Novel peptoids and their use in the prevention or treatment of chronic pain

Abstract

The present invention relates to novel peptidomimetic (or peptoid) molecules for the prevention and / or treatment of chronic pain, in particular chronic pain resulting from peripheral neuropathy.

Description

[Technical Field]

[0001] The present invention relates to a novel peptide mimetic molecule (also called a peptoid, peptide-peptoid hybrid, or peptomer) for the prevention and / or treatment of chronic pain, particularly chronic pain resulting from peripheral neuropathy (e.g., induced by chemotherapy).

[0002] In the following explanation, references enclosed in square brackets ([]) refer to the bibliography provided at the end of the text. [Background technology]

[0003] Recent technologies Chronic pain affects 1.5 billion people worldwide, and treatment responses are clearly insufficient for them (Finnerup NB et al., 2015)[1]. Recent epidemiological studies estimate that chronic pain affects approximately 20% of adults and 50% of older adults (von Hehn et al., 2012)[2]. Globally, 60% of people with pain report reduced ability to work, and 20% report job loss due to pain. At the same time, 20 billion units of analgesics were sold in 2010, up 9.3% from 14 billion units in 2005. In 2017, the global pain market reached $61.05 billion and is projected to reach $77.13 billion by 2023, growing at an annual rate of 4% from 2017 to 2023.

[0004] Neuropathic pain affects approximately 7% of the general population and 25% of patients with chronic pain. There are two types of neuropathic pain: peripheral neuropathic pain (which may involve nerve plexuses, roots, or trunks depending on its origin, or may be more diffuse in cases of polyneuropathy) and central neuropathic pain (lesions affecting sensory pathways or pain control). Peripheral neuropathy induced by chemotherapy is a frequent adverse effect of many anticancer drugs, characterized by significant chronic pain with a persistent impact on the patient's quality of life and leading to dose adjustments with a risk of reduced clinical efficacy. The increasingly limited range of analgesics (withdrawal of Di-antalvic in 2011) is characterized by outdated and often poorly tolerated treatments.

[0005] Innovation has stalled for more than 50 years, with the majority of proposed new drugs being simple re-formulations or combinations of existing drugs. Recent meta-analyses have shown that, with the exception of gabapentin and duloxetine, treatment for neuropathic pain has not been truly effective to date, and efficacy scores remain low (Hershman et al., 2014)[3].

[0006] The HCN ion channel family (Ludwig et al., Nature 1998; Santoro et al., 1998; Seifert et al., 1999)[4-6] includes four HCN 1-4 members and may offer excellent opportunities for the development of novel heart rate lowering agents and novel analgesics. HCN channels are widely distributed in the pain pathway and play an important role in the onset and maintenance of neuropathic pain (Dunlop et al., 2009; Lewis et al., 2011)[7,8]. Non-selective HCN blockers have been shown to reduce pain symptoms in rodent models of neuropathic pain (Descoeur et al., 2011; Young et al., 2014)[9,10]. However, their cardiac and visual side effects (e.g., pan-HCN blockers) limit the clinical interpretation of their use for treating neuropathic pain.

[0007] Interestingly, HCN channel function is tightly regulated by a co-existing subunit, Rab8b-interacting protein (TRIP8b), which is not expressed in the heart. This cytoplasmic protein binds to the C-terminus of the HCN channel subunit via two contact sites (Lewis et al., 2009)

[11] . Furthermore, the atomic structure of the tetratricopeptide (TPR) region of TRIP8b in a complex with six amino acid peptides representing the C-terminus of HCN2 was determined by X-ray crystallography (Bankston et al., 2012)

[12] .

[0008] Given that much of the current research focuses on developing inhibitors that directly act on HCN channels, targeting TRIP8b and / or TRIP8b-HCN interactions may offer a new possibility for modulating HCN activity in pain states without the risk of affecting cardiac and visual function. [Overview of the Initiative] [Means for solving the problem]

[0009] Description of the present invention This invention is based on the hypothesis that disrupting the TRIP8b-HCN interaction, which results in a decrease in HCN channel-induced current (Ih), may have an analgesic effect in rodent models of oxaliplatin-induced acute and chronic peripheral neuropathy, faithfully reproducing human symptomatology.

[0010] The effectiveness of this strategy was first validated using NUCC-5953, a molecule from a 20,000-molecule library screening capable of disrupting the TRIP8b-HCN interaction (Han et al., 2015)

[13] . Thus, we demonstrated that oxaliplatin-induced acute cold hypersensitivity was dose-dependently reversed by intrathecal (Figure 2) or systemic (Figure 3) injection of NUCC-5953. No changes in walking motor activity were observed. Preparatory data also showed that administration of NUCC-5953 reduced membrane addressing and, consequently, reduced HCN1 and HCN2 activity observed in animals with neurological disorders (Figure 5B). Furthermore, in vitro application of NUCC-5953 showed no effect on HCN activation-induced currents in cardiac cells (in contrast to nonspecific blockers of HCN channels, Figure 6).

[0011] Next, the inventors decided to design a specific blocker of the TRIP8b-HCN interaction based on the interaction mode of the two proteins. However, protein-protein interactions (PPIs) play an essential role at all levels of cellular function and are undeniably therapeutic targets, making the design of potent PPI inhibitors a real challenge. While small molecular weight molecules have been widely screened, they are not well-suited to competing with interactions involving large surfaces (>800 Å). Furthermore, in order to design effective inhibitors, it is crucial that the protein / protein recognition segment can be mimicked in its bioactive "hot segment" conformation. Peptides are compounds of choice for designing these novel inhibitors because they can thereby solve not only the problems of metabolic and conformational stability but also the problem of cell permeability to reach intracellular targets.

[0012] Next, the inventors turned their attention to peptide-mimicking chemicals via the synthesis of peptoids (N-substituted glycine oligomers), which are particularly suited to the development of inhibitors targeting protein-protein interactions and may also be called peptomers (i.e., oligomers combining a "peptoid" monomer and an "amino acid" monomer, Ostergaard et al 1997

[17] —i.e., peptide-peptoid hybrids).

[0013] These are synthetic peptide mimetic compounds, and their oligomeric nature allows for great modularity in terms of size and chemical diversity.

[0014] These allow for finer structural design and possess better bioavailability than peptides (protease resistance, better membrane permeability, and a non-immunogenic skeleton).

[0015] Based on this, the inventors designed a novel peptoid that targets the interaction between the TPR domain of TRIP8b and the C-terminus of HCN, and can exert an analgesic effect in a model of oxaliplatin-induced acute neuropathy without harmful cardiac or visual side effects.

[0016] Therefore, the present invention relates to the following general formula (I): [ka] [In the formula, R 1 These are independently CH3(L-Ala), CH(CH3)2(L-Val), CH(CH3)CH2CH3(L-Ile), CH2CH(CH3)2(L-Leu and D-Leu), CH2C(CH3)3, C(CH3)3, and CH2(cyclobutyl). R 2 , R 3 , and R 4 These are side chains of peptoid units that are sequentially incorporated during the synthesis from primary amines. R 2 and R 3relates to a peptoid independently selected from CH2CH(CH3)2, CH2CH2NH2, CH2CH2OH, CH2CH2CH2OH, CH2CH(OH)CH3, CH2CONH2, CH2CH2CONH2. Side chains having an alcohol or amine functional group are incorporated into the oligomer in a protected form, i.e., silyl ether for the alcohol functional group and tert-butyl carbamate (Boc) for the amine functional group, R 4 is one of the following:

Chemical formula

[0017] Advantageously, R4 is one of the following:

Chemical formula

[0018] Advantageously, R 5 is independently CH3, CH2C6H5, C6H4X (where X at the ortho, meta or para position is independently H, OMe, CF3, CH3, CH2CH3, F, Br, Cl), 3-indolyl, 3-quinolyl.

[0019] Advantageously, the peptoid of the present invention has the general formula (I): [wherein, R 1These are independently CH3(L-Ala), CH(CH3)2(L-Val), CH(CH3)CH2CH3(L-Ile), CH2CH(CH3)2(L-Leu and D-Leu), CH2C(CH3)3, C(CH3)3, and CH2(cyclobutyl). R 2 , R 3 , and R 4 These are side chains of peptoid units that are sequentially incorporated during the synthesis from primary amines. R 2 and R 3 The peptoids are independently selected from CH2CH2NH2, CH2CH2OH, CH2CH2CH2OH, CH2CH(OH)CH3, CH2CONH2, and CH2CH2CONH2. The side chains having alcohol or amine functional groups are incorporated into the oligomer in a protected form, i.e., silyl ether for alcohol functional groups and tert-butylcarbamate (Boc) for amine functional groups. R 4 The following: [ka] It is an aliphatic or aromatic group selected from the following. R 5 These are independently CH3, CH2C6H5, C6H4X (wherein X at the ortho, meta, or para positions is independently H, OMe, CF3, CH3, CH2CH3, F, Br, or Cl), 3-indolyl, and 3-quinolinyl.

[0020] Preferably, the peptoid of the present invention of general formula (I) has two carbon atoms on the second unit (starting from the C-terminus).

[0021] Advantageously, the peptoid of the present invention has the following general formula (I'): [ka] [In the formula, R 1 , R 2 , R3 , R 4 and R 5 It is selected from the peptoids as defined above.

[0022] According to a particular embodiment, the peptoid of the present invention has the following formula: [ka] [ka]

[0023] The present invention also relates to an intermediate for the synthesis of a peptoid of general formula (I), wherein the peptoid comprises 3 units. The synthesis intermediate is of the following general formula (II): [ka] [In the formula, R 1 , R 2 and R 3 This is as defined above, R 6 H, R 3 COR 5 (R in the formula 5 (is as defined above) or R 7 And, R 7 Independently, [ka] It is a peptoid of the base of [the substance].

[0024] In particular, the present invention is R 6 This relates to the peptoid of equation (II) where H is.

[0025] In particular, the present invention is R 6 COR 5 This relates to the peptoid of equation (II).

[0026] In particular, the present invention is R 6 R 7 This relates to the peptoid of equation (II).

[0027] In particular, the present invention is R 6 R 3 This relates to the peptoid of equation (II).

[0028] Advantageously, the peptoid according to the present invention has the following general formulas (IIa), (IIb), (IIc), and (IId): [ka] [ka] [ka] [ka] [In the formula, R 1 , R 2 , R 3 , R 5 and R 7 It can be selected from the peptoids as defined above.

[0029] Preferably, the peptoids of the present invention of formula (II), (IIa), (IIb), (IIc), or (IId) have two carbon atoms on the second unit (starting from the C-terminus). Advantageously, the peptoids of the following formulas are (II'), (II'a), (II'b), (II'c), or (II'd): [ka] [ka] [ka] [ka] [ka] [In the formula, R 1 , R 2 , R 3 , R 5 , R 6 and R 7 It is selected from the peptoids as defined above.

[0030] The present invention also relates to a pharmaceutical composition comprising a peptoid according to the present invention and a pharmaceutically acceptable carrier.

[0031] The present invention also relates to a peptoid or pharmaceutical composition according to the present invention for use as a pharmaceutical.

[0032] The present invention also addresses chronic pain, particularly chronic pain of neurological origin, and more specifically, chronic pain of neuropathy, for example, chemotherapy. The present invention relates to a peptoid or pharmaceutical composition for use in the prevention or treatment of peripheral neuropathic pain induced by law. [Brief explanation of the drawing]

[0033] [Figure 1] The images show the expression of HCN1 (A), HCN2 (B), and TRIP8b (C) proteins in DRG neurons 4 days after administration of vehicle or oxaliplatin (6 mg / kg, intraperitoneal). TRIP8b protein expression in the dorsal horn of the spinal cord 4 days after administration of vehicle or oxaliplatin (6 mg / kg, intraperitoneal) (D) or after repeated administration of vehicle or oxaliplatin (twice a week for 3 weeks) (E), N=3 / group, Mann-Whitney U test. [Figure 2]The following shows (A) the study design, (B) the temporal changes in the mean ± SEM of the foot immersion latency threshold (seconds), and (C) the area under the curve (AUC, seconds.minutes) for dose-dependent analgesia in OIPN (oxaliplatin-induced neurological disorder model mice) (n=8) treated with vehicle or NUCC5953 (0.2 μg, 1 μg, or 5 μg, intrathecal). *p<0.05, **p<0.01, ***p<0.001, vs. group, two-way ANOVA and Tukey post-hoc test. [Figure 3] The following shows (A) the study design, (B) the time course of the mean ± SEM of the foot immersion latency (seconds), and (C) the area under the curve (AUC, sec.min) for dose-dependent analgesia in OIPN mice (n=8) treated with vehicle, NUCC5953 (4 mg / kg, subcutaneous), or duloxetine (30 mg / kg, subcutaneous). *p<0.05, **p<0.01, ***p<0.001, vs. vehicle group, two-way ANOVA and Tukey post-hoc test. [Figure 4] This histogram shows the latency to tipping over (A) and the tipping speed (B) of mice treated with a vehicle or NUCC5953 (4 mg / kg, subcutaneously) and subjected to a rotorod test at increasing speed (from 4 rpm to 40 rpm over 10 minutes). [Figure 5] This shows (A) the design of the study and (B) the intracellular (membrane) expression of HCN1, HCN2, and TRIP8b in animal-derived DRGs treated with vehicle (V), oxaliplatin (O), or oxaliplatin + NUCC5953 (5 nmol, intrathecal) (N). [Figure 6] In contrast to ivabradine (VIA), this demonstrates the lack of effect of NUCC5953 on the If current amplitude. [Figure 7] Mouse heart rate (HR) is represented by: (A) baseline value, (B) mean heart rate 30 minutes after injection of NUCC5953 (4 mg / kg, subcutaneous), and (C) mean value at the end of 30 minutes. Values ​​are mean ± SEM (n=3). [Figure 8] This diagram illustrates the molecular dynamics of TRIP8b interacting with SNL peptides. [Figure 9]The following shows (A) the study design, (B) the mean ± SEM time course of foot immersion latency (seconds), and (C) the area under the curve (AUC, seconds.minutes) for dose-dependent analgesia in OIPN mice (n=8) treated with vehicle or peptoid YC55 (0.2, 1, or 5 μg, intrathecal). *p<0.05, **p<0.01, ***p<0.001, vs. vehicle group, two-way ANOVA and Tukey post-hoc test. [Figure 10] The following shows (A) the study design, (B) the temporal changes in the mean ± SEM of the foot immersion latency threshold (seconds), and (C) the area under the curve (AUC, seconds.minutes) for dose-dependent analgesia in OIPN mice (n=8) treated with vehicle or peptoid YC55 (2.5 mg / kg, 5 mg / kg, or 10 mg / kg, subcutaneously). *p<0.05, **p<0.01, ***p<0.001, vs. vehicle group, two-way ANOVA and Tukey post-hoc test. [Figure 11] This histogram shows the latency to tipping over (A) and tipping speed (B) of mice treated with a vehicle, YC55 (2.5 mg / kg, 5 mg / kg, or 10 mg / kg, subcutaneously), or barium (2.5 mg / kg, subcutaneously) and subjected to an accelerated rotarad test (from 4 rpm to 40 rpm over 10 minutes). ***p<0.001 Student's t-test of T(■) vs T0(□) 30 minutes after administration for a given molecule. [Figure 12] The following shows (A) the study design, (B) the mean ± SEM time course of the mechanical threshold (g) and foot immersion latency threshold (seconds), and (C) the area under the curve (AUC, seconds.minutes) for PTX rats (n=7-9) treated with vehicle or peptoid YC55 (0.2, 1, or 5 μg, intrathecal). *p<0.05, **p<0.01, ***p<0.001, vs. vehicle group, two-way ANOVA and Tukey post-hoc test. [Figure 13] This shows (A) the test design and (B) the temporal changes of the mean ± SEM of the foot immersion latency threshold (in seconds) in neuropathic rats (n=7-9) treated with vehicle (●) or peptoid YC55 (■) (5 μg, intrathecal). ***p<0.001, vs. vehicle group, two-way ANOVA and Tukey post-hoc test. [Figure 14] This table shows (A) the study design, (B) the mean ± SEM time course of the foot immersion latency threshold (in seconds), and (C) the area under the curve (AUC, seconds.minutes) for analgesic effect in OIPN mice (n=7-9) treated with a vehicle or peptoid MP208, MP405, or YC55 (5 μg, intrathecal). *p<0.05, **p<0.01, ***p<0.001, vs. vehicle group, two-way ANOVA and Tukey post-hoc test. [Figure 15] This report shows (A) the study design, (B) the time course of the mean ± SEM of the foot immersion latency threshold (seconds), and (C) the area under the curve (AUC, seconds.minutes) for analgesic effect in OIPN mice (n=7-9) treated with a vehicle or peptoid MP376, MP354, or MP341 (10 mg / kg, subcutaneously). *p<0.05, **p<0.01, ***p<0.001, vs. vehicle group, two-way ANOVA and Tukey post-hoc test. [Figure 16] This table shows (A) the study design, (B) the mean ± SEM time course of the foot immersion latency threshold (in seconds), and (C) the area under the curve (AUC, seconds.minutes) for analgesic effect in OIPN mice (n=7-9) treated with a vehicle or peptoid LF108, LF306, or LF188 (5 μg, intrathecal). *p<0.05, **p<0.01, ***p<0.001, vs. vehicle group, two-way ANOVA and Tukey post-hoc test. [Figure 17] This table shows (A) the study design, (B) the temporal changes in the mean ± SEM of the foot immersion latency threshold (in seconds), and (C) the area under the curve (AUC, seconds.minutes) for analgesic effect in OIPN mice (n=7-9) treated with a vehicle or peptoid LF295, LF400, or LF329 (5 μg, intrathecal). *p<0.05, **p<0.01, ***p<0.001, vs. vehicle group, two-way ANOVA and Tukey post-hoc test. [Figure 18]This table shows (A) the test design, (B) the mean ± SEM time course of the foot immersion latency threshold (in seconds), and (C) the area under the curve (AUC, seconds.minutes) for analgesic effect in OIPN mice (n=7-9) treated with vehicle or peptoid LF126, LF261, or LF275 (5 μg, intrathecal). *p<0.05, **p<0.01, ***p<0.001, vs. vehicle group, two-way ANOVA and Tukey post-hoc test. [Figure 19] This table shows (A) the test design, (B) the temporal changes in the mean ± SEM of the foot immersion latency threshold (in seconds), and (C) the area under the curve (AUC, seconds.minutes) for analgesic effect in OIPN mice (n=7-9) treated with a vehicle or peptoid LF222, LF239, or LF176 (5 μg, intrathecal). *p<0.05, **p<0.01, ***p<0.001, vs. vehicle group, two-way ANOVA and Tukey post-hoc test. [Figure 20] This table shows (A) the study design, (B) the mean ± SEM time course of the foot immersion latency threshold (seconds), and (C) the area under the curve (AUC, seconds.minutes) for analgesic effect in OIPN mice (n=7-9) treated with either a vehicle or peptoid LF369 (5 μg, intrathecal). *p<0.05, **p<0.01, ***p<0.001, vs. vehicle group, two-way ANOVA and Tukey post-hoc test. [Modes for carrying out the invention]

[0034] Given the stagnation in therapeutic innovation in the field of analgesics, the peptoids of the present invention and their therapeutic potential can have a significant impact on improving the quality of life and, in some cases, the survival of patients suffering from anticancer drug-induced neuropathic pain. Furthermore, since HCN channels are also involved in the pathogenesis of various neuropathic and inflammatory pain conditions, the peptoids of the present invention pave the way for a new class of analgesics that are more broadly active against various types of chronic pain. [Examples]

[0035] Example 1: Materials and Method animal The experiments were conducted using 20-24g male C57Bl / 6JRj mice or 150-175g male Sprague-Dawley rats provided by Janvier Laboratories (France), maintained in a 12-hour light / dark cycle, with feeding and watering provided as needed. Behavioral experiments were conducted blindly in a quiet room by the same experimenter for each given test, with careful attention paid to minimizing or avoiding animal discomfort. All animal husbandry procedures were approved by the local animal husbandry ethics committee, and the experiments followed the guidelines (Directives) for the management and use of animal husbandry provided by the European Community. It was implemented in accordance with 2010 / 63 / EU).

[0036] Behavioral Test Von Frey Test: Mechanical sensitivity was evaluated using calibrated Von Frey filaments of 0.07g, 0.6g, or 1.4g. The filaments were applied five times perpendicularly to the surface of the hind paw's sole, in increasing order of stiffness, and pressed until they bent. The number of responses to a given filament force was counted.

[0037] Immersion test: The tail or leg was immersed in a water bath set at 10°C or 46°C until contraction was observed (cutoff time: 30 seconds). The average of two separate measurements during escape lurking was calculated (Janssen, Nimegeers & Dony, 1963)

[14] .

[0038] Foot pressure test in rats: Rats were subjected to the foot pressure test previously described by Randall and Selitto (1957)

[15] . The nociceptive threshold, expressed in grams, was measured using a Ugo Basile analgesic (Apelex, probe tip diameter 1 mm, weight 30 g) by increasing pressure applied to the rat's right hind paw until signs of pain (squeal threshold) were obtained (cutoff was 750 g). Prior to the procedure, the rats were accustomed to the test by handling them without exposing them to foot pressure. The therapeutic effect was then evaluated at 15, 30, 45, 60, 90, and 120 minutes after obtaining two consecutive stable squeal thresholds. The results are expressed in grams as squeal thresholds. To investigate the overall effect, the area under the time-course curve of the analgesic effect (AUC, g.min) was calculated from the individual scores at each time point using the trapezoidal method. The data were analyzed using two-way ANOVA followed by the Bonferroni test, during which the time course of the effect was examined. The effects of the different treatments determined by CSA were analyzed using one-way ANOVA followed by the Student-Newman-Keuls test. The level of statistical significance was set at p<0.05.

[0039] Rotarod Test: For the rotorod test (Bioseb), the animals were first accustomed to remaining on a fixed cylinder for 10 minutes, then switched to a rotation speed of 4 rpm for another 10 minutes. The animals were administered a drug immediately after training, and the test was performed 30 minutes later. The test consisted of measuring the latency until the animal fell over, while linearly increasing the rotation speed from 4 rpm to 40 rpm over a 5-minute period. The average of the two closest values ​​over three trials was calculated. Ta.

[0040] In vivo heart rate recording: ECG telemetry recording was performed using a subcutaneous pouch-based TA10EA-F20 telemetry transmitter (DSI) with a pair of wire electrodes positioned on the thoracic cavity, as previously published (Mesirca et al., 2014)

[27] .

[0041] Animal pain models Oxaliplatin-induced acute neuropathy in mice: Mice were intraperitoneally (IP) injected with 6 mg / kg oxaliplatin dissolved in 5% glucose immediately after the initial behavioral assessment. A second assessment of cold sensitivity and mechanical sensitivity was performed 90 hours after drug administration (Descoeur et al., 2011) [9], corresponding to the time required to reach peak hyperalgesia.

[0042] Acute paclitaxel-induced neuropathy in rats: Rats were intraperitoneally injected with dimethyl sulfoxide (DMSO; solvent) or 1 mg / kg / 1 mL of paclitaxel (Sigma-Aldrich, Lyon, France) on days 0 (D0), 2 (D2), 4 (D4), and 7 (D7). Behavioral tests were performed on the day of the first paclitaxel injection (D0) and on days 10 (D10) and 14 (D14) after the first paclitaxel injection.

[0043] Spinal nerve ligation-induced neuropathy in rats: Following the procedure of Kim and Chung (1992)

[16] , rats were placed in a supine position and anesthetized with 10 mg / kg xylazine and 75 mg / kg ketamine (ip). The hair on the left side of the spine was then shaved to expose the skin. The skin was cut parallel to the spine and the paravertebral muscles were separated. The L5 transverse epiphysis was removed until the L5 spinal nerve was exposed. The L5 nerve was isolated and ligated with silk sutures. Finally, the muscle and skin incisions were sutured and the wounds were disinfected with iodine and 75% ethyl alcohol. In the placebo group, rats underwent the same surgical procedure without nerve ligation.

[0044] Ex vivo testing RNA isolation and quantification: DRG pairs were excised and pooled from lumbar vertebral segments L4-L6. RNA was extracted using TRIzol® reagent (Nitrogen) according to the manufacturer's (Thermofisher) instructions. RNA quality was assessed using Agilent. Samples with an RNA Integrity Number (RIN) greater than 8 were evaluated using a 2100 Bioanalyzer (Agilent Technologies) and retained for further analysis. Reverse transcription was performed using SuperScript II reverse transcriptase according to the protocol provided by Invitrogen. Amplification was performed using the Light Cycler Fast Start DNA Kit (Roche Diagnostics) for real-time quantitative PCR, and the amount of DNA was measured in real time using a Mastercycler Realplex thermal cycler (Eppendorf). The relative amounts of cDNA in each triplicate were averaged using a standard concentration curve and normalized to the GUS gene cDNA.

[0045] The amplification conditions were as follows: pre-incubation at 95°C for 7 minutes, followed by 50 cycles of 20 seconds at 95°C and 20 seconds at the specific hybridization temperature of the primer pair, followed by 20 seconds at 72°C.

[0046] RNA sequencing: The inventors used RNA sequencing analysis on L4-L5-L6 DRGs from mice treated with oxaliplatin (3 mg / kg, intraperitoneal) or vehicle twice weekly for 3 weeks. At the end of the experiment (D21), mice were anesthetized and killed, L4-L5-L6 DRGs were rapidly collected, frozen in liquid nitrogen, and stored at -80°C until use. Total RNA from L4-L5-L6 DRGs was analyzed using RNeasy according to the manufacturer's protocol. Extraction was performed using a Microkit (Qiagen), and ribosomal RNA was removed using a Ribo-0 Plus rRNA depletion kit (Illumina). RNA concentration was determined using a Take3 Nucleic Acid Quantification Plate (Epoch, BioTek, USA). RNA integrity was analyzed by capillary electrophoresis (Fragment Analyser, Agilent®), and RNA sequencing was performed using Fasteris (https: / / www.fasteris.com). Libraries were prepared using the Illumina TruSeq protocol, and SBS-based sequencing was performed using the HiSeq 2500 platform (Illumina). The analysis was performed in various steps. Splice site mapping (TopHat2), counting (HTSeq-count), filtering, normalization (edgeR, DESeq, and DESeq2), and differential analysis (edgeR, DESeq, and DESeq2) were performed by Benjamin Bertin and Yoan Renaud (GREd, Clermont-Ferrand).

[0047] Membrane fractionation: Tissue was honodinated at 4°C in HEPES buffer containing a protease inhibitor (20 mM HEPES pH 7.4, 320 mM sucrose, 5 mM EDTA, 5 mM EGTA), sonicated, and centrifuged at 1000 g for 15 minutes at 4°C. The pellet was discarded. For organelle spins, the supernatant was centrifuged at 15,000 g for 10 minutes at 4°C, and the organelle pellet was retained. For cytoplasmic spins, the supernatant was centrifuged at 100,000 g for 1 hour at 4°C. The cytoplasmic fraction was in the supernatant, and the membrane fraction was in the pellet. Next, the membrane pellet was resuspended in SB buffer (20 mM HEPES pH 7.4, 1 M KI, 5 mM EDTA, 5 mM EGTA, 4°C) and centrifuged at 100,000 g for 1 hour at 4°C to remove weakly bound membrane proteins. Then, the membrane fraction was resuspended in RB buffer (20 mM HEPES pH 7.4, 100 mM NaCl, 5 mM EDTA, 5 mM EGTA) containing Triton (1%) and a protease inhibitor. After sonication in an ice bath, the membrane fraction was gently stirred at 4°C for 2 hours. The solubilized membrane fraction was centrifuged at 100,000 g for 1 hour at 4°C. The membrane fraction was found in the supernatant.

[0048] Western blotting: L4-L6 DRGs from three mice were pooled and honogenized in lysis buffer containing Triton (1%) and a protease inhibitor. The homogenates were centrifuged at 18,000 g for 20 minutes at 4°C, and the supernatant was collected. 50 μg of total protein was separated by SDS-PAGE on a 7.5% gel. Primary antibodies were transferred to a nitrocellulose membrane, saturated with BSA (5%), and then used at dilutions of 1 / 500 for HCN1 and HCN2 (Neuromab), 1 / 1000 for TRIP8b (Neuromab), and 1 / 5000 for actin (Sigma-Aldrich). After washing three times consecutively with TBST, the corresponding HRP-conjugated secondary antibody (ThermoFisher) was incubated at a 1:10,000 dilution and detected with chemiluminescent substrate (SuperSignal West Pico Chemiluminescent Substrate, ThermoFisher).

[0049] Patch-clamp recording: Electrophysiological experiments were performed on isolated pacemaker cardiac cells as previously described (Marger et al., 2011)

[28] . If currents generated by HCN channels were recorded using an Axopatch 200A amplifier (Axon Instruments Inc., Foster USA). Recording electrodes with a resistance of approximately 5 mMΩ were filled with an intracellular solution containing mM / L: K+-aspartate, 130 mM / L; NaCl, 10 mM / L; ATP-Na+ salt, 2 mM / L; creatine phosphate, 6.6 mM / L; GTP-Mg2+, 0.1 mM / L; CaCl2, 0.04 mM / L (PCa=7); Hepes-KOH, 10 mM / L; (pH=7.2 with KOH). IK1 current Tyrode extracellular solution containing 5 mM BaCl2 was used to block the current. The effects of NUCC5953 (5 μM) on if current amplitude and dynamics were compared with those of ivabradine (3 μM), both diluted in extracellular solution. Data acquisition was performed using pClamp software (ver. 9, Axon Instruments Inc.).

[0050] in vitro assay Fluorescence polarization assay: Using a fluorescence polarization (FP)-based assay, we identified compounds that can disrupt the interaction between HCN and the TRIP8b tail, as published (Han et al., 2015)

[13] .

[0051] Example 2: Results For the first time, the presence of TRIP8b mRNA (RNA-seq data obtained from DRG mice showed the presence of splice variants 2-5: NM_01163516, NM_01163517, NM_01289505, NM_01310460 (FPKM>1)) and the protein were observed in DRG and TG neurons of mice. In addition, the expression of HCN1-2 channels and TRIP8b transcripts and proteins was significantly increased in peripheral neurons of oxaliplatin-treated animals compared to controls, suggesting the involvement of these two proteins in the development of OIPN (oxaliplatin-induced neurological disorder model mouse) (Figure 1 A-C). Analysis of TRIP8b protein expression in the spinal cord of animals treated with a single dose of oxaliplatin versus vehicle (Figure 1D) or after repeated doses (Figure 1E) shows a clear increase in TRIP8b protein under the action of oxaliplatin.

[0052] Using NUCC5953 (Han et al., 2015)

[13] , a molecule capable of disrupting the TRIP8b-HCN interaction, it was shown that oxaliplatin-induced acute cold hypersensitivity can be dose-dependently reversed either intrathecal (0.2); 15 μg / mouse; Figure 2) or systemically (4 mg / kg; Figure 3).

[0053] When mice were treated with subcutaneous administration of NUCC5953 (4 mg / kg), no changes in walking activity were observed in the rotarod test (Figure 4).

[0054] Simultaneously, the preparatory data also suggest that administration of NUC5953 reduces the overexpression of HCN1 and 2, as well as TRIP8b proteins, in the membrane fraction of DRG neurons derived from OIPN animals (Figure 5).

[0055] Patch-clamp experiments on isolated cardiac pacemaker cells showed no effect of NUCC5953 on If current amplitude, in contrast to the pan-HCN blocker ivabradine (Figure 6).

[0056] When administered at an analgesic dose (4 mg / kg, intraperitoneal), NUCC5953 is detected in heart rate. It has no effect in vivo (Figure 7). The results show that the mean heart rate did not change in three mice before (A), at the time of (B), and 30 minutes after (C) injection of NUCC5953.

[0057] Example 3: Design of a peptide mimetic Design of peptide mimetic molecules Tetrameric peptide mimetic We designed and synthesized peptide mimetic compounds that mimic the carboxyl-terminal sequence of the HCN2 channel. The design of these peptide mimetic compounds was based on extensive X-ray imaging of cocrystals of TRIP8b with the C-terminal region of HCN2, available in the protein database (pdb:4 EQF). The model was based on structural testing. Preparatory modeling work was performed to complete and improve this structure. The missing region was modeled by sequence homology from different PEX5 structures (PDB: 1FCH, 3R9A, 4KXK, 4KYO, 2J9Q, 2C0M, and 2C0L) that exhibit 30–60% sequence identity with TRIP8b. The resulting model identified the contacts between residues involved in the formation of the HCN-TRIP8b complex. In particular, this study identified two hydrophobic pockets in the TPR domain of TRIP8b, which are essential for good ligand affinity to this protein domain (Figure 8).

[0058] The hydrophobic pocket 1 of TRIP8b provides a side chain that anchors the C-terminal residue of the ligand. Effective interaction between the ligand and the hydrophobic pocket 2, located near the central region of TRIP8b and identified as the binding site for the CNBD domain of HCN, also enhances inhibitor affinity. Several four-residue peptide-peptoid hybrid oligomers with various side chains capable of targeting these hydrophobic pockets were designed and evaluated by molecular dynamics (MD).

[0059] Structural design identified peptide-mimicking ligands that interact with the SNL peptide binding site (TPR domain) of TRIP8b. These ligands are represented by the formula of LF108 above or the following general formula (I): [ka] [In the formula, R 1 , R 2 , R 3 , R 4 and R 5 It has an expression represented by [as defined above].

[0060] This compound is a peptide-peptoid hybrid oligomer (peptomer) (Ostergaard et al., 1997)

[17] . These consist of four residues containing an α-amino acid located at the C-terminus of the sequence, and three consecutive peptoid residues. The third residue (from the N-terminus) is either a β-peptoid unit or an α-peptoid unit. Generally referred to as "peptoids," α-peptoids refer to N-substituted glycine oligomers (Simon et al., 1992)

[18] . Compared to peptides, the side chain is bonded to the nitrogen atom of the amide bond rather than to the α-carbon atom. β-peptoids are oligomers composed of N-substituted β-aminopropionic acid units (Hinder et al., 1998)

[19] . Therefore, β-peptoid residues contain an additional methylene group in the backbone compared to α-peptoid residues. Peptoids have characteristics that are advantageous for the development of bioactive compounds. These are suitable for solid-phase synthesis using a stepwise “submonomer” approach in which each novel unit is constructed in two steps (Zuckermann et al., 1992)

[20] . Peptoids can incorporate a wide variety of side chains, both protein-constitutive and unnatural side chains (Culf et al., 2010)

[21] . In “submonomer” synthesis, simple starting molecules are used, for example, bromoacetic acid and a primary amine for the construction of α-units, and acryloyl chloride and a primary amine for the construction of β-peptoid units. Hundreds of primary amines into which side chains are incorporated are commercially available. They are prepared or, where appropriate, can be prepared before use. Peptoid skeletons based on N,N-disubstituted amides (tertiary amides) have been shown to be resistant to proteases (Miller et al., 1995)

[22] . The presence of a tertiary amide bond also gives them better cell permeability (Kwon et al., 2007; Vollrath et al., 2013)[23, 24], and some studies have shown that peptoids tend to be less immunogenic than peptides (Li et al., 2010; Chongsiriwatana et al., 2008)[25, 26].

[0061] The synthesis was carried out on a 2-chlorotrityl chloride (2-CTC) resin, and a carboxylic acid was added to the C-terminus of the peptomer after cleavage. The first residue immobilized on the support, the α-amino acid, was mainly selected from the group of hydrophobic protein constituent amino acids, including L-leucine, which corresponds to the C-terminus of the SNL peptide. The α-amino acid was immobilized on the resin in a protected N-Fmoc form. After removal of the Fmoc group, chain elongation by the peptoid residue was achieved by a "submonomer chemistry" approach. Although known protocols were followed, several technical adjustments were necessary for the synthesis of the β-peptoid monomer. After acylation of the N-terminus, cleavage was performed using hexafluoroisopropanol (HFIP) under acidic conditions, and the side chain was deprotected using trifluoroacetic acid (TFA) in dichloromethane.

[0062] Crude peptomer compounds were analyzed by LC-MS and then purified using a C18 flash chromatography cartridge with UV detection at 214 nm and 220 nm. The pure fraction was collected and lyophilized. The pure compounds were analyzed by reverse-phase HPLC on a Nucleodur® C4 column (5 μm, 300 Å, 250 mm × 4.6 mm). Table 2 below shows the HRMS and HPLC data for the compounds. [Table 1] a LC-MS. UHPLC Ultimate 3000 RSLC chain (ThermoScientific), Kinetex EVO C18 column (100×2.1mm, 1.7μm) (Phenomenex), flow rate: 0.45mL / min, column oven: 30℃, H2O gradient (0.1% formic acid / ACN (0.1% formic acid): t=0 min 95:5; t=7.5 min 1:99, t=8.5 min 1:99; t=9 min 95:5; t=11 min 95:5). bHPLC. Series 1100 Agilent chain, Nucleodur® C4 column (5 μm, 300 Å, 250 mm x 4.6 mm), flow rate 0.5 mL / min. H2O / ACN gradient (0.1% TFA): t=0 min 95:5; t=5 min 95:5; t=25 min 5:95; t=35 min 5:95, t=40 min 95:5, t=50 min 95:5. c UV 214nm detection.

[0063] side chain R 1 These are independently CH3(L-Ala), CH(CH3)2(L-Val), CH(CH3)CH2CH3(L-Ile), CH2CH(CH3)2(L-Leu and D-Leu), CH2C(CH3)3, C(CH3)3, and CH2(cyclobutyl). R 2 , R 3 , and R 4 These are side chains of peptoid units that are sequentially incorporated during the synthesis from primary amines. R 2 and R 3 These are independently selected from CH2CH(CH3)2, CH2CH2NH2, CH2CH2OH, CH2CH2CH2OH, CH2CH(OH)CH3, CH2CONH2, and CH2CH2CONH2. The side chains having alcohol or amine functional groups are incorporated into the oligomer in a protected form, i.e., silyl ether for alcohol functional groups and tert-butylcarbamate (Boc) for amine functional groups. R 4 The following: [ka] It is an aliphatic or aromatic group selected from the following. R 5 These are independently CH3, CH(CH3)2, CH2C6H5, C6H4X (wherein X at the ortho, meta, or para positions is independently H, OMe, CF3, CH3, CH2CH3, F, Br, Cl, NO2), 3-indolyl, and 3-quinolinyl.

[0064] A typical synthetic scheme on 2-chlorotrityl chloride resin for the synthesis of N-acylated peptomers incorporating an α-amino acid at the C-terminus, followed by β- and two α-peptoid residues. [ka] DIEA: Diisopropylethylamine; Fmoc: Fluorenylmethyloxycarbonyl; DMF: N,N-Dimethylformamide; DIC: Diisopropylcarbodiimide; HFIP: 1,1,1,3,3,3-Hexafluoroisopropanol; ta: Room temperature; TFA: Trifluoroacetic acid

[0065] Synthesized molecules [ka] [ka]

[0066] synthesis Compounds corresponding to general formula (I) were synthesized on 2-chlorotrityl chloride resin (Novabiochem 100-200 mesh) in a 5 mL plastic syringe equipped with a sinter and stopcock. During the reaction, the syringe was heated in an orbital shaker. It was placed on the form.

[0067] In a typical synthesis described for the C-terminal L-leucine and the β-peptoid residue preceding this amino acid, 2-chlorotrityl chloride resin (1.33 mmol 1, 0.100 g, 0.133 mmol) was swollen in 2 mL of dry CH2Cl2 (2 × 20 min). After filtration, 0.12 mL (2.0 equivalents) of a 2.2 M CH2Cl2 solution of Fmoc-Leu-OH was added, followed by 2.0 mL (6.0 equivalents) of a 0.4 M CH2Cl2 solution of DIEA. The mixture was stirred at 25°C for 40 minutes. The resin was drained and washed with CH2Cl2 (2 × 2 mL), and the bonding of Fmoc-leucine to the resin was repeated under the same conditions. The resin was washed with CH2Cl2 (5 × 2 mL). The Fmoc group was removed by adding 2.0 mL of a 20% piperidine DMF solution and stirring at room temperature for 15 minutes. The procedure was repeated under the same conditions, and then the resin was washed with DMF (5 × 2 mL) and drained.

[0068] Introduction of β-peptoid residues. The resin was cooled by washing twice with -CH2Cl2 at -20°C. 1.21 mL (2.0 equivalents) of a 0.22 M CH2Cl2 solution of acryloyl chloride cooled to -20°C was added, followed by 0.8 mL of a 0.5 M CH2Cl2 solution of Et3N (3.0 equivalents) cooled to -20°C. The resin was stirred at 25°C for 1 hour, discharged, and then treated again with acryloyl chloride under the same conditions. The resin was filtered, washed with CH2Cl2 (5 × 2 mL), and discharged. The resulting acrylamide was treated with amine R 2 The NH2 was treated with a 2.0 M isopropanol solution (12.0 equivalents) at 50°C for 24 hours. After filtering the resin, the Aza-Michael reaction was repeated under the same conditions. After washing the resin with isopropanol (5 × 2 mL), the two α-peptoid residues were synthesized as submonomers.

[0069] Synthesis of α-peptoid residues. 2.0 mL (6.0 equivalents) of a 0.4 M DMF solution of bromoacetamide was added to the resin, followed by 2.13 mL (8.0 equivalents) of a 0.5 M DMF solution of CID. After stirring at 25°C for 5 minutes, the resin was drained and washed with DMF (5 × 2 mL). The resulting bromoacetamide was then treated with amine R 3The mixture was treated with a 2.0 M DMF solution of NH2 (20.0 equivalents) and left on a stirring platform at 40°C for 1 hour. The resin was filtered and washed with DMF (5 × 2 mL). The last two bromoacetylation and substitution steps were repeated under the same conditions to obtain the desired R 4 A fourth residue with a side chain was constructed. The N-terminus of the peptomer was then acylated according to one of the general procedures described below, and subsequently removed from the resin.

[0070] For cutting, the resin was treated with 1.5 mL of HFIP 2.4 M CH2Cl2 solution. After stirring at 25°C for 40 minutes, the resin was filtered and washed with CH2Cl2 (5 × 2 mL). The solvent was removed under reduced pressure. Finally, the peptomer was treated with 25% (v / v) TFA in CH2Cl2. The silylation protecting group or Boc protecting group of the side chain was removed by treating with 2 mL at room temperature for 10 minutes. The reaction mixture containing the final product was diluted with CH2Cl2 (5 mL) and then completely evaporated under vacuum. To remove all trace amounts of TFA, the product dissolved in CH2Cl2 was evaporated under vacuum, and this procedure was repeated five times.

[0071] General procedure for N-terminus acylation of resin-bound peptomers. Acetylation: 1.56 mL (20.0 equivalents) of a 1.7 M DMF solution of acetic anhydride was added to the resin, followed by 0.8 mL (12.0 equivalents) of a 2.0 M DMF solution of DIEA. After stirring at 25°C for 90 minutes, the resin was filtered and washed with DMF (5 × 2 mL).

[0072] Benzoylation: 0.8 mL (6.0 equivalents) of a 1.0 M CH2Cl2 solution of benzoyl chloride was added to the resin, followed by 0.8 mL (12.0 equivalents) of a 2.0 M CH2Cl2 solution of DIEA. After stirring at 25°C for 40 minutes, the resin was drained and CH2Cl2 (5 × 2 mL) was added. It was washed.

[0073] Acylation from carboxylic acids: In a typical coupling reaction described herein from phenylacetic acid, 2.0 mL (6.0 equivalents) of a 0.4 M DMF solution of phenylacetic acid and 2.13 mL (8.0 equivalents) of a 0.5 M DMF solution of DIC were successively added to the resin. After stirring at 25°C for 1 hour, the resin was discharged and washed with DMF (5 × 2 mL).

[0074] Of the synthesized compounds (YC55, YC-60), YC55 showed the same potency as NUCC5953 in disrupting the TRIP8b-HCN interaction (Table 1: IC50 of peptoids). [Table 2]

[0075] The analgesic effect of novel peptide mimetic compounds, administered either intrathecally or systemically, was demonstrated in mice with oxaliplatin-induced acute neuropathic pain.

[0076] Thus, when the lead compound YC55 was administered intrathecally, it showed a dose-dependent analgesic effect in OIPN mice (Figure 9).

[0077] Compound YC55 also showed dose-dependent analgesic effects in OIPN mice when administered subcutaneously (Figure 10).

[0078] When mice were treated with YC55 subcutaneously, no changes in walking activity were observed in the rotarod test (Figure 11).

[0079] YC55 also showed a dose-dependent analgesic effect when administered intrathecally in a rat model of paclitaxel-induced neuropathy (Figure 12).

[0080] Intrathecal administration of YC55 demonstrated an anti-hyperalgesic effect in a model of neuropathic injury induced by traumatic sciatic nerve injury (Figure 13), suggesting that strategies disrupting TRIP8b interactions may have analgesic effects in several neuropathic pain conditions.

[0081] Compounds MP208, MP405 (5 μg, intrathecal) and MP341, MP354, and MP376 (10 mg / kg, subcutaneous) showed analgesic effects in a mouse oxaliplatin-induced neuropathy model (Figures 14 and 15, respectively).

[0082] Similarly, compounds LF108, LF306, LF188 (Figure 16), LF400, LF329, and LF295 (Figure 17), LF275, LF261, and LF126 (Figure 18), LF239, LF222, and LF176 (Figure 19) showed analgesic effects (5 μg, intrathecal) in a mouse model of oxaliplatin-induced acute neurological injury.

[0083] β-trimer peptomime Trimeric peptomer compounds are also mimics of the carboxyl-terminal sequence of the HCN2 channel. They were designed based on extensive X-ray structural analysis of cocrystals of TRIP8b and the C-terminal region (bp:4EQF) of HCN2.

[0084] These ligands are represented by the following formula LF369 or the following general formula (II): [ka] [In the formula, R 1 , R 2 and R 3 As defined above, R 6 H, R 3 COR 5 (R in the formula 5 (As defined above), or R 7 And, R 7 It is, independently, one of the following: [ka] It has an expression represented by "[is]".

[0085] This compound is a peptide-peptoid hybrid oligomer (peptomer) (Ostergaard et al., 1997)

[17] . These consist of three units, each containing an α-amino acid located at the C-terminus of the sequence, followed by two peptoid residues.

[0086] The synthesis was carried out in solution from α-amino acids protected as tert-butyl esters (Xaa-tBu, L-Leu-tBu in the case of LF369 synthesis). These synthesis can also be carried out on a resin, as previously described for the tetrameric peptide mimetic. From this unit, extensions corresponding to the molecule of general formula (I) were performed using a "submonomer chemistry" approach. When the core unit is of the β-peptoid type, the two steps required for its construction are the reaction of Xaa-tBu with acryloyl chloride, followed by the reaction of the desired side chain (R 2 This is the Aza-Michael reaction for introducing ) into the skeleton. The third α-peptoid unit is constructed by acyling the dimer with bromoacetic acid bromide, followed by substituting the bromine atom with an amine. 6 If is H, the amine used is a primary amine R 3 It is NH2. 6 COR 5 If R 3 The secondary amine obtained after treatment with NH2 is acylated with an acid chloride, acid anhydride, or carboxylic acid under coupling conditions. 6 =R 7 The compound is given an N-terminal bromoacetylated dimer, which is a secondary amine R 3 R 7 Obtained by processing with NH. Then, NHR is applied to the N-terminus. 3 It can also be obtained by reductive amination of compounds containing [the specified compound]. The final step is to deprotect the side chain and C-terminus by treatment with TFA.

[0087] The precise experimental protocol for the synthesis of LF369 is as follows: In a flask, dissolve L-leu-tBu (1.74 g, 7.8 mmol) in 45 mL of anhydrous THF. Cool the flask to 0°C under an inert argon atmosphere. Add triethylamine (2.3 mL, 31.4 mmol, 4 equivalents), followed by acryloyl chloride (0.76 mL, 9.4 mmol, 1.2 equivalents). After stirring at 0°C for 3 hours, complete conversion was observed by thin-layer chromatography (50 / 50 cyclohexane / AcOEt). The formed salt is removed by filtration, the filtrate is evaporated under vacuum, and the resulting product is purified by silica gel chromatography (50 / 50 cyclohexane / AcOEt). Acrylamide is isolated as a white solid in 93% yield (1.76 g, 7.27 mmol).

[0088] Aza-Michael reaction. The previously formed acrylamide (1.76 g, 7.27 mmol) is dissolved in 20 mL of ethanol. Ethanolamine (2.68 g, 15.28 mmol, 2.1 equivalents) having an alcohol functional group protected as tert-butyldimethylsilyl ether is added, and the reaction mixture is heated under reflux for 60 hours. The solvent is then evaporated, and the reaction crude is purified by silica gel chromatography (90 / 10 / 0.1 AcOEt / MeOH / Et3N) to obtain the dimer as a yellowish oil in 90% yield (2.74 g, 6.58 mmol).

[0089] Dimer acylation reaction. The dimer (2.74 g, 6.58 mmol) is dissolved in 10 mL of THF, the reactor is inactivated with argon, and then cooled to -20°C. Triethylamine (1.1 mL, 7.89 mmol, 1.2 equivalents) is added to the solution, followed by 2-bromoacetyl bromide (0.62 mL, 7.89 mmol, 1.2 equivalents). After stirring at -20°C for 3 hours, the reaction mixture is filtered. The resulting filtrate is evaporated, and the crude reaction product is chromatographically treated with silica gel (cyclohexane / AcOEt / Et3N in 60 / 40 / 0.1). The bromoacetylated dimer is isolated as a yellowish oil in 67% yield (2.36 g, 4.4 mmol).

[0090] Substitution reaction. The product obtained previously (2.36 g, 4.4 mmol) is dissolved in 10 mL of THF. Triethylamine (1.2 mL, 8.8 mmol, 2 equivalents), followed by protected ethanolamine H2NCH2CH2OTBDMS (2.31 g, 13.2 mmol, 3 equivalents), is added to the solution. The reaction medium under an argon atmosphere is stirred at room temperature overnight. Then, the reaction mixture is filtered and the resulting filtrate is evaporated. The crude product is purified by silica gel chromatography (95 / 5 / 0.1 AcOEt / MeOH / Et3N) to obtain the trimer as an oil in a yield of 83% (2.30 g, 3.64 mmol).

[0091] Acetylation of the trimer. The amine obtained (2.3 g, 3.64 mmol) is dissolved in 20 mL of ethyl acetate. Triethylamine (2.02 mL, 14.5 mmol, 4 equivalents), followed by acetic anhydride (2.75 mL, 29 mmol, 8 equivalents), is added to the solution. The reaction is placed under argon at room temperature for 60 hours. After the solvent is evaporated, the resulting crude product is chromatographed on silica gel (90 / 10 / 0.1 AcOEt / cyclohexane / Et3N) to obtain the N-acetylated trimer as a colorless oil in a yield of 92% (2.27 g, 3.36 mmol).

[0092] Deprotection of the side chain and C-terminus. The trimer compound (502 mg, 0.776 mmol, 1 equivalent) is dissolved in 4 mL of a TFA / CH2Cl2 / H2O (47.5:47.5:5) mixture. After stirring at room temperature for 2 hours, 10 mL of dichloromethane is added. The reaction medium is evaporated under vacuum and then subjected to 5 co-evaporations with dichloromethane. Then, the product is redissolved in 5 mL of distilled water and left under stirring at room temperature for 2 hours. The solvent is evaporated and then subjected to 5 co-evaporations with toluene to obtain LF369 (362 mg) as an oil.

[0093] Side chain R 1 、R 2 and R 3 are as defined previously, R 6 is R 3 、H、COR 5 (where R5 is as previously defined), or R 7 is R 7 is independently one of the following:

Chem.

[0094] Typical synthetic scheme for solution synthesis of peptoids of general formula (II), such as LF369.

Chem.

[0095] Typical synthetic scheme for solid-phase synthesis of peptoids of general formula (II), such as LF369.

Chem.

[0096] Synthesized molecule

Chem.

[0097] Synthesis Compound LF369 (5 μg, intrathecal) showed analgesic effect in a mouse model of oxaliplatin-induced neuropathy (Figure 20).

[0098] List of references 1. Finnerup et al., Lancet Neurol., 14(2):162 - 173, 2015 2. von Hehn et al., Neuron, 73(4):638 - 652 , 2012 3. Hershman et al., J. Clin. Oncol., 3(18):1941 - 1967, 2014 4. Ludwig et al.,Nature,393(6685):587-591,1998 5. Santoro et al.,Cell,93(5):717-729,1998 6. Seifert et al.,Proc.Natl.Acad.Sci.USA,96(16):9391-9396,1999 7. Dunlop et al.,Curr.Pharm.Des.,15(15):1767-1772,2009 8. Lewis et al.,Mol.Cell Neurosci.,46(2):357-367,2011 9. Descoeur et al.,EMBO Mol.Med.,3(5):266-278,2011 10. Young et al.,Pain,155(9):1708-1719,2014 11. Lewis et al.,J.Neurosci.,29(19):6250-6265,2009 12. Bankston et al.,Proc.Natl.Acad.Sci.USA,109:7899,2012 13. Han et al.,J.Biomol.Screen,20(9):1-8,2015 14. Janssen et al.,Arzneimittelforschung,13:502-507,1963 15. Randall et Selitto,Arch.Int.Pharmacodyn.Ther.,111(4):409-419,1957 16. Kim et Chung,Pain,50(3):355-363,1992 17. Ostergaard et al.,Molecular Diversity,3:17-27,1997( 18. Simon et al.,Proc.Natl.Acad.Sci USA,89:9367-9371,1992 19. Hamper et al., J.Org.Chem.,63:708-718,1998 20. Zuckermann et al., J.Am.Chem.Soc.,114:10646-10647,1992 21. Culf et al.,Molecules,15:5282-5335,2010 22. Miller et al., Drug Dev. Res., 35:20-32, 1995 23. Kwon et al., J.Am.Chem.Soc.,129:1508-1509,2007 24. Vollrath et al.,Organic&Biomolecular Chemistry,11:8497-8201,2013 25. Li et al.,Cellular&Molecular Immunol.,7:133-142,2010 26. Chongsiriwatana et al.,Proc.Natl.Acad Sci.USA,105:2794-2799,2008 27. Mesirca et al., Nat. Commun., 5:4664.d oi:10.1038 / ncomms5664,2014 28. Marger et al., Channels (Austin), 5(3):241-250, 2011

Claims

1. The following general formula (I): 【Chemistry 1】 [In the formula, R １ is independently CH ３ (L-Ala), CH(CH ３ ) ２ (L-Val), CH(CH ３ )CH ２ CH ３ (L-Ile), CH ２ CH(CH ３ ) ２ (L-Leu and D-Leu), CH ２ C(CH ３ ) ３ , C(CH ３ ) ３ , CH ２ (cyclobutyl), and R ２ and R ３ CH is independent. ２ CH ２ NH ２ ,CH ２ CH ２ OH, CH ２ CH ２ CH ２ OH, CH ２ CH(OH)CH ３ ,CH ２ CONH ２ ,CH ２ CH ２ CONH ２ Selected from, R ４ The following: 【Chemistry 2】 an aliphatic or aromatic group selected from, R ５ CH is independent. ３ , CH (CH ３ ) ２ ,CH ２ C ６ H ５ , C ６ H ４ X (wherein the formula, the ortho, meta, or para X are independently H, OMe, CF) ３ ,CH ３ , F, NO ２ Peptoid (is) is]

2. The following formula: 【Transformation 3】 【Chemistry 4】 The peptoid according to claim 1, wherein the peptoid is as described above.

3. The following general formula (II): 【Transformation 5】 [In the formula, R1 is as defined in claim 1, R2 and R3 are CH2CH2OH, R ６ It is a peptoid of COCH-3.

4. A pharmaceutical composition comprising a peptoid according to any one of claims 1 to 3 and a pharmaceutically acceptable carrier.

5. A pharmaceutical composition for use as a pharmaceutical, comprising a peptoid according to any one of claims 1 to 3, or the pharmaceutical composition according to claim 4.

6. A pharmaceutical composition comprising a peptoid according to any one of claims 1 to 3, or the pharmaceutical composition according to claim 4, for use in the prevention or treatment of chronic pain.

7. The pharmaceutical composition according to claim 6, wherein the chronic pain is of neurological origin.

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