrogénio ou um aminoácido selecionado do grupo que consiste de F, W, L, I, V, P, G; AA is an amino acid selected from the group consisting of F, W, L, I, V, P, G; AA 2 is hydrogen or an amino acid selected from the group consisting of F, W, L, I, V, P, G;
Ri é hidrogénio, o aminoácido V, ou dipeptídeo PV; e  R 1 is hydrogen, the amino acid V, or dipeptide PV; and
R2 é ausente quando AA2 é hidrogénio ou é hidrogénio, o aminoácido L, ou o dipeptídeo LS quando Ri é hidrogénio, R 2 is absent when AA 2 is hydrogen or is hydrogen, the amino acid L, or the dipeptide LS when R 1 is hydrogen,
e/ou formas modificadas, cíclicas dos mesmos, versões amidadas, metiladas,and / or modified, cyclic forms thereof, amidated, methylated,
PEGiladas, seus sais; e/ou combinações dos mesmos. PEGylated, their salts; and / or combinations thereof.
[0037] Em uma concretização, AA é F, W ou L.  In one embodiment, AA is F, W, or L.
[0038] Em uma concretização, Ri e R2 são ambos hidrogénio. [0038] In one embodiment, Ri and R2 are both hydrogen.
[0039] Concretizações do composto peptídico da invenção úteis em seus vários objetos incluem um ou mais peptídeos selecionados do grupo que consiste de: NFK, NWK, NLK, NFKF, NWKF, NLKF, NFKW, NWKW, NLKW, NFKL, NWKL, NLKL, VNFK, VNWK, VNLK, NFKFL, NWKFL, NLKFL, NFKWL, NWKWL, NLKWL, NFKLL, NWKLL, NLKLL, VNFKF, VNWKW, VNLKL, VNFKFL, VNWKWL, VNLKLL, PVNFKF, PVNWKW, PVNLKL, NFKFLS, NWKWLS, NLKLLS, bem como formas modificadas, cíclicas, amidadas, metiladas, PEGiladas dos mesmos; seus sais ou combinações dos mesmos. Embodiments of the peptidic compound of the invention useful in the various objects thereof include one or more peptides selected from the group consisting of consists of: NFK, NWK, NLK, NWK, NLKF, NWKF, NWKF, NWKWL, NWKW, NLKW, NLKW, NLKW, NWKL, NWKLL, NWKLL, NWKLL, NLKLL, , VNFKF, VNWKW, VNLKL, VNFKFL, VNWKWL, VNLKLL, PVNFKF, PVNWKW, PVNLKL, NFKFLS, NWKWLS, NLKLLS, as well as modified, cyclic, amidated, methylated, PEGylated forms thereof; their salts or combinations thereof.
[0040] Em uma concretização, o composto peptídico da invenção é selecionado do grupo que compreende o tripeptídeo, NFK, o tetrapeptídeo NFKF, tetrapeptídeo NFKL, ou combinações dos mesmos. In one embodiment, the peptidic compound of the invention is selected from the group comprising tripeptide, NFK, NFKF tetrapeptide, NFKL tetrapeptide, or combinations thereof.
[0041] O composto da invenção proporciona melhor estabilidade e/ou facilidade de manipulação, sendo particularmente útil em preparações farmacêuticas e de medicamentos. The compound of the invention provides improved stability and / or ease of handling, being particularly useful in pharmaceutical and drug preparations.
[0042] O composto da invenção proporciona também a administração oral a um animal mamífero.  The compound of the invention also provides oral administration to a mammalian animal.
[0043] O composto da invenção, entre outras, proporciona vantagens na administração, biodisponibilização e/ou ação terapêutica em um animal quando comparado a outros peptídeos, como por exemplo a hemopressina e variantes conhecidos, usualmente de maior tamanho.  The compound of the invention, among others, provides advantages in the administration, bioavailability and / or therapeutic action in an animal when compared to other peptides, such as hemopressin and known variants, usually of larger size.
[0044] É um outro objeto da invenção proporcionar um composto substituto ao canabidiol e que proporciona vantagens na produção, manipulação, uso e segurança, podendo substituí-lo em todas ou quase todas as aplicações já descritas para o mesmo.  It is a further object of the invention to provide a substitutable compound to cannabidiol and which provides advantages in the production, handling, use and safety and can replace it in all or almost all applications already described therefor.
[0045] É um outro objeto da invenção proporcionar um composto útil para modular receptores muscarínicos e/ou o sistema canabinóide, seja pela modulação do receptor CB1 , do receptor CB2, de ambos concomitantemente, pela modulação da ligação ou da ação de outras substâncias que interagem no sistema canabinóide, pela modulação de proteases ou peptidases que levam à geração ou degradação de peptídeos ativos no sistema canabinóide, ou combinações dos mesmos.  It is a further object of the invention to provide a compound useful for modulating muscarinic receptors and / or the cannabinoid system, either by modulating the CB1 receptor, the CB2 receptor, both concomitantly, by modulating the binding or action of other substances interact in the cannabinoid system by the modulation of proteases or peptidases that lead to the generation or degradation of active peptides in the cannabinoid system, or combinations thereof.
[0046] É outro dos objetos da presente invenção um intermediário de síntese na preparação de compostos de interesse farmacêutico que compreendem o composto descrito acima e podem incluir modificações químicas, substituições, inclusão de outros grupos funcionais. Another object of the present invention is a synthetic intermediate in the preparation of compounds of pharmaceutical interest which comprise the compound described above and may include chemical modifications, substitutions, inclusion of other functional groups.
[0047] É outro dos objetos da presente invenção o uso do composto descrito acima para a preparação de outros compostos de interesse farmacêutico, referidos compostos compreendendo o composto descrito acima e modificações químicas, substituições, outros grupos funcionais.  Another object of the present invention is the use of the compound described above for the preparation of other compounds of pharmaceutical interest, said compounds comprising the compound described above and chemical modifications, substitutions, other functional groups.
[0048] É outro dos objetos da presente invenção o uso do composto definido acima para a preparação de uma composição farmacêutica moduladora de receptores canabinóides, sendo útil na modulação de funções metabólicas de um mamífero. It is another object of the present invention to use the compound defined above for the preparation of a pharmaceutical composition modulating cannabinoid receptors, being useful in modulating the metabolic functions of a mammal.
[0049] É um outro objeto da presente invenção o uso do composto definido acima para a preparação de um medicamento neuromodulador, neuroprotetor e/ou para o tratamento curativo ou profilático de convulsões em um mamífero.  It is another object of the present invention to use the compound defined above for the preparation of a neuromodulatory, neuroprotective medicament and / or for the curative or prophylactic treatment of seizures in a mammal.
[0050] A administração do composto da invenção a um mamífero proporciona atividade neuromoduladora, neuroprotetora e/ou anticonvulsivante; viabiliza administração oral em animais; não tem ou acarreta os inconvenientes decorrentes da produção, armazenamento, transporte e uso de substâncias canabinóides, além de proporcionar adicionais vantagens na preparação de produtos terapêuticos para mamíferos, em sua administração e/ou efeitos. Administration of the compound of the invention to a mammal provides neuromodulatory, neuroprotective and / or anticonvulsant activity; enables oral administration in animals; does not have or entails the drawbacks arising from the production, storage, transport and use of cannabinoid substances, in addition to providing additional advantages in the preparation of therapeutic products for mammals in their administration and / or effects.
[0051] Além disso, a administração do composto da invenção a um animal proporciona importantes e surpreendentes vantagens técnicas, incluindo superior atividade anticonvulsivante em relação à hemopressina, cujo uso como anticonvulsivante é objeto do pedido de patente co-pendente dos mesmos inventores. In addition, administration of the compound of the invention to an animal provides important and surprising technical advantages, including superior anticonvulsant activity over hemopressin, the use of which as an anticonvulsant is the subject of the co-pending co-pending application of the same inventors.
[0052] É um outro objeto da presente invenção um método terapêutico para modulação metabólica e/ou neuromodulação compreendendo a administração a um animal do peptídeo descrito acima.  Another object of the present invention is a therapeutic method for metabolic modulation and / or neuromodulation comprising administering to an animal the peptide described above.
[0053] É um outro objeto da invenção uma composição farmacêutica neuromoduladora, neuroprotetora e/ou para o tratamento curativo ou profilático de convulsões em um mamífero, referida composição compreendendo um veículo farmaceuticamente aceitável; e, como princípio ativo, o composto descrito acima. Another object of the invention is a neuromodulatory, neuroprotective pharmaceutical composition and / or for the curative or prophylactic treatment of seizures in a mammal, said composition comprising a pharmaceutically acceptable carrier; and, as active principle, the compound described above.
[0054] Em uma concretização, a composição farmacêutica se apresenta na forma de tablete, comprimido, gel, líquido oral ou xarope, cápsula, supositório, solução injetável ou formas inaláveis ou em adesivo, podendo opcionalmente compreender outros princípios ativos.  In one embodiment, the pharmaceutical composition is in the form of a tablet, tablet, gel, oral liquid or syrup, capsule, suppository, injectable solution or inhalable or adhesive forms, and may optionally comprise other active principles.
[0055] Estes e outros objetos do presente pedido de patente serão imediatamente valorizados pelos versados na arte e pelas empresas com interesses no segmento, e serão descritos em detalhes suficientes para sua reprodução na descrição a seguir.  These and other objects of the present application will be immediately appreciated by those skilled in the art and by companies having interests in the art and will be described in sufficient detail for their reproduction in the following description.
Breve Descrição das Figuras Brief Description of the Figures
[0056] A descrição detalhada a seguir e as figuras em anexo ilustram as principais características e concretizações da presente invenção, apresentadas detalhadamente para dar melhor suporte ao técnico no assunto e para que o mesmo possa entender e reproduzir o conceito inventivo da invenção em qualquer de suas concretizações. Tais detalhes ou figuras não devem ser entendidos de forma limitativa e servem somente para ilustrar algumas das concretizações da presente invenção.  The following detailed description and the accompanying figures illustrate the main features and embodiments of the present invention, set forth in detail to provide further support to the person skilled in the art and so that the same may understand and reproduce the inventive concept of the invention in either of their achievements. Such details or figures are not to be construed as limiting and serve only to illustrate some of the embodiments of the present invention.
[0057] A figura 1 mostra os resultados de testes comparativos de estabilidade do composto da invenção vs Hp (PVNFKFLSH). Os dados do gráfico mostram os resultados da medida de concentração de cada um destes ativos por HPLC, após congelamento por 24h ou, separadamente, após aquecimento a 100°C por 10 minutos (n=2). Os dados de significância estatística são também mostrados, os asteriscos indicando: (*) P<0.05 vs Controle (***) P<0.005 vs Controle; (****) P<0.0001 vs Controle. Figure 1 shows the results of comparative stability tests of the compound of the invention vs Hp (PVNFKFLSH). The data on the graph shows the results of the concentration measurement of each of these actives by HPLC after freezing for 24 hours or separately after heating at 100 ° C for 10 minutes (n = 2). Statistical significance data are also shown, the asterisks indicating: ( * ) P <0.05 vs Control ( *** ) P <0.005 vs Control; ( **** ) P <0.0001 vs Control.
[0058] A figura 2 apresenta os resultados de testes de neuroproteção com o com o composto da invenção NFKF no modelo de pilocarpina, sendo indicado o perfil de sobrevivência/morte dos animais mediante a administração do peptídeo da invenção. Em A) é mostrado o perfil de sobrevivência dos animais aos quais foi administrado o controle (apenas salina); Em B) é mostrado o perfil de sobrevivência dos animais aos quais foi administrado o canabidiol 30mg/kg; Em C) é mostrado o perfil de sobrevivência dos animais aos quais foi administrado o composto da invenção NFKF 50C^g/kg; Em D) são mostrados todos os perfis em um só gráfico. Interessante notar que no grupo tratado com o neuromodulador da invenção NFKF 500 μg/kg apenas dois animais que morreram. Os demais animais do grupo tratado com o peptídeo da invenção, no total de 3, seguiram vivos por mais de uma semana, enquanto praticamente todos os demais morreram em menos de 30 minutos. Consequentemente, o tempo médio de sobrevida para este grupo é significativamente diferente e muito maior do que o dos demais. Ademais, quando se compara o grupo ao qual foi administrado o composto invenção NFKF (500 μg/kg) com o grupo ao qual foi administrado o canabidiol (30mg/kg) destaca-se que a sobrevivência é três vezes maior no primeiro grupo, ou seja, no grupo ao qual foi administrado o composto invenção NFKF 500 μg/kg, mesmo usando uma concentração relativa de composto 60 vezes menor. Figure 2 shows the results of neuroprotection tests with the compound of the invention NFKF in the pilocarpine model, the survival / death profile of the animals being indicated by administration of the peptide of the invention. In A) the survival profile of the animals to which the control was administered (saline only) is shown; In B) the profile is shown survival rate of the animals administered cannabidiol 30mg / kg; In C) the survival profile of the animals to which the compound of the invention was administered NFKF 50C g / kg; In D) all profiles are shown in a single graph. It is interesting to note that in the group treated with the neuromodulator of the invention NFKF 500 μg / kg only two animals died. The remaining animals in the group treated with the peptide of the invention, totaling 3, remained alive for more than a week, while practically all the others died in less than 30 minutes. Consequently, the mean survival time for this group is significantly different and much higher than the others. In addition, when comparing the group to which the invention compound NFKF (500 μg / kg) was administered with the group to which cannabidiol (30 mg / kg) was administered, it is noted that survival is three times greater in the first group, or in the group to which the compound invention was administered NFKF 500 μg / kg, even using a 60-fold lower relative concentration of compound.
[0059] A figura 3 apresenta os resultados de testes do composto da invenção NFKF no modelo de pilocarpina, sendo indicado o tempo para a ocorrência da primeira salivação com a administração de controle, canabidiol (30mg/kg), ou o composto da invenção NFKF (500μg/kg). Os dados entre controle e demais compostos teste não apresentem significância estatística entre si nas condições testadas.  Figure 3 shows the test results of the compound of the invention NFKF in the pilocarpine model, indicating the time for the first salivation to occur with the administration of cannabidiol (30mg / kg), or the compound of the invention NFKF (500μg / kg). The data between control and other test compounds do not present statistical significance among themselves under the conditions tested.
[0060] A figura 4 apresenta os resultados de testes do composto da invenção NFKF como anticonvulsivante no modelo de pilocarpina, sendo indicado o tempo para a ocorrência da primeira convulsão com a administração de canabidiol (30mg/kg) ou o peptídeo da invenção NFKF (500μg/kg). Os asteriscos indicam a significância estatística: (**) P<0.02 vs Controle; (***) P<0.002 vs Controle. Figure 4 shows the test results of the compound of the invention NFKF as an anticonvulsant in the pilocarpine model, indicating the time for the first seizure to occur with the administration of cannabidiol (30mg / kg) or the peptide of the invention NFKF ( 500μg / kg). The asterisks indicate the statistical significance: ( ** ) P <0.02 vs Control; ( *** ) P <0.002 vs Control.
[0061] A figura 5 apresenta os resultados de testes do composto da invenção NFKF comparativamente aos resultados de testes da hemopressina, ambos no modelo de pilocarpina. São apresentados os tempos para a ocorrência da primeira salivação com a administração das seguintes doses de tratamento: o controle (salina); hemopressina (Hp ou PVNFKFLSH, 0,551334 μιηοΙ/kg); hemopressina (0,91889 μιηοΙ/kg); o composto da invenção NFKF (0,540882 μηποΙ/kg); NFKF (0,901469 μηποΙ/kg); o composto PEP-19 (DIIADDEPLT, 0,9081 17 μιηοΙ/kg). Os asteriscos indicam a significância estatística: (*) P<0.05 vs Controle; (**) P<0.01 vs Controle; o sinal + indica P<0.05 vs Hp 0,91889 μιηοΙ/kg. Figure 5 shows the test results of the compound of the invention NFKF compared to the results of hemopressin tests, both in the pilocarpine model. The times for the first salivation occur with administration of the following treatment doses: control (saline); hemopressin (Hp or PVNFKFLSH, 0.551334 μιηοΙ / kg); hemopressin (0.91889 μιηοΙ / kg); the compound of the invention NFKF (0.540882 μηποΙ / kg); NFKF (0.901469 μηποΙ / kg); the compound PEP-19 (DIIADDEPLT, 0.9081 17 μl / kg). The asterisks indicate the statistical significance: ( * ) P <0.05 vs Control; ( ** ) P <0.01 vs Control; the + sign indicates P <0.05 vs Hp 0.91889 μιηοΙ / kg.
[0062] A figura 6 apresenta os resultados de testes do composto da invenção NFKF como anticonvulsivante comparativamente aos resultados de testes da hemopressina como anticonvulsivante, ambos no modelo de pilocarpina. São apresentados os porcentuais do tempo (em relação ao controle) para a ocorrência da primeira convulsão com a administração das seguintes doses de tratamento: o controle (salina); hemopressina (Hp ou PVNFKFLSH, 0,551334 μιηοΙ/kg); hemopressina (0,91889 μιηοΙ/kg); o composto da invenção NFKF (0,540882 μη-iol/kg); NFKF (0,901469 μη-ιοΙ/kg); o composto PEP-19 (DIIADDEPLT, 0,9081 17 μιηοΙ/kg). Os asteriscos indicam a significância estatística: (*) P<0.05 vs Controle; (**) P<0.01 vs Controle; o sinal + indica P<0.05 vs Hp 0,91889 μη-ιοΙ/kg. Figure 6 shows the test results of the compound of the invention NFKF as an anticonvulsant compared to the results of tests of hemopressin as an anticonvulsant, both in the pilocarpine model. The percentage of time (relative to the control) for the first seizure is presented with the administration of the following treatment doses: control (saline); hemopressin (Hp or PVNFKFLSH, 0.551334 μιηοΙ / kg); hemopressin (0.91889 μιηοΙ / kg); the compound of the invention NFKF (0.540882 μl-iol / kg); NFKF (0.901469 μη-ιοΙ / kg); the compound PEP-19 (DIIADDEPLT, 0.9081 17 μl / kg). The asterisks indicate the statistical significance: ( * ) P <0.05 vs Control; ( ** ) P <0.01 vs Control; the + sign indicates P <0.05 vs Hp 0.91889 μη-ιοΙ / kg.
[0063] A figura 7 apresenta uma visão geral da estrutura tridimensional de um GPCR, neste caso, o receptor canabinóide do subtipo 1 . Em destaque as sete hélices transmembranares (l-VII), as alças intracelulares (ICL1 e ICL2) e, as alças extracelulares (ECL2 e ECL3).  Figure 7 shows an overview of the three-dimensional structure of a GPCR, in this case, the cannabinoid receptor of subtype 1. The seven transmembrane helices (l-VII), the intracellular loops (ICL1 and ICL2) and the extracellular loops (ECL2 and ECL3) were highlighted.
[0064] A figura 8 mostra a sobreposição da estrutura de AM6538 da estrutura cristalográfica (PDB 5TGZ), e o resultado obtido após a validação pela função Goldscore (quando a figura aparece colorida, a estrutura cristalográfica está em roxo e o resultado da função Goldscore em azul/ciano, embora para fins deste pedido de patente tais cores sejam irrelevantes).  Figure 8 shows the overlap of the AM6538 structure of the crystallographic structure (PDB 5TGZ), and the result obtained after validation by the Goldscore function (when the figure appears colored, the crystal structure is purple and the result of the Goldscore function in blue / cyan, although for the purposes of this application such colors are irrelevant).
[0065] A figura 9 mostra as principais interações observadas para AM6538 no sítio de ligação do receptor CB1 .  Figure 9 shows the main interactions observed for AM6538 at the CB1 receptor binding site.
[0066] A Figura 10 mostra as principais interações observadas para o rimonabanto no sítio de ligação do receptor CB1 . [0067] A Figura 1 1 mostra as principais interações observadas para o canabidiol no sítio de ligação do receptor CB1 . Figure 10 shows the major interactions observed for rimonabant at the CB1 receptor binding site. Figure 11 shows the major interactions observed for cannabidiol at the CB1 receptor binding site.
[0068] A Figura 12 mostra as principais interações observadas para o peptídeo da invenção NFKF no sítio de ligação do receptor CB1 .  Figure 12 shows the major interactions observed for the peptide of the invention NFKF at the binding site of the CB1 receptor.
[0069] A Figura 13 mostra os resultados do processo de obtenção de estrutura do receptor CB2. Em A) é mostrada a estrutura tridimensional do receptor CB2 obtida; em B) é mostrado o Gráfico de Ramachandran para o modelo de CB2 humano obtido.  Figure 13 shows the results of the process of obtaining CB2 receptor structure. In A) the three-dimensional structure of the obtained CB2 receptor is shown; in B) the Ramachandran Graph for the obtained human CB2 model is shown.
[0070] A figura 14 mostra as principais interações entre o receptor CB2 e o ligante AM6538.  Figure 14 shows the major interactions between the CB2 receptor and the AM6538 linker.
[0071] A Figura 15 mostra as principais interações entre o receptor CB2 e o rimonabanto.  Figure 15 shows the major interactions between the CB2 receptor and rimonabant.
[0072] A Figura 16 mostra as principais interações entre o receptor CB2 e o canabidiol.  Figure 16 shows the major interactions between the CB2 receptor and cannabidiol.
[0073] A Figura 17 mostra as principais interações entre o receptor CB2 e o tetrapeptídeo NFKF.  Figure 17 shows the major interactions between the CB2 receptor and the NFKF tetrapeptide.
Descrição Detalhada da Invenção Detailed Description of the Invention
[0074] O composto da invenção demonstrou diversas utilidades e resultados surpreendentes em testes in vitro e in vivo. O composto da invenção é útil para a preparação de outros compostos de interesse farmacêutico. O composto da invenção é útil para a preparação de uma composição farmacêutica. A composição farmacêutica da invenção é moduladora de receptores canabinóides e/ou muscarínicos, sendo particularmente útil na modulação de funções metabólicas e/ou na neuromodulação de um animal mamífero.  The compound of the invention has demonstrated several surprising uses and results in in vitro and in vivo tests. The compound of the invention is useful for the preparation of other compounds of pharmaceutical interest. The compound of the invention is useful for the preparation of a pharmaceutical composition. The pharmaceutical composition of the invention is cannabinoid and / or muscarinic receptor modulator, being particularly useful in modulating metabolic functions and / or neuromodulation of a mammalian animal.
[0075] O conceito inventivo comum aos diversos objetos da invenção é o uso de um composto com até 6 aminoácidos de fórmula: The inventive concept common to the various objects of the invention is the use of a compound having up to 6 amino acids of the formula:
R1-N-AA1-K-AA2-R2 R 1 -N-AA 1 -K-AA 2 -R 2
onde: at where:
AAi é um aminoácido selecionado do grupo que consiste de F, W, L, I, V, P, G; AA2 é hidrogénio ou um aminoácido selecionado do grupo que consiste de F, W, L, I, V, P, G; AAi is an amino acid selected from the group consisting of F, W, L, I, V, P, G; AA 2 is hydrogen or an amino acid selected from the group consisting of F, W, L, I, V, P, G;
Ri é hidrogénio, o aminoácido V, ou dipeptídeo PV; e  R 1 is hydrogen, the amino acid V, or dipeptide PV; and
R2 é ausente quando AA2 é hidrogénio ou é hidrogénio, o aminoácido L, ou o dipeptídeo LS quando Ri é hidrogénio, e/ou formas modificadas, cíclicas dos mesmos, versões amidadas, metiladas, PEGiladas, seus sais; e/ou combinações dos mesmos para a preparação de uma composição de interesse farmacêutico. R 2 is absent when AA 2 is hydrogen or is hydrogen, the amino acid L, or the dipeptide LS when R 1 is hydrogen, and / or modified, cyclic forms thereof, amidated, methylated, PEGylated versions, salts thereof; and / or combinations thereof for the preparation of a composition of pharmaceutical interest.
[0076] Para fins da presente invenção as seguintes definições são utilizadas:  For purposes of the present invention the following definitions are used:
[0077] Composto de interesse farmacêutico Compound of pharmaceutical interest
[0078] No contexto do presente pedido de patente, deve-se entender "composto de interesse farmacêutico" qualquer entidade molecular que compreenda o composto descrito como conceito inventivo comum ao presente pedido de patente, incluindo também entidades moleculares obtidas mediante derivatização química do mesmo, com a inclusão de outros grupos funcionais, cadeias laterais lineares ou ramificadas, alteração de hidrofilicidade ou hidrofobicidade, entre outras, desde que compreendam como núcleo a entidade R1-N-AA1-K-AA2-R2 conforme definido acima, excetuando-se as entidades naturais e já conhecidas.  In the context of the present application, "compound of pharmaceutical interest" means any molecular entity comprising the compound described as inventive concept common to the present application, including also molecular entities obtained by chemical derivatization thereof, with the inclusion of other functional groups, linear or branched side chains, alteration of hydrophilicity or hydrophobicity, among others, provided that they comprise as nucleus the entity R1-N-AA1-K-AA2-R2 as defined above, with the exception of entities natural and already known.
[0079] Composição farmacêutica Pharmaceutical composition
[0080] No contexto do presente pedido de patente, deve-se entender "composição farmacêutica" como toda e qualquer composição que contenha um princípio ativo, com fins profiláticos, paliativos e/ou curativos, atuando de forma a manter e/ou restaurar a homeostase, podendo ser administrada de forma oral, tópica, parenteral, enteral e/ou intratecal.  In the context of the present patent application, "pharmaceutical composition" is to be understood as any and all compositions containing an active principle, for prophylactic, palliative and / or curative purposes, acting in a manner to maintain and / or restore the homeostasis, and may be administered orally, topically, parenterally, enterally and / or intrathecally.
[0081] Formulação farmaceuticamente aceitável [0081] Pharmaceutically acceptable formulation
[0082] No contexto do presente pedido de patente, deve-se entender "formulação farmaceuticamente aceitável" uma formulação contendo excipientes e carreadores farmaceuticamente aceitáveis bem conhecidos por técnicos no assunto, como é o desenvolvimento de doses e tratamentos convenientes para uso em composições particulares que podem ser descritas em uma série de regimentos de tratamento, incluindo oral, parenteral, intravenoso, intranasal, intravítreo e intramuscular, intracerebral, intracerebroventricular e intraocular e sua administração e/ou formulação. In the context of the present application, a "pharmaceutically acceptable formulation" is meant a formulation containing pharmaceutically acceptable excipients and carriers well known to those skilled in the art, such as the development of convenient doses and treatments for use in particular compositions which can be described in a series of treatment regimens, including oral, parenteral, intravenous, intranasal, intravitreal and intramuscular, intracerebral, intracerebroventricular and intraocular and their administration and / or formulation.
[0083] Peptídeo modificado [0083] Modified peptide
[0084] No contexto do presente pedido de patente, deve-se entender "peptídeo modificado" como um peptídeo não natural, modificado artificialmente ou obtido por síntese, incluindo os ciclizados, amidados, metilados, PEGilados, ou suas formas de sal, bem como um peptídeo compreendendo um ou mais aminoácido não natural, como as formas d-aminoácido. O composto peptídico pode ser peguilado utilizando as técnicas conhecidas pelos versados na arte, como, por exemplo, a peguilação com reagentes contendo o grupamento succinimidil, os quais reagem preferencialmente com aminas primárias presentes na região N- terminal do peptídeo.  In the context of the present application, "modified peptide" is to be understood as a non-naturally occurring, artificially modified or synthesized peptide, including cyclized, amidated, methylated, PEGylated, or salt forms thereof, as well as a peptide comprising one or more unnatural amino acid, such as d-amino acid forms. The peptidic compound may be pegylated using techniques known to those skilled in the art, such as, for example, pegylation with reagents containing the succinimidyl group, which preferentially react with primary amines present in the N-terminal region of the peptide.
[0085] Peptídeo cíclico ou circular [0085] Cyclic or circular peptide
[0086] No contexto do presente pedido de patente, deve-se entender "peptídeo cíclico, ciclizado ou "circular" como um peptídeo que teve uma ligação covalente entre as duas extremidades de uma molécula linear de peptídeo através de qualquer método conhecido da técnica, particularmente pela atividade de enzimas. O peptídeo cíclico pode ser utilizado em substituição ao peptídeo linear devido ao fato de ser mais difícil de ser degradado, já que suas extremidades ou zonas de ataque por enzimas hidrolisantes não estão tão expostas quanto em um peptídeo linear.  In the context of the present application, "cyclic, cyclized or" circular peptide "is to be understood as a peptide which has a covalent bond between the two ends of a linear peptide molecule by any method known in the art, particularly by the activity of enzymes.Cyclic peptide can be used instead of the linear peptide because it is more difficult to be degraded, since its ends or zones of attack by hydrolyzing enzymes are not as exposed as in a linear peptide.
[0087] Aqonista [0087] Aqonist
[0088] No contexto do presente pedido de patente, deve-se entender "agonista" como um fármaco, droga, hormônio, neurotransmissor ou outra molécula sinalizadora que forma um complexo com um sítio receptor, dessa forma acionando uma resposta ativa de uma célula.  In the context of the present application, "agonist" is to be understood as a drug, drug, hormone, neurotransmitter or other signaling molecule which forms a complex with a receptor site, thereby triggering an active response of a cell.
[0089] Aqonista inverso / antagonista [0089] Aqonista inverse / antagonist
[0090] No contexto do presente pedido de patente, deve-se entender "agonista inverso ou antagonista" como agente(s) (por exemplo: fármacos, drogas, hormônios ou enzimas) que se liga(m) aos receptores dos agonistas e produz(em) efeitos farmacológicos opostos aos dos agonistas, de tal forma que a ação de um inibe parcialmente ou totalmente o efeito da outra. Particularmente, um composto é um agonista inverso quando atua na presença de um agonista, mas reduzindo sua atividade; um antagonista é um composto que bloqueará totalmente a atividade do agonista. In the context of the present application, "inverse agonist or antagonist" as agent (s) (for example, drugs, drugs, hormones or enzymes) which binds to agonist receptors and produces pharmacological effects opposite to agonists, in such a way that the action of one partially or totally inhibits the effect of the other. Particularly, a compound is an inverse agonist when it acts in the presence of an agonist, but reducing its activity; an antagonist is a compound that will totally block the activity of the agonist.
[0091] Dose Equivalente em Humanos [0091] Equivalent Dose in Humans
[0092] Na presente invenção, o conceito de "dose equivalente em humanos" é a dose na qual, em humanos, se espera a mesma magnitude de efeitos observada em animais numa dada dose, conforme preconiza o "Guidance for Industry Estimating the Maximum Safe Starting Dose in Initial Clinicai Trials for Therapeutics in Adult Healthy Volunteers" publicado pelo U.S. Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and Research (CDER), July 2005 Pharmacology and Toxicology. No referido guia, a conversão de dose observada em animais (mg/kg) para Dose Equivalente em Humanos (mg/kg) implica na divisão do resultado obtido em ratos por 6,2 e do resultado obtido em camundongos por 12,3. Esse valores são aplicáveis a um humano com 60 kg de peso padrão. Para outras espécies ou para pesos fora das faixas de peso padrão, a Dose Equivalente em Humanos (DEH) pode ser calculada pela fórmula: DEH = dose no animal em mg/kg x (peso do animal em kg/peso do humano em kg)0 33. O referido Guidance considera adequada uma faixa de segurança de 10 vezes os limites de concentração testados. In the present invention, the concept of "equivalent dose in humans" is the dose at which, in humans, the same magnitude of effects observed in animals at a given dose is expected, as advocated in Guidance for Industry Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers "published by the US Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and Research (CDER), July 2005 Pharmacology and Toxicology. In said guide, the conversion of the dose observed in animals (mg / kg) to Equivalent Dose in Humans (mg / kg) implies dividing the result obtained in rats by 6.2 and the result obtained in mice by 12.3. These values apply to a human with 60 kg of standard weight. Equivalent Human Dose (DEH) can be calculated by the formula: DEH = dose in animal in mg / kg x (animal weight in kg / human weight in kg) for other species or for weights outside the standard weight ranges 0 33 . This Guidance considers a safety range of 10 times the concentration limits tested to be adequate.
[0093] Modular a função de receptor CB ou do sistema canabinóide  Modulating the CB receptor function or the cannabinoid system
[0094] No contexto do presente pedido de patente, deve-se entender "modular a função de receptor CB" como uma interação que acarreta na alteração da atividade bioquímica do receptor CB, particularmente CB1 ou CB2. Entende-se que a alteração é positiva quando ocorre um efeito antagonista ou agonista inverso nos receptores CB e que a alteração é negativa quando ocorre um efeito agonista nos receptores CB. Os testes apresentados no presente pedido de patente sugerem que o composto da invenção interage com e/ou modula o receptor CB1 e/ou o receptor CB2, provavelmente como um modulador alostérico do receptor CB1 e/ou CB2. Assim o composto da invenção é útil para modular o sistema canabinóide, seja pela modulação do receptor CB1 , do receptor CB2, de ambos concomitantemente, pela modulação da ligação ou da ação de outras substâncias que interagem no sistema canabinóide, pela modulação de proteases ou peptidases que levam à geração ou degradação de peptídeos ativos no sistema canabinóide, ou combinações dos mesmos. In the context of the present application, it is meant to "modulate the CB receptor function" as an interaction that entails in altering the biochemical activity of the CB receptor, particularly CB1 or CB2. It is understood that the change is positive when an antagonist or inverse agonist effect occurs at CB receptors and that the change is negative when an agonist effect occurs at CB receptors. The tests presented in the present patent application suggest that the compound of the invention interacts with and / or modulates the CB1 receptor and / or CB2 receptor, probably as a modulator allosteric ratio of CB1 and / or CB2 receptor. Thus the compound of the invention is useful for modulating the cannabinoid system, either by modulating the CB1 receptor, the CB2 receptor, both concomitantly, by modulating the binding or action of other substances interacting in the cannabinoid system, by modulating proteases or peptidases which lead to the generation or degradation of active peptides in the cannabinoid system, or combinations thereof.
[0095] Modular a função de receptores muscarínicos [0095] Modulating the function of muscarinic receptors
[0096] No contexto do presente pedido de patente, deve-se entender "modular a função de receptores muscarínicos" como uma interação que acarreta alterações neuronais, incluindo receptor muscarínico de acetilcolina (mAChRs), que desempenha um papel importante nas funções cognitivas, tais como a aprendizagem e memória, controle da liberação de dopamina, modulação da atividade locomotora, sua modulação sendo também útil no controle da doença de Alzheimer e/ou controle da dependência ou vício de drogas de abuso. Entende-se que a alteração é positiva quando ocorre um efeito antagonista ou agonista inverso nos receptores muscarínicos e que a alteração é negativa quando ocorre um efeito agonista nos receptores muscarínicos. Os testes apresentados no presente pedido de patente sugerem que o composto da invenção interage com e/ou modula receptores muscarínicos.  In the context of the present application, the term "modulating muscarinic receptor function" should be understood as an interaction leading to neuronal changes, including muscarinic acetylcholine receptor (mAChRs), which plays an important role in cognitive functions, such as such as learning and memory, control of dopamine release, modulation of locomotor activity, its modulation being also useful in the control of Alzheimer's disease and / or control of addiction or addiction to drugs of abuse. It is understood that the change is positive when an antagonist or inverse agonist effect occurs at muscarinic receptors and that the change is negative when an agonist effect occurs at muscarinic receptors. The tests presented in the present patent application suggest that the compound of the invention interacts with and / or modulates muscarinic receptors.
[0097] Modulação de funções metabólicas [0097] Modulation of metabolic functions
[0098] No contexto do presente pedido de patente, deve-se entender esta expressão como incluindo a modulação de: metabolismo de energia e/ou de lipídeos; hipertensão arterial, regulação da motilidade intestinal; sistema imune; equilíbrio do ciclo do cálcio, condições associadas à glândula tiroidal, órgãos periféricos e tecidos, incluindo órgãos reprodutivos, tecido adiposo, fígado, músculos e trato gastrointestinal, sendo útil no tratamento de obesidade, diabetes, doenças ou distúrbios imunes/inflamatórios, osteopenia, osteoporose, câncer.  In the context of the present application, this term is understood to include modulation of: energy and / or lipid metabolism; arterial hypertension, regulation of intestinal motility; Imune system; balance of the calcium cycle, conditions associated with the thyroid gland, peripheral organs and tissues, including reproductive organs, adipose tissue, liver, muscles and gastrointestinal tract, being useful in the treatment of obesity, diabetes, diseases or immune / inflammatory disorders, osteopenia, osteoporosis , cancer.
[0099] Neuromodulador  [0099] Neuromodulator
[0100] No contexto do presente pedido de patente, deve-se entender por "neuromodulador" ou "neuromoduladora" a função de modular a função neuronal/neurológica, incluindo a modulação da atividade cerebral, do córtex, hipocampo, amígdala, pituitário, hipotálamo; glândula adrenal. A neuromodulação inclui a modulação benéfica de neuroproteção contra agentes ou condições que levam a processos fisiopatológicos. Agentes ou compostos neuroprotetores são preferencialmente usados antes da (ou durante a) fase prodrômica das doenças, o que muitas vezes tem início muitos anos antes do surgimento dos sintomas. Na presente invenção, um neuromodulador é potencialmente útil no tratamento curativo ou profilático de diversas condições ou doenças neurológicas, incluindo tremor essencial, enxaqueca, dor neuropática, transtornos psiquiátricos como ansiedade, esquizofrenia ou transtorno bipolar, Alzheimer, Parkinson, autismo, sendo também potencialmente útil na modificação dos processos fisiopatológicos envolvidos na ocorrência de convulsões e/ou epilepsia, bem como em outras condições clínicas relacionadas a distúrbios da excitabilidade neuronal ou de lesões neuronais decorrentes de isquemia, hipoxia ou outras condições danosas. [0100] In the context of the present patent application, the term "neuromodulator" or "neuromodulator" is understood as the function of modulating the function neuronal / neurological, including modulation of brain activity, cortex, hippocampus, amygdala, pituitary, hypothalamus; adrenal gland. Neuromodulation includes the beneficial modulation of neuroprotection against agents or conditions leading to pathophysiological processes. Neuroprotective agents or compounds are preferably used prior to (or during) the prodromal stage of disease, which often begins many years before the onset of symptoms. In the present invention, a neuromodulator is potentially useful in the curative or prophylactic treatment of a variety of neurological conditions or diseases, including essential tremor, migraine, neuropathic pain, psychiatric disorders such as anxiety, schizophrenia or bipolar disorder, Alzheimer's, Parkinson's, autism, and is also potentially useful in modifying the pathophysiological processes involved in the occurrence of seizures and / or epilepsy, as well as in other clinical conditions related to disorders of neuronal excitability or neuronal lesions due to ischemia, hypoxia or other harmful conditions.
[0101] Embora nesta invenção seja demonstrado que o referido composto se liga e/ou modula receptores canabinóides, a surpreendente ação farmacêutica da invenção pode estar ligada à ação sobre CB1 e/ou CB2 e/ou muscarínicos ou eventualmente estar ligada à modulação do uptake de adenosina, do GGPR55, de receptores PPARy, do nível de cálcio intracelular ou combinações destes. Assim, qualquer indicação terapêutica relacionada a estes alvos pode se beneficiar da presente invenção. Although in this invention it is shown that said compound binds and / or modulates cannabinoid receptors, the surprising pharmaceutical action of the invention may be linked to action on CB1 and / or CB2 and / or muscarinic or possibly linked to uptake modulation adenosine, GGPR55, PPARγ receptors, intracellular calcium level, or combinations thereof. Thus, any therapeutic indication related to these targets may benefit from the present invention.
[0102] A presente invenção é também definida pelas seguintes cláusulas.  [0102] The present invention is also defined by the following clauses.
[0103] Uso do composto descrito acima. [0103] Use of the compound described above.
[0104] Uso conforme descrito acima em que AAi e/ou AA2 é F, W ou L. [0104] Use as described above wherein AAi and / or AA 2 is F, W or L.
[0105] Uso conforme descrito acima em que o referido peptídeo é selecionado do grupo consiste de: NFK, NWK, NLK, NFKF, NWKF, NLKF, NFKW, NWKW, NLKW, NFKL, NWKL, NLKL, VNFK, VNWK, VNLK, NFKFL, NWKFL, NLKFL, NFKWL, NWKWL, NLKWL, NFKLL, NWKLL, NLKLL, VNFKF, VNWKW, VNLKL, VNFKFL, VNWKWL, VNLKLL, PVNFKF, PVNWKW, PVNLKL, NFKFLS, NWKWLS, NLKLLS, bem como formas modificadas, cíclicas, amidadas, metiladas e/ou PEGiladas dos mesmos; seus sais; ou combinações dos mesmos. The use as described above wherein said peptide is selected from the group consisting of: NFK, NWK, NLK, NFKF, NWKF, NLKF, NFKW, NWKW, NLKW, NFKL, NWKL, NLKL, VNFK, VNWK, VNLK, NFKFL , NWKFL, NLKLL, NWKLL, NWKLL, NWKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, NLKLL, amidated, methylated and / or PEGylated derivatives thereof; their salts; or combinations thereof.
[0106] Intermediário de síntese na preparação de compostos de interesse farmacêutico compreendendo o composto descrito acima.  Synthesis Intermediate in the preparation of compounds of pharmaceutical interest comprising the compound described above.
[0107] Uso do composto descrito acima como intermediário de síntese na preparação de outros compostos de interesse farmacêutico.  Use of the above-described compound as the synthesis intermediate in the preparation of other compounds of pharmaceutical interest.
[0108] O composto descrito acima, com modificado.  The compound described above, is modified.
[0109] Uso do composto descrito acima para preparação de uma composição farmacêutica moduladora do sistema canabinóide e/ou de receptores muscarínicos.  Use of the compound described above for the preparation of a pharmaceutical composition modulating the cannabinoid system and / or muscarinic receptors.
[0110] Uso do composto descrito acima para preparação de uma composição farmacêutica moduladora de funções metabólicas.  [0110] Use of the compound described above for the preparation of a pharmaceutical composition modulating metabolic functions.
[0111] Uso do composto descrito acima para a preparação de um medicamento modulador de funções metabólicas, neuromodulador, neuroprotetor e/ou curativo ou profilático de convulsões em um mamífero em um mamífero.  Use of the compound described above for the preparation of a metabolising, neuromodulatory, neuroprotective and / or curative or prophylactic modulatory drug of convulsions in a mammal in a mammal.
[0112] Uso do composto descrito acima para a preparação de um ligante de interesse diagnóstico em um mamífero.  [0112] Use of the compound described above for the preparation of a binder of diagnostic interest in a mammal.
[0113] Composição farmacêutica moduladora de funções metabólicas em um mamífero compreendendo um veículo farmaceuticamente aceitável; e, como princípio ativo, o composto descrito acima.  A pharmaceutical composition modulating metabolic functions in a mammal comprising a pharmaceutically acceptable carrier; and, as active principle, the compound described above.
[0114] Composição farmacêutica neumoduladora curativa ou profilática em um mamífero compreendendo um veículo farmaceuticamente aceitável; e, como princípio ativo, o composto descrito acima.  [0114] A curative or prophylactic pneumodulatory pharmaceutical composition in a mammal comprising a pharmaceutically acceptable carrier; and, as active principle, the compound described above.
[0115] Composição farmacêutica para o tratamento curativo ou profilático de convulsões em um mamífero compreendendo um veículo farmaceuticamente aceitável; e, como princípio ativo, o composto descrito acima.  A pharmaceutical composition for the curative or prophylactic treatment of seizures in a mammal comprising a pharmaceutically acceptable carrier; and, as active principle, the compound described above.
[0116] Composição farmacêutica neuromoduladora compreendendo o tripeptídeo NFK, o tetrapeptídeo NFKF, o tetrapeptídeo NFKL, ou combinações dos mesmos. [0117] Composição farmacêutica conforme descrito acima na forma de tablete, comprimido, gel, líquido oral ou xarope, cápsula, supositório, solução injetável, forma inalável ou em adesivo. [0116] Neuromodulatory pharmaceutical composition comprising the NFK tripeptide, the NFKF tetrapeptide, the NFKL tetrapeptide, or combinations thereof. Pharmaceutical composition as described above in the form of a tablet, tablet, gel, oral liquid or syrup, capsule, suppository, solution for injection, inhalable form or in adhesive.
[0118] Método terapêutico modulador de funções metabólicas de um animal, compreendendo a administração do peptídeo descrito acima. Particularmente para esta aplicação, o peptídeo da invenção é preferencialmente modificado para que seu peso molecular seja maior, minimizando ou impedindo a passagem do mesmo pela barreira hematoencefálica.  A therapeutic modulator of metabolic functions of an animal, comprising administering the peptide described above. Particularly for this application, the peptide of the invention is preferably modified so that its molecular weight is greater, minimizing or preventing the passage thereof through the blood-brain barrier.
[0119] Método terapêutico para a neuromodulação, neuroproteção e/ou tratamento curativo ou profilático de convulsões compreendendo a administração, a um animal, do composto descrito acima.  [0119] Therapeutic method for neuromodulation, neuroprotection and / or curative or prophylactic treatment of seizures comprising administering, to an animal, the compound described above.
[0120] Testes in vitro e in silico mostraram que o composto da invenção proporciona vantagens na preparação de composições farmacêuticas, em sua estabilidade, na ligação a receptores canabinóides CB1 e/ou CB2.  [0120] In vitro and in silico tests have shown that the compound of the invention provides advantages in the preparation of pharmaceutical compositions, in their stability, in binding to CB1 and / or CB2 cannabinoid receptors.
[0121] O composto da invenção mostrou-se muito mais estável que a hemopressina, que ademais acarreta os problemas técnicos de formação de fibra ou fibrilas, assim como ocorre com seus variantes maiores conhecidos.  The compound of the invention has been shown to be much more stable than hemopressin, which in addition causes the technical problems of fiber formation or fibrils, as it does with its known larger variants.
[0122] Testes in vivo em mamíferos mostraram excelente efeito terapêutico a baixas dosagens, sem indícios de efeitos colaterais importantes.  [0122] In vivo mammalian tests showed excellent therapeutic effect at low dosages, with no evidence of significant side effects.
[0123] O efeito neuromodulador/neuroprotetor é evidenciado pela ausência sintomas, danos cerebrais e mortes associadas à administração de substâncias reconhecidamente danosas, como é o caso da pilocarpina.  The neuromodulator / neuroprotective effect is evidenced by the absence of symptoms, brain damage, and deaths associated with the administration of substances known to be harmful, such as pilocarpine.
[0124] Em uma concretização, a invenção proporciona o uso do referido composto para a preparação de um medicamento neuromodulador, neuroprotetor e/ou para o tratamento curativo ou profilático de convulsões em um mamífero. A administração, a um animal, do composto da invenção proporciona atividade neuromoduladora, neuroprotetora e/ou anticonvulsivante; viabiliza administração oral; não tem ou acarreta os inconvenientes decorrentes da produção, armazenamento, transporte e uso de substâncias canabinóides, além de proporcionar adicionais vantagens na preparação de produtos terapêuticos para mamíferos, em sua administração e/ou efeitos. Além disso, a administração do composto da invenção a um animal proporciona importantes e surpreendentes vantagens técnicas, incluindo superior atividade anticonvulsivante em relação à hemopressina, cujo uso como anticonvulsivante é objeto do pedido de patente co-pendente dos mesmos inventores. In one embodiment, the invention provides the use of said compound for the preparation of a neuromodulatory, neuroprotective medicament and / or for the curative or prophylactic treatment of seizures in a mammal. Administration, to an animal, of the compound of the invention provides neuromodulatory, neuroprotective and / or anticonvulsant activity; makes oral administration feasible; does not have or entails the drawbacks arising from the production, storage, transport and use of cannabinoid substances, in addition to providing additional advantages in the preparation of therapeutic products for mammals in their administration and / or effects. In addition administering the compound of the invention to an animal provides important and surprising technical advantages, including superior anticonvulsant activity with respect to hemopressin, the use of which as an anticonvulsant is the subject of the co-pending co-pending application of the same inventors.
[0125] A presente invenção descreve o uso de um composto para a preparação de composições farmacêuticas úteis para um variado conjunto de condições médicas, incluindo aquelas relacionadas ao sistema nervoso central. Surpreendentemente, embora o composto ativo da invenção seja peptídico ou predominantemente peptídico, a administração oral proporcionou efeitos cerebrais em animais. The present invention describes the use of a compound for the preparation of pharmaceutical compositions useful for a varied set of medical conditions, including those related to the central nervous system. Surprisingly, although the active compound of the invention is peptidic or predominantly peptidic, oral administration has provided brain effects in animals.
[0126] Em uma concretização, testes in vivo com a composição da invenção demonstraram resultados surpreendentes quanto à sua atividade neuroprotetora. A composição da invenção, quando administrada previamente a animais, proporcionou surpreendente, potente e longa proteção contra danos decorrentes da subsequente administração de substâncias sabidamente danosas aos neurónios. Assim sendo, a neuroproteção proporcionada pelo composto da invenção é particularmente útil como alternativa terapêutica para diversas condições médicas, incluindo aquelas relacionadas a distúrbios de excitabilidade neuronal, como, por exemplo, as convulsões.  In one embodiment, in vivo tests with the composition of the invention demonstrated surprising results regarding its neuroprotective activity. The composition of the invention, when administered previously to animals, provided surprising, potent and long protection against damage resulting from the subsequent administration of substances known to be harmful to neurons. Accordingly, the neuroprotection provided by the compound of the invention is particularly useful as a therapeutic alternative for various medical conditions, including those related to disorders of neuronal excitability, such as seizures.
[0127] Em várias concretizações, testes in vivo com o composto da invenção demonstraram resultados surpreendentes quanto à sua atividade anticonvulsivante. O composto da invenção, quando usado como anticonvulsivante, adicionalmente proporciona a vantagem de ser um bom candidato a substituto dos compostos canabinoides conhecidos por sua atuação como anticonvulsivantes, como é o caso o Canabidiol. O Canabidiol, apesar de seus efeitos comprovados como anticonvulsivante, tem enfrentando problemas regulatorios devido à sua origem, a planta Cannabis sativa. A presente invenção proporciona uma adicional abordagem terapêutica para os pacientes que sofrem de convulsões e que têm dificuldade de obter medicamentos, sendo baseada em um peptídeo, ou seja, não usa derivados da Cannabis sativa. Além disso, os resultados mostraram que o composto da invenção, quando usado como anticonvulsivante, proporciona outras surpreendentes vantagens técnicas no uso, incluindo maior efeito terapêutico, uso oral, menor dosagem, menor ocorrência de efeitos colaterais como a prostração e sangramento nasal, dentre outras vantagens técnicas. In various embodiments, in vivo tests with the compound of the invention demonstrated surprising results regarding its anticonvulsant activity. The compound of the invention, when used as an anticonvulsant, additionally provides the advantage of being a good candidate to substitute the cannabinoid compounds known for their acting as anticonvulsants, such as Canabidiol. Cannabidiol, despite its proven effects as an anticonvulsant, has been facing regulatory problems due to its origin, the Cannabis sativa plant. The present invention provides an additional therapeutic approach for patients suffering from seizures and having difficulty in obtaining drugs, being based on a peptide, ie, does not use derivatives of Cannabis sativa. In addition, the results showed that the compound of invention, when used as an anticonvulsant, provides other surprising technical advantages in use, including greater therapeutic effect, oral use, lower dosage, less occurrence of side effects such as prostration and nasal bleeding, among other technical advantages.
[0128] Nos testes com modelo experimental de convulsão, a administração da composição farmacêutica da invenção a mamíferos resultou em excelente efeito terapêutico em baixas dosagens, quando comparado às dosagens de um composto de referência, o canabidiol. Além disso, os resultados de testes mostraram que a composição da invenção proporciona surpreendentes vantagens técnicas no uso, incluindo maior efeito terapêutico, viabilidade de uso oral, desnecessidade de usar óleo como veículo (que em muitos casos causa efeitos colaterais), menor dosagem e menor ocorrência de efeitos colaterais como a prostração e sangramento nasal, entre outros. In tests with an experimental seizure model, administration of the pharmaceutical composition of the invention to mammals resulted in excellent therapeutic effect at low dosages when compared to dosages of a reference compound, cannabidiol. In addition, test results have shown that the composition of the invention provides surprising technical advantages in use, including greater therapeutic effect, viability of oral use, lack of use of carrier oil (which in many cases causes side effects), lower dosage and lower occurrence of side effects such as prostration and nasal bleeding, among others.
[0129] Testes comparativos in vivo demonstram que o composto da invenção tem atividade superior e/ou requer menor dosagem que a hemopressina, cujo uso como anticonvulsivante é objeto do pedido de patente co-pendente dos mesmos inventores. In vivo comparative tests demonstrate that the compound of the invention has superior activity and / or requires less dosage than hemopressin, the use of which as an anticonvulsant is the subject of the co-pending co-pending application of the same inventors.
[0130] A composição farmacêutica da invenção é também útil para o tratamento de doenças associadas à modulação da atividade do sistema canabinóide, de receptores canabinóides (CB) e/ou muscarínicos - com diversas vantagens técnicas e sem que ocorram efeitos indesejáveis conhecidos dos congéneres disponíveis no estado da técnica.  The pharmaceutical composition of the invention is also useful for the treatment of diseases associated with modulation of the activity of the cannabinoid system, cannabinoid (CB) and / or muscarinic receptors - with various technical advantages and without known undesirable effects of the available congeners in the state of the art.
[0131] Na presente invenção, o composto conforme descrito acima é útil para a preparação de um medicamento modulador de funções metabólicas.  In the present invention, the compound as described above is useful for the preparation of a medicament modulator of metabolic functions.
[0132] Na presente invenção, o composto conforme descrito acima é útil para a preparação de um medicamento neuromodulador, neuroprotetor e/ou de um medicamento para o tratamento de convulsões.  In the present invention, the compound as described above is useful for the preparation of a neuromodulator, neuroprotective medicament and / or a medicament for the treatment of seizures.
[0133] Adicionalmente, conforme será demonstrado nos exemplos a seguir, o uso do composto da invenção na preparação de um medicamento neuromodulador, neuroprotetor e/ou anticonvulsivante proporciona a obtenção de um medicamento administrável oralmente a um mamífero. Os resultados dos testes revelaram importante ação cerebral, sugerindo que a administração do composto da invenção proporciona que o elemento ativo ultrapasse a barreira hematoencefálica. Assim, os resultados mostram/suportam o uso do composto da invenção independentemente de o composto de invenção ser o ativo que atua diretamente no alvo, ou seja, não se degradando durante a ingestão oral, ou de o composto ser um precursor que, após modificação química pós-administração, atua no alvo - neste caso, se caracterizando como pró-droga. Essa característica de proporcionar importantes efeitos mesmo mediante administração oral é particularmente desejável, uma vez que as enzimas naturais de um mamífero em geral degradam peptídeos e proteínas quando no trato digestivo e raramente um medicamento com um ativo peptídico se mostra viável. Entretanto, de forma surpreendente, o composto da invenção - mesmo quando administrado pela via oral - proporciona forte ação terapêutica, neste caso ainda mais surpreendentemente, atuando no cérebro. In addition, as will be demonstrated in the following examples, the use of the compound of the invention in the preparation of a neuromodulatory, neuroprotective and / or anticonvulsant drug provides for the delivery of a medicament orally administrable to a mammal. The results of the tests revealed significant brain action, suggesting that administration of the compound of the invention provides that the active element crosses the blood brain barrier. Thus, the results show / support the use of the compound of the invention regardless of whether the compound of the invention is the active which acts directly on the target, ie, does not degrade during oral ingestion, or the compound is a precursor which, upon modification chemistry, acts on the target - in this case, being characterized as a pro-drug. This feature of providing important effects even by oral administration is particularly desirable, since the natural enzymes of a mammal in general degrade peptides and proteins while in the digestive tract and rarely a drug with a peptidic active is shown to be viable. However, surprisingly, the compound of the invention - even when administered orally - provides strong therapeutic action, in this case even more surprisingly, acting in the brain.
[0134] Portanto, independentemente do mecanismo de ação, o qual não é objeto do presente pedido de patente, o fato de a administração oral da composição farmacêutica da invenção ter proporcionado importante ação neuromoduladora, neuroprotetora e anticonvulsivante e mesmo evitar mortes, mostra com clareza a surpreendente magnitude e relevância dos problemas técnicos resolvidos. Therefore, regardless of the mechanism of action, which is not the subject of the present application, the fact that oral administration of the pharmaceutical composition of the invention has provided important neuromodulatory, neuroprotective and anticonvulsive action and even preventing deaths, shows clearly the surprising magnitude and relevance of the technical problems solved.
[0135] O presente pedido de patente revela uma composição farmacêutica que compreende o composto descrito acima. A referida composição farmacêutica compreende também um veículo farmaceuticamente aceitável, opcionalmente compreendendo também outros ativos e/ou sais farmaceuticamente aceitáveis dos mesmos. O composto da invenção é o ou um componente ativo da composição farmacêutica da invenção, que é administrada na forma de tablete, gel, cápsula, líquido oral ou xarope, supositório, solução injetável ou outras formas de administração adequadas para fins farmacêuticos e médicos.  [0135] The present application discloses a pharmaceutical composition comprising the compound described above. Said pharmaceutical composition also comprises a pharmaceutically acceptable carrier, optionally also comprising other pharmaceutically acceptable actives and / or salts thereof. The compound of the invention is one or an active component of the pharmaceutical composition of the invention, which is administered in the form of a tablet, gel, capsule, oral liquid or syrup, a suppository, an injectable solution or other suitable forms of administration for pharmaceutical and medical purposes.
[0136] Os exemplos a seguir mostrados têm o intuito somente de exemplificar algumas das diversas maneiras de se concretizar a invenção, contudo, sem limitar o escopo da mesma. [0137] Testes comparativos mostraram que o composto da invenção mostrou superior estabilidade in vitro em extremos de temperatura quando comparado à hemopressina. The following examples are only intended to exemplify some of the various ways of practicing the invention, however, without limiting the scope thereof. Comparative tests showed that the compound of the invention showed superior in vitro stability at temperature extremes when compared to hemopressin.
[0138] Em alguns exemplos, foram avaliados in vivo os efeitos neuromoduladores/neuroprotetores/anticonvulsivantes da composição da invenção mediante a administração oral a animais. A composição farmacêutica da invenção foi administrada a mamíferos {Mus musculus ou camundongo) com dose oral de tratamento com diferentes concretizações do composto da invenção, comparativamente a outros compostos ou ao controle com salina. Nestes experimentos, os compostos-teste foram administrados oralmente 10 minutos antes da administração (intraperitoneal) da pilocarpina. O hidrocloreto de pilocarpina (320 mg/kg, Merck), dissolvido em 0,9% de salina estéril, foi administrado de forma intraperitoneal para indução do SE {status epileticus) (Turski et al., 1983). No modelo de Turski, os efeitos neurotóxicos iniciam-se cerca de 15-25 minutos após a injeção de Pilo, com a ocorrência de crises convulsivas motoras e límbicas, os animais evoluindo para um estado de crises contínuas (clônicas) que caracterizam o SE (Sanabria e Cavalheiro, 2000).  In some instances, the neuromodulatory / neuroprotective / anticonvulsive effects of the composition of the invention have been evaluated in vivo by oral administration to animals. The pharmaceutical composition of the invention was administered to mammals (Mus musculus or mouse) with an oral dose of treatment with different embodiments of the compound of the invention, as compared to other compounds or the saline control. In these experiments, the test compounds were administered orally 10 minutes prior to (intraperitoneal) administration of pilocarpine. Pilocarpine hydrochloride (320 mg / kg, Merck), dissolved in 0.9% sterile saline, was administered intraperitoneally for induction of SE (status epileticus) (Turski et al., 1983). In the Turski model, the neurotoxic effects begin about 15-25 minutes after the injection of Pilo, with the occurrence of motor and limbic seizures, the animals evolving to a state of continuous (clonic) seizures that characterize SE Sanabria and Cavalheiro, 2000).
[0139] Testes comparativos mostraram também que o composto desta invenção mostrou superior atividade terapêutica in vivo quando comparado à hemopressina, cujo uso como anticonvulsivante é objeto do pedido de patente co-pendente dos mesmos inventores. Comparative tests have also shown that the compound of this invention showed superior in vivo therapeutic activity when compared to hemopressin, the use of which as an anticonvulsant is the subject of the co-pending co-pending application of the same inventors.
[0140] Diante dos surpreendentes resultados obtidos mediante a administração in vivo de diferentes concretizações do composto da invenção, em outros exemplos foram avaliadas in vitro e in silico as formas de ligação do composto da invenção aos receptores CB1 e CB2.  In view of the surprising results obtained by the in vivo administration of different embodiments of the compound of the invention, in other examples the binding forms of the compound of the invention were evaluated in vitro and in silico to CB1 and CB2 receptors.
Exemplos Examples
[0141] Exemplo 1. Testes de estabilidade em condições extremas  [0141] Example 1. Stability tests under extreme conditions
[0142] Nesta concretização, a estabilidade do composto da invenção foi comparada à da hemopressina (Hp, PVNFKFLSH) em condições extremas. A Hp sabidamente tem o problema de formação de fibrilas, assim como os variantes da mesma que têm um número maior de aminoácidos. Amostras de NFKF e de Hp foram submetidas a dois testes separados, o da estabilidade mediante congelamento por 24h e mediante aquecimento a 100°C por 10 minutos. In this embodiment, the stability of the compound of the invention was compared to that of hemopressin (Hp, PVNFKFLSH) under extreme conditions. Hp is known to have the problem of fibril formation, as well as variants thereof which have a greater number of amino acids. Samples of NFKF and Hp were subjected to two separate tests, that of stability by freezing for 24 hours and heating at 100 ° C for 10 minutes.
[0143] Os resultados da figura 1 mostram que, mediante congelamento por 24h, significante parte da Hp é perdida ou degradada (de 140 para 97, ou seja, aproximadamente 31 %), conforme evidenciam as medidas por HPLC (coluna analítica de 2.1 mm, com corrida em gradiente de 10-60%B. O solvente A é água/0.1 %TFA e o solvente B é acetonitrila/0,075%TFA). O composto da invenção na concretização NFKF, por outro lado, seguiu muito mais estável e sofreu substancialmente menos degradação (de 120 para 100, ou seja, 16,7%). Os resultados da figura 1 mostram também que mediante aquecimento a 100°C por 10 minutos, ainda mais significante parte da Hp é perdida ou degradada (de 140 para 50, ou seja, aproximadamente 64,3%), conforme evidenciam as medidas por HPLC. O composto da invenção na concretização NFKF, por outro lado, seguiu estável não sofreu degradação estatisticamente significante. The results of figure 1 show that, by freezing for 24h, a significant part of Hp is lost or degraded (from 140 to 97, ie approximately 31%), as evidenced by measurements by HPLC (analytical column of 2.1 mm , with gradient running of 10-60% B. Solvent A is water / 0.1% TFA and solvent B is acetonitrile / 0.075% TFA). The compound of the invention in the NFKF embodiment, on the other hand, remained much more stable and suffered substantially less degradation (from 120 to 100, ie, 16.7%). The results of Figure 1 also show that upon heating at 100 ° C for 10 minutes, even more significant part of the Hp is lost or degraded (from 140 to 50, ie approximately 64.3%), as evidenced by measurements by HPLC . The compound of the invention in the otherwise stable NFKF embodiment did not suffer statistically significant degradation.
[0144] Em conjunto, esses dados mostram que o composto da invenção proporciona muito maior estabilidade e menor degradação em condições extremas de temperatura, o que o favorece na manipulação, galênica, preparações medicamentosas e na estabilidade do produto farmacêutico tanto na fase pós-obtenção do princípio ativo, quanto na produção industrial de medicamentos e, não menos importante, na cadeia logística de transporte. Os dados sugerem que o tempo de prateleira do produto farmacêutico contendo o composto da invenção devem ser maiores que os congéneres contendo Hp ou outros ativos com problemas de instabilidade. Together, these data show that the compound of the invention provides much greater stability and less degradation under extreme temperature conditions, which favors it in the manipulation, pharmaceutical, pharmaceutical preparations and stability of the pharmaceutical both in the post-acquisition phase of the active principle, and in the industrial production of medicines and, not least, in the transport logistics chain. The data suggest that the shelf life of the pharmaceutical containing the compound of the invention should be greater than the congeners containing Hp or other actives with instability problems.
[0145] Exemplo 2. Uso do composto R -N-AA -K-AA2-R2 para a preparação de composição farmacêutica  Example 2. Use of compound R -N-AA -K-AA2-R2 for the preparation of pharmaceutical composition
[0146] Nesta concretização, o composto R -N-AA -K-AA2-FÍ2 é o tetrapeptídeo NFKF, que foi sintetizado por síntese química. Referido peptídeo foi utilizado na preparação de uma composição farmacêutica líquida de uso oral compreendendo entre 2,7x10"4 Molar do referido peptídeo e um veículo farmaceuticamente aceitável. Nesta concretização, o referido veículo é solução salina, a composição farmacêutica sendo uma solução de uso oral. Referida composição foi utilizada para a administração oral in vivo a mamíferos conforme exemplos 3 -6 a seguir. In this embodiment, the compound R-N-AA -K-AA 2- F 2 is the NFKF tetrapeptide, which has been synthesized by chemical synthesis. Said peptide was used in the preparation of a liquid pharmaceutical composition for oral use comprising between 2,7x10 "4 Molar said peptide and a pharmaceutically acceptable carrier. In this embodiment, said carrier solution is saline, the pharmaceutical composition being a solution for oral use. Said composition was used for oral in vivo administration to mammals according to examples 3-6 below.
[0147] Em outras concretizações, a composição farmacêutica se apresenta na forma de tablete, gel, líquido oral ou xarope, cápsula, supositório, solução injetável ou formas inaláveis ou em adesivo, opcionalmente compreendendo outros princípios ativos.  In other embodiments, the pharmaceutical composition is in the form of a tablet, gel, oral liquid or syrup, capsule, suppository, injectable solution or inhalable or adhesive forms, optionally comprising other active principles.
[0148] Exemplo 3. Composição farmacêutica neuromoduladora compreendendo o composto NFKF - resultados de testes in vivo  [0148] Example 3. Neuromodulatory pharmaceutical composition comprising compound NFKF - in vivo test results
[0149] Nesta concretização, o efeito neuromodulador da composição da invenção preparada de acordo com o exemplo 2 foi avaliado mediante prévia administração da composição invenção e subsequente administração de pilocarpina a animais. Outros compostos-teste também foram avaliados conforme descrito a seguir. A administração de pilocarpina leva a lesões encefálicas severas, neurotoxicidade e normalmente culmina na morte dos animais. Esta substância foi usada nos experimentos descritos a seguir, porém seus efeitos neuronais/encefálicos danosos foram inibidos pela prévia administração da composição da presente invenção: a vasta maioria dos animais submetidos a estes experimentos não apresentou sintomas relacionados às lesões cerebrais e sobreviveu sem danos aparentes, em contraste aos grupos de animais tratados com outras substâncias conhecidas. In this embodiment, the neuromodulatory effect of the composition of the invention prepared according to example 2 was evaluated by prior administration of the inventive composition and subsequent administration of pilocarpine to animals. Other test compounds were also evaluated as described below. Administration of pilocarpine leads to severe brain injury, neurotoxicity and usually culminates in the death of the animals. This substance was used in the experiments described below but its harmful neuronal / encephalic effects were inhibited by the prior administration of the composition of the present invention: the vast majority of the animals subjected to these experiments had no symptoms related to brain lesions and survived without apparent damage, in contrast to groups of animals treated with other known substances.
[0150] A composição da invenção preparada de acordo com o Exemplo 2 foi administrada previamente aos animais. A figura 2 apresenta os resultados de testes de neuroproteção com o com o composto da invenção NFKF no modelo de pilocarpina, sendo indicado o perfil de sobrevivência/morte dos animais mediante a administração do peptídeo da invenção. Em A) é mostrado o perfil de sobrevivência dos animais aos quais foi administrado o controle (apenas salina); Em B) é mostrado o perfil de sobrevivência dos animais aos quais foi administrado o canabidiol 30mg/kg; Em C) é mostrado o perfil de sobrevivência dos animais aos quais foi administrado o peptídeo da invenção NFKF 500^g/kg; Em D) são mostrados todos os perfis em um só gráfico. Interessante notar que no grupo tratado com o composto da invenção NFKF 500 μg/kg apenas dois animais que morreram. Os demais animais do grupo tratado com o peptídeo da invenção, no total de 3, seguiram vivos por mais de uma semana, enquanto praticamente todos os demais morreram em menos de 30 minutos. Consequentemente, o tempo médio de sobrevida para este grupo é significativamente diferente e muito maior do que o dos demais. Ademais, quando se compara o grupo ao qual foi administrado o composto invenção NFKF (500 μg/kg) com o grupo ao qual foi administrado o canabidiol (30mg/kg) destaca-se que a sobrevivência é três vezes maior no primeiro grupo, ou seja, no grupo ao qual foi administrado o composto invenção NFKF 500 μg/kg, mesmo usando uma concentração relativa de composto 60 vezes menor. The composition of the invention prepared according to Example 2 was pre-administered to the animals. Figure 2 shows the results of neuroprotection tests with the compound of the invention NFKF in the pilocarpine model, the survival / death profile of the animals being indicated by administration of the peptide of the invention. In A) the survival profile of the animals to which the control was administered (saline only) is shown; In B) the survival profile of the animals administered cannabidiol 30mg / kg is shown; In C) the survival profile of the animals to which the peptide of the invention is administered NFKF 500 Âμg / kg; In D) all profiles are shown in a single graph. Interesting note that in the group treated with the compound of the invention NFKF 500 μg / kg only two animals died. The remaining animals in the group treated with the peptide of the invention, totaling 3, remained alive for more than a week, while practically all the others died in less than 30 minutes. Consequently, the mean survival time for this group is significantly different and much higher than the others. In addition, when comparing the group to which the invention compound NFKF (500 μg / kg) was administered with the group to which cannabidiol (30 mg / kg) was administered, it is noted that survival is three times greater in the first group, or in the group to which the compound invention was administered NFKF 500 μg / kg, even using a 60-fold lower relative concentration of compound.
[0151] Além dos resultados mostrados na figura 2 acima, importantes efeitos técnicos adicionais incluem a observação de não ocorrência de prostração ou sangramento nasal nos animais submetidos ao tratamento com o composto da invenção NFKF, enquanto que no grupo dos animais tratados com canabidiol tanto a prostração quanto o sangramento nasal foram observados. Estes, portanto, são adicionais problemas técnicos resolvidos pelo composto da invenção. In addition to the results shown in figure 2 above, important additional technical effects include observation of non-occurrence of prostration or nasal bleeding in animals undergoing treatment with the compound of the invention NFKF, while in the group of animals treated with both cannabidiol prostration and nasal bleeding were observed. These, therefore, are additional technical problems solved by the compound of the invention.
[0152] Exemplo 4. Composição farmacêutica anticonvulsivante compreendendo o composto NFKF - resultados de testes in vivo  Example 4. Anticonvulsive pharmaceutical composition comprising the compound NFKF - in vivo test results
[0153] Nesta concretização, o efeito anticonvulsivante da composição da invenção preparada de acordo com o exemplo 2 foi avaliado mediante prévia administração da composição invenção e subsequente administração de pilocarpina a animais. A figura 3 apresenta os resultados de testes com o modelo de pilocarpina, sendo indicado o tempo para a ocorrência da primeira salivação com a administração de controle, canabidiol (30mg/kg) do composto da invenção R -N-AA -K-AA2-FÍ2, em concretização na qual é o tetrapeptídeo NFKF (500μg/kg). Os dados entre controle e demais compostos teste não apresentem significância estatística entre si nas condições testadas. In this embodiment, the anticonvulsive effect of the composition of the invention prepared according to example 2 was evaluated by prior administration of the inventive composition and subsequent administration of pilocarpine to animals. Figure 3 presents the results of the pilocarpine model, indicating the time for the first salivation to occur with the administration of cannabidiol (30mg / kg) of the compound of the invention R-N-AA-K-AA 2 -F 2, in which embodiment it is the NFKF tetrapeptide (500μg / kg). The data between control and other test compounds do not present statistical significance among themselves under the conditions tested.
[0154] A figura 4 apresenta os resultados de testes com o modelo de pilocarpina, sendo indicado o tempo para a ocorrência da primeira convulsão com a administração de canabidiol (30mg/kg) e o peptídeo da invenção NFKF (50C^g/kg). Os asteriscos indicam: (**) P<0.02 vs Controle; (***) P<0.002 vs Controle. [0154] Figure 4 presents the results of tests with the pilocarpine model, indicating the time for the occurrence of the first convulsion with the administration of cannabidiol (30mg / kg) and the peptide of the invention NFKF (50C gg / kg). The asterisks indicate: ( ** ) P <0.02 vs Control; ( *** ) P <0.002 vs Control.
[0155] Além dos resultados mostrados nas figuras 3 e 4, importantes efeitos técnicos adicionais incluem a observação de não ocorrência de prostração ou sangramento nasal nos animais submetidos ao tratamento com o composto da invenção NFKF, enquanto que no grupo dos animais tratados com canabidiol tanto a prostração quanto o sangramento nasal foram observados. Estes, portanto, são adicionais problemas técnicos resolvidos pelo composto da invenção.  In addition to the results shown in Figures 3 and 4, important additional technical effects include observation of non-occurrence of prostration or nasal bleeding in animals undergoing treatment with the compound of the invention NFKF, while in the group of animals treated with both cannabidiol prostration and nasal bleeding were observed. These, therefore, are additional technical problems solved by the compound of the invention.
[0156] Exemplo 5. Comparativo da composição farmacêutica compreendendo o composto NFKF com a composição farmacêutica compreendendo Hp - resultados de testes in vivo  Example 5. Comparative pharmaceutical composition comprising the compound NFKF with the pharmaceutical composition comprising Hp - in vivo test results
[0157] Neste exemplo, foram comparados os efeitos da composição da invenção em relação à composição farmacêutica contendo Hemopressina (Hp ou PVNFKFLSH), cujo uso como anticonvulsivante é objeto de pedido de patente co-pendente dos mesmos inventores.  In this example, the effects of the composition of the invention with respect to the pharmaceutical composition containing Hemopressin (Hp or PVNFKFLSH), whose use as an anticonvulsant is the subject of co-pending patent application of the same inventors, were compared.
[0158] A figura 5 apresenta os resultados de testes do composto da invenção NFKF como comparativamente aos resultados de testes da hemopressina, ambos no modelo de pilocarpina. São apresentados os porcentuais de tempo em relação ao controle para a ocorrência da primeira salivação com a administração das seguintes doses de tratamento: o controle (salina) ; hemopressina (Hp ou PVNFKFLSH, 0,551334 μιτιοΙ/kg); hemopressina (0,91889 μη-iol/kg); o peptídeo da invenção NFKF (0,540882 μη-ιοΙ/kg); NFKF (0,901469 μηποΙ/kg); o PEP-19 (DIIADDEPLT, 0,9081 17 μηποΙ/kg). Os asteriscos indicam a significância estatística: (*) P<0.05 vs Controle; (**) P<0.01 vs Controle; o sinal + indica P<0.05 vs Hp 0,91889 μπιοΙ/kg. Figure 5 shows the test results of the compound of the invention NFKF as compared to the results of hemopressin tests, both in the pilocarpine model. The percentages of time in relation to the control for the occurrence of the first salivation are presented with the administration of the following treatment doses: control (saline); hemopressin (Hp or PVNFKFLSH, 0.551334 μιτιοΙ / kg); hemopressin (0.91889 μ-iol / kg); the peptide of the invention NFKF (0.540882 μη-ιοΙ / kg); NFKF (0.901469 μηποΙ / kg); or PEP-19 (DIIADDEPLT, 0.9081 17 μηποΙ / kg). The asterisks indicate the statistical significance: ( * ) P <0.05 vs Control; ( ** ) P <0.01 vs Control; the + sign indicates P <0.05 vs Hp 0.91889 μπιοΙ / kg.
[0159] De um lado, a salivação induzida pela administração de pilocarpina é indicativa de alterações nos receptores muscarínicos. Consequentemente, a substancial alteração no perfil de tempo para a ocorrência da primeira salivação observada com a administração prévia do composto da invenção, sugere modulação, direta ou indireta, de receptores muscarínicos. [0159] On the one hand, salivation induced by administration of pilocarpine is indicative of changes in muscarinic receptors. Consequently, the substantial change in the time profile for the occurrence of the first salivation observed with prior administration of the compound of the invention suggests modulation, either directly or indirectly, of muscarinic receptors.
[0160] Além disso, os resultados da figura 5 mostram claramente que a administração oral do composto da presente invenção proporciona atividade moduladora do tempo de ocorrência da primeira salivação substancialmente maior que aquela observada com a administração oral da Hp.  Furthermore, the results of Figure 5 clearly show that oral administration of the compound of the present invention provides substantially higher salivation time modulating activity substantially greater than that observed with oral administration of Hp.
[0161] Exemplo 6. Comparativo da composição farmacêutica anticonvulsivante compreendendo o composto NFKF com a composição farmacêutica anticonvulsivante compreendendo Hp - resultados de testes in vivo  Example 6. Comparative anticonvulsant pharmaceutical composition comprising the compound NFKF with the anticonvulsant pharmaceutical composition comprising Hp - in vivo test results
[0162] Neste exemplo, foram comparados os efeitos anticonvulsivantes da composição da invenção em relação à composição farmacêutica contendo Hemopressina (Hp ou PVNFKFLSH), cujo uso como anticonvulsivante é objeto de pedido de patente co-pendente dos mesmos inventores.  In this example, the anticonvulsive effects of the composition of the invention were compared to the pharmaceutical composition containing Hemopressin (Hp or PVNFKFLSH), the use of which as an anticonvulsant is co-pending co-pending application by the same inventors.
[0163] A figura 6 apresenta os resultados de testes do composto da invenção NFKF como anticonvulsivante comparativamente aos resultados de testes da hemopressina como anticonvulsivante, ambos no modelo de pilocarpina. São apresentados os porcentuais do tempo (em relação ao controle) para a ocorrência da primeira convulsão com a administração das seguintes doses de tratamento: hemopressina (Hp ou PVNFKFLSH, 0,551334 μιηοΙ/kg); hemopressina (0,91889 μιηοΙ/kg); o peptídeo da invenção NFKF (0,540882 μηποΙ/kg); NFKF (0,901469 μηποΙ/kg); o PEP-19 (DIIADDEPLT, 0,9081 17 μιηοΙ/kg) e o controle (salina). Os asteriscos indicam: (*) P<0.05 vs Controle; (**) P<0.01 vs Controle; o sinal + indica P<0.05 vs Hp 0,91889 μπιοΙ/kg. Figure 6 shows the test results of the compound of the invention NFKF as an anticonvulsant compared to the results of tests of hemopressin as an anticonvulsant, both in the pilocarpine model. The percentages of time (relative to the control) for the first seizure occur with administration of the following treatment doses: hemopressin (Hp or PVNFKFLSH, 0.551334 μιηοΙ / kg); hemopressin (0.91889 μιηοΙ / kg); the peptide of the invention NFKF (0.540882 μηποΙ / kg); NFKF (0.901469 μηποΙ / kg); PEP-19 (DIIADDEPLT, 0.9081 17 μιηοΙ / kg) and control (saline). The asterisks indicate: ( * ) P <0.05 vs Control; ( ** ) P <0.01 vs Control; the + sign indicates P <0.05 vs Hp 0.91889 μπιοΙ / kg.
[0164] Além disso, os resultados da figura 6 mostram claramente que a administração oral do composto da presente invenção proporciona atividade moduladora da ocorrência da primeira convulsão substancialmente maior que aquela observada com a administração oral da Hp. Observa-se, por exemplo, o mesmo efeito com metade da dose do composto da invenção quando comparado com a Hp. [0165] Os resultados dos testes realizados nos exemplos 3 a 6 acima mostram importantes e significativos resultados in vivo, em faixas de dosagem da ordem de 500 a 1000 μg de composto da invenção por kg de animal. Considerando se tratar de testes em camundongos e a conversão para a Dose Equivalente em Humanos referida acima, efeitos de mesma magnitude são esperados em humanos na faixa de dosagem de 40 a 80 μg de composto da invenção por kg de humano e, considerando-se as faixas de segurança cabíveis, concentrações entre 4 a 800 μg/kg para administração em humanos são consideradas na presente invenção. Furthermore, the results of Figure 6 clearly show that oral administration of the compound of the present invention provides substantially higher seizure-modulating activity substantially greater than that observed with oral administration of Hp. For example, the same effect is observed with half the dose of the compound of the invention as compared to Hp. The results of the tests performed in examples 3 to 6 above show significant and significant in vivo results, in dosage ranges of the order of 500 to 1000 μg of compound of the invention per kg of animal. Considering the tests in mice and the conversion to the Human Equivalent Dose mentioned above, effects of the same magnitude are expected in humans in the dosage range of 40 to 80 μg of compound of the invention per kg of human and, Safety ranges, concentrations between 4 to 800 μg / kg for administration to humans are considered in the present invention.
[0166] Exemplo 7. Testes in silico de liqação/interação do composto NFKF ao receptor CB1  Example 7. In silico liqation tests / interaction of compound NFKF to CB1 receptor
[0167] O receptor canabinóide 1 (CB1 ) corresponde ao GPCR (do inglês, Receptores Acoplados à Proteína G) mais expresso no cérebro humano e é encontrado em elevados níveis no Sistema Nervoso Central de modo geral (Figura 7). É ativado por endocanabinóides e tem sido apontado como um alvo terapêutico promissor para o tratamento de diversas doenças como dor e inflamação, esclerose múltipla e doenças neurodegenerativas (efeito agonista) e ainda, obesidade, fibrose hepática e dependência à nicotina (efeito antagonista) (HUA et al., 2016).  [1] The cannabinoid receptor 1 (CB1) corresponds to GPCR (Expressed Receptors to Protein G) most expressed in the human brain and is found at high levels in the Central Nervous System in general (Figure 7). It is activated by endocannabinoids and has been indicated as a promising therapeutic target for the treatment of various diseases such as pain and inflammation, multiple sclerosis and neurodegenerative diseases (agonist effect) and obesity, liver fibrosis and nicotine dependence (antagonistic effect) et al., 2016).
[0168] A figura 7 mostra uma visão geral da estrutura tridimensional de um GPCR, neste caso, o receptor canabinóide do subtipo 1 . Em destaque as sete hélices transmembranares (l-VII), as alças intracelulares (ICL1 e ICL2) e, as alças extracelulares (ECL2 e ECL3).  Figure 7 shows an overview of the three-dimensional structure of a GPCR, in this case, the cannabinoid receptor of subtype 1. The seven transmembrane helices (l-VII), the intracellular loops (ICL1 and ICL2) and the extracellular loops (ECL2 and ECL3) were highlighted.
[0169] Através da utilização de ferramentas de Modelagem Molecular foi avaliado neste exemplo o modo de interação, no sítio de ligação de CB1 , do composto da invenção R1-N-AA1-K-AA2-R2, em concretização na qual é o tetrapeptídeo NFKF. Além disso, foi comparado o perfil observado com o modo de interação de outros ligantes conhecidos, como o canabidiol, rimonabanto e AM6538. Through the use of Molecular Modeling tools we have evaluated in this example the mode of interaction at the CB1 binding site of the compound of the invention R1-N-AA1-K-AA2-R2, in which embodiment it is the tetrapeptide NFKF. In addition, the profile observed was compared with the mode of interaction of other known binders, such as cannabidiol, rimonabant and AM6538.
Figure imgf000035_0001
Figure imgf000035_0001
[0170] Os testes in silico do perfil de interação do tetrapeptídeo NFKF com o receptor CB1 foram feitos a partir do desenho e otimização das estruturas dos ligantes. Inicialmente, foi utilizado o programa Percepta para a verificação do estado de ionização desses ligantes em pH plasmático (pH = 7,4). Posteriormente, a construção das estruturas dos ligantes foi realizada no Programa Spartan v. 16 (Wavefunction, Inc.) seguida de minimização de energia por mecânica molecular. [0170] In silico tests of the interaction profile of the NFKF tetrapeptide with the CB1 receptor were made from the design and optimization of the ligand structures. Initially, the Percepta program was used to verify the ionization status of these ligands at plasma pH (pH = 7.4). Subsequently, the construction of ligand structures was performed in the Spartan v. 16 (Wavefunction, Inc.) followed by energy minimization by molecular mechanics.
[0171] A seleção da estrutura cristalográfica CB1 para os estudos de docking foi feita a partir dos dados disponíveis no Protein Data Bank (PDB), código PDB 5TGZ, pertencendo à espécie Homo sapiens e apresentando uma resolução de 2,8 Á. A referida estrutura corresponde ao único cristal de CB1 disponível no banco de dados e foi disponibilizada após a data de prioridade deste pedido de patente.  The selection of the CB1 crystallographic structure for the docking studies was done from data available in the PDB code 5TGZ, belonging to the species Homo sapiens and presenting a resolution of 2.8 Á. Said structure corresponds to the only available CB1 crystal in the database and was made available after the priority date of this patent application.
[0172] Em seguida, foi feita a validação das funções do Programa GOLD v. 5.1 e realização dos estudos de docking. Para identificação do complexo mais estável, foram consideradas três funções de pontuação presentes no Programa GOLD (ChemScore, Goldscore e ChemPLP). Para a realização do ancoramento molecular no Programa GOLD v. 5.5 (CCDC) foi necessária a localização do sítio ativo utilizando-se como referência o próprio ligante co- cristalizado (AM6538) e todos os aminoácidos a 6 Á de distância do mesmo. Posteriormente, foi realizado o ancoramento molecular e análise dos resultados no Programa PyMOL v. 1 .8.6.2 (Schrodinger, LCC). [0172] The validation of the functions of the GOLD v. 5.1 and conducting docking studies. To identify the most stable complex, three scoring functions were considered in the GOLD Program (ChemScore, Goldscore and ChemPLP). For the realization of molecular anchoring in the GOLD v. 5.5 (CCDC) the location of the active site was required using the co-crystallized binder (AM6538) itself and all amino acids at 6 Âμ apart. Subsequently, molecular anchoring and analysis of the results in the PyMOL v. 1 .8.6.2 (Schrodinger, LCC).
[0173] Inicialmente, as funções de pontuação do programa GOLD v. 5.5 foram avaliadas segundo a capacidade de prever complexos proteína-ligante compatíveis com os resultados obtidos experimentalmente. Para isso, três funções de pontuação {Goldscore, Chemscore e ChemPLP) foram validadas através do redocking do ligante de referência, AM6538, no sítio de ligação do receptor CB1 (PDB 5TGZ). Primeiramente, realizou-se o docking na ausência de moléculas de água e, posteriormente, buscando-se estimar o quanto as moléculas de água presentes no sítio de ligação poderiam influenciar na orientação do ligante, a mesma avaliação foi feita na presença de moléculas de água. Os resultados alcançados por desvio quadrático médio {Root Mean Square Deviation RMSD), considerando os complexos de maior pontuação, podem ser observados na tabela 1 .  [0173] Initially, the scoring functions of the GOLD v. 5.5 were evaluated according to the ability to predict protein-binding complexes compatible with experimental results. For this, three scoring functions (Goldscore, Chemscore and ChemPLP) were validated by redocking the reference ligand, AM6538, at the CB1 receptor binding site (PDB 5TGZ). Firstly, docking was performed in the absence of water molecules and, afterwards, trying to estimate how much the water molecules present in the binding site could influence the orientation of the binder, the same evaluation was made in the presence of water molecules . The results obtained by Root Mean Square Deviation RMSD, considering the complexes with higher scores, can be observed in table 1.
[0174] Para o cálculo do RMSD são considerados e comparados pares de átomos do resultado dos experimentos de modelagem molecular e dos experimentos de estrutura cristalográfica. Assim, a raiz quadrada da média entre as distâncias desses átomos é calculada, obtendo-se os valores para o desvio. Quanto menor é este desvio, maior é a aproximação entre os resultados, indicando portanto a função mais adequada a ser utilizada (MAIOROV; CRIPPEN, 1994; HILDEBRANDT et al., 2013).  [0174] For the calculation of the RMSD are considered and compared pairs of atoms of the result of the molecular modeling experiments and the experiments of crystallographic structure. Thus, the square root of the mean between the distances of these atoms is calculated, obtaining the values for the deviation. The smaller the deviation, the greater the approximation between the results, thus indicating the most adequate function to be used (MAIOROV; CRIPPEN, 1994; HILDEBRANDT et al., 2013).
[0175] Tabela 1 : Valores de RMSD obtidos a partir da validação das funções de pontuação do programa GOLD v. 5.5 para o redocking de AM6538 no receptor CB1 . [0175] Table 1: RMSD values obtained from the validation of the scoring functions of the GOLD v. 5.5 for the redocking of AM6538 in the CB1 receptor.
Função RMSD  RMSD function
Com água Sem água  With water Without water
CHEMPLP 0,9543 0,9134  CHEMPLP 0.9543 0.9134
CHEMSCORE 1 ,1762 1 ,1578  CHEMSCORE 1, 1762, 1, 1578
GOLDSCORE 0,9384 0,8864 [0176] Como indicado na tabela 1 , a função que apresentou menores valores de RMSD, e que demonstrou ser a mais adequada para os experimentos de docking, foi a Goldscore na ausência de moléculas de água no sítio de ligação, indicando que tais moléculas não influenciam no modo de interação do ligante. GOLDSCORE 0.9384 0.8864 As indicated in Table 1, the function which showed the lowest RMSD values, and which proved to be most suitable for the docking experiments, was Goldscore in the absence of water molecules at the binding site, indicating that such molecules did not influence the mode of interaction of the binder.
[0177] A Figura 8 mostra de forma visual a sobreposição entre o complexo da estrutura cristalográfica (PDB 5TGZ) e o complexo resultante do experimento de redocking de AM6538 utilizando a metodologia validada. Figure 8 shows visually the overlap between the crystallographic structure complex (PDB 5TGZ) and the resulting complex from the AM6538 redocking experiment using the validated methodology.
[0178] O complexo resultante do redocking de AM6538 em CB1 obteve uma pontuação de 87,39 e o perfil de interação observado foi coerente com o descrito na literatura (HUA et al., 2016), sendo observadas diversas interações hidrofóbicas com resíduos de aminoácidos como: Phe-102, Phe-170, Phe-174 e Phe-379, bem como resíduos de leucina, metionina, cisteína e valina (ver Figura 9). The complex resulting from the redocking of AM6538 in CB1 obtained a score of 87.39 and the interaction profile observed was consistent with that described in the literature (HUA et al., 2016), with several hydrophobic interactions with amino acid residues such as: Phe-102, Phe-170, Phe-174 and Phe-379, as well as leucine, methionine, cysteine and valine residues (see Figure 9).
[0179] Por outro lado, para o rimonabanto e canabidiol, os perfis de interação observados foram bastante semelhantes ao do AM6538, apresentando, respectivamente, pontuações de 73,43 e 54,91 (Figuras 10 e 1 1 , respectivamente).  [0179] On the other hand, for the rimonabant and cannabidiol, the interaction profiles observed were very similar to the AM6538, respectively, presenting scores of 73.43 and 54.91 (Figures 10 and 11, respectively).
[0180] No experimento do docking de uma concretização do composto da invenção (tetrapeptídeo NFKF) sua estrutura foi analisada quanto a seu estado de ionização em pH plasmático (pH = 7,4) no programa Percepta, que indicou a estrutura apresentada abaixo, a ual foi utilizada para os estudos de docking.  In the docking experiment of one embodiment of the compound of the invention (NFKF tetrapeptide) its structure was analyzed for its ionizing state at plasma pH (pH = 7.4) in the Percepta program, which indicated the structure shown below, ual was used for docking studies.
Figure imgf000037_0001
Figure imgf000037_0001
[0181] O complexo resultante do docking do tetrapeptídeo NFKF em CB1 obteve uma pontuação de 100,66, substancialmente maior que a pontuação alcançada pelos ligantes de referência. Este fato pode ser explicado por ligações de hidrogénio adicionais observadas entre o tetrapeptídeo e os resíduos Ser-123, Thr-197, Ser-167 e Ser-383. Além disso, as interações hidrofóbicas observadas para AM6538 também estão presentes no modo de interação de NFKF (Figura 12). The resulting docking complex of the NFKF tetrapeptide in CB1 obtained a score of 100.66, substantially greater than the score achieved by the reference ligands. This fact can be explained by additional hydrogen bonds observed between the tetrapeptide and residues Ser-123, Thr-197, Ser-167 and Ser-383. In addition, the hydrophobic interactions observed for AM6538 are also present in the NFKF interaction mode (Figure 12).
[0182] Em conjunto, os resultados destes experimentos mostram que o perfil de interação observado para NFKF, com interações semelhantes aos ligantes de referência, bem como interações adicionais que resultaram em uma maior pontuação obtida nos estudos de docking, sugerem que o tetrapeptídeo NFKF é um potencial ligante de receptores canabinóides do subtipo 1 .  Together, the results of these experiments show that the observed interaction profile for NFKF, with similar interactions to the reference ligands, as well as additional interactions that resulted in a higher score obtained in the docking studies, suggest that the NFKF tetrapeptide is a potential binding of cannabinoid receptors of subtype 1.
[0183] Exemplo 8. Testes in silico de liqação/interação do composto NFKF ao receptor CB2 Example 8. In silico liqation tests / interaction of compound NFKF to CB2 receptor
[0184] Para viabilizar experimentos in silico da interação do composto da invenção R -N-AA -K-AA2-FÍ2, em concretização na qual é o tetrapeptídeo NFKF, com o receptor CB2, foi primeiramente construído um modelo tridimensional do receptor CB2, para subsequentes experimentos de docking. In order to enable in silico experiments of the interaction of the compound of the invention R-N-AA-K-AA 2- F 2 , in which the NFKF tetrapeptide with the CB 2 receptor is made, a three-dimensional model of the CB2 receptor was first constructed , for subsequent docking experiments.
[0185] Nesta concretização, a construção do modelo 3D de CB2 foi feita a partir de uma busca no banco de dados UniProt da sequência de aminoácidos deste receptor. O critério utilizado para a seleção da sequência foi a espécie {Homo sapiens). Assim, a sequência selecionada para a realização dos estudos de modelagem molecular foi a de código P34972. [0185] In this embodiment, the construction of the 3D model of CB2 was done from a search in the UniProt database of the amino acid sequence of this receptor. The criterion used for sequence selection was the species (Homo sapiens). Thus, the sequence selected for carrying out the molecular modeling studies was that of code P34972.
[0186] Em seguida, foi utilizada a ferramenta Template Identification do servidor SwissModel para identificação e seleção da proteína molde. A sequência apontada pelo servidor como a de maior identidade estrutural com a sequência de CB2 humana foi a de código PDB 5TGZ (HUA et al., 2016) pertencente à espécie Homo sapiens, correspondente ao receptor CB1 . [0186] Next, the Template Identification tool of the SwissModel server was used to identify and select the protein template. The sequence identified by the server as having the highest structural identity to the human CB2 sequence was that of PDT 5TGZ code (HUA et al., 2016) belonging to the species Homo sapiens, corresponding to the CB1 receptor.
[0187] As sequências alvo e molde foram então alinhadas, utilizando-se o software ClustalW2 vinculado ao banco de dados UniProt, visando a comparação entre as sequências para estabelecer a porcentagem de identidade entre elas. O modelo 3D por homologia de CB2 humana foi construído através da ferramenta Automated Mode disponível na página do servidor Swiss-model e, para a validação do modelo gerado, foram analisadas a qualidade estrutural global e a qualidade estereoquímica do modelo através do valor apresentado para o parâmetro GMQE e da análise do gráfico de Ramachandran. The target and template sequences were then aligned using ClustalW2 software linked to the UniProt database to compare the sequences to establish the percentage of identity between them. The 3D homology model of human CB2 was constructed using the Automated Mode tool available on the Swiss-model server page and, for the validation of the generated model, were analyzed the overall structural quality and the stereochemical quality of the model through the value presented for the GMQE parameter and the analysis of the Ramachandran graph.
[0188] Após a criação do modelo 3D para CB2, foram realizados os estudos de docking para o tetrapeptídeo NFKF bem como para o canabidiol, rimonabanto e AM6538.  After the creation of the 3D model for CB2, the docking studies for NFKF tetrapeptide as well as for cannabidiol, rimonabant and AM6538 were performed.
[0189] Após a identificação e seleção da sequência do receptor CB2 humano no UniProt e da identificação da proteína-molde (código PDB 5TGZ) a partir da ferramenta Template Identification no servidor Swiss-Model, foi feito o alinhamento entre ambas as sequências no programa ClustalW2. A identidade entre as duas sequências foi de 46,18%, o que viabiliza a utilização da modelagem comparativa como ferramenta de obtenção da estrutura 3D do receptor CB2 humano, conforme preconizado em estudos previamente publicados (MORGAN; HURLEY, 2015), os quais afirmam ser necessária a identidade de no mínimo 30% entre sequências primárias para a criação de modelos por homologia.  Following the identification and selection of the human CB2 receptor sequence in UniProt and the identification of the template protein (PDB code 5TGZ) from the Template Identification tool on the Swiss-Model server, alignment was made between the two sequences in the program ClustalW2. The identity between the two sequences was 46.18%, which makes possible the use of comparative modeling as a tool to obtain the 3D structure of the human CB2 receptor, as recommended in previously published studies (MORGAN and HURLEY, 2015), which affirm identity of at least 30% between primary sequences for the creation of homology models is required.
[0190] A qualidade estereoquímica do modelo foi analisada através do gráfico de Ramachandran (figura 13B). O gráfico de Ramachandran corresponde a um modelo matemático, o qual relaciona os ângulos diedros Φ (ângulo C-N-Ca-C) no eixo-x e Ψ (ângulo N-Ca-C-N) no eixo-y. Este gráfico é dividido em regiões capazes de representar a probabilidade de uma combinação entre os ângulos Φ e Ψ (RAMACHANDRAN; RAMAKRISHNAN; SASISEKHARAN, 1963). Essas regiões são classificadas em: favoráveis, permitidas, generosamente permitidas e proibidas.  [0190] The stereochemical quality of the model was analyzed using the Ramachandran graph (Figure 13B). The Ramachandran graph corresponds to a mathematical model, which relates the dihedral angles Φ (angle C-N-Ca-C) on the x-axis and Ψ (angle N-Ca-C-N) on the y-axis. This graph is divided into regions capable of representing the probability of a combination between the angles Φ and Ψ (RAMACHANDRAN; RAMAKRISHNAN; SASISEKHARAN, 1963). These regions are classified as: favorable, permitted, generously permitted and prohibited.
[0191] O gráfico de Ramachandran construído para o modelo de CB2 humano mostrou a presença de aproximadamente 96% dos resíduos de aminoácidos nas regiões mais favoráveis e mais de 4% dos resíduos em regiões aceitáveis. Nenhum resíduo de aminoácido foi localizado em regiões proibidas, indicando a validação estereoquímica do modelo criado. Após a validação do modelo foram realizados os estudos de docking em CB2 do tetrapeptídeo NFKF, do antagonista AM6538, do ligante endógeno canabidiol e do antagonista rimonabanto. The Ramachandran graph constructed for the human CB2 model showed the presence of approximately 96% of the amino acid residues in the most favorable regions and more than 4% of the residues in acceptable regions. No amino acid residue was found in forbidden regions, indicating the stereochemical validation of the model created. After the validation of the model, the docking studies on CB2 of the NFKF tetrapeptide, AM6538 antagonist, the cannabidiol endogenous ligand and the rimonabant antagonist.
[0192] Para todos os compostos analisados a pontuação obtida nos estudos foi inferior às atingidas para CB1 , sendo de 61 ,53 para AM6538, 72,33 para o tetrapeptídeo NFKF, 43,31 para o canabidiol e finalmente, 57,40 para o rimonabanto.  [0192] For all the compounds analyzed, the scores obtained in the studies were lower than those achieved for CB1, being 61, 53 for AM6538, 72.33 for NFKF tetrapeptide, 43.31 for cannabidiol and finally 57.40 for rimonabant.
[0193] Como, de maneira simplista, os valores de pontuação podem ser considerados como um somatório das interações identificadas entre a proteína e o ligante, as pontuações menores podem então ser explicadas por um menor número de interações no sítio de ligação, ainda que as principais interações hidrofóbicas observadas em CB1 sejam mantidas em CB2 como pode ser observado nas Figuras 14-17.  Since, in a simplistic way, punctuation values can be considered as a sum of the interactions identified between the protein and the linker, the lower scores can then be explained by a lower number of interactions at the binding site, although the major hydrophobic interactions observed in CB1 are maintained in CB2 as can be seen in Figures 14-17.
[0194] Também é importante ressaltar que para o tetrapeptídeo NFKF as interações por ligação de hidrogénio apontadas em CB1 não ocorrem em CB2, contribuindo para sua pontuação inferior em CB2 (Figura 17), ainda que este peptídeo tenha apresentado uma outra ligação de hidrogénio com o resíduo de Lys-192.  [0194] It is also important to note that for the NFKF tetrapeptide the hydrogen bonded interactions indicated in CB1 do not occur in CB2, contributing to its lower score in CB2 (Figure 17), although this peptide showed another hydrogen bond with the Lys-192 residue.
[0195] A Tabela 2 apresenta de forma resumida a comparação entre as pontuações obtidas pelos estudos de docking em ambos os receptores (CB1 e CB2) nos exemplos 7 e 8 acima:  Table 2 summarizes the comparison between the scores obtained by the docking studies on both receptors (CB1 and CB2) in examples 7 and 8 above:
[0196] Tabela 2: Comparação entre as pontuações obtidas nos estudos de docking para os receptores CB1 e CB2.  [0196] Table 2: Comparison between the scores obtained in the docking studies for CB1 and CB2 receptors.
Figure imgf000040_0001
Figure imgf000040_0001
[0197] Os resultados dos testes experimentais descritos nos exemplos 7 e 8 acima permitiram reproduzir, para os demais compostos testados, o modo de interação do antagonista AM6538 no sítio de ligação do receptor CB1 , conforme descrito na literatura por Hua e colaboradores. Além disso, os resultados proporcionaram a validação da metodologia de docking para os experimentos com o tetrapeptídeo NFKF comparativamente ao canabidiol e ao antagonista rimonabanto, sendo possível, portanto, a previsão de seus perfis de interação em CB1 . [0197] The results of the experimental tests described in examples 7 and 8 above allowed reproducing, for the other compounds tested, the mode of interaction of the AM6538 antagonist at the CB1 receptor binding site, as described in the literature by Hua et al. In addition, the results provided validation of the docking methodology for experiments with NFKF tetrapeptide compared to cannabidiol and rimonabant antagonist, and it is therefore possible to predict their interaction profiles in CB1.
[0198] A construção e validação de um modelo 3D por homologia do receptor CB2 resultou também na visualização do comportamento de todos os ligantes citados no sítio de ligação de tal receptor, proporcionando uma análise comparativa com os resultados obtidos com CB1 .  The construction and validation of a 3D model by homology of the CB2 receptor also resulted in visualization of the behavior of all of the binders mentioned at the binding site of such receptor, providing a comparative analysis with the results obtained with CB1.
[0199] Em resumo, os resultados dos experimentos in silico dos exemplos 7 e 8 indicam que o tetrapeptídeo NFKF é um ligante de receptores CB1 . Além disso, interações menos favoráveis desse composto foram evidenciadas em receptores CB2, indicando um eventual perfil de seletividade. Além disso, os resultados dos experimentos dos exemplos 7 e 8, notadamente os mostrados nas das figuras 12 e 17, mostram que os aminoácidos NFK participam muito mais fortemente das ligações com CB1 e com CB2 do que o aminoácido F da posição C-terminal, indicando que este tripeptídeo é um potencial candidato a ligante/modulador de tais receptores.  [0199] Briefly, the results of the in silico experiments of examples 7 and 8 indicate that the NFKF tetrapeptide is a CB1 receptor linker. In addition, less favorable interactions of this compound were evidenced in CB2 receptors, indicating a possible selectivity profile. Furthermore, the results of the experiments of examples 7 and 8, especially those shown in Figures 12 and 17, show that NFK amino acids participate much more strongly in the binding of CB1 and CB2 than the amino acid F in the C-terminal position, indicating that this tripeptide is a potential candidate for ligand / modulator of such receptors.
[0200] Exemplo 9. Testes in vitro de ligação do composto NFKF ao receptor CB1  [0200] Example 9. In vitro binding tests of compound NFKF to CB1 receptor
[0201] Neste exemplo, foi avaliado o perfil de afinidade in vitro do composto da invenção R -N-AA -K-AA2-FÍ2, em concretização na qual é o tetrapeptídeo NFKF, com o receptor canabinóide CB1 . In this example, the in vitro affinity profile of the compound of the invention R-N-AA-K-AA 2- F 2 , in which is the NFKF tetrapeptide, was evaluated with the cannabinoid receptor CB1.
[0202] Para tanto, foram usadas de técnicas de binding para a medição da afinidade do tetrapeptídeo NFKF (em pó com 100% de pureza e preparado como solução estoque 10mM em DMSO) pelo receptor canabinóide CB1 nas concentrações de 1 e 10 μΜ. Esta avaliação foi feita usando o receptor canabinóide CB1 recombinante humano, associado ao radioligante agonista CP 55940 (IC 50(M)=1 .1 nM; Ki(M)=0.94 nM; nH=1 ). Os ensaios in vitro de binding foram realizados com o uso de células CHO recombinantes expressando o receptor CB1 humano e do ligante [3H]CP 55940 (concentração 0.5nM; Kd 3.5nM) em incubação por 1 20 minutos a 37°C, usando o composto não específico WIN 5521 2-2 (1 0mM), sendo a detecção por contagem de cintilografia (Munro et al, 1 993). For this purpose, binding techniques were used to measure the affinity of the NFKF tetrapeptide (powder in 100% purity and prepared as stock solution 10mM in DMSO) by the cannabinoid receptor CB1 at the concentrations of 1 and 10 μΜ. This evaluation was done using the human recombinant CB1 cannabinoid receptor, associated with the radioligand agonist CP 55940 (IC 50 (M) = 1.1 nM; Ki (M) = 0.94 nM; nH = 1). In vitro binding assays were performed using recombinant CHO cells expressing the human CB1 receptor and the ligand [3H] CP 55940 (0.5Nm concentration, Kd 3.5nM) in incubation for 1 20 minutes at 37 ° C using the non - specific compound WIN 5521 2-2 (1 0mm), and detection by counting scintigraphy (Munro et al, 1993).
[0203] Os resultados são expressos como o porcentual do binding do controle específico de acordo com a fórmula:  [0203] Results are expressed as the percentage of specific control binding according to the formula:
Binding específico medido X 1 00  Specific Binding X 1 00
Binding específico do controle  Specific Binding Control
[0204] E como a inibição porcentual do binding do controle específico obtida na presença do composto da invenção: [0204] And as the percent inhibition of specific control binding obtained in the presence of the compound of the invention:
1 00 - ( Binding específico medido X 1 00)  1 00 - (specific Binding measured X 100)
Binding específico do controle  Specific Binding Control
[0205] Os valores de IC50 (concentração que causa metade de máxima inibição do binding do controle específico) e coeficientes de Hill (nH) foram determinados por regressão não linear das curvas de competição geradas com os valores médios das replicatas usando a equação de Hill. IC50 values (concentration that causes half of maximum inhibition of specific control binding) and Hill (nH) coefficients were determined by nonlinear regression of the competition curves generated with the mean values of the replicates using the Hill equation .
Y = D + ( A - D )  Y = D + (A-D)
1 + (C/C50)nH 1 + (C / C 50 ) nH
[0206] Onde Y = binding específico, A = assintótica esquerda da curva, D = assintótica direita da curva, C= concentração do composto, CscHCso e nH= coeficiente da curva/s/ope fator. Esta análise foi realizada usando o Hill software a validada por comparação com dados gerados pelo software Sigmaplot 4.0. As constantes de inibição (K,) foram calculadas usando a equação de Chen Prusoff:
Figure imgf000042_0001
[0206] Where Y = specific binding, A = asymptotic left curve, D = asymptotic right curve, C = compound concentration, CscHCso and nH = curve coefficient / s / ope factor. This analysis was performed using the Hill software validated by comparison with data generated by Sigmaplot 4.0 software. Inhibition constants (K1) were calculated using the Chen Prusoff equation:
Figure imgf000042_0001
[0207] Onde L=concentração do radioligante no ensaio; KD=afinidade do radioligante pelo receptor. Um gráfico é usado para determinar o KD. [0208] Resultados mostrando inibição ou estimulação superior a 50% são considerados como representativos de efeito significativo do composto teste. Resultados mostrando inibição ou estimulação entre 25% e 50% são indicativos de baixo a moderado efeito do composto teste. Resultados mostrando inibição ou estimulação inferior a 25% podem ser considerados como pouco significativos. Resultados mostrando inibição maior ou igual a 50% são indicativos efeitos não específicos ou de efeito alostérico do composto- teste. [0207] Where L = concentration of the radioligand in the assay; K D = affinity of the radioligand for the receptor. A graph is used to determine the KD. [0208] Results showing inhibition or stimulation greater than 50% are considered to represent significant effect of the test compound. Results showing inhibition or stimulation between 25% and 50% are indicative of low to moderate effect of the test compound. Results showing inhibition or stimulation of less than 25% can be considered as minor. Results showing inhibition greater than or equal to 50% are indicative non-specific or allosteric effects of the test compound.
[0209] Os resultados obtidos em duplicata para cada concentração testada (1 e 10 μΜ) são mostrados na tabela 3.  [0209] The results obtained in duplicate for each concentration tested (1 and 10 μΜ) are shown in table 3.
[0210] Tabela 3 - Dados de inibição in vitro de binding do agonista radioligante CP55940 ao receptor CB1 pelo NFKF.  [0210] Table 3 - In vitro inhibition data of binding of the radioligand agonist CP55940 to the CB1 receptor by NFKF.
% de inibição do binding do controle específico  % inhibition of specific control binding
Figure imgf000043_0001
Figure imgf000043_0001
[0211] Ainda que fosse desconsiderada a primeira medida a 1 mM, em função do valor surpreendentemente alto, o dado da segunda medida a 1 mM tomado sozinho ainda seria considerado indicativo de efeito. Although the first measurement at 1 mM was disregarded, as a function of the surprisingly high value, the data from the second measurement at 1 mM taken alone would still be considered indicative of effect.
[0212] Os resultados indicam que o NFKF tem maior poder de modular o binding do respectivo radioligante quando em baixas concentrações, sugerindo que na faixa de nM os efeitos são maiores.  [0212] The results indicate that NFKF has greater power to modulate the binding of the respective radioligand when in low concentrations, suggesting that in the range of nM the effects are greater.
[0213] Estes resultados apontam que o provável mecanismo de ação associado ao perfil terapêutico evidenciado para o tetrapeptídeo NFKF envolve a modulação alostérica de receptores canabinóides CB1 , o que é consistente com os surpreendentes/inesperados resultados dos experimentos in silico (exemplo 7) e experimentos in vivo (exemplos 3, 4, 5, 6).  These results indicate that the likely mechanism of action associated with the therapeutic profile evidenced for the NFKF tetrapeptide involves the allosteric modulation of CB1 cannabinoid receptors, consistent with the surprising / unexpected results of the in silico experiments (example 7) and experiments in vivo (examples 3, 4, 5, 6).
[0214] Estes dados experimentais indicam que o composto da invenção interage com e/ou modula a atividade do receptor CB1 e/ou seus ligantes endógenos. Assim o composto da invenção é potencialmente em diversas funções metabólicas, incluindo o controle de ingestão de alimentos, o metabolismo de energia e/ou de lipídeos, na regulação da motilidade intestinal, no sistema imune, no equilíbrio do ciclo do cálcio, entre outros. Os receptores canabinóides são amplamente expressos no cérebro, incluindo córtex, hipocampo, amígdala, pituitário, hipotálamo, glândula adrenal. Os receptores CB, particularmente CB1 , já foram identificados em inúmeros órgãos periféricos e tecidos, incluindo glândula tiroidal, órgãos reprodutivos, tecido adiposo, fígado, músculos e trato gastrointestinal. Estes resultados suportam, portanto, o uso do composto da invenção na modulação de funções metabólicas e/ou na neumodulação. [0214] These experimental data indicate that the compound of the invention interacts with and / or modulates the activity of the CB1 receptor and / or its endogenous linkers. Thus the compound of the invention is potentially in various metabolic functions, including control of food intake, metabolism of energy and / or lipids, regulation of intestinal motility, immune system, calcium cycle balance, among others. Cannabinoid receptors are widely expressed in the brain, including cortex, hippocampus, amygdala, pituitary, hypothalamus, adrenal gland. CB receptors, particularly CB1, have already been identified in numerous peripheral organs and tissues, including the thyroid gland, reproductive organs, adipose tissue, liver, muscle, and gastrointestinal tract. These results therefore support the use of the compound of the invention in the modulation of metabolic functions and / or pneumodulation.
[0215] Exemplo 10. Testes in vitro de ligação do composto NFKF ao receptor CB2  [0215] Example 10. In vitro in vitro binding of the compound NFKF to the CB2 receptor
[0216] Neste exemplo, foi avaliado o perfil de afinidade in vitro do composto da invenção R -N-AA -K-AA2-R2, em concretização na qual é o tetrapeptídeo NFKF, com o receptor canabinoide CB2. In this example, the in vitro affinity profile of the compound of the invention R-N-AA-K-AA 2 -R 2 , in which is the NFKF tetrapeptide, is evaluated with the cannabinoid receptor CB2.
[0217] Para tanto, foram usadas de técnicas de binding para a medição da afinidade do tetrapeptídeo NFKF (em pó com 100% de pureza e preparado como solução estoque 10mM em DMSO) pelo receptor canabinoide CB2 nas concentrações de 1 e 10 μΜ. Esta avaliação foi feita usando o receptor canabinoide CB2 recombinante humano, associado ao radioligante agonista WIN 55212-2 (IC 50(M)=1 .5 nM; Ki(M)=0.96 nM; nH=0.9). Os ensaios in vitro de binding foram realizados com o uso de células CHO recombinantes expressando o receptor CB2 humano e do ligante [3H]WIN55212-2 (concentração 0.8nM; Kd 1 .5nM) em incubação por 120 minutos a 37°C, usando o composto não específico WIN 55212-2 (5mM), sendo a detecção por contagem de cintilografia (Rinaldi-Carmona et al, 1996). For this purpose, binding techniques were used to measure the affinity of the NFKF tetrapeptide (powder in 100% purity and prepared as stock solution 10mM in DMSO) by the cannabinoid receptor CB2 at concentrations of 1 and 10 μΜ. This evaluation was done using the human recombinant cannabinoid receptor CB2, associated with the radioligand agonist WIN 55212-2 (IC 50 (M) = 1.5 nM; Ki (M) = 0.96 nM; nH = 0.9). In vitro binding assays were performed with the use of recombinant CHO cells expressing the human CB2 receptor and [ 3 H] WIN55212-2 (0.8 nM concentration; 1.5 nM Kd) linker in incubation for 120 minutes at 37 ° C, using the non-specific compound WIN 55212-2 (5 mM), the detection being by scintigraphy counting (Rinaldi-Carmona et al, 1996).
[0218] Os resultados são expressos conforme indicado no exemplo 9 acima. [0218] The results are expressed as indicated in example 9 above.
[0219] Os resultados obtidos em duplicata para cada concentração testada (1 e 10 μΜ) são mostrados na tabela 4. [0219] The results obtained in duplicate for each concentration tested (1 and 10 μΜ) are shown in table 4.
[0220] Tabela 4 - Dados de inibição in vitro de binding do agonista radioligante WIN 55212-2 ao receptor CB2 pelo NFKF. % de inibição do binding do controle específico Table 4 - In vitro inhibition data of binding of radioligand agonist WIN 55212-2 to CB2 receptor by NFKF. % inhibition of specific control binding
Figure imgf000045_0001
Figure imgf000045_0001
[0221] Os resultados indicam que o NFKF não proporcionou significativa modulação no binding do respectivo radioligante nas condições testadas. [0221] The results indicate that NFKF did not provide significant modulation in the binding of the respective radioligand under the conditions tested.
[0222] Tomando estes resultados em conjunto com os resultados dos experimentos descritos no exemplo 8, pode-se chegar a interessantes conclusões. A despeito do peptídeo NFKF ter se demonstrado um ligante seletivo de receptores canabinóides CB2, conforme indicam os resultados dos experimentos de docking para este receptor, os resultados dos teste in vitro indicam que este tetrapeptídeo não é capaz de se ligar a este receptor canabinoide de forma significativa. Entretanto, não se pode descartar uma interação do composto da invenção com o receptor CB2 do tipo alostérica. Assim, os resultados de testes indicaram uma modulação positiva de receptor CB2, ainda que moderada nas concentrações testadas, sugerem um efeito combinado de modulação (alostérica) negativa para CB1 e positiva para CB2. [0222] Taking these results together with the results of the experiments described in example 8, one can reach interesting conclusions. Although the NFKF peptide has demonstrated a selective binder of CB2 cannabinoid receptors, as indicated by the results of the docking experiments for this receptor, in vitro test results indicate that this tetrapeptide is not able to bind to this cannabinoid receptor in a manner significant. However, an interaction of the compound of the invention with the allosteric type CB2 receptor can not be ruled out. Thus, the test results indicated a positive modulation of CB2 receptor, although moderate at the concentrations tested, suggest a combined negative (allosteric) modulation effect for CB1 and CB2 positive.
[0223] Além disso, os resultados dos testes in vitro (exemplos 9 e/ou 10) demonstram que o composto da invenção pode ser usado como ligante de interesse diagnóstico, como por exemplo a marcação radioativa ou com cromóforos para subsequente identificação dos locais de ligação em células e/ou tecidos. In addition, in vitro test results (examples 9 and / or 10) demonstrate that the compound of the invention can be used as a binder of diagnostic interest, for example radioactive labeling or with chromophores for subsequent identification of the sites of binding in cells and / or tissues.
[0224] O conceito inventivo ora revelado e exemplificado de uma ou mais formas foi tratado como segredo industrial e não foi previamente revelado até o momento do depósito deste pedido de patente. Este segredo industrial é ativo imaterial do depositante. A eventual futura publicação do pedido de patente não constitui, em si, autorização de uso por terceiros, servindo apenas como: (i) cientificação a terceiros da existência do referido segredo industrial na data do depósito; (ii) indicação inequívoca de seu detentor; e (iii) estímulo ao desenvolvimento de novas melhorias a partir do conceito ora revelado, para evitar o reinvestimento no desenvolvimento do mesmo bem já detido pelo depositante. The inventive concept herein disclosed and exemplified in one or more ways was treated as an industrial secret and was not previously disclosed until the filing of this patent application. This industrial secret is immaterial asset of the depositor. The possible future publication of the patent application does not in itself constitute authorization for use by third parties, serving only as: (i) scientific knowledge to third parties of the existence of said industrial secret at the time of filing; (ii) unequivocal indication of its holder; and (iii) stimulating the development of new improvements based on the concept to avoid reinvestment in the development of the same asset already held by the depositor.
[0225] Desde logo se adverte que eventual uso comercial requer autorização do detentor e que o uso não autorizado enseja sanções previstas em Lei. Neste contexto, desde logo se esclarece que a partir da revelação do presente conceito inventivo, os versados na arte poderão considerar outras formas de concretizar a invenção não idênticas às meramente exemplificadas acima, mas que na hipótese de pretensão de uso comercial tais formas poderão ser consideradas como estando dentro do escopo das reivindicações anexas.  [0225] It is pointed out that any commercial use requires permission from the holder and that unauthorized use imposes penalties provided for by Law. In this context, it is first clarified that from the disclosure of the present inventive concept, those skilled in the art may consider other embodiments of the invention not identical to those merely exemplified above, but that in the hypothesis of claim to commercial use such forms may be considered to be within the scope of the appended claims.

Claims

Reivindicações Claims
1 . Uso do composto peptídico com até seis aminoácidos de fórmula: 1 . Use of the peptidic compound with up to six amino acids of the formula:
R1-N-AA1-K-AA2-R2 R 1 -N-AA 1 -K-AA 2 -R 2
onde: at where:
AA é um aminoácido selecionado do grupo que consiste de F, W, L, I, V, P, G; AA2 é hidrogénio ou um aminoácido selecionado do grupo que consiste de F, W, L, I, V, P, G; AA is an amino acid selected from the group consisting of F, W, L, I, V, P, G; AA 2 is hydrogen or an amino acid selected from the group consisting of F, W, L, I, V, P, G;
Ri é hidrogénio, o aminoácido V, ou o dipeptídeo PV; e  R 1 is hydrogen, the amino acid V, or the dipeptide PV; and
R2 é ausente quando AA2 é hidrogénio ou é hidrogénio, o aminoácido L, ou o dipeptídeo LS quando Ri é hidrogénio, R 2 is absent when AA 2 is hydrogen or is hydrogen, the amino acid L, or the dipeptide LS when R 1 is hydrogen,
e/ou formas modificadas, cíclicas, amidadas, metiladas e/ou PEGiladas dos mesmos; seus sais; e/ou combinações dos mesmos caracterizado por ser para a preparação de um produto modulador do sistema canabinóide e/ou de receptores muscarínicos. and / or modified, cyclic, amidated, methylated and / or PEGylated forms thereof; their salts; and / or combinations thereof characterized in that it is for the preparation of a modulator product of the cannabinoid system and / or muscarinic receptors.
2. Uso de acordo com a reivindicação 1 caracterizado pelo fato de que AA ou AA2 é F, W ou L e/ou Ri e R2 são ambos hidrogénio. 2. Use according to claim 1 , characterized by the fact that AA or AA 2 is F, W or L and / or R and R 2 are both hydrogen.
3. Uso de acordo com a reivindicação 1 ou 2 caracterizado pelo fato de ser selecionado do grupo que consiste de: NFK, NWK, NLK, NFKF, NWKF, NLKF, NFKW, NWKW, NLKW, NFKL, NWKL, NLKL, VNFK, VNWK, VNLK, NFKFL, NWKFL, NLKFL, NFKWL, NWKWL, NLKWL, NFKLL, NWKLL, NLKLL, VNFKF, VNWKW, VNLKL, VNFKFL, VNWKWL, VNLKLL, PVNFKF, PVNWKW, PVNLKL, NFKFLS, NWKWLS, NLKLLS, bem como formas modificadas, cíclicas dos mesmos, versões amidadas, metiladas, PEGiladas; seus sais; ou combinações dos mesmos.  Use according to claim 1 or 2 characterized in that it is selected from the group consisting of: NFK, NWK, NLK, NFKF, NWKF, NLKF, NFKW, NWKW, NLKW, NFKL, NWKL, NLKL, VNFK, VNWK VNLKL, VNLKL, VNFKFL, VNWKL, VNWKWL, VNLKLL, PVNFKF, PVNWKW, PVNLKL, NFKFLS, NWKWLS, NLKLLS, as well as modified forms, cyclic, amidated, methylated, PEGylated versions; their salts; or combinations thereof.
4. Uso de acordo com a reivindicação 1 , 2 ou 3 caracterizado pelo fato de ser selecionado do grupo que consiste do tripeptídeo NFK, o tetrapeptídeo NFKF, tetrapeptídeo NFKL, ou combinações dos mesmos.  Use according to claim 1, 2 or 3 characterized in that it is selected from the group consisting of the NFK tripeptide, the NFKF tetrapeptide, the NFKL tetrapeptide, or combinations thereof.
5. Uso de acordo com uma das reivindicações 1 a 4 caraterizado por ser para a preparação de um medicamento modulador de função metabólica em um mamífero. Use according to one of Claims 1 to 4, characterized in that it is for the preparation of a metabolic function modulating medicament in a mammal.
6. Uso de acordo com uma das reivindicações 1 a 4 caraterizado por ser para a preparação de um medicamento neuromodulador curativo ou profilático em um mamífero. Use according to one of Claims 1 to 4, characterized in that it is for the preparation of a curative or prophylactic neuromodulator medicament in a mammal.
7. Uso de acordo com uma das reivindicações 1 a 5 caraterizado por ser para a preparação de um medicamento para o tratamento de distúrbios do metabolismo de energia e/ou de lipídeos; hipertensão arterial, distúrbios da motilidade intestinal; do sistema imune; o equilíbrio do ciclo do cálcio, distúrbios da glândula tiroidal, de órgãos reprodutivos, obesidade, diabetes, hipertensão arterial, doenças ou distúrbios imunes/inflamatórios, osteopenia, osteoporose, câncer.  Use according to one of Claims 1 to 5, characterized in that it is for the preparation of a medicament for the treatment of disorders of energy metabolism and / or lipids; arterial hypertension, intestinal motility disorders; of the immune system; calcium cycle balance, thyroid gland disorders, reproductive organs, obesity, diabetes, hypertension, immune / inflammatory diseases or disorders, osteopenia, osteoporosis, cancer.
8. Uso de acordo com uma das reivindicações 1 a 4 ou 6 caraterizado por ser para a preparação de um medicamento para o tratamento de doença neurológica, tremor essencial, enxaqueca, dor neuropática, transtornos psiquiátricos, ansiedade, esquizofrenia ou transtorno bipolar, Alzheimer, Parkinson, autismo, epilepsia.  Use according to one of claims 1 to 4 or 6 for the preparation of a medicament for the treatment of neurological disease, essential tremor, migraine, neuropathic pain, psychiatric disorders, anxiety, schizophrenia or bipolar disorder, Alzheimer's, Parkinson's, autism, epilepsy.
9. Uso de acordo com uma das reivindicações 1 a 4 ou 6 caracterizado por ser para a preparação de um medicamento para o tratamento curativo ou profilático de convulsões em um mamífero.  Use according to one of claims 1 to 4 or 6, characterized in that it is for the preparation of a medicament for the curative or prophylactic treatment of seizures in a mammal.
10. Uso de acordo com uma das reivindicações 1 a 4 caracterizado por ser para a preparação de um ligante de interesse diagnóstico em um mamífero. Use according to one of Claims 1 to 4, characterized in that it is for the preparation of a binder of diagnostic interest in a mammal.
1 1 . Intermediário de síntese na preparação de compostos de interesse farmacêutico caracterizado por consistir do composto peptídico descrito em uma das reivindicações 1 a 4. 1 1. Synthesis intermediary in the preparation of compounds of pharmaceutical interest which comprises the peptidic compound described in one of claims 1 to 4.
12. Composto de fórmula:  A compound of the formula:
R1-N-AA1-K-AA2-R2 R 1 -N-AA 1 -K-AA 2 -R 2
onde: at where:
AA é um aminoácido selecionado do grupo que consiste de F, W, L, I, V, P, G; AA2 é hidrogénio ou um aminoácido selecionado do grupo que consiste de F, W, L, I, V, P, G; AA is an amino acid selected from the group consisting of F, W, L, I, V, P, G; AA 2 is hydrogen or an amino acid selected from the group consisting of F, W, L, I, V, P, G;
Ri é hidrogénio, o aminoácido V, ou o dipeptídeo PV; e R2 é ausente quando AA2 é hidrogénio ou é hidrogénio, o aminoácido L, ou o dipeptídeo LS quando Ri é hidrogénio, caracterizado pelo fato de ser modificado, ciclizado, amidado, metilado ou PEGilado, bem como seus sais e/ou combinações dos mesmos. R 1 is hydrogen, the amino acid V, or the dipeptide PV; and R 2 is absent when AA 2 is hydrogen or is hydrogen, the amino acid L, or the dipeptide LS when R 1 is hydrogen, characterized in that it is modified, cyclized, amidated, methylated or PEGylated, as well as salts and / or combinations thereof. themselves.
13. Composição farmacêutica para a modulação da função do sistema canabinoide e/ou de receptores muscarínicos em um mamífero caracterizada por compreender um veículo farmaceuticamente aceitável; e, como princípio ativo, o composto conforme definido em uma das reivindicações 1 a 4.  A pharmaceutical composition for the modulation of the function of the cannabinoid system and / or muscarinic receptors in a mammal characterized in that it comprises a pharmaceutically acceptable carrier; and, as active principle, the compound as defined in one of claims 1 to 4.
14. Composição farmacêutica para a modulação da função metabólica em um mamífero caracterizada por compreender um veículo farmaceuticamente aceitável; e, como princípio ativo, o composto conforme definido em uma das reivindicações 1 a 4.  A pharmaceutical composition for the modulation of metabolic function in a mammal characterized in that it comprises a pharmaceutically acceptable carrier; and, as active principle, the compound as defined in one of claims 1 to 4.
15. Composição farmacêutica para a neuromodulação curativa ou profilática em um mamífero caracterizada por compreender um veículo farmaceuticamente aceitável; e, como princípio ativo, composto conforme definido em uma das reivindicações 1 a 4.  A pharmaceutical composition for curative or prophylactic neuromodulation in a mammal which comprises a pharmaceutically acceptable carrier; and, as active principle, compound as defined in one of claims 1 to 4.
16. Composição farmacêutica para o tratamento curativo ou profilático de convulsões em um mamífero caracterizada por compreender um veículo farmaceuticamente aceitável; e, como princípio ativo, o composto conforme definido em uma das reivindicações 1 a 4.  A pharmaceutical composition for the curative or prophylactic treatment of seizures in a mammal characterized in that it comprises a pharmaceutically acceptable carrier; and, as active principle, the compound as defined in one of claims 1 to 4.
17. Composição farmacêutica de acordo com qualquer uma das reivindicações 13 a 16 caracterizada por se apresentar na forma de tablete, gel, líquido oral ou xarope, cápsula, supositório, solução injetável, forma inalável ou em adesivo.  Pharmaceutical composition according to any one of claims 13 to 16, characterized in that it is in the form of a tablet, gel, oral liquid or syrup, a capsule, a suppository, an injectable solution, an inhalable form or an adhesive.
18. Método terapêutico modulador de função metabólica caracterizado por compreender a administração, a um animal, do composto descrito em uma das reivindicações 1 a 4.  A metabolic metabolic modulating therapeutic method comprising administering to an animal the compound described in one of claims 1 to 4.
19. Método terapêutico neuromodulador caracterizado por compreender a administração, a um animal, do composto descrito em uma das reivindicações 1 a 4. A neuromodulatory therapeutic method comprising administering to an animal the compound described in one of claims 1 to 4.
20. Método terapêutico de acordo com a reivindicação 18 ou 19 caracterizado pelo fato de que a dose do composto administrada em humanos é de 4 a 800 μ§/\(§. A therapeutic method according to claim 18 or 19, characterized in that the dose of the compound administered in humans is 4 to 800 μg / μg.
PCT/BR2017/050314 2016-10-13 2017-10-11 Use of a compound, synthetic intermediate, pharmaceutical composition, and neuromodulatory therapeutic method WO2018068120A1 (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018209415A1 (en) * 2017-05-15 2018-11-22 Remer Consultores Assessoria Empresarial Ltda. Compound, synthetic intermediate, use, pharmaceutical composition, and neuromodulatory therapeutic method

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070213302A1 (en) * 2006-03-10 2007-09-13 Jenrin Discovery Cannabinoid receptor antagonists/inverse agonists useful for treating obesity
WO2011011847A2 (en) * 2009-07-31 2011-02-03 Sociedade Beneficiente De Senhoras Hospital Sirio Libanes Pharmaceutical composition for treating medical conditions and a method for treating alimentary disorders and related diseases
WO2013021196A2 (en) * 2011-08-08 2013-02-14 University Of Coimbra Oligodendrocyte differentiation
WO2014008567A1 (en) * 2012-07-13 2014-01-16 Proteimax Biotecnologia Ltda Modified peptide, cb receptor ligand, kit, in vitro process for evaluating cb receptor bonds, uses, pharmaceutical composition for modulating cb receptor activity

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070213302A1 (en) * 2006-03-10 2007-09-13 Jenrin Discovery Cannabinoid receptor antagonists/inverse agonists useful for treating obesity
WO2011011847A2 (en) * 2009-07-31 2011-02-03 Sociedade Beneficiente De Senhoras Hospital Sirio Libanes Pharmaceutical composition for treating medical conditions and a method for treating alimentary disorders and related diseases
US20150297669A1 (en) * 2009-07-31 2015-10-22 Sociedade Beneficente De Senhoras Hospital Sirio Libanes Pharmaceutical composition for treating medical conditions and a method for treating alimentary disorders and related diseases
WO2013021196A2 (en) * 2011-08-08 2013-02-14 University Of Coimbra Oligodendrocyte differentiation
WO2014008567A1 (en) * 2012-07-13 2014-01-16 Proteimax Biotecnologia Ltda Modified peptide, cb receptor ligand, kit, in vitro process for evaluating cb receptor bonds, uses, pharmaceutical composition for modulating cb receptor activity

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
GOMES I ET AL.: "Novel endogenous peptide agonists of cannabinoid receptors", FASEB J., vol. 23, no. 9, 2009, pages 3020 - 3029, XP055186735 *
IVONE GOMES ET AL.: "Hemoglobin-derived Peptides as Novel Type of Bioactive Signaling Molecules", AAPS J., vol. 12, no. 4, 2010, pages 658 - 669, XP035719135 *
MELISA J WALLACE ET AL.: "Assessment of the role of CB1 receptors in cannabinoid anticonvulsant effects", EUROPEAN JOURNAL OF PHARMACOLOGY, vol. 428, no. 1, 2001, pages 51 - 57, XP055475315 *
MELISA J. WALLACE ET AL.: "The Endogenous Cannabinoid System Regulates Seizure Frequency and Duration in a Model of Temporal Lobe Epilepsy", JOURNAL . OF PHARMACOLOGY AND EXPERIMENTAL . THERAPEUTIC., vol. 307, no. 1, 2003, pages 129 - 137, XP055360835 *
RIOLI ET AL.: "Novel natural peptide substrates for endopeptidase 24.15, neurolysin, and angiotensin-converting enzyme", J BIOL CHEM., vol. 278, no. i 0, 2003, pages 8547 - 8555, XP002353088 *
SZLAVICZ ET AL.: "Further Characterization of Hemopressin Peptide Fragments in the Opioid and Cannabinoid Systems", ANESTH ANALG., vol. 121, no. 6, 2015, pages 1488 - 1494, XP055476031 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018209415A1 (en) * 2017-05-15 2018-11-22 Remer Consultores Assessoria Empresarial Ltda. Compound, synthetic intermediate, use, pharmaceutical composition, and neuromodulatory therapeutic method
CN110891963A (en) * 2017-05-15 2020-03-17 雷默咨询顾问有限公司 Compounds, synthetic intermediates, uses, pharmaceutical compositions and neuromodulation therapy methods
CN110891963B (en) * 2017-05-15 2024-02-20 雷默咨询顾问有限公司 Compounds, synthetic intermediates, uses, pharmaceutical compositions and methods of neuromodulation therapy

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