Novel central ghrelin agonists and medical uses thereof

Abstract

The novel compound 3-(1-(2,3-dichloro-4-methoxyphenyl)ethyl)-1-methyl-1-(1,3,3-trimethylpiperidin-4-yl)urea monohydrochloride has high permeability, is able to cross the blood-brain barrier and shows consistent ghrelin agonist activity at the level of the central nervous system; the compound is effective in the treatment and / or prevention of medical conditions mediated by ghrelin receptors in the central nervous system. In particular, in experimental tests, the compound shows high efficacy in the treatment of neurotoxic injuries and has an effective neuroprotective action combination pattern at both central and peripheral levels. The compound is also useful in the treatment of diseases requiring a reduction in heart rate. The compound has pharmacological activity at low to moderate doses, thus showing a good therapeutic index.

Description

[0001] This application is a divisional application of the invention application filed on March 14, 2019, with application number 201980021301.3 and entitled "Novel Central Growth Hormone Releasing Peptide Agonist and Its Medical Use". Background Technology

[0002] Ghrelin is a naturally occurring peptide hormone produced by ghrelinergic cells in the gastrointestinal tract, acting as a neuropeptide in the central nervous system. The biological target of ghrelin is the G protein-coupled ghrelin receptor (GHSR), first cloned in 1996 (1). Two receptor subtypes, 1a and 1b, are known, but only the former can activate signal transduction (2). GHSR is mainly expressed in the nervous system and in multiple non-neural organs involved in various physiological processes (2-4). Ghrelin peptides are primarily involved in the regulation of appetite and also play an important role in regulating energy distribution and utilization. Ghrelin peptides act on hypothalamic cells, increasing hunger sensation, gastric acid secretion, and gastrointestinal motility, preparing the body for food intake. Ghrelin peptides also play an important role in regulating reward perception in dopamine neurons, which connect the ventral tegmental area to the nucleus accumbens (a site that plays a role in processing libido, reward and reinforcement, and addiction) via their colocalized receptors and their interactions with dopamine and acetylcholine. Clinical trials have evaluated the therapeutic potential of growth hormone-releasing peptides in a variety of disease states, including anorexia nervosa (5), cancer cachexia (6,7), sleep-wake regulation (8), chronic heart failure (9), and gastrointestinal motility disorders (10). In animals, growth hormone-releasing peptides promote neuronal proliferation and neurogenesis (11). Ghrelin peptides have also been shown to have neuroprotective properties and prevent apoptosis (12). Ghrelin receptors (GHSRs) are located in several different regions of the central nervous system (CNS). Exogenous administration of growth hormone-releasing peptides has improved experimental encephalomyelitis (15), Parkinson's disease (16), and Alzheimer's disease (17) in preclinical models. Furthermore, ghrelin peptides have been proposed to possess direct neuroreparative properties following central or peripheral nervous system injury (18). Neuropathic pain has a significant inflammatory component, characterized by persistent activation of glial cells and increased production of pro-inflammatory cytokines. Additionally, ghrelin peptides have been reported to have therapeutic effects in rodent models of diabetes (20), chronic contractile injury (21), and chemotherapy-induced neurotoxicity (CIPN) (22), as well as acute pain (23) and chronic arthritis (24). However, the limited brain penetration of ghrelin peptides (25), lack of oral bioavailability, and short half-life of only 8–24 minutes in rodents (26) and 37 minutes in humans (27) limit their efficacy as a clinical drug. In practice, preclinical efficacy demonstrations typically require multiple systemic injections or intrathecal / intraventricular administration, emphasizing the need for continuous infusion of ghrelin peptides or the use of agonists with longer half-lives and enhanced neuropathic penetration.

[0003] The identification of the auxin-releasing peptide receptor has spurred research aimed at identifying novel compounds with binding affinity to the receptor that exhibit similar activity to auxin-releasing peptides, in order to seek potential therapeutic advantages relative to the original peptide. Non-peptide small molecules may bypass the metabolic inactivation pathways of peptides and are therefore of particular interest here. However, at the same time, significant structural differences from the original peptide may alter the original activity profile of auxin-releasing peptides and auxin-releasing peptide-like molecules.

[0004] Patent application WO2012 / 116176 describes an asymmetric urea of ​​general formula (I).

[0005]

[0006] It possesses GHSR1a receptor regulatory properties; the application contains experimental data on the compound's GHSR1a receptor affinity and in vivo activity in mice after food intake; the compound is proposed for the treatment of a variety of diseases, including obesity, overweight, eating disorders, metabolic syndrome, emaciation due to aging or AIDS, gastrointestinal diseases, stomach ailments, etc.

[0007] Patent application WO2015 / 134839 further describes the following auxin-releasing peptide regulator:

[0008]

[0009] The application contains experimental data on the compound's affinity for the GHSR1a receptor and its in vivo activity in mouse models of food intake and alcohol abuse.

[0010] Despite significant efforts, recently discovered small-molecule agonists have not yet been approved for therapeutic use. Their clinical utility is often limited by unsatisfactory safety profiles and / or poor central nervous system absorption. In particular, the CNS impermeability of ghrelin agonists is a serious limitation, given that most ghrelin receptors are expressed in the central nervous system and that many ghrelin-dependent diseases are at least partially mediated by the central nervous system. On the other hand, creating molecules capable of effectively crossing the blood-brain barrier is a complex task: successful crossing requires overcoming various key steps, particularly: the uptake of drug molecules by brain endothelial cells from systemic circulation; the efficient internalization of the drug into these cells; and the ability of these cells to release unmetabolized forms of the drug into the central nervous system compartments in amounts sufficient to elicit a pharmacological response. Only fine-tuning / synergistic effects of the above mechanisms can lead to the flow of the active form of the drug across the barrier to reach the central nervous system target: in reality, after systemic administration, only a small fraction of known drug molecules are detected in the central nervous system in detectable amounts. This is not surprising, as the functional structure of the blood-brain barrier isolates the CNS compartments from potentially dangerous exogenous organisms in the blood.

[0011] Moreover, for medical conditions requiring treatment at both the central and peripheral levels, discovering molecules capable of crossing the blood-brain barrier is not an ideal solution. In fact, they face the further challenge of achieving / maintaining a balance between the active form and concentration of the drug in both regions across the barrier, ensuring that any pharmacokinetic bias towards central accumulation does not impair useful effects at the peripheral level. This problem is particularly acute in the field of neurological diseases, which often involve damage at both the central and peripheral levels. Furthermore, while the ability to cross the blood-brain barrier opens pathways for desired central treatment, it also introduces new problems related to the potential for excessive drug accumulation in the brain. Therefore, ideal neurotransmitter drugs should be effective at very low doses to ensure a balance between accumulation and elimination processes, thus preventing the risk of significant brain accumulation.

[0012] Therefore, there is still no small molecule that meets the requirements, which needs to be a synthetic auxin-releasing peptide agonist with a strong affinity for auxin-releasing peptide receptors in the brain, capable of crossing the blood-brain barrier in large quantities in a non-metabolic form and establishing pharmacologically active concentrations; in addition, there is a need for auxin-releasing peptide agonists that are particularly effective in the nervous system, showing therapeutic effects at both the peripheral and central levels; at the same time, there is also a need for auxin-releasing peptide agonists that can reduce the risk of unwanted brain accumulation. Invention Overview

[0013] This application identifies the compound 3-(1-(2,3-dichloro-4-methoxyphenyl)ethyl)-1-methyl-1-(1,3,3-trimethylpiperidin-4-yl)urea-hydrochloride as a ghrelin agonist with a high capacity to penetrate the blood-brain barrier and significant ghrelin agonist activity at the central nervous system level. Therefore, this compound is effective in treating and / or preventing ghrelin-releasing peptide receptor-mediated medical conditions in the central nervous system. In particular, experimental testing has demonstrated its high efficacy in treating neurotoxic injuries and a useful combination of neuroprotective effects at both the central and peripheral levels. The compound also produces centrally mediated bradycardia, which is previously unknown in ghrelin-releasing peptide-like molecules, making it particularly suitable for the treatment of cardiovascular diseases requiring heart rate reduction. The compound also exhibits a non-linear dose / efficacy ratio, maximizing at moderate rather than maximum dose levels: this allows for maximizing the intended effect while administering moderate doses, thereby limiting the risk of unwanted brain accumulation in the brain or other organs, as well as any other general toxicity issues. Attached Figure Description

[0014] Figure 1Effects of Compound A treatment on body weight, food intake, and mechanosensitive hyperalgesia induced by cisplatin injection in rats. Cisplatin (0.5 mg / kg), administered intraperitoneally once daily for 3 consecutive days (day 0 to day 2), resulted in reduced body weight and daily food intake, and induced mechanosensitive hyperalgesia. A) One-way ANOVA and post-hoc Tukeys multiple tests revealed a significant overall effect of cisplatin on body weight (p<0.05), with increased body weight after treatment with Compound A at 3, 10 (both p<0.01), and 30 mg / kg (p<0.05). B) Cisplatin significantly reduced food intake (p<0.05), which was improved by treatment with Compound A at 10 and 30 mg / kg (p<0.05). C) Cisplatin induced mechanosensitive hyperalgesia, which was significantly improved by treatment with Compound A at 10 and 30 mg / kg (p<0.05), but not by treatment with Compound A at 3 mg / kg.

[0015] Figure 2 Effects of Compound A on Nerve Conduction Velocity (NCV) and Potential Amplitude in Oxaliplatin-treated Mice. All parameters were measured before and 24 hours after completion of the dosing regimen, which consisted of daily oral administration of the excipient or Compound A (10 or 30 mg / kg) followed by intraperitoneal injection of oxaliplatin (6 mg / kg) or the excipient (8 doses every 4 days). The last dose of Compound A was administered 1 hour before recording. (* indicates p = <0.05 relative to veh / oxaliplatin; # = p <0.05 relative to veh / veh). Administration of Compound A at 10 mg / kg and 30 mg / kg in A) and C) significantly prevented the decrease in finger and tail NCV observed in oxaliplatin-treated mice. In B) Oxaliplatin administration reduced potential amplitude in the fingers, and treatment with Compound A tended to normalize the potential amplitude, but administration of Compound A alone at 30 mg / kg normalized this data. D) Oxaliplatin treatment had no significant effect on the amplitude of the tail nerve potential.

[0016] Figure 3 Compound A reversed the decrease in intradermal nerve fiber density (IENFD). IENFD was measured 24 hours after the last administration of compound A. The significant reduction in the number of fibers produced by oxaliplatin could be completely normalized by treatment with compound A at 10 or 30 mg / kg daily (p<0.01). A) Administration of compound A at 10 mg / kg and 30 mg / kg normalized IENFD relative to mice treated with the excipient. B) and C) Long-term treatment with bortezomib caused a statistically significant reduction in IENFD; in the prophylactic setting but not the therapeutic setting, all doses of compound A significantly prevented this reduction.

[0017] Figure 4 Plasma, DRG, and sciatic nerve concentrations of compound A. Following single oral doses of 10 and 30 mg / kg of compound A in A) and B), the compound was readily absorbed and distributed in plasma and nerve tissues at peak concentrations within 0.25–0.5 hours. The terminal half-life in plasma was short, but the terminal half-life in the sciatic nerve and DRG was significantly longer. The compound exhibited excellent tissue penetration in both the sciatic nerve and DRG. In C), after 30 days of daily administration, the plasma concentration of compound A was very low, but increased in the sciatic nerve and DRG, indicating significant accumulation of compound A in these tissues.

[0018] Figure 5 Compound A reverses abnormal pain induced by bortezomib. Preventive research. A) At the end of treatment, no abnormal pain was observed in the groups treated with bortezomib in combination with all doses of compound A, while abnormal pain was observed in rats treated with bortezomib alone. Therapeutic research. B) Abnormal pain was observed in all animals treated with bortezomib 4 weeks prior to co-administration with compound A. Abnormal pain was observed in the group treated with bortezomib alone one week after the start of co-treatment and at the end of treatment, while all groups treated with compound A were protected.

[0019] Figure 6 Effects of compound A on nerve conduction velocity (NCV) and potential amplitude in rats treated with bortezomib. Preventive research. A) Statistically significant reductions in digital NCV values ​​were observed with bortezomib alone or in combination with compound A at doses of 3 and 10 mg / kg, while no change was observed in the group treated with the highest dose of compound A compared to the control group. B) No changes in digital potential amplitude were observed in any group. C) and D) Tail NCV and potential amplitude were significantly reduced in all groups treated with bortezomib alone or in combination with all doses of compound A. Therapeutic research. Animals treated with bortezomib alone (E) and (F) showed a significant reduction in numerical NCV, while a reduction in potential amplitude was observed only in the group treated with the highest dose of compound A. As shown in the figures for G) and H), in terms of prophylaxis, both tail NCV and potential amplitude were significantly reduced in all groups treated with bortezomib alone or in combination with all doses of compound A.

[0020] Figure 7 Mean heart rate – absolute value (mean equal to the average of three readings) in human patients treated with 0.1, 0.3, 1, or 10 mg of compound A compared to placebo.

[0021] Figure 8Mean heart rate – the change from baseline in human patients treated with 0.1, 1, 0.3, or 10 mg of compound A compared to placebo (mean equal to the average of three readings).

[0022] Figure 9 XRPD data for compound A.

[0023] Figure 10 Compound A 1 H-NMR.

[0024] Figure 11 Compound A 13 C-NMR.

[0025] Figure 12 Mean concentrations of compound A in plasma (ng / mL) and brain (ng / g) after a single intravenous injection of 10 mg / kg into male Sprague Dawley rats. Invention Details

[0026] The compound 3-(1-(2,3-dichloro-4-methoxyphenyl)ethyl)-1-methyl-1-(1,3,3-trimethylpiperidin-4-yl)urea hydrochloride of the present invention is referred to herein as "Compound A". It has the following chemical structural formula:

[0027]

[0028] As used in this article, the term "medical condition" refers to a disease or disturbance; the term "disease" refers to an established medical syndrome; and the term "disturbance" refers to any damage or dysfunction of a particular body part, organ, or tissue that may or may not cause a complete pathological syndrome.

[0029] The term "medical condition mediated by auxin-releasing peptide receptor in the central nervous system" refers to those typical or at least partially typical central nervous system diseases / disorders that respond to treatment with auxin-releasing peptide; these can be encephalomyelitis, Parkinson's disease, Alzheimer's disease, and cognitive impairment; in particular, compound A is highly effective against neuropathy, neuropathic pain, and / or neurodegeneration; furthermore, compound A also exhibits unexpected bradycardia activity and can be used for diseases requiring a reduction in heart rate (tachycardia), such as in chemotherapy-induced cardiovascular toxicity; tachycardia is part of a central nervous system disease because it responds to vagal control. When used to treat neuropathy, it is preferred to be chemotherapy-induced neuropathy present at the central and / or peripheral levels. Chemotherapy agents are well known in the art: generally, but not limited to, they are alkylating agents or proteasome inhibitors (32). Among alkylating agents, platinum complexes, such as cisplatin, carboplatin, etc., may be mentioned. Among the proteasome inhibitors that can be mentioned are bortezomib, carfizomib, ixazomib, oprozomib, delanzomib, marizomib, MG-132, ONX-0914, VR-23, celastrol, and epoxomicin.

[0030] The term “growth hormone-releasing peptide receptor” is well known in the art and also has alternative names such as “growth hormone secretagogue receptor” or “GHS receptor”. All these synonyms are equivalent and can be used interchangeably herein; the term growth hormone-releasing peptide receptor and its synonyms are extended to all its possible forms (e.g., the GHS1 form) and all its isoforms (e.g., isoforms GHS1a, GHS1b, etc.).

[0031] When used herein, the term "treatment" refers to the medical administration of a patient with a disease, pathological condition, symptom, or disorder aimed at curing, improving, stabilizing, or preventing such disease, pathological condition, or disorder. This term includes active treatment, which is treatment specifically aimed at improving a disease, pathological condition, symptom, or disorder, and also includes causal treatment, which is treatment aimed at eliminating the cause of the associated disease, pathological condition, or disorder. Additionally, the term includes palliative treatment, which is treatment aimed at relieving symptoms, pathological condition, or disorder rather than curing the disease; preventative treatment, which is treatment aimed at minimizing or partially or completely suppressing the development of the associated disease, pathological condition, or disorder; and supportive treatment, which is treatment used to complement another specific treatment aimed at improving the associated disease, pathological condition, or disorder.

[0032] The present invention provides a method for treating or preventing one or more medical conditions (i.e. diseases or disturbances) mediated by auxin-releasing peptide receptors in the central nervous system, characterized by administering compound A to the patient in need.

[0033] Another object of the present invention is to use compound A for the treatment or prevention of one or more medical conditions (diseases or disturbances) mediated by ghrelin receptors in the central nervous system.

[0034] Another object of the present invention is the use of compound A in the preparation of a medicament for treating or preventing one or more medical conditions (diseases or disturbances) mediated by ghrelin receptors in the central nervous system.

[0035] Another object of the present invention is a method for promoting the absorption of a growth hormone-releasing peptide agonist in the central nervous system in patients in need, characterized in that a therapeutically effective amount of compound A, which is a growth hormone-releasing peptide agonist, is administered to the patient.

[0036] Another object of the present invention is a method for establishing a therapeutically effective concentration of a growth hormone-releasing peptide agonist in the central nervous system of a patient in need, characterized in that the patient is given a therapeutically effective amount of compound A as a growth hormone-releasing peptide agonist.

[0037] As used herein, the term “pharmaceutical acceptable” means that a pharmaceutical composition is generally safe, non-toxic and does not have undesirable biological or other effects, and includes those pharmaceutically acceptable for veterinary and human use.

[0038] Compound A, namely 3-(1-(2,3-dichloro-4-methoxyphenyl)ethyl)-1-methyl-1-(1,3,3-trimethyl-piperidin-4-yl)urea hydrochloride, is a novel compound; the corresponding free base is described in patent application WO2012 / 116176. Therefore, the present invention includes compound A itself, its use in medicine, and pharmaceutical compositions comprising it; the present invention also includes a method for preparing compound A, characterized by the reaction of its free base with hydrochloric acid, as shown in the Examples section.

[0039] As further illustrated in the Examples section, compound A possesses interesting purity, stability, and solubility, making it particularly suitable for formulation in various pharmaceutical forms under different manufacturing conditions without a significant decrease in purity and potency. Therefore, the invention extends to compound A in crystalline form, particularly those exhibiting... Figure 9The XRDP peak pattern shown is a specific crystalline form. Also due to these properties, compound A can be freely formulated in any pharmaceutical form according to the needs of the selected therapeutic treatment. A form suitable for systemic administration is preferred. For example, it can be formulated as tablets, pills, capsules, microcapsules, granules, microgranules, pellets, micropellets, powders, lyophilized powders, solutions, suspensions or emulsions, gels, creams, transdermal or percutaneous drug delivery systems, etc.

[0040] Based on the weight of the free base, compound A is preferably administered at a dose of about 0.03 mg to about 10 mg, more preferably about 0.1 mg to about 2 mg. For typical adult patients, these doses are daily doses. They can be modified and / or adjusted according to the severity of the disease, specific patient condition, chosen route of administration, etc.

[0041] The route of administration for compound A is systemic: due to its ability to penetrate the blood-brain barrier, it is not necessary to inject it directly into the central nervous system or brain to target the central nervous system. In fact, it is sufficient for the drug to reach systemic circulation via a conventional systemic route, such as oral, oral, inhalation, rectal, etc. Once circulating in the bloodstream, compound A is absorbed by the endothelial cells of the central nervous system and released from there into the central nervous system in its active form. Therefore, it is possible to treat central nervous system diseases mediated by auxin-releasing peptide receptors without having to resort to invasive routes of administration that directly enter the central nervous system (e.g., intrathecal, intraspinal, etc.). Advantageously, according to the present invention, invasive routes of administration that directly enter the central nervous system can be avoided. Therefore, the intended method of administration of the present invention can also be characterized as "around the central nervous system" or "outside the central nervous system."

[0042] Various pharmaceutical compositions utilizing compound A of the present invention can be developed. These compositions are suitable for administration via any suitable route, such as oral, parenteral, or intravenous administration in liquid or solid form. Preferred routes of administration are injection and / or oral. These compositions will typically contain an inert diluent or edible excipient. They can be encapsulated in gelatin capsules (for oral administration), compressed into tablets (for oral or buccal use), or formulated into lozenges (for buccal use). For these purposes, the active compound can be combined with excipients and used in the form of tablets, lozenges, or capsules. Pharmaceutically compatible binders and / or adjuvant materials can be included as part of the composition.

[0043] Tablets, pills, capsules, lozenges, etc., may contain any of the following ingredients or compounds with similar properties: binders, such as microcrystalline cellulose, astragalus gum, or gelatin; excipients, such as starch or lactose; disintegrants, such as alginic acid, Primogel, or corn starch; lubricants, such as magnesium stearate; slippers, such as colloidal silica; sweeteners, such as sucrose or saccharin; or flavorings, such as peppermint, methyl salicylate, or orange flavorings. When the dosage unit form is a capsule, it may also contain a liquid carrier, such as fatty oil, in addition to the materials of the types described above. Furthermore, the dosage unit form may contain various other materials that alter the physical form of the dosage unit, such as sugar, shellac, or other enteric coating layers.

[0044] The compound can be administered as an elixir, suspension, syrup, tablet, orally disintegrating film, orally disintegrating tablet, or chewing gum component. In addition to the active compound, the syrup may also contain sucrose as a sweetener and certain preservatives, dyes, colorants, and flavorings.

[0045] Solutions or suspensions intended for injection may include the following components: sterile diluents, such as water for injection, saline solutions, non-volatile oils, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; antimicrobial agents, such as benzyl alcohol or methylparaben; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers, such as acetates, citrates, or phosphates; and agents for toning, such as sodium chloride, mannitol, and glucose. Injectable formulations may be packaged in glass or plastic ampoules, disposable syringes, or multi-dose vials.

[0046] The following is an exemplary implementation scheme:

[0047] 1. Compound 3-(1-(2,3-dichloro-4-methoxyphenyl)ethyl)-1-methyl-1-(1,3,3-trimethylpiperidin-4-yl)urea hydrochloride, for the treatment and / or prevention of medical conditions of the central nervous system mediated by auxin-releasing peptide receptors.

[0048] 2. The compound used as described in embodiment 1, wherein the medical condition is selected from nerve root hyperplasia, neuropathy, neuropathic pain, encephalomyelitis, Parkinson's disease, Alzheimer's disease, cognitive impairment, vagus nerve hyperstimulation, and tachycardia.

[0049] 3. The compound used as described in embodiment 2, wherein the neurosis is a chemotherapy-induced neurosis.

[0050] 4. The compound used as described in embodiment 3, wherein the chemotherapeutic induced neurosis is induced by a proteasome inhibitor or an alkylating agent.

[0051] 5. The compound for use as described in Embodiment 4, wherein the proteasome inhibitor is selected from bortezomib, carfilzomib, ixazomib, aprozomib, dilanzomib, malizomib, MG-132, ONX-0914, VR-23, sennatoxin, and epothilone.

[0052] 6. The compound used as described in embodiment 4, wherein the alkylating agent is selected from cisplatin or carboplatin.

[0053] 7. The compound used as described in embodiment 2, wherein the tachycardia is chemotherapy-induced tachycardia.

[0054] 8. The compound used as described in embodiments 1-7, wherein the dosage of the compound, expressed as a free base, is 0.03 to 10 mg.

[0055] 9. A compound used as described in embodiments 1-8, wherein the compound is applied externally to the central nervous system.

[0056] 10. A compound for use as described in embodiment 9, wherein the compound is administered via a route selected from: oral, oral, buccal, sublingual, ocular, percutaneous, dermal, intravenous, intramuscular, inhalation, or rectal.

[0057] 11. Compound 3-(1-(2,3-dichloro-4-methoxyphenyl)ethyl)-1-methyl-1-(1,3,3-trimethylpiperidin-4-yl)urea hydrochloride.

[0058] 12. Compound 3-(1-(2,3-dichloro-4-methoxyphenyl)ethyl)-1-methyl-1-(1,3,3-trimethylpiperidin-4-yl)urea hydrochloride, for treatment.

[0059] 13. A pharmaceutical composition comprising 3-(1-(2,3-dichloro-4-methoxyphenyl)ethyl)-1-methyl-1-(1,3,3-trimethylpiperidin-4-yl)urea hydrochloride in the presence of one or more pharmaceutically acceptable excipients.

[0060] 14. The composition of embodiment 13, wherein the compound is in an applicable form selected from: tablets, pills, lozenges, chewing gum, capsules, microcapsules, powders, lyophilized forms, pellets, micropellets, granules, microparticles, gels, creams, ointments, films, patches, suppositories, solutions, suspensions, syrups, elixirs, or sheets.

[0061] The invention will now be described with reference to the following non-limiting embodiments. Example

[0062] Example 1

[0063] Synthesis and characterization of (3-(1-(2,3-dichloro-4-methoxyphenyl)ethyl)-1-methyl-1-(1,3,3-trimethylpiperidin-4-yl)urea monohydrochloride (Compound A).

[0064] At 22℃±3℃, 1.63 kg of dried (3-(1-(2,3-dichloro-4-methoxyphenyl)ethyl)-1-methyl-1-(1,3,3-trimethylpiperidin-4-yl)urea product obtained by the synthesis method previously described in WO2012 / 116176 was dissolved in 11.2 kg of acetone. The solution was filtered through a 1-micron filter bag, polished, and the filter was washed with acetone (1.6 kg). Maintaining the internal temperature at 22℃±3℃, 4M (1.2 kg) hydrochloric acid aqueous solution was added... The resulting suspension was stirred at 22℃±3℃ for at least 2 hours in the filtered solution, and then the product was separated by centrifugation and washing with acetone (1.6 kg) to obtain 1.5 kg of wet product. The wet product was dried under vacuum at 60℃±5℃ for at least 18 hours to obtain 1.4 kg of (3-(1-(2,3-dichloro-4-methoxyphenyl)ethyl)-1-methyl-1-(1,3,3-trimethylpiperidin-4-yl)urea monohydrochloride (compound A), as a white to off-white crystalline powder. Minimum purity (HPLC) 98%.

[0065] XRPD of compound A 1 H-NMR, 13 C-NMR characterization is shown in [reference needed]. Figure 9-11 .

[0066] Example 2 Water solubility test

[0067] Compound A has a solubility of approximately 33 mg / mL in water at room temperature. For reference, the solubility of the corresponding fumarate is only 10 mg / mL, and the solubility of the corresponding free base is much less than 2 mg / mL.

[0068] Example 3 Stability Test

[0069] The stability of compound A was analyzed by HPLC. The table below shows the data collected during wet storage and laboratory-scale drying. Additionally, a 3-year stability study was conducted on industrial batches of the dried material.

[0070] Stability of compound A in crude wet material at 2-8°C and room temperature in air:

[0071]

[0072] Based on the results, no degradation of compound A was observed in the wet powder after storage at 2-8°C for more than 4 weeks. Similarly, samples stored at room temperature for 24 days after being placed in a refrigerator for 36 days showed no degradation in HPLC.

[0073] Stability of dried crude compound A at 25°C / 60% RH (relative humidity):

[0074]

[0075] Based on the results, we did not observe any degradation of compound A after the dried product was stored at 25°C / 60% RH for 3 years.

[0076] Stability of dried crude compound A during drying at 60°C

[0077]

[0078] Based on the results, we observed that the material was stable during drying at 60°C for 74 hours.

[0079] Example 4: Evaluation of the ghrelin agonist activity of compound A

[0080] HEK293 cells stably expressing the human GHSR1a receptor were used for FLIPR analysis. Cells were maintained using standard procedures. The day before the test, cells were cultured at 1.5 x 10⁻⁶ cells / day. 4 / wells were inoculated at a density of / wells in 30 μl of DMEM complete medium The sample was coated in a 384-well plate and incubated at 37°C in a 5% CO2 environment for 22–26 hours. On the day of testing, 4× loading dye was added to each well (10 μl per well for a 384-well plate). The plate was incubated in the dark at 37°C for 30 minutes. Then, the dye was removed by centrifugation at 300 rpm for 30 seconds. 40 μl of HBSS / Hepes containing 1 mM insecticide was added using a Platemate Matrix (low speed setting, Thermo). The plate was then placed in a FLIPR Tetra (Molecular Device), and 5× working concentration of agonist was added via FLIPR. Fluorescence signal was detected by FLIPR at room temperature according to standard settings.

[0081] Compound A exhibited strong agonist activity in FLIPR analysis, EC 100%. 50 It is 1.25 ± 0.42 nM.

[0082] In another binding study, compound A was dissolved in DMSO and diluted with water at different concentration ranges. Membrane proteins prepared from BHK cells stably expressing the human GHSR1 receptor were used for the binding assay. The membranes were diluted in analytical buffer to obtain 20 μg / well, 120 μl per well. The binding assay was performed in 96-well plates as follows: 120 μl [ 125 [I] Ghrelin (final concentration 1 nM) and 15 μl of compound A (10x) were diluted in assay buffer. The reaction mixture was incubated at room temperature for 30 min, then rapidly filtered through a GF / B filter plate pre-soaked in 0.3% PEI using a cell collector (Perkin Elmer) to terminate the reaction. The filter membrane was washed three times and dried overnight at 37°C. The radioactivity bound to the filter membrane was measured using MicroBeta Trilux (Perkin Elmer). Compound A in [ 125 The GHrelin binding assay showed a strong affinity for the GHSR1 A receptor, with a Ki value of 1.42 ± 0.35 nM.

[0083] Example 5

[0084] Pharmacokinetic studies and brain penetration assessment of compound A

[0085] Following a single intravenous injection of 10 mg / kg of compound A into Sprague Dawley rats, the compound was rapidly cleared from systemic circulation and distributed to tissues and organs. Bioanalysis of compound A in plasma and brain was performed using LC-MS-MS, and pharmacokinetic analysis was performed using standard non-compartmental methods. In this assay, the concentrations of compound A measured in the brain at 1, 2, and 8 hours after administration at a dose of 10 mg / kg were found to be 1.5 to 1.9 times higher than in plasma, and both curves showed parallel decay to the corresponding plasma concentration curves. Figure 12 This study shows the changes in the mean concentrations of compound A in plasma (ng / mL) and brain (ng / g) after a single intravenous injection of 10 mg / kg of compound A into male Sprague Dawley rats. Interestingly, the higher concentrations of compound A in the brain decreased in parallel with the plasma concentration: this indicates that despite its high affinity for the brain, the compound does not accumulate there, thus avoiding the risk of local neurotoxicity.

[0086] In detail, after single oral administration at doses of 3, 10, and 30 mg / kg, the bioavailability of compound A was high, exceeding 80%. Following single intravenous and oral administration, the exposure to compound A increased with increasing dose within the measured dose range. A summary of pharmacokinetic data for rats administered compound A orally or intravenously is reported below:

[0087]

[0088]

[0089] In further research, the pharmacokinetics of compound A were compared with those of two reference compounds previously described in patent application WO2012 / 116176. The data obtained are as follows:

[0090] Reference 1

[0091]

[0092] Reference 2

[0093]

[0094] Compound A (Invention)

[0095]

[0096] The data above indicate that, two hours after intravenous administration, the tested reference compounds failed to reach therapeutically significant concentrations in the brain. In contrast, compound A showed a 2- to 3-fold increase in concentration (172 ng / g). Particularly interesting is the brain / blood ratio, which, compared to the fraction remaining in the blood, indicates how the administered dose selectively and directionally crosses the entire blood-brain barrier and accumulates in the brain: both reference compounds showed only a 0.1 / 0.3 brain / blood ratio, indicating poor brain permeability, with the administered dose essentially retained in the blood. In contrast, compound A had a brain / blood ratio of 1.4: compound A has a stronger affinity for the ventricles, thus a large fraction of the administered dose was found in the brain only 2 hours after systemic administration; brain absorption was not yet complete, allowing the drug to partially exhibit useful activity at the peripheral level. This is particularly useful in neurological disorders involving peripheral nerve degeneration in addition to damage at the CNS level, requiring repair at this level. Unbound by theory, it seems that the high brain permeability of compound A may be due to its ability not to act as a P-gp substrate, which is an efflux pump system that prevents molecules from crossing the blood barrier.

[0097] The following table lists other pharmacokinetic data:

[0098] Pharmacokinetics of Compound A after acute and chronic administration

[0099]

[0100] * NR (Not Reported) when the correlation coefficient for terminal elimination is <0.9

[0101] SN = sciatic nerve; DRG = dorsal root ganglion

[0102] Example 6

[0103] Effects of compound A on in vivo neurotoxicity models in animals

[0104] Drugs and preparations

[0105] Compound A was prepared as a suspension in 0.5% carboxymethyl cellulose solution and administered to rats and mice at doses of 1 and 10 mL / kg, respectively, orally at doses of 3, 10, or 30 mg / kg. Cisplatin was administered intraperitoneally at 0.5 mg / kg in physiological saline (1 mL / kg). Oxaliplatin was prepared at a concentration of 0.6 mg / mL in 5% glucose solution and administered intraperitoneally (10 mL / kg). Bortezomib was prepared at a concentration of 0.2 mg / kg in 10% Tween 80, 10% EtOH 100%, and 80% saline solutions and administered intravenously (1 mL / kg). Fresh solutions of all doses were prepared on each day of administration.

[0106] Cisplatin Research

[0107] At the start of the experiment, fifty male Wistar rats weighing 250-300g were randomly divided into five groups of ten each. Group 1 received compound A (orally) for six days and cisplatin (intraperitoneally) for three days daily. Group 2 received compound A (orally) and 0.5 mg / kg cisplatin (intraperitoneally) for three days, followed by another three days of compound A. Group 3 received 3 mg / kg compound A (orally) and 0.5 mg / kg cisplatin (intraperitoneally) for three days daily, followed by another three days of compound A. Group 4 received 10 mg / kg compound A (orally) and 0.5 mg / kg cisplatin (intraperitoneally) for three days, followed by another three days of compound A. Group 5 received 30 mg / kg compound A (orally) and 0.5 mg / kg cisplatin (intraperitoneally) for three days daily, followed by another three days of compound A. In each case, cisplatin or cisplatin excipient was administered 30 minutes after compound A or compound A excipient, and 1 hour before the von Frey filament test. Individual body weight and 24-hour food intake were measured daily from the start of administration.

[0108] Oxaliplatin Studies

[0109] At the start of the experiment, 60 female Balb / C mice weighing 20-25g were used and divided into 4 groups of 15 mice each. Group 1 received compound A (orally) once daily, 60-90 minutes before intraperitoneal injection of oxaliplatin excipient, twice weekly for 4 weeks. Group 2 received compound A (orally) once daily, 60-90 minutes before intraperitoneal injection of oxaliplatin excipient, twice weekly for 4 weeks. Group 3 received 10mg / kg of compound A (orally) twice daily, 60-90 minutes before intraperitoneal injection of 6mg / kg oxaliplatin, twice weekly for 4 weeks. Group 4 received 30mg / kg of compound A (orally) twice daily, 60-90 minutes before intraperitoneal injection of 6mg / kg oxaliplatin, twice weekly for 4 weeks. Body weight was measured immediately before the start of each administration. Nerve conduction velocity (NCV) and potential amplitude were measured 24 h after the last dose of oxaliplatin (or excipient) and 1 h after the last dose of compound A (or excipient). Blood and tissue samples (DRG, sciatic nerve [SN], and footpad) were then collected for pharmacokinetic and IENFD assessment.

[0110] Bortezomib Research

[0111] Prevention example: At the start of the experiment, 56 female Wistar rats weighing 200-225g were randomly divided into 5 experimental groups. Group 1 received no treatment (CTRL, n=10); Group 2 received bortezomib 0.2 mg / kg intravenously three times a week for 8 weeks (BTZ, n=10); Group 3 received bortezomib 0.2 mg / kg intravenously three times a week for 8 weeks in combination with oral compound A 3 mg / kg 90 minutes before daily intravenous bortezomib injection (BTZ+compound A3, n=12); Group 4 received bortezomib 0.2 mg / kg intravenously three times a week for 8 weeks in combination with oral compound A 10 mg / kg 90 minutes before daily intravenous bortezomib injection (BTZ+compound A10, n=12); Group 5 received bortezomib 0.2 mg / kg intravenously three times a week for 8 weeks in combination with oral compound A 30 mg / kg 90 minutes before daily intravenous bortezomib injection (BTZ+compound A30, n=12). At baseline and 8 weeks after treatment, coccygeal and digital nerve conduction and potential amplitude studies, behavioral tests (dynamic), and blood collection were performed to investigate proteasome inhibition. After 8 weeks of treatment, tissue samples (sciatic and coccygeal nerves, DRG, and footpads) were collected and analyzed to investigate morphological parameters and intraepidermal nerve fiber density (IENFD).

[0112] Treatment Example: At the start of the experiment, 56 female Wistar rats weighing 200-225g were randomly divided into 5 experimental groups. Group 1 received no treatment (CTRL, n=10); Group 2 received bortezomib 0.2mg / kg intravenously three times a week for 8 weeks (BTZ, n=10); Group 3 received bortezomib 0.2mg / kg intravenously three times a week for 4 weeks, followed by bortezomib 0.2mg / kg intravenously three times a week and compound A 3mg / kg orally daily 90 minutes before bortezomib intravenous injection for 4 weeks (BTZ+compound A 3, n=12); Group 4 received bortezomib 0.2mg / kg intravenously three times a week for 4 weeks, followed by bortezomib 0.2mg / kg intravenously three times a week and compound A 10mg / kg orally daily 90 minutes before bortezomib intravenous injection for 4 weeks (BTZ+compound A). Group 10 (n=12); Group 5 received bortezomib 0.2 mg / kg intravenously three times a week for 4 weeks, followed by combination therapy of bortezomib 0.2 mg / kg intravenously three times a week and compound A 30 mg / kg orally daily 90 minutes before bortezomib intravenous injection for 4 weeks (BTZ + compound A 30, n=12). At baseline and 8 weeks after treatment, caudal and digital nerve conduction and potential amplitude studies, as well as blood collection, were performed to investigate proteasome inhibition. Behavioral tests (dynamic) were performed 4 and 5 weeks after bortezomib treatment. At 8 weeks after treatment, sciatic nerve, caudal nerve, DRG, and footpads were collected from all animals and analyzed to study morphological parameters and IENFD.

[0113] Evaluate

[0114] von Frey test

[0115] In the cisplatin study, rats were housed and acclimatized to the treatment and Von Frey testing equipment a few days before the start of the experiment. Baseline measurements were obtained prior to administration. Behavioral tests were performed according to the previously described procedure (28). Each filament was tested five times, and testing continued until three exit responses to a particular filament were recorded.

[0116] Dynamic anesthesia device test

[0117] In the bortezomib study, the mechanical nociceptive threshold was assessed using a dynamic anesthesia testing device (model 37450, Ugo Basile Biological Instruments, Comerio, Italy). The mechanical threshold was assessed three times on each side every 2 minutes to obtain an average. The results represent the maximum stress the animal could withstand.

[0118] Nerve Conduction Studies (NCS)

[0119] As previously described in mice (29) and rats (30), the NCV and potential amplitude measurements of the caudal and digital nerves were determined. Baseline NCS measurements were performed prior to drug administration. Animals were then randomly assigned to one of the study treatment groups with similar mean NCS values. NCS measurements were performed again 24 hours after administration of the antitumor drug and 1 hour after the last administration of compound A. During all recordings, animals were anesthetized with 2% isoflurane and placed prone on a warm heating pad, with rectal temperature monitored and maintained between 37.0 and 41.0 °C. Each nerve segment was stimulated at least 3 times and up to 6 times with increasing voltage until a maximal response was achieved. Latency was scored from the onset of stimulation and the potential amplitude from baseline.

[0120] DRG and sciatic nerve morphology

[0121] At the end of the oxaliplatin and bortezomib experiments, animals were euthanized under deep anesthesia. SN fragments and L5 and L6 DRGs were dissected from 5 animals / groups and embedded for analysis as described above (31).

[0122] IENFD analysis

[0123] IENFD analysis was performed on specimens collected and processed as described above (31). Each biopsy was divided into four parts. Unmyelinated axons within the epidermis were counted in a blinded manner, and the fiber density per millimeter of skin fibers passing through the dermal / epidermal junction was determined as described above.

[0124] Statistical analysis

[0125] For all statistical analyses, one or two ANOVAs were used to compare the data to compare the mean group responses, followed by Tukey or Dunnet post-hoc comparisons using Prism Graphpad software version 4.03 (GraphPad Inc, La Jolla, CA), with significance defined as p < 0.05.

[0126] result

[0127] Cisplatin Research

[0128] Food intake and weight change

[0129] Compared with rats treated with excipients, cisplatin treatment caused a significant decrease in daily food intake and body weight (p<0.05). Figure 1 (A, 1B). Compared with cisplatin alone, treatment with compound A at doses of 3, 10, and 30 mg / kg increased total food intake and body weight (p<0.05).

[0130] abnormal pain

[0131] Compared with excipient-treated rats, cisplatin treatment reduced the paw withdrawal threshold (p<0.01), indicating the development of mechanosensitive hyperalgesia. Pretreatment with compound A at 10 and 30 mg / kg significantly alleviated this hyperalgesia. From day 3 to day 5, treatment with compound A at a dose of 30 mg / kg actually resulted in a significantly higher overall paw withdrawal threshold compared with excipient-treated rats (p<0.05). Compound A at 3 mg / kg had no significant effect on cisplatin-induced hyperalgesia.

[0132] Oxaliplatin Studies

[0133] weight change

[0134] Mice treated with oxaliplatin showed significant weight loss from day 10 to the end of the study compared to mice treated with the excipient (p<0.01). Animals co-treated with oxaliplatin and compound A at 10 or 30 mg / kg showed less weight loss compared to mice treated with oxaliplatin (p<0.01). This effect reached statistical significance on multiple test days (p<0.05), with compound A at 10 mg / kg showing the strongest effect (data not shown).

[0135] Nerve conduction research

[0136] Compared with excipient-treated mice, oxaliplatin administration caused a significant reduction in finger and tail NCV (reductions of 10.6 ± 1.6% and 8 ± 1.1%, respectively, p < 0.01). Co-treatment with compound A at 10 mg / kg and 30 mg / kg significantly prevented both types of damage. Figure 2 Oxaliplatin (13.5+ / - 6.4%) also reduced digital potential amplitude, although its effect did not reach statistical significance given the variability. Treatment with compound A tended to normalize the reduction in potential amplitude, thus significantly improving digital potential amplitude at 30 mg / kg compared to oxaliplatin-treated mice (an increase of 33+ / - 9%; p < 0.01). Figure 2 B). Oxaliplatin treatment had no significant effect on the amplitude of the tail nerve potential. Figure 2 D).

[0137] Pathological examination

[0138] No degenerative changes were observed in the DRG neuron cell bodies or satellite cells or sciatic nerve of mice treated with oxaliplatin, and treatment with compound A itself did not cause any degenerative changes (data not shown).

[0139] In the footpad, oxaliplatin treatment significantly reduced IENFD (-31.7 ± 3.5%, p < 0.01 compared to excipient-treated mice). Parallel compound A treatment completely normalized IENFD to values ​​in mice treated with excipients at 10 and 30 mg / kg (p < 0.01). Figure 3 A).

[0140] Pharmacokinetics of Compound A after acute and chronic administration

[0141] Compound A exhibited very high dose-dependent sciatic nerve and DRG exposures. Following single oral doses of 10 and 30 mg / kg, compound A was readily absorbed and distributed in plasma and nerve tissue, with peak concentrations occurring within 0.25–0.5 hours post-administration. The terminal half-life of compound A in plasma was short (approximately 1 h), but significantly longer in the sciatic nerve (approximately 4.7 h). Extrapolation to infinity (AUC) under the concentration-time curves... 0-∞ The concentration of compound A was lowest in plasma and almost 3-4 times higher in the sciatic nerve and DRG, and this was dose-dependent, indicating that compound A has an enhanced ability to penetrate the peripheral nervous system (Table 1). Figure 4 (AB). After 30 days of daily administration, the tissue permeability index (tissue / plasma ratio) of compound A further increased to approximately 9-12 times that of SN and 18-19 times that of DRG, indicating significant accumulation of compound A in these tissues (Table 1 and ). Figure 4 C).

[0142] Bortezomib Research

[0143] weight change

[0144] In a prophylactic study, animals treated with bortezomib and different doses of compound A in combination had significantly higher body weights at several time points during the first few weeks up to day 21 compared to the CTRL and bortezomib groups. However, this difference was not significant at the end of treatment (data not shown).

[0145] In therapeutic studies, animals treated with bortezomib and compound A for 10 days showed a significant increase in body weight compared to the CTRL and bortezomib groups. At the end of treatment, animals treated with BTZ + compound A 30 alone showed a significant increase in body weight compared to the CTRL and bortezomib groups (p<0.01, data not shown).

[0146] Mechanical threshold

[0147] In both bortezomib studies, the use of the antitumor drug lowered the mechanical threshold. At the end of prophylactic treatment, the group treated with bortezomib in combination with compound A did not show abnormal pain compared to CTRL, while this phenomenon was observed in the bortezomib group (p<0.001). Figure 5 A).

[0148] In therapeutic studies ( Figure 5 (B) At 4 weeks, all groups receiving bortezomib showed a shortened latency of abnormal pain compared to CTRL until withdrawal (p<0.001 vs CTRL). At 5 weeks (i.e., 1 week of combined BTZ and compound A treatment) and at the end of treatment, compared to CTRL, the group receiving bortezomib alone showed hyperalgesia, while all groups treated with compound A were protected (p<0.001).

[0149] Nerve conduction research

[0150] In the preventative study of digital NCV, the groups treated with bortezomib alone or in combination with compound A3 at 10 mg / kg showed a statistically significant reduction (p < 0.01 vs CTRL), while the group treated with BTZ + compound A30 showed no change relative to CTRL. Figure 6 A). At the end of treatment, no change in digital potential amplitude was observed relative to CTRL in any group ( Figure 6 B).

[0151] In prophylactic studies, all groups treated with bortezomib alone or in combination with all doses of compound A showed tail NCV (p<0.05) and tail potential amplitude (p<0.01 vs CTRL). Figure 6 The statistical significance of C and 6D was significantly reduced.

[0152] In therapeutic studies, the reduction in digital nerve NCV was statistically significant in animals treated with bortezomib alone (p<0.05 vs. CTRL). Figure 6 E), while a potential reduction in amplitude was observed in the group receiving the highest dose of compound A in combination (p < 0.05 vs CTRL). Figure 6 F). All groups treated with bortezomib alone or in combination with compound A at all doses showed a statistically significant reduction in tail NCV and potential amplitude (p<0.001 vs CTRL). Figure 6 G, 6H).

[0153] Pathological examination

[0154] Occasionally, mice treated with bortezomib exhibited DRG sensory neuron degeneration and satellite cell cytoplasmic vacuolation. In a prophylactic study, co-administration of compound A provided incomplete protection against these changes. In the sciatic nerve, mild axonal changes were observed in all bortezomib-treated groups, and co-administration of different doses of compound A did not alter these changes.

[0155] In the caudal nerve, animals treated with bortezomib alone showed decreased fiber density, axonal and Schwann cell degeneration, and animals treated with bortezomib in combination with compound A showed a mild, non-dose-dependent reduction in these pathological changes at different doses.

[0156] These protective effects were not present when compound A was delivered in therapeutic studies.

[0157] Compared with CTRL, chronic treatment with bortezomib resulted in a statistically significant reduction in IENFD (p<0.001). Although an inhibitory trend was observed, all doses of compound A were in the preventative formulation ( Figure 3 B) significantly prevented the reduction of IENFD induced by bortezomib, but in the treatment setting ( Figure 3 C) is not available.

[0158] Proteasome inhibition research

[0159] Chronic treatment with bortezomib induced statistically significant inhibition of proteasome activity. In prevention or treatment studies, co-administration of different doses of compound A did not impair bortezomib-induced proteasome inhibition (data not shown).

[0160] The following table summarizes the assessment results from various neurotoxicity models:

[0161]

[0162] + Compound A has a significant effect on anti-tumor drugs; + / - Compound A is partially effective against anti-tumor drugs; - Compound A has no effect on anti-tumor drugs; = Anti-tumor drugs have no effect.

[0163] discuss

[0164] Using well-established acute and chronic preclinical CIPN models, we tested the effects of compound A administration. Furthermore, in a bortezomib model, we demonstrated that the drug's antitumor mechanism of action was not interfered with. These animal models have been widely used to test the efficacy of hypothetical neuroprotective therapies, thus providing a theoretical basis for our study.

[0165] A reduction in mechanical abnormal pain was consistently observed across all test conditions. These results are particularly significant in the bortezomib model, as it is an antitumor drug that induces the most painful, dose-limited CIPN. As the results from the cisplatin study suggest, the antinociceptive effect observed in our study was very rapid and likely centrally mediated due to the high permeability of compound A to the brain.

[0166] However, results obtained using long-term administration indicate that compound A also possesses certain peripheral nutritional effects, as it effectively prevented drug-induced reductions in IENFD in both oxaliplatin and bortezomib models. Notably, in therapeutic studies, the reduction in abnormal pain remained significant but was unrelated to IENFD, further supporting the hypothesis of a separation between central analgesia and peripheral neuroprotection, the latter of which can only occur if compound A treatment is initiated before small fiber atrophy.

[0167] In addition to this effect, compound A alleviated some neurophysiological deficits induced by chronic, repeated administration of oxaliplatin, and to a lesser extent, also alleviated some neurophysiological deficits induced by bortezomib. This supports the additional activity of compound A in the myelin fibers of peripheral branches of sensory neurons in the nervous system. In fact, this significant neurophysiological effect, measured directly in the peripheral nerves and primarily reflecting the activity of large myelinated fibers, cannot be attributed to central events, but is likely due to the preservation of the number of peripheral axonal responses.

[0168] A dose-response relationship of 10 mg / kg was also observed for the bell-shaped compound A.

[0169] Compound A’s remarkable ability to penetrate the CNS, along with its peripheral neurotrophic activity, non-drug-specific activity, and potential use in treating cancer cachexia, make it a unique neuroprotective agent in cancer patients undergoing neurotoxic-induced chemotherapy.

[0170] Example 7

[0171] Effects of compound A on the cardiovascular system of human volunteers (Phase 1 study)

[0172] This study was conducted as a single-center, randomized, double-blind, placebo-controlled phase 1 trial in which healthy male subjects received a single escalating dose of compound A orally. The dose-escalation design was deemed suitable for investigating the safety and tolerability of this novel compound in humans with a first single dose. In each cohort, six subjects received a single oral dose of compound A on day 1, and two subjects received a placebo.

[0173] Human Batch 1:10mg Compound A

[0174] Batch 2 for human subjects: 0.1 mg of compound A

[0175] Batch 3 for human subjects: 0.3 mg of compound A

[0176] Batch 4: 1.0 mg Compound A

[0177] Each subject was randomly assigned to receive a single oral dose of either compound A or a placebo. Administration of the study drug was performed on the morning of Day 1. Proper administration of the study drug was supervised by the investigator or a representative. This included checking the oral cavity and buccal cavity. The study drug or placebo was provided in gelatin capsules containing powder; they were taken orally with 240 ml of water. Subjects were administered the drug on a fasting basis (no food for at least 8 hours prior to administration). Fasting was not lifted until 4 hours after administration.

[0178] Record a 12-lead electrocardiogram (ECG) one day before administration (Day -1) and from Day 1 to 72 hours after administration. Additionally, record Holter-ECGs from at least approximately 12 hours before planned administration until 24 hours after administration. When taking paper ECGs, take three samples at the same time points (1 minute apart). Measure vital signs (blood pressure, pulse rate, temperature [auricular measurement]) on Day -1, and then from Day 1 before administration to 72 hours after administration. Use an ECG system ( Marquette Hellige recorded 12-lead ECGs as planned in the study flowchart. Triple ECGs were recorded at 36, 48, and 72 hours post-drug administration. ECGs were recorded in a supine position at the time of extraction from the Holter-ECG (to allow sufficient time) after a period of sitting still for at least 5 minutes or at least 15 minutes. ECGs were drawn at a paper speed of 50 mm / s and an amplitude of 10 mm / mV, with a recording time of 10 seconds for all leads and at least 3 waves per lead, but preferably 5 waves. The following ECG parameters were evaluated: heart rate, PR interval, QRS interval, QT interval, QTcB (using the Bazett correction formula), and QTcF (using the Fridericia correction formula).

[0179] There were no clinically relevant changes or differences in mean systolic and diastolic blood pressure and body temperature among the treatment groups after administration. A decrease in pulse rate was observed in the group treated with compound A.

[0180] Figure 7 and 8 The results were shown. They exhibited a significant dose-related decrease in heart rate, reaching its maximum 1 to 3 hours after administration. At 1 to 3 hours after administration of 0.1, 0.3, 1.0, and 10 mg of compound A, the maximum decrease in mean heart rate (Holter-ECG data, average of triple readings) was 5.1 bpm, 6.8 bpm, 15.9 bpm, and 18.3 bpm, respectively, compared to a maximum decrease of 6.9 bpm after placebo administration. Heart rate returned to pre-administration levels at 6 hours post-administration. The lowest heart rate values ​​were observed in the 1.0 and 10 mg dose groups.

[0181] Quotation

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Claims

1. A pharmaceutical composition comprising a compound 3-(1-(2,3-dichloro-4-methoxyphenyl)ethyl)-1-methyl-1-(1,3,3-trimethylpiperidin-4-yl)urea hydrochloride in the presence of one or more pharmaceutically acceptable excipients, wherein the compound has ghrelin agonist activity and is permeable to the blood-brain barrier.

2. The composition of claim 1, wherein the compound is suitable for external application to the central nervous system.

3. The composition of claim 2, which is suitable for administration by means of a route selected from: oral, oral, buccal, sublingual, ocular, percutaneous, dermal, intravenous, intramuscular, inhalation, or rectal.

4. The composition of claim 3, wherein the applicable form comprises: Tablets, pills, lozenges, chewing gum, capsules, microcapsules, powders, lyophilized preparations, pellets, micro-pellets, granules, microparticles, gels, creams, ointments, films, patches, suppositories, solutions, suspensions, syrups, elixirs, or sheets.

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