Synthesis of novel tetralone derivatives as GPER pathway modulators

Abstract

The invention relates to novel tetralone derivatives as GPER pathway modulators, and to their use in particular for nociception control.

Description

[0001] SYNTHESIS OF NEW TETRALONE DERIVATIVES AS MODULATORS OF THE GPER PATHWAY

[0002] DESCRIPTION

[0003] Technical field of the invention

[0004] The present invention relates to novel tetralone derivatives as modulators of the GPER (G protein-coupled estrogen receptor) pathway.

[0005] The present invention finds particular application in the control of nociception, especially in the prevention or treatment of cancer-related pain (cancer, metastatic, chemotherapy-induced, and inflammatory pain), particularly in the context of triple-negative breast cancer (TNBC). The novel derivatives also find application as anticancer agents, particularly in the context of triple-negative breast cancer (TNBC), but also in the prevention of chemotherapy-induced peripheral neuropathies as well as other neuropathies (diabetes) and neurodegenerative diseases.

[0006] In the description below, references in brackets ([ ]) refer to the list of references presented at the end of the text.

[0007] Prior art

[0008] Since 2020, pain has been defined by the IASP (International Association for the Study of Pain) as "an unpleasant sensory and emotional experience associated with, or resembling that associated with, actual or potential tissue damage" (Raja et al., 2020) [1]. In terms of time frame, pain can evolve in two ways, namely: 1) acute, constituting an alarm signal for the body and 2) chronic (lasting more than at least three months), thus indicating a true pathology whose prevalence, in terms of the general population, is far from trivial (International Classification of Diseases ICD-11, 2019).

[0009] Currently, according to the IASP terminology, three types of chronic pain are identified based on the underlying neurobiological mechanisms that cause them: 1) nociceptive pain (corresponding to local tissue damage, mechanical or inflammatory: inflammatory pain, osteoarthritis, etc.), 2) neuropathic pain (related to a lesion of the central or peripheral nervous system), and 3) nociplastic pain, also called psychogenic pain (due to a disruption of pain modulation at the cerebral and / or peripheral level, and until 2017 referred to as dysfunctional pain, such as fibromyalgia, irritable bowel syndrome, and complex regional pain syndrome type 1). Cancer pain is a special case because it frequently combines nociceptive and neuropathic pain. The prevalence of chronic pain is significant, estimated at 31.7% in France.These pains represent a significant factor in impairing quality of life and can be associated with comorbidities such as depression, anxiety, insomnia, etc. They result in a substantial economic and social burden. Individuals with chronic pain are generally more prone to absenteeism than presenteeism at work, and the medical expenses incurred for these patients are generally higher.

[0010] Many patients report that their pain is not under control. Indeed, the various treatments indicated for chronic pain conditions have limited effectiveness or significant side effects.

[0011] Given this situation, the management of chronic pain is not always optimal due to an aging pharmacopoeia and an often unsatisfactory benefit-risk ratio.

[0012] It is therefore important to have access to treatment solutions for this pain with an improved benefit-risk ratio, or alternative solutions for pain management. This major public health problem calls for significant innovation.

[0013] Since 2004, cancer has been the leading cause of premature death in France, surpassing cardiovascular diseases. It is the leading cause of death in men and the second leading cause in women. An estimated 3.8 million people in France are currently living with a cancer diagnosis. Globally, according to the latest estimates from the International Agency for Research on Cancer (IARC), 19.3 million new cases of cancer were diagnosed worldwide in 2020, with 9.9 million deaths, primarily from triple-negative breast cancer (TNBC).

[0014] Cancer treatment often relies on chemotherapy, which can cause chemotherapy-induced neuropathies. These neuropathies are particularly common, debilitating, and difficult to treat. Moreover, they are dose-dependent. Peripheral nerve damage accounts for the majority of neurological complications related to chemotherapy toxicity. It results from direct toxic damage to the axon or its demyelination and represents the most frequent limiting factor after hematological toxicity.

[0015] Thus, when chemotherapy-induced neuropathies appear, chemotherapy doses are reduced, or even treatment is stopped, which constitutes a real loss of opportunity for the patient.

[0016] This explains why neuropathies have been observed following treatment with alkaloids (vincristine, vinblastine, vinorelbine), affecting small fibers, as well as, for example, with platinum derivatives (oxaliplatin, cisplatin, carboplatin), topoisomerase inhibitors (VP16), proteasome inhibitors (bortezomib, carfilzomib), thalidomide derivatives such as lenalidomide, or taxanes such as Taxol or Taxotere, which primarily affect large fibers. Neuropathies also occur after treatment with immunotherapy, such as with anti-CD20, anti-CD30, or anti-CD38 agents.

[0017] In patients with cancer, in addition to these peripheral neuropathic pains, bone metastatic pain may also occur, at least in cases of advanced TNBC.

[0018] TNBC is an aggressive form of breast cancer that is heterogeneous and presents a variety of clinical signs and morphological and molecular characteristics (Won and Spruck, 2020) [2]. TNBCs represent approximately 15 to 20% of breast cancers (Yin et al., 2020; Almansour, 2022) [3,4]. Their immunohistochemical profile reveals an absence of expression of estradiol receptors (ER) and progesterone receptors (PR), and overexpression of HER2 (human epidermal growth factor receptor 2) receptors (Yin et al., 2020) [3]. TNBCs often occur in premenopausal women under 40 years of age (Yin et al., 2020) [3], The median survival rate for TNBC is approximately 10.2 months with current treatments and rises to 5 years in approximately 65% ​​of cases of local tumors and 11% in cases of metastatic tumors (Won and Spruck, 2020) [2]. The clinical profile of TNBC shows a particularly aggressive type of cancer with a high risk of local and distant recurrence, as half of patients with early-stage TNBC experience a recurrence within 3 to 5 years of diagnosis (Li et al., 2022) [5]. Approximately 45% of patients with TNBC have metastases (Okorafor et al., 2024) [6], particularly bone metastases. Breast cancer management relies on a multidisciplinary approach involving medical oncology, surgical oncology, radiation oncology, and drug therapy.The absence of hormone receptors or HER2 overexpression limits therapeutic options despite the recent development of personalized therapies such as PARP (Poly(ADP-ribose) polymerase) inhibitors and immunotherapy. Consequently, aggressive chemotherapies remain the standard pharmacological treatment for patients with TNBC. These include alkylating agents (e.g., cyclophosphamide), anthracyclines (e.g., doxorubicin, a topoisomerase blocker and DNA intercalating agents), antimetabolites (e.g., fluorouracil) and microtubule-stabilizing agents (e.g., taxane) such as paclitaxel (Almansour, 2022) [4]. Used as first-line treatment for various cancers (Alves et al., 2018) [7], paclitaxel causes chemotherapy-induced neuropathic pain (CINP) in 80% of patients with TNBC (Molassiotis et al., 2019) [8], which are managed with analgesics, antidepressants or anticonvulsants (Staff et al., 2017; Moisset et al., 2020) [9,10], However, there is no effective drug to prevent their development to which may be added bone metastatic pain (Zajqczkowska et al., 2019)

[0011] for which no treatment is available.

[0019] The G protein-coupled estrogen receptor (GPER) is a seven-transmembrane-domain protein identified as binding Ie17[3-estradiol and structurally distinct from classical estrogen receptors α and β (ORα and ORβ) (Wnuk et al., 2023)

[0012] . GPER regulates numerous physiological functions such as metabolism, cell proliferation, and pain perception, and is implicated in multiple pathologies such as cancer, autoimmune diseases, and visceral hypersensitivity (Prossnitz and Barton, 2023)

[0013] . Studies concerning GPER have notably highlighted its important role in pain processes (Deliu et al., 2012; An et al., 2014; Zielihska et al., 2017; Xu et al., 2021; Jiao et al., 2023). [14-18], tumor growth and metastasis formation (Wang et al., 2010; Lappano et al., 2014; Jacenik et al., 2016; Xu et al., 2019; Tirado-Garibay et al., 2024) [19-23], The Inventors highlighted that the membrane protein GPER could be used as an innovative therapeutic target in the control of chronic and cancer pain (Mallet, 2021)

[0028] . In the context of TNBC, modulation of GPER, which exhibits a higher level of expression than in other breast cancer subtypes (Steiman et al., 2013)

[0025] , particularly by an inverse agonist (e.g., peptide ERa17p, ROa residues 295-311; PLMI motif "Pro Leu Met Ile" (SEQ ID NO: 1), ROa residues 295-298), induces both anti-tumor (Lappano et al., 2019; Kampa et al., 2023) [26,27] and analgesic (Mallet et al., 2021; Jouffre et al., 2023) [28,29], and this at the same low doses. However, this peptide has several limitations: it is not bioavailable orally, it forms aggregates at high concentrations, and it has a short half-life.

[0020] Description of the invention

[0021] The present invention provides a solution to all or part of the problems of the prior art. The inventors have thus developed new tetralone derivatives targeting the GPER pathway. These new derivatives have been tested on an animal model (mouse) of chronic neuropathic pain induced by chronic paclitaxel injections and on tumor growth (cell assay). The anti-allodynic effects were evaluated by measuring the 50% paw withdrawal threshold and using an adaptation of Dixon's up-down method by applying von Frey filaments. These original tetralone derivatives targeting GPER have shown their usefulness as antitumor and analgesic agents, particularly in the treatment of triple-negative breast cancer (TNBC) and associated chronic pain (e.g., metastatic bone pain). The present invention therefore relates to a tetralone derivative with the following general formula (I):

[0022] Ri is H or CH3;

[0023] R2 is H, OH or OCH3;

[0024] R3is H or F, OCH3, CH3OU Br;

[0025] R4 and R5 are H, OCH3 or together form a 1,3-dioxolane;

[0026] Rest H or Br.

[0027] According to a particular embodiment of the present invention, said derivative has a formula (I) where R1, R2, R3 and R6 are as defined above and where R4 and R5 are H.

[0028] According to a particular embodiment of the present invention, said derivative has a formula (I) where Ri, R2, R3 and R6 are as defined above and where R4 and R5 together form a 1,3-dioxolane.

[0029] According to a particular embodiment of the present invention, said derivative has a formula (I) where Ri is H or CH3; R2 is H or OCH3; R3 is H or F; R4 and R5 are H or together form a 1,3-dioxolane; R6 is Br; whereas at least one among Ri, R2 and R3 is not H.

[0030] According to a particular embodiment of the present invention, said derivative has a formula (I) where Ri, R2, R3 and R6 are as defined above and where R4 and R5 are H.

[0031] According to a particular embodiment of the present invention, said derivative has a formula (I) where Ri, R2, R3 and R6 are as defined above and where R4 and R5 together form a 1,3-dioxolane.

[0032] According to a particular embodiment of the present invention, said derivative has one of the following formulas:

[0033]

[0034] According to a particular embodiment of the present invention, said derivative has one of the following formulas:

[0035] derived is E3, E4 or E5.

[0036] The present invention also relates to a pharmaceutical composition comprising at least one derivative according to the invention, and a pharmaceutically acceptable excipient.

[0037] According to a particular embodiment of the present invention, the pharmaceutical composition comprises at least one tetralone derivative of the following general formula (I): Ri is H or CH3;

[0038] R2 is H or OCH3;

[0039] R3est H or F;

[0040] R4 and R5 are H or together form a 1,3-dioxolane;

[0041] Rest H or Br.

[0042] According to a particular embodiment of the present invention, the pharmaceutical composition comprises at least one tetralone derivative where R1, R2, R3, R4, R5 are as defined above and where R6 is Br.

[0043] According to a particular embodiment of the present invention, the pharmaceutical composition comprises at least one derivative having a formula (I) where Ri is H or CH3; R2 is H or OCH3; R3 is H or F; R4 and R5 are H or together form a 1,3-dioxolane; R6 is Br; whereas at least one among Ri, R2 and R3 is not H.

[0044] According to a particular embodiment of the present invention, the pharmaceutical composition comprises at least one derivative having a formula (I) where Ri, R2, R3 and Re are as defined above and where R4 and R5 are H.

[0045] According to a particular embodiment of the present invention, the pharmaceutical composition comprises at least one derivative having a formula (I) where R1, R2, R3 and R6 are as defined above and where R4 and R5 together form a 1,3-dioxolane.

[0046] According to a particular embodiment of the present invention, the pharmaceutical composition comprises at least one derivative selected from:

[0047]

[0048] According to a particular embodiment of the present invention, the pharmaceutical composition further comprises at least one anti-tumor agent and / or one pain-relieving (analgesic) agent, different from the derivatives according to the invention.

[0049] According to a particular embodiment of the present invention, said at least one anti-tumor agent is paclitaxel, oxaliplatin or bortezomib and / or said at least one analgesic is a morphine-based pain reliever, for example an agent selected from codeine, tramadol, morphine, oxycodone, or a non-morphine-based pain reliever, in particular a non-steroidal anti-inflammatory agent (NSAID), a pain reliever (e.g. paracetamol) or a local anesthetic agent.

[0050] The present invention also relates to a derivative or composition according to the invention, for use as a medicinal product.

[0051] According to a particular embodiment of the present invention, the drug is an analgesic and / or an anti-tumor agent. In particular, the drug is intended for the control of nociception.

[0052] The present invention also relates to a derivative or composition according to the invention, for use in the prevention or treatment of pain, for example, acute or chronic pain, skin pain, muscle pain, joint pain, osteoarticular pain, visceral pain, tendon pain, postoperative pain, dental pain, cancer pain (e.g., metastatic bone pain), or post-traumatic pain. This may include pain associated with arthritis, osteoarthritis, inflammatory bowel disease, irritable bowel syndrome, migraine, headache, myalgia, tendinitis, back pain, or lower back pain. In particular, it relates to neuropathic or arthritic pain. More specifically, pain is associated with cancer or its treatments, particularly the treatment of triple-negative breast cancer (TNBC).

[0053] According to a particular embodiment of the present invention, the use according to the invention (i.e. as an analgesic drug; or for the prevention or treatment of pain) of a derivative or pharmaceutical composition involves at least one derivative represented by one of the following formulas:

[0054] the invention (i.e. as an analgesic drug; or for the prevention or treatment of pain) of a derivative or pharmaceutical composition implements at least one derivative selected from E2, E3, E4 and E5, preferably from E3, E4 and / or E5.

[0055] The present invention also relates to a derivative or composition according to the invention, for use in the prevention or treatment of cancer, in particular triple negative breast cancer (TNBC).

[0056] Brief description of the figures

[0057] Antiproliferative effect

[0058] Figure 1 represents the percentage of cell viability in the presence of the different compounds according to the invention (AC: E 1-E 3) and comparative (DK: EC 1-EC 6, EC 8, EC 9), at different concentrations (i.e. 0.1 pM, 1 pM, 5 pM or 10 pM) on human triple negative breast cancer cells (MDA-MB-231).

[0059] Figure 2 illustrates the inhibition of cell growth in 4T1-Luc2 triple-negative breast cancer (TNBC) cells by compound E2 and paclitaxel. Cell viability and Cl 50 Evaluated by measuring bioluminescence in 4T1-Luc2 cells treated with the indicated concentrations of paclitaxel (0.01 to 1000 nM) and E2 (0.5 to 100 pM) for 24 hours. Data are expressed as mean ± SEM (n = 6) in quadruplicate. *p < 0.05, **p < 0.01, ***p < 0.001, relative to the vehicle dose by one-way ANOVA followed by Dunnett's test for the dose-response of E2 and Kruskal-Wallis followed by Dunn's test for paclitaxel.

[0060] Figure 3 illustrates the synergistic antiproliferative effect of compound E2 and paclitaxel on 4T1-Luc2 triple-negative breast cancer (TNBC) cells. (A) Dose-response curve of the E2-paclitaxel combination. (B) Dose reduction index (DRI) of the molecules. Favorable dose reduction occurs when DRI > 1. The dashed lines show the DRI of paclitaxel (8.6) and E2 (2.39) when 50% of viable cells are inhibited. (C) Combination index showing that the interaction between E2 and paclitaxel is synergistic because IndC < 1. Isobologram of IC25 (D) and Cl2 50 (E) of the combination. Data are expressed as mean ± CI95 (n = 6) in quadruplicate.

[0061] Figure 4 illustrates the apoptotic effect of the combination of compound E2 and paclitaxel in 4T1-Luc2 triple-negative breast cancer (TNBC) cells. (A) Representative flow cytometry data for double labeling with Annexin V / IP after 24 h of treatment with E2 (15.13 pM), paclitaxel (117.5 nM), or the combination in the murine 4T1-Luc2 TNBC cell line. (B) Frequency of apoptotic and (C) necrotic cells. Data are expressed as mean ± SEM (n = 4) in quadruplicate. **p < 0.01, compared to the control group by Kruskal-Wallis test followed by Dunn's test.

[0062] Analgesic effect (neuropathic pain)

[0063] Figure 5 shows the evaluation of the analgesic effect of compound E2 (administered intraperitoneally, ip) using the von Frey test in a murine model of paclitaxel-induced neuropathic pain. Top: effect kinetics. Bottom: area under the curve.

[0064] Figure 6 shows the evaluation of the analgesic effect of E2 (administered orally) using the von Frey test in a paclitaxel-induced mouse model of neuropathic pain. Left: effect kinetics. Right: area under the curve.

[0065] Figure 7 represents the evaluation of the GPER-dependent analgesic effect of compound E 2 (15 mg / kg, ip) by the von Frey test in a murine model of paclitaxel-induced neuropathic pain after administration of vehicle or G-15 (specific GPER antagonist ip).

[0066] Figure 8 illustrates the effect of GPER deletion specifically in primary afferent fibers on paclitaxel-induced neuropathic pain. Figure 9 shows the evaluation of the peripheral analgesic effect of compound E2 by the von Frey test in a murine model of paclitaxel-induced neuropathic pain. (A) E2 injected intraplantarly (IPL) into the animal's paw. (B) Vehicle or G-15 injection (IPL) 10 min prior to vehicle or E2 injection (IPL). (C) Vehicle or G-15 injection (IPL) 10 min prior to vehicle or E2 injected intraperitoneally (IPL). Left: effect kinetics. Right: area under the curve.

[0067] Figure 10 illustrates the effect of E2 and PLMI(ip) on a paclitaxel-induced neuropathic pain model following specific deletion of the GPER in primary afferent fibers. Top: effect kinetics. Bottom: area under the curve.

[0068] Figure 11 depicts the effect of GPER deletion specifically in the spinal cord on paclitaxel-induced neuropathic pain.

[0069] Figure 12 shows the evaluation of the spinal analgesic effect of compound E2 using the von Frey test in a paclitaxel-induced murine model of neuropathic pain. (A) E2 injected intrathecally (it). (B) Vehicle or G-15 injection (it) 10 min before vehicle or E2 injection (it). (C) Vehicle or G-15 injection (it) 10 min before vehicle or E2 injection via the intraperitoneal (ip) route. Left: effect kinetics. Right: area under the curve.

[0070] Figure 13 shows the kinetics of E2 analogs in female mice where pain was assessed before baseline pain induction, and then after receiving four intraperitoneal (IP) injections of paclitaxel (4 mg / kg) at least 10 days before the first kinetic study. All molecules were dissolved in dimethyl sulfoxide (DMSO) stock solution, then diluted in DMSO, Tween 80, and saline to a final solution of 1 mg / mL in 5% DMSO, 2% Tween, and saline. The administered dose was 10 mg / kg IP for each group and each molecule. The kinetics studies were performed in a randomized, blinded block design, with each mouse being reused four times (with a washout period of more than 48 hours).Each group consists of eight mice. The statistical tests used are ANOVA for repeated measures followed by Tukey's post-hoc test for kinetics, and the Kruskal-Wallis test (4k, 4h, and 4g did not pass the normality tests) followed by Dunn's post-hoc test. *vs vehicle.

[0071] Figure 14 represents the evaluation of the effect of repeated treatment with compound E 2 or PLMI (three PIs per day from D-1 to D 10) on the paclitaxel-induced neuropathic pain model.

[0072] Figure 15 illustrates the in vivo effect of combining compound E2 and paclitaxel on paclitaxel-induced neuropathic pain reduction and tumor growth. (A) Study overview. Nineteen days after tumor implantation, mice received compound E2 at 10 mg / kg three times daily, followed by paclitaxel injections of 4 mg / kg every other day starting on day 20. A von Frey test was performed before each paclitaxel injection and on day 30 at the end of the experiment. Tumor growth was measured with calipers three times weekly. (B) Monitoring tumor growth over time using calipers. (C) Tumor mass of mice collected on day 31 that received vehicle, paclitaxel 4 mg / kg, compound E 2 (10 mg / kg) or co-administration, (D) Evaluation of mechanical allodynia according to the different treatment groups over ten days. Data are expressed as mean ± SEM (n = 8 per group).*p < 0.05, **p < 0.01, ***p < 0.001 compared to the vehicle group and ##p < 0.01 compared to the paclitaxel 4 mg / kg group. Kruskal-Wallis tests followed by Dunn's test (tumor weight measurement), 2-way ANOVA followed by Dunnett's test (analysis of 50% of the withdrawal thresholds obtained by von Frey, tumor volume).

[0073] Figure 16 shows the evaluation of the analgesic effect of compound E2, administered intraperitoneally, using the von Frey test in a murine model of neuropathic pain induced by oxaliplatin (top, 3 mg / kg, twice weekly for four weeks) and bortezomib (bottom, 0.4 mg / kg, three times weekly for four weeks). Left: effect kinetics. Right: area under the curve.

[0074] Analgesic effect (arthritic pain)

[0075] Figure 17 illustrates the analgesic effect of compound E2 or PLMI in a murine model of osteoarthritic pain induced by an intra-articular injection of monoiodoacetate (MIA). Top: effect kinetics. Bottom: area under the curve.

[0076] Figure 18 depicts the effect of GPER deletion specifically in primary afferent fibers on pain in a murine model of osteoarthritic pain induced by destabilization of the medial meniscus.

[0077] Figure 19 shows the effect of compound E2(ip) after specific deletion of the GPER of primary afferent fibers in a murine model of osteoarthritic pain induced by destabilization of the medial meniscus. Top: effect kinetics. Bottom: area under the curve.

[0078] Figure 20 represents the effect of viral deletion of GPER specifically in the spinal cord on pain in a murine model of osteoarthritic pain induced by destabilization of the medial meniscus.

[0079] Figure 21 shows the effect of compound E2(ip) on a model of osteoarthritis pain after specific deletion of GPER in the spinal cord in a murine model of osteoarthritis pain induced by destabilization of the medial meniscus. Top: effect kinetics. Bottom: area under the curve.

[0080] EXAMPLES EXAMPLE 1: MATERIALS AND METHODS

[0081] The reactions leading to the expected molecules were carried out in an organic medium (with variable reaction times) using reflux setups. After returning to room temperature, the synthesized molecules were purified by silica gel chromatography, precipitation, crystallization, and / or recrystallization. The physicochemical characteristics of the expected molecules were determined by thin-layer chromatography (TLC, measurement of Rf values) and by measuring melting points on a Kofler hot stage (melting point, mp, °C). Their color and appearance (amorphous or crystalline powder, oil, turbidity) were recorded. Once purified, the molecules were characterized by high-resolution mass spectrometry (HRMS), and their structure (including their stereochemistry) was elucidated by Fourier transform infrared spectroscopy (FTIR), 1D and 2D nuclear magnetic resonance (NMR). 1 H, 13C-RMN, NOESY, COSY, HMBC, HSQC, DEPT135) and X-ray diffraction.

[0082] A- Chemistry

[0083] 1- Purification by silica gel column chromatography

[0084] The crude reaction products were purified, when necessary, by silica gel column chromatography (40-63 pm). The eluents were, as appropriate, composed of the solvent pairs: cyclohexane (CHX) / ethyl acetate (AE) or dichloromethane (CH2Cl2) / MeOH, in varying proportions.

[0085] 2- Thin-layer chromatography (TLC)

[0086] TLC was performed on Kieselgel 60 F254 silica gel aluminum plates (0.20 mm thick, Macherey-Nagel, Düren, Germany) and developed under a 254 nm UV lamp. Rf values ​​were calculated using the equation Rf = h / H, where h and H correspond respectively to the distance between the application line and the center of the spot of interest (h) and the distance between the application line and the solvent front. Ideally, 0.25 < Rf < 0.80.

[0087] 3- Melting points (T f )

[0088] Melting points were measured after calibration on a Kofler bench (REICHERT-JUNG, 220 V, measuring range: 50-260°C, accuracy: ±1°C). The values ​​obtained were not corrected.

[0089] 4- Fourier Transform Infrared (FTIR) Spectroscopy FTIR analyses were performed on a PERKIN ELMER Spectrum 65FT-IR Fourier transform instrument (measurement range: 650 - 4000 cm⁻¹) -1) equipped with Spectrometer 10.02 software. Wavenumbers (on the x-axis) are expressed in cm -1 and the transmittances (on the y-axis) as a percentage (%).

[0090] 5- Nuclear magnetic resonance (NMR) spectroscopy

[0091] 1D NMR spectra ( 1 H, 13 C, DEPT) and 2D (COSY, HSQC, HMBC, NOESY, ROESY) were recorded on a Bruker 300 MHz spectrometer equipped with a QNP direct probe ( 1 H, 13 C, 19 F, 31 P) and the Avance I console, or on a Bruker 400 MHz spectrometer equipped with a BBFO probe ( 1 H, 13 C, 15 N, 19 F, 31 P) and the Avance III console, or on a Bruker 600 MHz spectrometer equipped with a BBI inverse probe ( 1 H, 13 C, 15 N, 31P) and the Avance I console. Samples were diluted in 0.5 mL of the appropriate deuterated solvent (CDCh or DMSO-d6) and then introduced into a Norell® NMR tube (length: 7 in (17.78 cm); OD x DL: 5 mm x 4.2 mm; Landisville, USA). Chemical shifts (δ) are expressed in ppm (parts per million) relative to the residual solvent peak ( 1 H CDCl3: 7.26 ppm; 1 H DMSO-d6: 2.50 ppm; 13 C CDCl3: 77.16 ppm; 13 C DMSO-d6: 39.52 ppm). Multiplicities are expressed as follows: s (singlet), d (doublet), t (triplet), q (quadruplet), m (multiplet). Coupling constants (J) are expressed in Hertz (Hz). Spectral data were acquired and processed using Bruker TopSpin software (Billerica, USA).

[0092] 6- High-resolution mass spectrometry (HRMS)

[0093] The samples to be analyzed were resuspended in a suitable solvent at a concentration of approximately 1 mg / ml and then diluted five hundredfold (final concentration = 2 ng / pL). Analyses were performed on a Bruker maXis Q-TOF instrument coupled to a Dionex Ultimate 3000 RSLC system. The analysis of the compounds of interest was carried out by continuous flow injection analysis (FIA) without a column. The solvent used was a 65 / 35 acetonitrile (ACN) / water (H2O) mixture at a flow rate of 200 pL / min. The injection volume was 0.2 pL. Analyses were performed in positive or negative mode by electrospray ionization (ESI). Positive mode analyses were performed in the presence of 0.1% formic acid (HCOOH). In negative mode analyses, the samples were free of HCOOH.

[0094] 7- Synthesis of compounds E according to the invention and comparative compounds EC Table 1 to the comparative compounds, respectively.

[0095] In a 100 mL round-bottom flask, 1.2 mmol of substituted tetralone and 1 equivalence point of either simple benzaldehyde or 6-bromobenzo[d][1,3]dioxole-5-carbaldehyde were dissolved in 10 mL of ethanol. 10 mL of a 40% NaOH ethanolic solution were added dropwise and the mixture was stirred vigorously for 15 h at room temperature. The reaction mixture was then neutralized with 6N aqueous HCl. The resulting precipitate was filtered and rinsed with cold ethanol. The purification of molecules E 1-E 4 and EC 1-EC 9 was carried out either by recrystallization in ethanol (compounds E 1 and EC 1-EC 2, E 2, E 3 and EC 3- EC 6), or by silica gel chromatography (eluent CHX / AE: 80 / 20, compounds E 4 and EC 7-EC 9).

[0096] In a 100 mL single-necked round-bottom flask, 1.4 mmol of 6-methoxy-1-tetralone (0.244 g) and 1.1 eq of 2-bromobenzaldehyde (0.18 mL) were dissolved in 10 mL of ethanol. 10 mL of a 40% NaOH ethanolic solution were added dropwise to the reaction mixture and stirred vigorously at room temperature for 24 h. The flask was then placed in an ice bath, and the reaction mixture was neutralized with 6N aqueous HCl. The resulting precipitate was filtered under vacuum. The E5 molecule was purified by recrystallization in ethanol.

[0097] - (E)-2-benzylidenyl-6-methoxy-3,4-dihydronaphthalenyl-1(2H)-one (E 1). Physicochemical characteristics: Ci8Hi6O2 (MW = 264.32 g / mol); yield = 79%. Yellow crystalline powder. T° f = 97 °C; TLC: R f = 0.50 (CHX / AE: 60 / 40). Spectral characteristics: FTIR (cm 1 ): 3013, 2938, 2906, 2843, 1657, 1601, 1583, 1490, 1440, 1138. NMR 1H (400 MHz, CDCI3) δ: 2.91 (2H, m, H-4); 3.1 1 (2H, m, H-3); 3.87 (3H, s, OCH3); 6.71 (1 H, d, 4 J= 2.4 Hz, H-5); 6.88 (1 H, dd, 3 J=8.7 Hz, 4 J= 2.5 Hz, H-7); 7.30 - 7.46 (5H, m, H-Ar); 7.84 (1 H, s, H-9); 8, 12 (1 H, d, 3 J=8.7 Hz, H-8). NMR 13 C (400 MHz, CDCI3) δ: 27.2; 29.3; 55.4; 1 12.2; 113.3; 127.0; 128.3; 128.4 (2C); 129.8 (2C); 130.7; 135.6; 135.9; 136.0; 145.7; 163.5; 186.7. HR-MS (m / z): 265 [MH] + .

[0098] - (E)-2-[6'-bromobenzo[d][T,3 , ]dioxolo-5 , -methylenyl]-3,4-dihydronaphthalenyl-1(2H)-one (E 2). Physicochemical characteristics: Ci8Hi3O3Br (MW = 357.20 g / mol); yield = 65%. Yellow crystalline powder. Melting point = 162 °C; TLC: Rf = 0.68 (CHX / AE: 70 / 30). Spectral characteristics: FTIR n (cm 1 ): 3024, 2908, 1654, 1588, 1496, 1239. NMR 1H (300 MHz, CDCI3) δ (ppm) : 2,97 (4H, s, H-3 et H-4) ; 6,03 (2H, s, H-2’) ; 6,80 (1 H, s, H-4’) ; 7,12 (1 H, s, H-7’) ; 7,25 (1 H, d, 3 J=7,5 Hz, H-5) ; 7,37 (1 H, m, 3 J=7,5 Hz, 4 J=0,9 Hz, H-7) ; 7,50 (1 H, m, 3 J=7,5 Hz, 4 J=1 ,3 Hz, H-6) ; 7,78 (1 H, s, H-9) ; 8,15 (1 H, dd, 3 J=7,9 Hz, 4 J=1 ,3 Hz, H-8). RMN 13 C (400 MHz, CDCh) δ (ppm) : 26,3 ; 27,9 ; 101 ,0 ; 108,8 ; 112, 1 ; 1 12,2 ; 1 15,6 ; 126,0 ; 127,2 ; 128,1

[0099] ; 132,3 ; 132,4 ; 134,3 ; 135,0 ; 142,3 ; 142,9 ; 147,5 ; 186,5. HR-MS (m / z) : 357 [M] + .

[0100] - (E)-2-[6'-bromobenzo[cf][T,3']dioxolo-5'-methylenyl]-4-methyl-3,4-dihydronaphtalenyl- 1(2H)-one (E 3). Caractéristiques physicochimiques : C 19 H 15 O3Br (MW = 371 ,23 g / mol) ; rdt = 75 % ; poudre cristalline orange. T° f = 162°C ; CCM : R f= 0,67 (CHX / AE : 70 / 30). Caractéristiques spectrales : FTIR n (cm 1 ) : 2965, 2918, 2875, 1665, 1600, 1586, 1498, 1479, 1240. RMN 1 H (400 MHz, CDCI3) δ (ppm) : 2,61 (3H, d, 3 J=7,0 Hz, CH3) ; 2,81 (1 H, m, H-3a) ; 3,00 (1 H, m, H-3b) ; 3,12 (1 H, m, 3 J=5,6 Hz, H-4) ; 6,03 (2H, s, H-2’) ; 6,77 (1 H, s, H-4’) ; 7, 1 1 (1 H, s, H-7’) ; 7,29 (1 H, d, 3 J=7,66 Hz, H-5) ; 7,37 (1 H, m, 3 J=7,7 Hz, 4 J=1 ,0 Hz, H-7) ; 7,53 (1 H, m, 3 J=7,7 Hz, 4 J=1 ,4 Hz, H-6) ; 7,83 (1 H, s, H-9) ; 8,15 (1 H, dd, 3 J=7,9 Hz, 4 J=1 ,3 Hz, H- 8). RMN 13 C (400 MHz, CDCI3) δ (ppm) : 21 ,8 ; 33,3 ; 34,5 ; 102,0 ; 109,8 ; 113, 1 ; 116,5 ; 127,0 ; 127,1 ; 128,3 ; 129,2 ; 132,3 ; 133,6 ; 134,6 ; 137,1 ; 147,0 ; 148,3 ; 148,5 ; 187,5. HR- MS (m / z) : 371 [M] +- (E)-2-[6'-bromobenzo[d][1',3']dioxolo-5'-methylenyl]-7-fluoro-3,4-dihydronaphthalenyl-1(2H)-one (E 4). Physicochemical characteristics: Ci8Hi2BrFO3(MW = 375.19 g / mol); yield = 16%; yellow crystalline powder. T° f = 208 °C; TLC: R f = 0.53 (CHX / AE: 70 / 30). Spectral characteristics: FTIR n (cm 1 ): 2913, 2841, 1656, 1582, 1493, 1418, 1243. NMR 1 H (400 MHz, CDCh) δ (ppm): 2.86 - 3.00 (4H, m, H-3 and H-4); 6.02 (2H, s, H-2'); 6.79 (1H, s, H-4'); 7.11 (1 H, s, H-7'); 7.15 - 7.26 (2H, m, H-5 and H-6); 7.75 - 7.82 (2H, m, H-8 and H-9). NMR 13 C (400 MHz, CDCh) δ (ppm): 27.5; 28.2; 102.1; 109.8; 113.1; 114.0; 116.8; 120.5; 128.9; 130.1; 134.8; 135.2; 136.5; 139.0; 147.0; 148.8; 163.0; 186.6. HR-MS (m / z): 375 [M] + .

[0101] (E)-2-(2-bromobenzylidene)-6-methoxy-3,4-dihydronaphthalen-1(2 / 7)-one (E5). Physicochemical characteristics: C 18 H 15BrO2(MW = 342.22.22 g / mol) ; rdt = 79 % ; yellow amorphous powder. T° f = 98 °C ; CCM : R f = 0.68 (CHX / AE : 70 / 30). Spectral characteristics: FTIR (cm 1 ) : 2936 (CH) ; 2886 (CH) ;2836 (CH) ; 1658 (C=O) ; 1607 (C=C) ; 1592 (C=C) ; 1255 (CO). NMR 1 H (400 MHz, CDCh) δ : 2.92 (4H, s, H-3 and H-4) ; 3.87 (3H, s, OCH3) ; 6.70 (1 H, s, H-5), 6.88 (1 H, dd, 3 J=8.7 Hz, 3 J=7.5 Hz, 4 J=2.5 Hz, H-7) ; 7.17 -7.23 (1 H, m, H-Ar, H- 4') ; 7.27 - 7.36 (2H, m, H-Ar, H-5' and H-6'); 7.65 (1 H, d, 3 J = 8.1 Hz, H-3') ; 7.80 (1 H, s, H-9) ; 8.14 (1 H, d, 3 J = 8.7 Hz, H-8). NMR 13 C (400 MHz, CDCh) δ : 27.3 ; 29.4 ; 55.4 ; 112.3 ; 113.4 ; 124.8 ; 126.9 ; 127.0 ; 129.5 ; 130.4 ; 130.8 ; 132.9 ; 134.9 ; 136.5 ; 137.0 ; 145.9 ; 163.7 ; 186.4. HR-MS (m / z) : 343 [MH] + .

[0102] - (E)-2-[2'-methoxybenzylidenyl]-6-methoxy-3,4-dihydronaphthalenyl-1(2 / 7)-one (EC 1). Physicochemical characteristics: CiH8O3(MW = 294.35 g / mol); yield = 83%. Yellow crystalline powder. T° f = 142°C; TLC: R f = 0.56 (CHX / AE: 60 / 40). Spectral characteristics: FTIR (cm 1 ): 3063, 3019, 2968, 2945, 2838, 1661, 1590, 1507, 1440, 1137. NMR 1 H (400 MHz, CDCh) 5: 2.92 (2H, m, H-4); 3.13 (2H, m, H-3); 3.85 (3H, s, OCH3); 3.87 (3H, s, OCH3); 6.70 (1 H, d, 4 J=2.5 Hz, H-5); 6.87 (1 H, dd, 3 J=8.8 Hz, 4 J= 2.6 Hz, H-7); 6.92 - 6.97 (2H, m, H-Ar); 7.38 - 7.44 (2H, m, H-Ar); 7.80 (1 H, s, H-9); 8.10 (1 H, d, 3 J=8.8 Hz, H-8). NMR 13 C (400 MHz, CDCh) 5:26.2; 28.2; 54.3; 54.4; 111.2; 112.2; 112.8 (2C); 126.1; 127.5; 129.6; 130.6 (2C); 132.7; 134.9; 144.5; 158.7; 162.4; 185.7. HR-MS (m / z): 295 [MH] + .

[0103] (E)-2-[2'-chlorobenzylidenyl]-7-methoxy-3,4-dihydronaphthalenyl-1(2H)-one (EC 2).

[0104] Physicochemical characteristics: C 18 H 15 O2Cl (MW = 298.77 g / mol); yield = 75%; yellow crystalline powder. T° f = 141 °C; TLC: R f = 0.60 (CHX / AE: 60 / 40). Spectral characteristics: FTIR (cm- 1 ): 3061, 2955, 2937, 2835, 1666, 1601, 1572, 1493, 1461, 1237. NMR 1 H (CDCI3, 400 MHz) 5: 2.87 - 2.92 (2H, m, H-4); 2.92 - 2.98 (2H, m, H-3); 3.88 (3H, s, OCH3); 7.08 (1 H, dd, 3 J=8.4 Hz, 4 J=2.8 Hz, H-6); 7.17 (1H, d, 3 J=8.4 Hz, H-5); 7.27 - 7.49 (4H, m, H-Ar); 7.65 (1 H, d, 4 J=2.8 Hz, H-8); 7.89 (1H,s,H-9); NMR 13 C (CDCI3, 400 MHz) 5: 26.5; 27.1; 54.5; 109.2; 120.7; 125.3; 128.5; 128.6; 128.7; 129.3; 132.4; 133.0; 133.5; 133.7; 135.1; 136.1; 157.6; 186.4. HR-MS (m / z): 299 [MH] + .

[0105] (E)-2-[6'-bromobenzo[c(][1',3']dioxolo-5'-methylenyl]-7-methoxy-3,4- dihydronaphtalenyl-1(2H)-one (EC 3). Caractéristiques physicochimiques : Ci9Hi5O4Br(MW = 387,23 g / mol) ; rdt = 43 % ; poudre amorphe jaune. T° f = 140 °C ; CCM : R f = 0,35 (CHX / AE : 80 / 20 AE). Caractéristiques spectrales : FTIR n (cm-1) : 2912, 2838, 1660, 1582, 1493, 1460, 1250. RMN 1 H (CDCb, 400 MHz) δ (ppm) : 2,86 - 3,00 (4H, m, H-3 et H-4) ; 3,87 (3H, s, OCH3) ; 6,02 (2H, s, H-2’) ; 6,79 (1 H, s, H-4’) ; 7,07 (1 H, dd, 3 J=8,4 Hz, 4 J=2,8 Hz, H-6) ; 7,10 (1 H, s, H-7’) ; 7,15 (1 H, d, 3 J=8,4 Hz, H-5) ; 7,63 (1 H, d, 4 J=2,8 Hz, H-8) ; 7,74 (1H, s, H-9). RMN 13 C (CDCI3, 400 MHz) δ (ppm) : 27,5 ; 28,1 ; 55,5 ; 102,0 ; 109,8 ; 110,2 ; 113,1 ; 116,6 ; 121 ,7 ; 129,2 ; 129,5 ; 134,1 ; 135,8 ; 136,0 ; 136,1 ; 147,0 ; 148,5 ; 158,7 ; 187,5. HR-MS (m / z) : 387 [M] + .

[0106] (E)-2-[6'-bromobenzo[d][1',3']dioxolo-5'-methylenyl]-5-methoxy-3,4- dihydronaphtalenyl-1(2H)-one (EC 4). Caractéristiques physicochimiques : C 19 H 15 O4Br (MW = 387,23 g / mol) ; rdt = 70 % ; poudre cristalline jaune. T° f = 195 °C ; CCM : R f = 0,61 (CHX / AE : 70 / 30). Caractéristiques spectrales : FTIR n (cm 1 ) : 2970, 2945, 2914, 2839, 1656, 1580, 1505, 1472, 1240. RMN 1 H (400 MHz, CDCb) δ (ppm) : 2,92 (4H, s, H-3 et H-4) ; 3,87 (3H, s, OCH3) ; 6,03 (2H, s, H-2’) ; 6,79 (1 H, s, H-4’) ; 7,04 (1H, dd, 3 J=8,1 Hz, 4 J=0,6 Hz, H-6) ; 7,11 (1 H, s, H-7’) ; 7,32 (1 H, m, 3 J=7,9 Hz, H-7) ; 7,72 (1 H, s, H-9) ; 7,78 (1 H, dd, 3 J=7,9 Hz, 4 J=0,9 Hz, H-8). RMN 13 C (400 MHz, CDCI3) δ (ppm) : 20,7 ; 25,6 ; 54,7 ; 100,9 ; 108,8 ; 112,0 ; 113,3 ; 115,6 ; 118,8 ; 126,2 ; 128,2 ; 131 ,4 ; 133,2 ; 134,4 ; 135,1 ; 145,9 ; 147,4 ; 155,3 ; 186,8. HR-MS (m / z) : 387 [M] + .

[0107] (E)-2-[6'-bromobenzo[d][1',3']dioxolo-5'-methylenyl]-5,7-dimethyl-3,4- dihydronaphtalenyl-1(2H)-one (EC 5). Caractéristiques physicochimiques : C 20 H 17 O3Br (MW = 385,26 g / mol) ; rdt = 78 % ; poudre amorphe jaune. T° f = 198 °C ; CCM : R f = 0,74 (CHX / AE : 70 / 30). Caractéristiques spectrales : FTIR n (cm 1 ) : 2901 , 1660, 1591 , 1499, 1472, 1246. RMN 1 H (400 MHz, CDCI3) δ (ppm) : 2,28 (3H, s, CH3) ; 2,36 (3H, s, CH3) ; 2,83 (2H, t, 3 J=6,4 Hz, H-4) ; 2,94 (2H, m, 3 J=6,4 Hz, 4 J=1 ,4 Hz, H-3) ; 6,02 (2H, s, H-2’) ; 6,79 (1 H, s, H-4’) ; 7,11 (1 H, s, H-7’) ; 7,20 (1 H, s, H-6) ; 7,70 (1 H, s, H-9) ; 7,85 (1 H, s, H-8). RMN 13 C (400 MHz, CDCb) 5 (ppm) : 19,3 ; 20,9 ; 25,2 ; 26,8 ; 102,0 ; 109,9 ; 113,1 ; 116,5 ; 126,2 ; 129,3 ; 133,3 ; 135,0 ; 135,6 ; 135,9 ; 136,1 ; 136,2 ; 138,9 ; 147,0 ; 148,4 ; 188,2. HR-MS (m / z) : 385 [M] + .

[0108] (E)-2[6'-bromobenzo[d][1',3']dioxolo-S'-methylenyl]-5-hydroxy-3,4- dihydronaphthalenyl-1(2H)-one (EC 6). Physicochemical characteristics: Ci8Hi3O4Br (MW = 373.20 g / mol); rdt = 47 % ; yellow crystalline powder. T° f = 228 °C ; CCM : R f = 0.38 (CHX / AE : 70 / 30). Spectral characteristics: FTIR of (cm -1 ) : 3266, 2968, 2917, 2834, 1656, 1603, 1587, 1560, 1500, 1466, 1243. NMR 1 H (300 MHz, DMSO-d6) δ (ppm) : 2.76 - 3.00 (4H, m, H-3 and H-4) ; 6.15 (2H, s, H-2') ; 7.08 (1 H, s, H-4') ; 7.08 (1 H, dd, 3 J=7.9 Hz, 4 J=1 .2 Hz, H-Ar) ; 7.25 (1 H, m, 3 J=7.9 Hz, H-7) ; 7.37 (1 H, s, H-7') ; 7.47 (1 H, dd, 3 J=7.9 Hz, 4 J=1 .2 Hz, H-Ar) ; 7.55 (1 H, s, H-9) ; 9.86 (1 H, s, OH). NMR 13 C (300 MHz, DMSO-d6,) δ (ppm) : 21 .7 ; 26.5 ;

[0109] 102.8; 110.4; 113.1; 116.4; 118.4; 119.8; 127.5; 128.7; 130.7; 134.3; 134.5; 136.6; 147.5; 149.1; 154.8; 187.3. HR-MS (m / z): 373 [M] + .

[0110] (E)-2-[6'-bromobenzo[d][T,3']dioxolo-5'-methylenyl]-6,7-dimethoxy-3,4-dihydronaphthalenyl-1(2 / 7)-one (EC 7). Physicochemical characteristics: C2OHi7BrO5(MW = 417.26 g / mol); yield = 3%; orange crystalline powder. T° f > 260 °C; TLC: R f = 0.37 (CHX / AE: 70 / 30). Spectral characteristics: FTIR n (cm 1 ): 2937, 1645, 1599, 1571, 1425, 1265. NMR 1 H (400 MHz, CDCI3) δ (ppm): 2.86 - 3.03 (4H, m, H-3 and H-4); 3.86 (6H, s, OCH3-6 and OCH3-7); 6.03 (2H, s, H-2'); 6.69 (1 H, s, H-4'); 6.84 (1 H, s, H-5); 7.11 (1 H, s, H-7'); 7.65 (1 H, s, H-8); 7.72 (1 H, s, H-9). NMR 13 C (400 MHz, CDCI3) δ (ppm): 27.6; 28.7; 56.1 (2C); 102.0; 109.5; 109.8; 109.9; 113.0; 116.5; 126.5; 129.4; 135.1; 136.0; 138.3;

[0111] 146.9; 148.3; 148.4; 153.6; 186.4. HR-MS (m / z): 417 [M] + .

[0112] (E)-2-[6'-bromobenzo[d][1',3']dioxolo-5'-methylenyl]-6-hydroxy-3,4-dihydronaphthalenyl-1(2H)-one (EC 8). Physicochemical characteristics: Ci8Hi3O4Br (MW = 373.20 g / mol); yield = 13%; yellow amorphous powder. T° f = 250 °C; TLC: R f = 0.50 (CHX / AE: 60 / 40). Spectral characteristics: FTIR n (cm 1 ): 3060, 2844, 1651, 1594, 1562, 1496, 1472, 1240. NMR 1 H (400 MHz, DMSO-d6) δ (ppm): 2.80 - 2.93 (4H, m, H-3 and H-4); 6.14 (2H, s, H-2'); 6.69 (1 H, dd, 3 J=8.0 Hz, 4 J=2.4 Hz, H-5); 6.78 (1 H, dd, 3 J=8.6 Hz, 4 J =2.4 Hz, H-7); 7.07 (1 H, s, H-4'); 7.36 (1 H, s, H-7'); 7.51 (1 H, s, H-9); 7.86 (1 H, d, 3 J =8.6 Hz, H-8); 10.44 (1 H, s, OH). NMR 13C (400 MHz, DMSO-d6) δ (ppm): 26.1; 27.6; 101.7; 109.4; 112, 1; 113.4; 114.2; 115.3; 124.3; 127.8; 129.7; 132.8; 135.8; 145.6; 146.4; 147.9; 161.8; 184.4. HR-MS (m / z): 373 [M] + (E)-2-[6'-bromobenzo[d][1',3']dioxolo-5'-methylenyl]-7-bromo-3,4-dihydronaphthalenyl-1(2 / 7)-one (EC 9). Physicochemical characteristics: Ci3Hi2Br2O3 (MW = 436.10 g / mol); yield = 13%; yellow crystalline powder. T° f = 186 °C; TLC: R f = 0.50 (CHX / AE: 90 / 10). Spectral characteristics: FTIR n (cm 1 ): 2920, 1653, 1578, 1498, 1473, 1217. NMR 1 H (400 MHz, CDCI3) δ (ppm): 2.85 - 3.00 (4H, m, H-3 and H-4); 6.02 (2H, s, H-2'); 6.77 (1 H, s, H-4'); 7.10 (1H, s, H-7'); 7.14 (1 H, d, 3 J=8.1 Hz, H-5), 7.59 (1 H, dd, 3 J=8.1 Hz, 4 J=2.1 Hz, H-6); 7.77 (1H, s, H-9); 8.24 (1 H, s, 4 J=2.1 Hz, H-8). NMR 13C (400 MHz, CDCI3) δ (ppm): 27.0; 28.4; 102.1; 109.8; 113.2; 116.8; 121.0; 128.8; 130.1; 130.9; 134.8; 135.2; 136.1; 136.7; 141.9; 147.0; 148.7; 186.3. HR-MS (m / z): 435 [M] + .

[0113] B- In vitro pharmacology

[0114] 1 - Reagents

[0115] Paclitaxel was obtained from LeanCare (Greenfield, UK), and compounds E1-E5 (compounds according to the invention) and EC1-EC9 (comparative compounds) were synthesized as described above. DMSO was obtained from Sigma Aldrich (France).

[0116] 2- Cell culture

[0117] MDA-MB-231 cells (a human triple-negative breast cancer cell line) were obtained from ATCC (Manassas, VA, USA) and maintained in DMEM / F12 culture medium (Dulbecco's modified Eagle's medium) containing phenol red and supplemented with 5% fetal bovine serum (FBS) and 1% of a penicillin / streptomycin mixture (Thermo Fisher Scientific, Monza, Italy). The cells were cultured at 37°C in an atmosphere composed of 95% air and 5% CO2.

[0118] 4T1-Luc2 cells (a murine triple-negative breast cancer cell line) were obtained from ATCC (Manassas, VA, USA) and maintained in Roswell Park Memorial Institute (RPMI) 1640 culture medium in the presence of GlutaMAX™ supplemented with 10% FBS, 8 pg / mL blasticidin, and 2 mM penicillin-streptomycin (Thermo Fisher Scientific, Waltham, MA, USA). The cells were cultured at 37°C in an atmosphere composed of 95% air and 5% CO2.

[0119] 3- Cell viability of MDA-MB-231

[0120] The viability of MDA-MB-231 cells was assessed using an automatic cell counter (Life Technologies, Milan, Italy) in accordance with the manufacturer's recommendations.

[0121] MDA-MB-231 cells were seeded onto 24-well plates containing standard growth medium. The plates were incubated with 2.5% FBS before being cultured with increasing concentrations of the compounds of interest for T2 hours. The treatment was repeated daily, and cells were counted on day 4. The number of live cells is expressed as a percentage of cells that received serum alone (100%, reference). Each point represents the mean of three experiments (n=3) ± standard deviation, with experiments performed in triplicate (N=3).

[0122] The x-axis represents the concentrations of the different products (in pM) and the y-axis represents cell viability (%).

[0123] 4- Cell viability of 4T1-Luc2

[0124] The viability of 4T1-Luc2 cells was measured by bioluminescence using 4T1-Luc2 cells expressing luciferase under the control of the elongation factor EF-1a promoter. 4T1-Luc2 cells were seeded onto 96-well plates (6 x 10⁻¹²). 3 / wells) containing 200 pL of standard growth medium. The 96-well plates were incubated for 24 h, then the cells were exposed to increasing concentrations of compound E2 (0.5, 1, 5, 7.5, 10, 25, 50, 75, 100 pM) or paclitaxel (0.01, 0.1, 1, 5, 10, 50, 100, 1000 nM) for 24 h. A 0.1% DMSO concentration was used as the vehicle.

[0125] After 24 hours of incubation with the different treatment concentrations (E2 or paclitaxel), luciferin (Promega) was added to each well (20 pL per well for a final concentration of 0.15 mg / mL). After 16 minutes of incubation in the dark, the bioluminescence (in RLU, Relative Light Unit) of each well was read by the Flexstation for 1 second per well, in quadruplicate (n=4) per group. The number of live cells is expressed as a percentage relative to the cells that received the vehicle (0.1% DMSO). Cell viability in the presence of the vehicle alone is considered to be 100%. Cell viability was calculated using the following formula: % cell viability

[0126] = (RLU treated cells / RLU vehicle cells) x 100.

[0127] Dose-response curves and their nonlinear regression curve were plotted using GraphPad Prism software (version 9.0) to determine the IC50 values ​​and the slope of the Hill curve. We then defined the affected fraction (Fa) and its counterpart, the unaffected fraction (Fu). The percentage of viable cells is defined by Fa, while 100 - Fa defines Fu. These data allowed us to calculate the concentrations leading to the inhibition percentages of 6.25, 12.5, 18.75, 25, 31.25, 37.5, and 50.

[0128] Compound E2 was then combined with paclitaxel at E2 and paclitaxel concentrations resulting in the same percentage of inhibition. The concentration of the single molecule yielded the same percentage of viability as that obtained with the combination (Fa combiné) is then calculated. The concentrations of individual molecules, inducing the same effect as the combination, and the concentration of molecules used in the combination allowed the calculation of the dose reduction index (DRI) and the combination index (IndC).

[0129] The DRI values ​​were used to assess whether the combination reduces the dose of molecule required to obtain an equivalent effect when DRI > 1. The combination index was used to determine the type of interaction between two molecules such as: antagonistic (IndC > 1), additive (IndC = 1) or synergistic (IndC < 1) using the Chou-Talalay method (Chou, 2010)

[0030] . The values ​​for the isobogram are presented in the form mean ± 95% CI.

[0130] 5- Flow cytometry

[0131] The apoptosis assay was performed using the FITC Annexin V Apoptosis Detection with PI kit (BioLegend), according to the manufacturer's instructions. 4T1-Luc2 cells were seeded onto a 6-well plate (250 x 10 3 cells / wells) and were allowed to attach for 24 h. The cells were then treated with an E2 concentration corresponding to its Cl 50(15, 17 pM) and a paclitaxel concentration corresponding to its IC50 (1, 17.5 nM), or a combination thereof, were incubated for 24 h. Cells were then collected by trypsinization and labeled for 15 minutes in the dark with FITC-conjugated Annexin V and propidium iodide before flow cytometry analysis. Data were acquired using a flow cytometer (BD LSR II) and analyzed using BD FACSDiva 9.0.1 software. Cells were classified according to physical parameters to select cells of interest (FSC / SSC, Forward SCatter / Side SCatter) and cell clusters (width / area).

[0132] C- In vivo pharmacology

[0133] 1- Reagents

[0134] Paclitaxel was obtained from LeanCare (Greenfield, UK), and compounds E1-E5 and EC1-EC9 were synthesized as previously described. For in vivo testing, paclitaxel was dissolved in a mixture of 10% 96% ethanol, 10% Cremophor (Sigma Aldrich, France), and 80% physiological saline. The reference compound E2 was dissolved in 5% DMSO, 2% TweenSO4 (Sigma Aldrich, France), and physiological saline. 2- Animals

[0135] Eight-week-old female C57BL6 / J and Balb / cJRj mice from Janvier (Le Genest-St-Isle, France) were housed with ad libitum access to food and water in an environment free of specific pathogens, at 22 ± 2°C and 20% humidity with a 12h / 12h day / night cycle. All experiments were conducted according to HASP recommendations and were approved by the Animal Experimentation Ethics Committee (CEMEA) of the Auvergne Region, in accordance with Directive 2010 / 63 / EU.

[0136] 3- Model of peripheral neuropathies induced by paclitaxel

[0137] The paclitaxel-induced peripheral neuropathy model was generated by four intraperitoneal injections, one every two days, of paclitaxel (LeanCare, Greenfield, United Kingdom) dissolved in 10% 96% ethanol, 10% Cremophor EL (Sigma-Aldrich, Saint-Quentin-Fallavier, France), and 80% saline. Neuropathies were considered chronic fourteen days after model initiation. Animals were randomly assigned to groups of eight mice per group using equal block randomization.

[0138] 4- Orthotopic breast tumor model

[0139] Cultured 4T1-luc2 cells were harvested and resuspended in phosphate-buffered saline (PBS) at various densities (250,000, 500,000, 1,000,000 cells / injection site) or in a mixture of PBS and 60% Matrigel® (Corning). Mice were anesthetized by isoflurane inhalation, and then the cells were implanted into the 4 ème left breast fat pad by subcutaneous injection of 50 pL of cell suspension using a Hamilton syringe and a 26G needle. Tumor volume was calculated using the formula (length x (width) 2 / 2) from the measurement with calipers of the length and width of the tumor.

[0140] To assess the optimal density of cells to be implanted, the tumor volume was assessed three times a week for thirty-five days.

[0141] To evaluate the effect of compound E2, alone or in combination with paclitaxel, tumor volume and pain sensitivity were measured three times a week after treatment initiation. Mice received the different treatments when tumors began their second growth phase (day 18).

[0142] 5- Murine model of osteoarthritis induced by monoiodoacetate (MIA) Mice are anesthetized by isoflurane gas anesthesia (Iso-Vet®, 5% induction, 6% maintenance). An intra-articular injection of monoiodoacetate (MIA, 5 mg / 10 pL) is administered to the left knee of the mice.

[0143] 6- Murine model of osteoarthritis induced by destabilization of the medial meniscus

[0144] Osteoarthritis is induced in mice by destabilization of the medial meniscus (MMD, (Glasson, 2007)

[0031] ). Mice are anesthetized by intraperitoneal (IP) injection of 10 mL / kg of a mixture of ketamine (Imalgene® 500, 100 mg / kg) and xylazine (Rompun®, 10 mg / kg) diluted in saline (0.9% NaCl). The surgical area is disinfected with 90% alcohol and then with Betadine® (povidone-iodine solution). A longitudinal incision of approximately 5 mm is made from the bottom of the tibial plateau to the top of the patella. The patella is then dislocated, the synovial capsule opened with a scalpel, and the infrapatellar fat pad divided to expose the medial meniscotibial ligament, which is then divided. The patella is then repositioning, then the skin is closed with Monocryl® 5.0. Control mice (Sham) undergo the same operation until the ligament is exposed, while keeping it intact.

[0145] 7- Evaluation of mechanical allodynia: the von Frey test

[0146] Mechanical pain sensitivity was assessed using the von Frey filament test. Nylon filaments (Bioseb) of varying diameters were applied to the animal's paw. Withdrawal or licking of the paw was considered a positive response. The measurement was then repeated with a finer or larger diameter filament if the animal did not react. The paw withdrawal threshold of 50% (PWT, in grams) was determined using the Dixon up-down method.

[0147] 8- Treatments

[0148] G-15 (a specific GPER antagonist, Tocris Bio-Techne, Noyal-Châtillon-sur-Seiche, France), at a dose of 0.3 mg / kg, dissolved in 5% DMSO (SigmaAldrich, Saint-Quentin-Fallavier, France), 5% Tween80 (Sigma-Aldrich, Saint-Quentin-Fallavier, France), and 90% physiological saline, and E2, dissolved in 5% DMSO and 95% physiological saline, were administered at a dose of 10 mL / kg intraperitoneally or orally. G-15 was injected 15 minutes before E2. Monoiodoacetate (MIA) was supplied by Sigma-Aldrich, Saint-Quentin-Fallavier, France.

[0149] EXAMPLE 2: RESULTS

[0150] A- Antiproliferative effect 1- In vitro evaluation of the antiproliferative effect of compounds according to the invention (E) and comparators (EC)

[0151] Cell viability studies performed on MDA-MB-231 TNBC cells show in all cases a concentration-dependent antiproliferative action (control: vehicle alone, 100%) (Figure 1).

[0152] Regarding compound E1, the antiproliferative action is visible at concentrations above 1 pM. It is visible from 5 pM for compound E2. As for compound E3, an antiproliferative action is observed at concentrations above 0.1 pM.

[0153] Regarding the EC series derivatives, an antiproliferative action is observed at concentrations above 0.1 pM for EC 3, EC 5, EC 6 and EC 8 derivatives. This activity is observed at concentrations above 1 pM for EC 1, EC 4 and EC 9 derivatives. For compound EC 2, this action is observed from 5 pM.

[0154] 2- Compound E 2 induces a cytotoxic effect on murine TNSC cells

[0155] The antiproliferative effects of compound E2 and paclitaxel (positive control) were evaluated in murine TNBC cells. 4T1-Luc2 cells were treated with increasing concentrations of compound E2 (0.5 to 100 pM) or paclitaxel (0.01 to 1000 nM) for 24 h, and cell viability was measured by bioluminescence. As shown in Figure 2, compound E2 and paclitaxel reduced cell viability with an inhibitory concentration of 50 (Cl₂). 50 ) of 15.1 pM (95% confidence interval (CI) 12.2 to 18.7 pM) and 117.5 nM (95% CI 63.3 to 242.9 nM), respectively. These results show that compound E2 and paclitaxel have an antiproliferative effect on murine TNBC cells.

[0156] 3- Compound E 2 and paclitaxel exhibit a synergistic cytotoxic effect in vitro

[0157] The effect of compound E2 was then evaluated by bioluminescence on the antiproliferative response of paclitaxel in 4T1-Luc2 cells. Co-treatment with compound E2 and paclitaxel, at concentrations corresponding to their percentage of inhibition (6.25, 12.5, 18.75, 25, 31.25, 37.5, and 50), induced an antiproliferative effect (Figure 3A). Indeed, the use of these two compounds at a Cl of 36.4% (95% CI 33.6–39.9%) reduced cell viability by 50%. These data allowed the determination of the dose reduction index for each molecule by calculating the ratio of the concentration of the single molecule, which produces the same effect as the combination, to the concentration of the molecule in the combination.The use of the combination allows the dose of paclitaxel used to be reduced by a factor of 8.60 (95% CI 4.84 to 13.80) and by a factor of 2.39 (95% CI 1.89 to 2.91) for compound E 2 to reduce cell viability by 50% (Figure 3B).

[0158] Since the combination index (IndC) of compound E2 and paclitaxel is less than 1, it is possible to conclude that these two molecules exhibit synergy at the different doses tested, although this index increases in a dose-dependent manner (IndC of 0.32 ± 0.05 for 75% viability, 0.46 ± 0.06 for 55% viability, and 0.87 ± 0.31 for 39% viability) (Figure 3C). These results therefore indicate a synergistic inhibitory effect of the paclitaxel-E2 combination at the tested doses on the growth of 4T1-Luc2 cells, as shown by the analysis of the isobologram at IC25 (Figure 3D) and Cl2. 50 (Figure 3E).

[0159] 4- The combination of compound E 2 and paclitaxel induces cellular apoptosis

[0160] To determine whether the cytotoxic effect of compound E2 and paclitaxel is due to the induction of cell death by apoptosis and / or necrosis, a dual labeling analysis with Annexin V and propidium iodide on 4T1-Luc2 cells was performed (Figure 4A). Flow cytometry analysis showed that compound E2 (at its IC50 of 15.13 pM) alone and paclitaxel (at its IC50 of 117.5 nM) alone induced apoptosis of 4T1-Luc2 cells in a non-significant manner (Figure 4B). Furthermore, the combination of the two compounds significantly increased induced apoptosis compared to cells treated with the vehicle (28.55 ± 1.45% apoptosis for the combination versus 12.05 ± 0.51% for the vehicle, p = 0.001). The different treatments did not induce necrosis (Figure 4C).

[0161] B- Analgesic effect: neuropathic pain

[0162] 1- Compound E 2 induces a dose-dependent analgesic effect in a murine model of paclitaxel-induced neuropathy

[0163] Repeated administration of paclitaxel induces neuropathic pain, observed by a decrease in pain thresholds (50% of the withdrawal threshold) between baseline (before paclitaxel administration) and time 0 (after paclitaxel treatment) (Figure 5). Intraperitoneal (IP) administration of compound E2 increases pain thresholds in animals in a dose-dependent manner. The positive control is duloxetine. At 20 mg / kg, E2 has an effect comparable to duloxetine at 30 mg / kg. At 10 mg / kg, E2 has an effect comparable to PLMI at the same dose. Similarly, a dose-dependent analgesic effect is observed when E2 is administered orally, highlighting its oral bioavailability (Figure 6).

[0164] 2- Compound E 2 induces a GPER-dependent analgesic effect on a murine model of paclitaxel-induced neuropathy. The analgesic effect of compound E 2 observed on a murine model of paclitaxel-induced neuropathic pain is suppressed by co-administration of G-15 (Figure 7).

[0165] 3- Compound E 2 induces an analgesic effect on a murine model of paclitaxel-induced neuropathy via a pool of GPERs whose functional localization is partly peripheral

[0166] The genetic deletion of GPER in primary afferent fibers (GPER mice) SNS KO) reduces paclitaxel-induced neuropathic pain (Figure 8) and shows that this receptor, when located peripherally, is involved in neuropathic pain.

[0167] Administered peripherally (subcutaneous injection in the animal's paw), compound E2 induces an analgesic effect (positive control: lidocaine) (Figure 9A). The analgesic effect of peripherally administered compound E2 is suppressed by peripheral co-administration of G-15 (Figure 9B). The analgesic effect of systemic administration of compound E2 (ip) is partially reduced by intraplantar co-administration of G-15 (Figure 9C).

[0168] The analgesic effect of compound E 2 or PLMI in a mouse model of paclitaxel-induced neuropathic pain is reduced in mice with GPER specifically deleted in primary afferent fibers (Figure 10).

[0169] All these results mean that the peripherally located GPER is at least partly responsible for the analgesic effect of compound E 2.

[0170] 4- Compound E 2 induces an analgesic effect on a murine model of paclitaxel-induced neuropathy dependent on GPER, the functional localization of which is partly spinal.

[0171] Viral deletion of GPER specifically in the spinal cord (GPER mice) DH KO) reduces paclitaxel-induced neuropathic pain (Figure 11) and shows that this receptor, when localized at the spinal level, is involved in neuropathic pain.

[0172] Administered intrathecally into the spinal cord, compound E2 induces an analgesic effect (positive control: morphine) (Figure 12A). The analgesic effect of compound E2 administered intrathecally is suppressed by intrathecal co-administration of G-15 (Figure 12B). The analgesic effect of systemic administration of compound E2 (intrathecally) is partially reduced by intrathecal co-administration of G-15 (Figure 12C).

[0173] These results indicate that the spinal GPER is at least partly responsible for the analgesic effect of compound E. 2.5 - Evaluation of the analgesic effect of compounds E on a murine model of paclitaxel-induced neuropathy

[0174] Repeated administration of paclitaxel induces neuropathic pain, as evidenced by the decrease in pain thresholds (50% of the withdrawal threshold) between baseline (before paclitaxel administration) and time 0 (after paclitaxel treatment) (Figure 13). The effect of the E compounds at a dose of 10 mg / kg (ip) is shown. The E compounds are more effective than the EC compounds.

[0175] 6- Repeated injection of compound E 2, in conjunction with paclitaxel, reduces neuropathy-induced pain

[0176] Chronic administration of compound E2 or PLMI (10 mg / kg, intraperitoneal) three times daily for 11 days (days 0 to 10) concurrently with paclitaxel (PIPN design, 4 injections on days 1, 3, 5, and 7) resulted in a significant reduction in tactile allodynia as assessed by the von Frey test on day 10 (Figure 14, top). This represents a residual analgesic effect. Daily pain thresholds were measured 6 hours after treatment with E2 or PLMI, a time point exceeding the duration of the acute effect of the compounds. It is noteworthy that the acute effect is still maintained after injection of E2 or PLMI, as demonstrated by the kinetics performed after the last injection of E2 or PLMI (Figure 14, bottom).

[0177] It should be noted that acute or chronic administration of E2 in animals does not induce weight loss or stereotypical behaviors that could be considered adverse effects of the molecule.

[0178] 7- The combination of compound E 2 and paclitaxel in vivo reduces neuropathic pain caused by paclitaxel and tumor growth

[0179] The antinociceptive and antitumor effect of compound E2 was evaluated on the murine model of primary TNBC tumor (implantation at J o ) treated or not with paclitaxel. After tumor establishment for 19 days, mice were treated with compound E 2 (10 mg / kg, administered ip 3 times daily from J19 to J3o) or its vehicle (physiological saline, 10 mL / kg) and with paclitaxel (4 mg / kg administered ip on J2o, J22, J24 and J2e) or its vehicle (physiological saline, 10 mL / kg) (Figure 15A).

[0180] After 30 days, tumor growth in animals treated with compound E2 was 35% lower than in animals treated with the vehicle (376.71 ±75.93 mm 3 for mice treated with E2 vs 577.98±54.317 mm 3for the vehicle group, p = 0.12) and 23% lower in paclitaxel-treated mice compared to vehicle mice (443.92 ± 38.73 mm 3 (Figure 15B). Furthermore, the paclitaxel-compound E2 combination resulted in the greatest inhibition of tumor growth, with a significant 42% reduction compared to the vehicle group (339.5 ± 55.1 mm²). 3, p = 0.02). These results are similar to those obtained when measuring tumor mass at the end of treatment, where a significant decrease in tumor mass was observed from the paclitaxel and combination-treated groups compared to the vehicle group (363.4±59.43 mg for the paclitaxel group vs 561,740.41 mg for the vehicle group, p = 0.0488, and 31855.37 mg for the combination-treated group, p = 0.0328 compared to the vehicle group) (Figure 15C). Paclitaxel-induced mechanical allodynia (0.13±0.05 g for paclitaxel vs 0.52±0.04 g for the vehicle group at D24, p = 0.0004, and 0.058±0.007 g for paclitaxel at D30 vs 0.41±0.05 g for the vehicle group, p = 0.002) is significantly reduced by co-administration of compound E 2 (0.29±0.07 g for the combination group vs 0.058±0.007 g for the paclitaxel alone group at D30, p = 0.002) (Figure 15D).

[0181] These results show that co-administration of compound E 2 and paclitaxel does not alter the antiproliferative effect of paclitaxel, and that it significantly reduces paclitaxel-induced mechanical allodynia.

[0182] 8- Compound E 2 induces an analgesic effect in oxaliplatin- and bortezomib-induced neuropathic pain models

[0183] Compound E 2 and PLMI (administered intraperitoneally) exhibit an analgesic effect in the von Frey test in a murine model of neuropathic pain induced by oxaliplatin (top, 3 mg / kg, twice weekly for 4 weeks) and by bortezomib (bottom, 0.4 mg / kg, three times weekly for 4 weeks).

[0184] C- Analgesic effect: arthritic pain

[0185] 1- Compound E 2 induces an analgesic effect on a murine model of osteoarthritic pain induced by an intra-articular injection of mono-iodo-acetate (MIA)

[0186] The onset of osteoarthritic pain is evidenced by the decrease in pain thresholds (50% of the withdrawal threshold) between baseline (before MIA injection) and time 0 (18 days after MIA injection) (Figure 17). Compound E2, administered intraperitoneally, increases pain thresholds in animals. The effect of PLMI is also demonstrated.

[0187] 2- Compound E2 induces an analgesic effect in a murine model of GPER-dependent osteoarthritic pain with a partially peripheral functional localization. The genetic deletion of GPER in primary afferent fibers (GPER mice) SNS KO) reduces surgery-induced osteoarthritic pain (Figure 18) and shows that this receptor, when located peripherally, is involved in osteoarthritic pain.

[0188] The analgesic effect of compound E 2 in a murine model of osteoarthritic pain is reduced in mice with GPER specifically deleted in primary afferent fibers (Figure 19, control: morphine 1 mg / kg, ip).

[0189] All these results mean that GPER, when localized in the periphery, is at least partly responsible for the analgesic effect of compound E 2.

[0190] 3- Compound E 2 induces an analgesic effect on a murine model of GPER-dependent osteoarthritic pain with a partly spinal functional localization

[0191] Viral deletion of GPER specifically in the spinal cord (GPER mice) DH KO) reduces arthritic pain (Figure 20) and shows that this receptor, when located at the spinal level, is involved in arthritic pain.

[0192] The analgesic effect of compound E2 in a murine model of osteoarthritic pain is reduced in GPER mice DH KO (positive control: morphine 1 mg / kg, ip) (Figure 21).

[0193] All these results mean that GPER, when localized to the spinal level, is at least partly responsible for the analgesic effect of compound E 2.

[0194] List of references

[0195] [1] Raja, SN, Carr, DB, Cohen, M., Finnerup, NB, Flor, H., Gibson, S., et al. (2020). The revised International Association for the Study of Pain definition of pain: concepts, challenges, and compromises. Bread 161 1976-1982.

[0196] [2] Won, K.-A., and Spruck, C. (2020). Triple-negative breast cancer therapy: Current and future perspectives (Review). Int. J. Oncol. 57:1245-1261.

[0197] [3] Yin, L., Duan, J. -J., Bian, X.-W., and Yu, S.-C. (2020). Triple-negative breast cancer molecular subtyping and treatment progress. Breast Cancer Res. BCR 22: 61 .

[0198] [4] Almansour, N.M. (2022). Triple-Negative Breast Cancer: A Brief Review About Epidemiology, Risk Factors, Signaling Pathways, Treatment and Role of Artificial Intelligence. Front. Mol. Biosci. 9: 836417.

[0199] [5] Li, Y., Zhang, H., Merkher, Y., Chen, L., Liu, N., Leonov, S., et al. (2022). Recent advances in therapeutic strategies for triple-negative breast cancer. J. Hematol. Oncol. J Hematol Oncol 75: 121.

[0200] [6] Okorafor, C.C., Shastri, S., Wen, K., Ebong, E.E. (2024). Mechanisms of triple-negative breast cancer extravasation: Impact of the physical environment and endothelial glycocalyx. FASEB J. 38: e23785.

[0201] [7] Alves, R.C., Fernandes, R.P., Eloy, J.O., Salgado, H.R.N., and Chorilli, M. (2018). Characteristics, Properties and Analytical Methods of Paclitaxel: A Review. Crit. Rev. Anal. Chem. 48: 110-118.

[0202] [8] Molassiotis, A., Cheng, H.L., Lopez, V., Au, J.S.K., Chan, A., Bandla, A., et al. (2019). Are we mis-estimating chemotherapy-induced peripheral neuropathy? Analysis of assessment methodologies from a prospective, multinational, longitudinal cohort study of patients receiving neurotoxic chemotherapy. BMC Cancer 79: 132.

[0203] [9] Staff, N.P., Grisold, A., Grisold, W., and Windebank, A.J. (2017). Chemotherapy-induced peripheral neuropathy: A current review. Ann. Neurol. 87: 772-781.

[0204]

[0010] Moisset, X., Bouhassira, D., Avez Couturier, J., Alchaar, H., Conradi, S., Delmotte, M.H., et al. (2020). Pharmacological and non-pharmacological treatments for neuropathic pain: Systematic review and French recommendations. Rev. Neurol. (Paris) 776: 325-352.

[0205]

[0011] Zajqczkowska, R., Kocot-Kçpska, M., Leppert, W., and Wordliczek, J. (2019). Bone Pain in Cancer Patients: Mechanisms and Current Treatment. Int. J. Mol. Sci. 20: E6047.

[0206]

[0012] Wnuk, A., Przepiôrska, K., Pietrzak, B.A., and Kajta, M. (2023). Emerging Evidence on Membrane Estrogen Receptors as Novel Therapeutic Targets for Central Nervous System Pathologies. Int. J. Mol. Sci. 24: 4043.

[0013] Prossnitz, E.R., and Barton, M. (2023). The G protein-coupled oestrogen receptor GPER in health and disease: an update. Nat. Rev. Endocrinol. 79: 407-424.

[0207]

[0014] Deliu, E., Brailoiu, G.C., Arterburn, J.B., Oprea, T.I., Benamar, K., Dun, N.J., et al. (2012). Mechanisms of G protein-coupled estrogen receptor-mediated spinal nociception. J. Pain Off. J. Am. Pain Soc. 73: 742-754.

[0208]

[0015] An, G., Li, W., Yan, T., and Li, S. (2014). Estrogen rapidly enhances incisional pain of ovariectomized rats primarily through the G protein-coupled estrogen receptor. Int. J. Mol. Sci. 75: 10479-10491.

[0209]

[0016] Zielihska, M., Fichna, J., Bashashati, M., Habibi, S., Sibaev, A., Timmermans, J.-P., et al. (2017). G protein-coupled estrogen receptor and estrogen receptor ligands regulate colonic motility and visceral pain. Neurogastroenterol. Motil. Off. J. Eur. Gastrointest. Motil. Soc. 29.

[0210]

[0017] Xu, J.-J., Gao, P., Wu, Y„ Yin, S.-Q., Zhu, L, Xu, S.-H., et al. (2021). G protein-coupled estrogen receptor in the rostral ventromedial medulla contributes to the chronification of postoperative pain. CNS Neurosci. Ther. 27: 1313-1326.

[0211]

[0018] Jiao, Y., Gao, P., Dong, L, Ding, X., Meng, Y., Qian, J., et al. (2023). Molecular identification of bulbospinal ON neurons by GPER, which drives pain and morphine tolerance. J. Clin. Invest. 133 e154588.

[0212]

[0019] Wang, D., Hu, L., Zhang, G., Zhang, L., and Chen, C. (2010). G protein-coupled receptor 30 in tumor development. Endocrine 38: 29-37.

[0213]

[0020] Lappano, R., Pisano, A., and Maggiolini, M. (2014). GPER function in breast cancer: An overview. Front. Endocrinol. 5: 66.

[0214]

[0021] Jacenik, D., Cygankiewicz, A.I., and Krajewska, W.M. (2016). The G protein-coupled estrogen receptor as a modulator of neoplastic transformation. Mol. Cell. Endocrinol. 429: 10— 18.

[0215]

[0022] Xu, S., Yu, S., Dong, D., and Lee, L.T.O. (2019). G Protein-Coupled Estrogen Receptor: A Potential Therapeutic Target in Cancer. Front. Endocrinol. 70: 725.

[0216]

[0023] Tirado-Garibay, A.C., Falcôn-Ruiz, E.A., Ochoa-Zarzosa, A., Lôpez-Meza, J.E. (2023). GPER: An Estrogen Receptor Key in Metastasis and Tumoral Microenvironments. Int.J. Mol. Sci. 24: 14993.

[0217]

[0025] Steiman, J., Peralta, E.A., Louis, S., and Kamel, O. (2013). Biology of the estrogen receptor, GPR30, in triple negative breast cancer. Am. J. Surg. 206: 698-703.

[0218]

[0026] Lappano, R., Mallet, C., Rizzuti, B., Grande, F., Galli, G.R., Byrne, C., et al. (2019). The peptide ERa17p is a GPER inverse agonist that exerts antiproliferative effects in breast cancer cells. Cells 8: 590.

[0027] Kampa, M., Lappano, R., Grande, F., Rizzuti, B., Maggiolini, M., Castanas, E., et al. (2023). Promising Perspectives of the Antiproliferative GPER Inverse Agonist ERa17p in Breast Cancer. Cells 12. 653.

[0219]

[0028] Mallet, C., Boudieu, L, Lamoine, S., Coudert, C., Jacquot, Y., and Eschalier, A. (2021). The antitumor peptide ERa17p exerts anti-hyperalgesic and anti-inflammatory actions through

[0220] GPER in mice. Front. Endocrinol. 12. 578250.

[0221]

[0029] Jouffre, B., Acramel, A., Belnou, M., Santolla, M.F., Talia, M., Lappano, R., et al. (2023). Identification of a human estrogen receptor a tetrapeptidic fragment with dual antiproliferative and anti-nociceptive action. Sci. Rep. 12. 1326.

[0030] Chou, T.-C. (2010). Drug combination studies and their synergy quantification using the

[0222] Chou-Talalay method. Cancer Res. 70: 440-446.

[0223]

[0031] Glasson, S.S. (2007). In vivo osteoarthritis target validation utilizing genetically-modified mice. Curr. Drug Targets 8: 367-376.

Claims

DEMANDS 1) Derivative of tetralone with the following general formula (I): Ri is H or CH3; R2 is H or OCH3; R3 is H or F; R4 and R5 are H or together form a 1,3-dioxolane; R6 is Br; given that at least one among Ri, R2 and R3 is not H. 2) A tetralone derivative according to claim 1, said derivative having one of the following formulas: 3) Derivative according to claim 1 or 2, where R1, R2, R3 and R6 are as defined in claim 1; and R4 and R5 are H. 4) A derivative according to claim 1 or 2, wherein R1, R2, R3, and R6 are as defined in claim 1; and R4 and R5 together form a 1,3-dioxolane. 5) A tetralone derivative according to any one of claims 1, 2, or 4; said derivative having one of the following formulas: one of The following general formula (I): Ri is H or CH3; R2 is H or OCH3; R3 is H or F; R4 and R5 are H or together form a 1,3-dioxolane; R6est Br. and a pharmaceutically acceptable excipient. 7) Pharmaceutical composition according to claim 6, further comprising at least one anti-tumor agent and / or an analgesic. 8) Pharmaceutical composition according to claim 7, wherein said at least one anti-tumor agent is paclitaxel, oxaliplatin or bortezomib. 9) Derivative of tetralone with the following general formula (I): Ri is H or CH3; R2est H or OCH3; R3est H or F; R4 and R5 are H or together form a 1,3-dioxolane; R6est H or Br; or pharmaceutical composition comprising said at least one derivative and one pharmaceutically acceptable excipient, for use as an analgesic drug. 10) Tetralone derivative or pharmaceutical composition for use according to claim 9, wherein the drug is intended for the control of nociception. 11) Derivative of tetralone with the following general formula (I): R1 is H or CH3; R2est H or OCH3; R3est H or F; R4 and R5 are H or together form a 1,3-dioxolane; R6est H or Br; or pharmaceutical composition comprising said at least one derivative and one pharmaceutically acceptable excipient, for use in the prevention or treatment of pain. 12) Tetralone derivative or pharmaceutical composition for use according to claim 11 or 12, where the pain is neuropathic or arthritic. 13) Tetralone derivative or pharmaceutical composition for use according to claim 11 or 12, where pain is associated with cancer treatment. 14) Tetralone derivative or pharmaceutical composition for use according to claim 13, where cancer is triple negative breast cancer (TNBC). 15) Tetralone derivative or pharmaceutical composition for use according to any one of claims 9 to 14, wherein said derivative has one of the following formulas:

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