C5AR1 inhibitors for treating hypersensitivity reactions to taxanes

Abstract

The present invention relates to the use of a C5aR1 inhibitor for the prevention or treatment of hypersensitivity reactions (HSR) to taxanes in an individual.

Description

[Technical field]

[0001] The present invention relates to the prevention and treatment of hypersensitivity reactions to taxanes. [Background technology]

[0002] Hypersensitivity reactions (HSRs) are unpredictable reactions characterized by clinical symptoms resembling allergies, even in the absence of evidence of an immunological mechanism, that may be life-threatening, require prolonged hospitalization, and require subsequent changes in therapy. In general, severe HSRs are defined as any reaction severe enough to justify discontinuing the drug. HSRs may be allergic, involving immune responses mediated by immunoglobulins and / or T cells, or nonallergic, not involving a specific immune response.

[0003] Clinically, HSR is generally classified as immediate or non-immediate / delayed, depending on the relationship of its onset to the treatment that caused it. Immediate HSRs usually occur during or within the first hour after drug exposure, are generally triggered by IgE-mediated mechanisms, and are usually non-allergic, whereas non-immediate HSRs occur more than one hour after the first drug administration and are usually associated with delayed T cell-dependent allergic mechanisms.

[0004] However, the above mechanistic classifications have limitations because other factors such as the route of administration, the role of drug metabolites, and the presence of cofactors or concomitant prescription medications may accelerate or delay the onset and / or progression of the response (Demoly et al., Allergy 2014, 69:420-437).

[0005] HSR to chemotherapy, especially taxanes and platin drugs, is very common and poses a serious threat to the treatment of cancer patients, in some cases preventing them from using first-line drugs that are the most effective drugs for their particular cancer.

[0006] Symptoms include skin symptoms such as facial flushing, chest, back and abdominal pain, and respiratory symptoms. More severe reactions are also characterized by oxygen desaturation and / or hypotension.

[0007] It is now established that the HSR to platins is IgE-mediated, but the mechanism of the HSR to taxanes remains unclear. The taxanes used in clinical practice are paclitaxel, docetaxel and cabazitaxel.

[0008] They exert their antineoplastic activity by inhibiting mitosis and inducing apoptosis in dividing cells by interfering with the dynamics of microtubules (components of the cytoskeleton).

[0009] Paclitaxel (Taxol®) is an albumin-bound paclitaxel (NAB TM -Paclitaxel; also available clinically as Abraxane®, is a natural compound originally isolated from the bark of the Pacific yew tree (Taxus brevifolia) in the 1960s. It is used to treat ovarian cancer, breast cancer, non-small cell lung cancer, and AIDS-related Kaposi's sarcoma.

[0010] Docetaxel (Taxotere®) is a semisynthetic taxane developed as a more soluble and easier to manufacture alternative to paclitaxel. It is used to treat breast, ovarian, prostate, head and neck, non-small cell lung and gastric adenocarcinoma.

[0011] Cabazitaxel (Jevtana®) is a semi-synthetic taxane developed to overcome tumor drug resistance to paclitaxel and docetaxel. Both of these molecules are poorly soluble in water, so their formulations require the addition of emulsifiers to enable intravenous administration, specifically Cremophor® EL (a mixture of polyoxyethylated triglycerides) for paclitaxel formulations and Polysorbate 80 (Tween 80) for docetaxel and cabazitaxel, which are more soluble than paclitaxel.

[0012] The majority of HSRs to taxanes occur during the first or second infusion of the drug and are usually immediate HSRs, occurring within minutes of the start of the infusion, suggesting that they are unrelated to prior sensitization (Picard et al., Clinic Rev Allerg Immunol 2015, 49:177-191).

[0013] It has been hypothesized that emulsifiers used to solubilize paclitaxel and other taxanes contribute to HSR through complement activation and production of anaphylatoxins and subsequent mast cell and basophil secretory responses. However, the solvent-free paclitaxel formulation, Nab TM Paclitaxel can still induce HSR in a significant proportion of treated patients, suggesting a direct sensitization mechanism.

[0014] Furthermore, recent findings raise the possibility that taxane-induced HSR may be IgE-mediated, supporting the use of skin prick testing to identify at-risk patients (Prieto et al., J Investig Allergol Clin Immunol 2010,20:170-171; Picard et al., J Allergy Clin Immunol 2016,137:1154-64), and suggesting that taxanes may induce basophil activation through non-IgE-mediated mechanisms (Picard et al., J Clinic Rev Allerg Immunol 2015,49:177-191).

[0015] This lack of clear understanding of the primary causes of taxane-induced HSR has led to a wide variety of management strategies in practice. To reduce the incidence of reactions, premedication with corticosteroids and antihistamine compounds is used as a prophylactic treatment. However, despite premedication, a high percentage of patients experience HSR, the incidence of which varies depending on the chemotherapy agent used (paclitaxel 10%, docetaxel 5%, Nab 10%), including severe and life-threatening HSR. TM -paclitaxel 4% and cabazitaxel approximately 1%) (Picard et al., Immunol Allergy Clin An Am 2017, 37:679-693).

[0016] During the past decade, the development of drug desensitization and graded challenging protocols has allowed the reintroduction of taxanes to some patients who experience HSR. However, there remains a great need for a better understanding of the mechanism of HSR to taxanes and for the development of safe clinical approaches to prevent HSR in patients treated with these drugs.

[0017] The anaphylatoxins (ATs) C3a, C5a, and C5a-desArg are proinflammatory polypeptides generated after proteolytic cleavage of C3 and C5 in response to complement activation. C5a exerts proinflammatory and immunoregulatory functions, acting primarily through C5a receptor 1 (C5aR1). Dysregulation of the C5a-C5aR1 axis is involved in various immune disorders.

[0018] C5aR1 is expressed on a wide range of cell types, including all cells of myeloid origin (neutrophils, eosinophils, monocytes, macrophages, dendritic cells, mast cells), lymphocytes, and non-myeloid cells, such as cells of the lung, liver, kidney, skin, and central nervous system (CNS). [Prior art documents] [Non-patent literature]

[0019] [Non-Patent Document 1] Demoly et al, Allergy 2014, 69: 420-437 [Non-Patent Document 2] Picard et al, Clinic Rev Allerg Immunol 2015, 49:177-191 [Non-Patent Document 3] Prieto et al, J Investig Allergol Clin Immunol 2010, 20: 170-171 [Non-Patent Document 4] Picard et al, J Allergy Clin Immunol 2016, 137:1154-64 [Non-Patent Document 5] Picard et al, J Clinic Rev Allerg Immunol 2015, 49:177-191 [Non-Patent Document 6] Picard et al, Immunol Allergy Clin An Am 2017, 37: 679-693 Summary of the Invention

[0020] The inventors have now surprisingly found that taxanes can bind to the C5a receptor, C5aR1, and thereby activate the same signaling pathway triggered by C5a binding to this receptor, thereby inducing the anaphylactic reactions responsible for the HSR associated with the administration of these drugs.

[0021] Thus, a first aspect of the invention is a C5aR1 inhibitor for use in the prevention or treatment of a hypersensitivity reaction (HSR) to a taxane in an individual. A second aspect of the invention is a pharmaceutical composition comprising a C5aR1 inhibitor and at least one inert pharma- ceutically acceptable excipient for use in the prevention or treatment of a hypersensitivity reaction to a taxane in an individual.

[0022] A third aspect of the invention is a method for preventing or treating a hypersensitivity reaction (HSR) to a taxane in an individual comprising administering to the individual a C5aR1 inhibitor. [Brief description of the drawings]

[0023] [Figure 1] Figure 1 shows the concentration over time, expressed in resonance units (RU), of C5aR1 bound to a sensor chip, obtained by C5aR1 capture via the c-Myc tag as described in Example 1a). The difference between the baseline RU before (start baseline) and after (final baseline) injection of C5AR1-cMyc onto the sensor chip corresponds to a stable binding of 77 RU of the receptor to the surface. [Diagram 2] FIG. 2 shows the concentration over time, expressed in resonance units (RU), of C5a bound to C5aR1 immobilized on a sensor chip as described in Example 1b). [Diagram 3] FIG. 3 shows the concentration, expressed in resonance units (RU), of paclitaxel bound to C5aR1 immobilized on a sensor chip as described in example 1c). [Figure 4] FIG. 4 shows the concentration over time, expressed in resonance units (RU), of paclitaxel bound to C5aR1 immobilized on a sensor chip, measured at various paclitaxel concentrations (4.06 μM to 37.5 μM) as described in Example 1c). [Diagram 5] FIG. 5 shows the steady-state concentration, expressed in resonance units (RU), of paclitaxel bound to C5aR1 immobilized on a sensor chip, measured as a function of paclitaxel concentration, as described in Example 1c). [Figure 6]Figure 6 shows the concentration over time, expressed in resonance units (RU), of C5aR2 bound to the microchip obtained by C5aR2 capture via the His tag as described in Example 2a). The difference between the baseline RU before (start baseline) and after (final baseline) injection of C5aR2-His onto the sensor chip corresponds to a stable binding of 569 RU of the receptor to the surface. [Figure 7] FIG. 7 shows the concentration over time, expressed in resonance units (RU), of C5a bound to C5aR2 immobilized on a sensor chip as described in example 2b). [Figure 8] Figure 8 shows the concentration over time, expressed in resonance units (RU), of paclitaxel bound to C5aR2 immobilized on a sensor chip, measured at various paclitaxel concentrations (4.06 μM to 37.5 μM) as described in Example 2c. Paclitaxel is unable to bind to the receptor at any of the concentrations tested. [Figure 9] Figure 9 shows the concentration over time, expressed in resonance units (RU), of paclitaxel bound to C5a on a sensor chip as described in Example 3b).Paclitaxel is unable to bind to C5a at the concentrations tested. [Figure 10] Figure 10 shows cytokine profiling in the supernatant of RAW264.7 treated with paclitaxel for 24 hours as described in Example 4. Numbers on the membrane indicate each cytokine reported in the histogram that was revealed in the assay (1: MCP-1 / CCL2; 2: C5a; 3: TNF alpha; 4: MIP-1 alpha / CCL3; 5: MI-1 beta / CCL4; 6: MIP2 / CXCL2; 7: IL-1ra; 8: Rantes / CCL5). [Figure 11] FIG. 11 shows cytokine levels, expressed as pixel density, in the supernatants of RAW264.7 cells treated for 24 hours with C5a or paclitaxel, measured as described in Example 4. [Figure 12]FIG. 12 shows the results of real-time PCR analysis of relative mRNA expression of IL-8 in F11 cells treated with 10 nM paclitaxel (PAC) or 50 nM C5a (50 nMC5a) for 20 hours. [Figure 13] FIG. 13 shows the average pixel density results of each cytokine released by mouse RAW264 macrophages (panels A and B) and human macrophages (panels C and D) that were untreated (CTR) or treated with C5a alone (C5a), or with C5A in combination with C5AR1 antibody (anti-C5AR+C5A), Avacopan (AV+C5A), or DF3966A (DF+C5A), or with paclitaxel alone (PAC), or with paclitaxel in combination with C5AR1 antibody (anti-C5AR+PAC), Avacopan (AV+PAC), or DF3966A (DF+PAC), measured as described in Example 6. [Figure 14] FIG. 14 shows membranes depicting each of the cytokines released by mouse RAW264 macrophages as determined by the assay described in Example 6 and reported in the histograms of FIG. [Figure 15] FIG. 15 shows the results of the average pixel density of each cytokine released in human macrophages that were untreated (CTR) or treated with C5a alone (C5a), or with C5A in combination with C5AR1 antibody (anti-C5AR+C5A), Avacopan (AV+C5A), or DF3966A (DF+C5A), or with paclitaxel alone (PAC), or with paclitaxel in combination with C5AR1 antibody (anti-C5AR+PAC), Avacopan (AV+PAC), or DF3966A (DF+PAC), measured as described in Example 6 (Panels A and B). [Figure 16] FIG. 16 shows membranes depicting each of the cytokines released by human macrophages as determined by the assay described in Example 6 and reported in the histograms of FIG. [Figure 17] FIG. 17 shows the results of permeability scores (Panel A) and vascular leakage (Panel B) calculated via spectrophotometer (OD) in mice treated with PEG / TWEEN alone (PEG / TWEEN), paclitaxel in DMSO / PEG / TWEEN (PAC in PEG / TWEEN), Cremophor EL alone (Cremophor) or paclitaxel in Cremophor EL (PAC in Cremophor). [Figure 18] Figure 18 shows the results of an in vivo experiment in mice treated with Cremophor alone (Cremophor) or paclitaxel dissolved in Cremophor in combination with the vehicle used for DF3966A (PAC+Veh DF in Cremophor), DF3966A (PAC+DF in Crem), Avacopan (PAC+Veh AVA in Cremophor), or Avacopan (PAC+AVA in Crem). Efficacy results are shown based on four different parameters measured as described in Example 8: permeability (panel A), vascular leakage (panel B), histamine, and SC5B-9 (panels C and D). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0024] definition According to the present invention, the term "prevention" refers to administration to an individual before the establishment or occurrence of a disorder or pathological event, in order to obtain partial or complete prevention thereof.

[0025] According to one aspect of the invention, the prevention is a total prevention, in which the disorder or pathological event is completely blocked. According to an alternative aspect of the invention, the prevention is partial prevention, in which the onset and / or severity of the disorder or pathological event is delayed.

[0026] According to the present invention, the term "treatment" refers to completely reversing or reducing the severity or progression of a disorder or pathological event after it has been established or has occurred. According to the present invention, the term "individual" refers to a human or an animal, preferably a human.

[0027] In accordance with the present invention, the term "hypersensitivity reaction" or "HSR" is used to refer to an unpredictable adverse reaction to a drug characterized by clinical symptoms resembling an allergic reaction, even in the absence of evidence of an immunological mechanism.

[0028] Detailed Description of the Invention A first aspect of the invention is a C5aR1 inhibitor for use in the prevention or treatment of a hypersensitivity reaction (HSR) to a taxane in an individual.

[0029] Preferably, the taxane is paclitaxel, Nab TM - Selected from paclitaxel, docetaxel and cabazitaxel. The individual is preferably a subject diagnosed with cancer whose treatment of choice is a taxane. Preferably, the cancer is selected from ovarian cancer, breast cancer, non-small cell lung cancer, prostate cancer, head and neck cancer, gastric adenocarcinoma, and AIDS-associated Kaposi's sarcoma.

[0030] According to a first aspect, the C5aR1 inhibitor is used for the prevention of a hypersensitivity reaction to a taxane in the individual, the inhibitor being administered one or more times prior to each administration of the taxane.

[0031] Preferably, the C5aR1 inhibitor is administered once or multiple times within a time frame of 30 minutes to 24 hours prior to administration of the taxane to the individual, more preferably 30 minutes to 2 hours prior to administration of the taxane to the individual.

[0032] In accordance with this embodiment, the individual is preferably a subject at risk of developing a hypersensitivity reaction. Preferably, the individual is a cancer patient with a previous episode of hypersensitivity reaction to a taxane, a positive skin prick test result for sensitivity to said taxane, and / or a history of atopy.

[0033] In accordance with this embodiment, the C5aR1 inhibitor may also be administered in combination with one or more corticosteroids and / or antihistamines. In some cases, the C5aR1 inhibitor and the corticosteroid and / or antihistamine are administered substantially simultaneously. Alternatively, the C5aR1 inhibitor and the corticosteroid and / or antihistamine are administered sequentially, all within a frame of 30 minutes to 24 hours prior to administration of the taxane to the individual.

[0034] Preferably, the corticosteroid and / or antihistamine are those commonly used for premedication with taxanes. Preferably, the C5aR1 inhibitor is administered in combination with dexamethasone, diphenhydramine and ranitidine. More preferably, the C5aR1 inhibitor is administered in combination with oral dexamethasone (8 mg) 12-6 hours prior to taxane injection and intravenous diphenhydramine (2 mg) and ranitidine (50 mg) 30 minutes prior to taxane injection.

[0035] According to an alternative embodiment, the C5aR1 inhibitor is for the acute treatment of a hypersensitivity reaction to a taxane in the individual, the inhibitor being administered at the onset of symptoms of the hypersensitivity reaction, in which case treatment of the individual with the taxane is also immediately discontinued.

[0036] According to this embodiment, the C5aR1 inhibitors may be administered in combination with other appropriate treatments, preferably corticosteroids, dopamine and / or antihistamines. In the most severe cases, epinephrine and / or supplemental oxygen may also be administered.

[0037] The term "C5aR1 inhibitor" according to the present invention means any compound that interacts with C5aR1 and prevents its binding to C5a or blocks the signaling of C5aR1 upon C5a binding. Preferably, said C5aR1 inhibitor is selected from C5aR1 competitive antagonists, anti-C5aR1 antibodies capable of blocking the C5a binding site on the receptor and C5aR1 non-competitive allosteric inhibitors.

[0038] Many different C5aR1 inhibitors conforming to the above definition have been developed and are known to those skilled in the art. According to one preferred embodiment, the C5aR1 antagonist is - (2R,3S)-2-[4-(cyclopentylamino)phenyl]-1-(2-fluoro-6-methylbenzoyl)-N-[4-methyl-3-(trifluoromethyl)phenyl]piperidine-3-carboxamide (Avacopan, Vynpenta®); - N-acetyl-L-phenylalanyl-L-ornityl-L-prolyl-3-cyclohexyl-D-alanyl-L-tryptophyl-L-arginine-N-5.2-C-1.6-lactam (PMX-53); - Acetylated phenylalanine-[Ornityl-Proline-(D)Cyclohexylalanine-Tryptophyl-Arginine]; - L-alanyl-L-seryl-glycyl-L-alanyl-L-prolyl-L-alanyl-L-prolyl-glycyl-L-prolyl-L-alanyl-glycyl-L-prolyl-L-alanyl-glycyl-L-prolyl-L-leucyl-L-arginyl-L-prolyl-L-methionyl-L-phenylalanine; - N,N-bis(1,3-benzodioxol-5-ylmethyl)-N-(1-butyl-2,4-diphenyl-1H-imidazol-5-ylmethyl)amine; - N-[2-(4-chlorophenyl)ethyl]-N-(1,4-dioxaspiro[4.5]dec-8-yl)-2-isobutylbenzamide; - N-[2-(4-chlorophenyl)ethyl]-N-(4-hydroxycyclohexyl)-1-benzothiophene-3-carboxamide; - N-[2-(4-chlorophenyl)ethyl]-N-(4-hydroxycyclohexyl)naphthalene-1-carboxamide; - 2-(2-ethyl-6-methylphenyl)-4-methoxy-N-(5-methoxy-2-methylphenyl)-5,6,7,8-tetrahydroquinolin-5-amine; - 2-(2,6-diethylphenyl)-N-ethyl-4-methoxy-N-(1-naphthyl)-5,6,7,8-tetrahydroquinolin-5-amine; - N-[2,6-dioxohexahydropyrimidin-4(S)-ylcarbonyl]-L-phenylalanyl-L-ornityl-L-prolyl-5-methyl-L-norleucyl-4-fluoro-L-phenylalanyl-L-phenylalaninamide (JPE-1375; JSM-1375); - N-(3-phenylpropionyl)-L-ornityl-L-prolyl-3-cyclohexyl-D-alanyl-L-tryptophyl-L-arginine N-5.1-C-1.5-lactam (PMX-205); - N,N'-bis(4-amino-2-methylquinolin-6-yl)urea (NSC12155); - N-[4-(dimethylamino)benzyl]-N-(4-isopropylphenyl)-7-methoxy-1,2,3,4-tetrahydronaphthalene-1-carboxamide hydrochloride (W54011); - L-phenylalanyl-L-ornithine-L-prolyl-3-cyclohexyl-D-alanyl-L-tryptophyl-L-arginine N-5.2-C-1.6-cyclic peptide; - (4aR,16aS)-6,18-dihydroxy-23(S)-[2(S)-hydroxy-2-[2(R)-hydroxy-6(R)-methyl-5(R)-[2(S)-methylbutyl]tetrahydro-2H-pyran-2-yl]propionamido]-22(S)-isopropyl-7(S),19(R)-dimethyldocosahydro-13H,22H-dipyridazino[6,1-f:6',1'-o][1,4,7,10,13,16]oxapentaazacyclononadecin-5,7,11,17,20,24-hexanone (L-156602) is selected from the group consisting of:

[0039] According to one preferred embodiment, the anti-C5aR1 antibody comprises - Avdoralimab (IPH-5401) - MOR-044254; - NOX-D20; - Anti-C5aR1ab-C5-SiRNA; - m20 / 70 mlgG2a.1; - 3C5 - 7F3 is selected from the group consisting of:

[0040] According to one preferred embodiment, the non-competitive allosteric inhibitor is (2R)-2-[3-(furan-2-carbonyl)phenyl]-N-[4-(trifluoromethyl)-1,3-thiazol-2-yl]propenamide (DF2427); and A compound having the general formula (I) or a pharma- ceutically acceptable salt thereof:

[0041] [ka]

[0042] [In the formula, i) R is - 2-thiazolyl or 2-oxazolyl (unsubstituted or substituted by a radical selected from the methyl, tert-butyl or trifluoromethyl radical); - C(Ra)=NW (W is linear or branched C1-C4 alkyl); - CORa, SORa, SO2Ra, PORa, PO2Ra Selected from In the above formula, Ra is, - C1-C5-alkyl, C3-C6-cycloalkyl, C2-C5-alkenyl, unsubstituted phenyl or substituted phenyl which is substituted by a group selected from halogen, C1-C4-alkyl, C1-C4-alkoxy, halo-C1-C4-alkoxy, hydroxy, C1-C4-acyloxy, phenoxy, cyano, nitro, amino; heteroaryl groups selected from pyridine, pyrimidine, pyrrole, thiophene, furan, indole, thiazole, oxazole, said heteroaryl being unsubstituted or substituted by groups selected from halogen, C1-C4-alkyl, C1-C4-alkoxy, halo-C1-C4-alkoxy, hydroxy, C1-C4-acyloxy, phenoxy, cyano, nitro, amino; - α or β carboxyalkyl residues (consisting of linear or branched C1-C6-alkyl, C3-C6-cycloalkyl, C2-C6-alkenyl, C1-C6-phenylalkyl, optionally substituted by further carboxy (COOH) groups); an ω-aminoalkylamino group of formula (II):

[0043] [ka]

[0044] {In the formula (II), X is - linear or branched C1-C6 alkylene, C4-C6 alkenylene, C4-C6 alkynylene, optionally substituted by a) a CO2R4 group, where R4 represents hydrogen or a linear or branched C1-C6 alkyl group or a linear or branched C2-C6 alkenylene group, or b) a CONHR5 group, where R5 represents hydrogen, a linear or branched C2-C6 alkyl group or an OR4 group, where R4 is as defined above; - (CH2) m -B-(CH2) n a) B is an oxygen, sulfur or nitrogen atom (optionally substituted with a C1-C4 alkyl group), m is 0 or an integer from 2 to 3, and n is an integer from 2 to 3, or b) B is a CO, SO or CONH group, m is an integer from 1 to 3, and n is an integer from 2 to 3. and; R2 and R3 are independently hydrogen, a straight or branched C1-C6 alkyl (optionally interrupted by oxygen or sulfur atoms), a C3-C7 cycloalkyl, a C3-C6 alkenyl, a C3-C6-alkynyl, an aryl-C1-C3-alkyl, a hydroxy-C2-C3-alkyl group; or R2 and R3, together with the N atom to which they are attached, form a group of formula (III):

[0045] [ka]

[0046] (In the above formula, Y is a single bond, CH2, O, S or a group N-R6, where R6 represents hydrogen, C1-C4 alkyl, C1-C4 acyl, unsubstituted phenyl or substituted phenyl, which is substituted by a group selected from halogen, C1-C4 alkyl, C1-C4 alkoxy, hydroxy, C1-C4 acyloxy, phenoxy, cyano, nitro, amino, and p represents an integer from 0 to 3; - residues of the formula SO2R7, where R7 is C1-C6-alkyl, C3-C6-cycloalkyl, C2-C6-alkenyl, aryl and heteroaryl; (represents or form a 3- to 7-membered nitrogen heterocyclic ring; or In the formula (II), X together with the nitrogen atom to which it is attached and the R2 group form a nitrogen-containing 3-7 membered heterocyclic, monocyclic or polycyclic ring, and R3 represents hydrogen, C1-C4 alkyl, C1-C4 acyl, unsubstituted phenyl or substituted phenyl substituted by a group selected from halogen, C1-C4 alkyl, C1-C4 alkoxy, hydroxy, C1-C4 acyloxy, phenoxy, cyano, nitro, amino. Selected from; ii) R1 is a linear or branched C1-C5 alkyl, C3-C5 cycloalkyl; iii) Ar is - an unsubstituted phenyl group or a substituted phenyl group, which is substituted by one or more groups independently selected from halogen, C1-C4-alkyl, C1-C4-alkoxy, hydroxy, C1-C4-acyloxy, phenoxy, cyano, nitro, amino, C1-C4-acylamino, halo-C1-C3-alkyl, halo-C1-C3-alkoxy, benzoyl, heteroarylcarbonyl, heteroaryl, linear or branched C1-C8-alkanesulfonate, linear or branched C1-C8-alkanesulfonamido, linear or branched C1-C8 alkylsulfonylmethyl; or - a heteroaryl ring selected from pyridine, pyrrole, thiophene, furan, and indole; is] Selected from.

[0047] Among the above compounds, particularly preferred are R, - 2-thiazolyl or 2-oxazolyl (unsubstituted or substituted by a radical selected from the methyl, tert-butyl or trifluoromethyl radical); - C(Ra)=NW (W is linear or branched C1-C4 alkyl); CORa, SORa or SO2Ra, where Ra is as defined above. Selected from; and Ar, 3'-Benzoylphenyl, 3'-(4-chloro-benzoyl)-phenyl, 3'-(4-methyl-benzoyl)-phenyl, 3'-acetyl-phenyl, 3'-propionyl-phenyl, 3'-isobutanoyl-phenyl, 4'-isobutyl-phenyl, 4'-trifluoromethanesulfonyloxy-phenyl, 4'-benzenesulfonyloxy-phenyl, 4'-trifluoromethanesulfonylamino-phenyl, 4'-benzenesulfonylamino-phenyl, 4'-benzenesulfonylmethyl-phenyl, 4'-acetoxyphenyl, 4'-propionyloxy-phenyl, 4'-benzoyloxy-phenyl, 4'-acetylamino-phenyl, 4'-propionylamino-phenyl, 4'-phenyloxy ... '-Benzoylamino-phenyl, 3'-(furan-2-carbonyl)-phenyl, 3'-(benzofuran-2-carbonyl)-phenyl, 3'-(thiophene-2-carbonyl)-phenyl, 3'-(pyridine-2-carbonyl)-phenyl, 3'-(thiazole-2-carbonyl)-phenyl, 3'-(oxazole-2-carbonyl)-phenyl, 3'-(2-furyl)-phenyl, 3'-(2-oxazolyl)-phenyl, 3'-(3-isoxazolyl)-phenyl, 3'-(2-benzoxazolyl)-phenyl, 3'-(3-benzoisoxazolyl)-phenyl, 3'-(2-thiazolyl)-phenyl, 3'-(2-pyridyl)-phenyl, 3'-(2-thiophenyl)-phenyl Selected from; or Ar is a heteroaryl ring selected from pyridine, pyrrole, thiophene, furan, or indole; The compound of formula (I) or a pharma- ceutically acceptable salt thereof.

[0048] Preferred compounds of formula (I) according to the invention are - 4-{(1R)-1-[(phenylsulfonyl)amino]ethyl}phenyl trifluoromethanesulfonate; - N-[(1R)-1-(3-benzoylphenyl)ethyl]benzenesulfonamide; - 4-{(1R)-1-[(pyridin-3-ylsulfonyl)amino]ethyl}phenyl trifluoromethanesulfonate; - N-[(1R)-1-(3-benzoylphenyl)ethyl]methanesulfonamide; - N-{(1R)-1-[3-(2-furoyl)phenyl]ethyl}thiophene-2-sulfonamide; - N-{(1R)-1-[3-(2-furoyl)phenyl]ethyl}methanesulfonamide; - 4-{(1R)-1-[(thien-2-ylsulfonyl)amino]ethyl}phenyl trifluoromethanesulfonate; - N-[(1R)-1-(3-benzoylphenyl)ethyl]thiophene-2-sulfonamide; - N-[(1R)-1-(3-benzoylphenyl)ethyl]-3-pyrrolidin-1-ylpropane-1-sulfonamide; - methyl 5-({[(1R)-1-(3-benzoylphenyl)ethyl]amino}sulfonyl)-2-furoate; - 5-({[(1R)-1-(3-benzoylphenyl)ethyl]amino}sulfonyl)-2-furoic acid; - 4-{(1R)-2-methyl-1-[(methylsulfonyl)amino]propyl}phenyl trifluoromethanesulfonate; - N-((1R)-1-{4-[1-methyl-1-(phenylsulfonyl)ethyl]phenyl}ethyl)methanesulfonamide; - 4-[(1R)-1-(isobutyrylamino)ethyl]phenyl trifluoromethanesulfonate; - 4-{[(1R)-1-(pyridin-3-ylcarbonyl)amino]ethyl]}phenyl trifluoromethanesulfonate; - N-[(1R)-1-(3-benzoylphenyl)ethyl]benzamide; - N-[(1R)-1-(3-benzoylphenyl)ethyl]-2-furamide; - N-[(1R)-1-(3-benzoylphenyl)ethyl]cyclobutanecarboxamide; - N-[(1R)-1-(4-trifluoromethanesulfonyloxy)phenylethyl]-4-piperidin-1-yl butanamide (DF2593Y); - 4-{(1R)-1-[(4-pyrrolidin-1-ylbutanoyl)amino]ethyl]}phenyl trifluoromethanesulfonate; - 3-{(1R)-1-[4-(4-trifluoromethyl-1,3-thiazol-2-yl)amino]ethyl}phenyl)(phenyl)methanone; - R(-)-2-[(4'-trifluoromethanesulfonyloxy)phenyl]-N-[3-(N'-pyrrolidinyl)propyl]propionamide (DF2297X) or its chloride salt (DF2297A); and - 5-[(1R)-1-(4-{[4-(trifluoromethyl)-1,3-thiazol-2-yl]amino}phenyl)ethyl]tetrazol-2-ide (DF3966Y) or its sodium salt (DF3966A) Selected from.

[0049] Particularly preferred compounds of formula (I) according to the invention are selected from N-[(1R)-1-(4-trifluoromethanesulfonyloxy)phenylethyl]-4-piperidin-1-yl butanamide (DF2593Y) and its pharma- ceutically acceptable salts, in particular its chloride salt (DF2593A), R(-)-2-[(4'-trifluoromethanesulfonyloxy)phenyl]-N-[3-(N'-pyrrolidinyl)propyl]propionamide (DF2297X) or its chloride salt (DF2297A), and 5-[(1R)-1-(4-{[4-(trifluoromethyl)-1,3-thiazol-2-yl]amino}phenyl)ethyl]tetrazol-2-ide (DF3966Y) or its sodium salt (DF3966A).

[0050] Compounds of formula (I) are disclosed in WO2007 / 060215, which also discloses their synthesis, their activity as C5aR inhibitors and their use in the treatment of diseases dependent on activation of the C5a pathway.

[0051] Preferred C5aR1 inhibitors according to the present invention are Avacopan, PMX-53, W54011, Avdoralimab, MOR-044254, PMX-205, DF2593A, DF2297A, DF2427 and DF3966A.

[0052] The C5aR1 antagonists or allosteric inhibitors of the present invention may form stable pharma- ceutically acceptable salts with pharma-ceutically acceptable organic or inorganic acids or bases, and in such cases it may be appropriate to administer the C5aR1 inhibitor compounds as salts.

[0053] Examples of acid addition salts are acetate, adipate, ascorbate, benzoate, benzenesulfonate, bicarbonate, hydrogensulfate, butyrate, camphorate, camphorsulfonate, choline, citrate, cyclohexylsulfamate, diethylenediamine, ethanesulfonate, fumarate, glutamate, glycolate, hemisulfate, 2-hydroxyethylsulfonate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, hydroxymaleate. Salts in the form of salts include, but are not limited to, phenylacetate, lactate, malate, maleate, methanesulfonate, meglumine, 2-naphthalenesulfonate, nitrate, oxalate, pamoate, persulfate, phenylacetate, phosphate, diphosphate, picrate, pivalate, propionate, quinate, salicylate, stearate, succinate, sulfamate, sulfanilate, sulfate, tartrate, tosylate (p-toluenesulfonate), trifluoroacetate, and undecanoate.

[0054] Examples of base addition salts are ammonium salts, alkali metal salts such as sodium, lithium and potassium salts, alkaline earth metal salts such as aluminum, calcium and magnesium salts, salts with organic bases such as dicyclohexylamine and N-methyl-D-glucamine, and salts with amino acids such as arginine, lysine, ornithine, etc. Basic nitrogen-containing groups may also be quaternized with agents such as lower alkyl halides, e.g., methyl, ethyl, propyl and butyl halides, dialkyl sulfates, e.g., dimethyl, diethyl, dibutyl, diamyl sulfate, long chain halides, e.g., decyl, lauryl, myristyl and stearyl halides, aryl alkyl halides, e.g., benzyl bromide and others. Non-toxic physiologically acceptable salts are preferred.

[0055] The salts can be formed by conventional means, such as by reacting the free product with one or more equivalents of the appropriate acid or base in a solvent or medium in which the salt is insoluble (e.g., water or ethanol), which is then removed under vacuum or by lyophilization.

[0056] The present invention also includes prodrugs, stereoisomers, isotopically labelled, eg deuterated, derivatives and enantiomers of the above C5aR1 inhibitors. As used herein, the term "prodrug" refers to an agent that is converted to the parent compound in vivo by some physiological process (e.g., the prodrug is converted to the desired drug form when exposed to physiological pH). Prodrugs are often useful because, in some situations, they may be easier to administer than the parent drug. For example, they may be bioavailable by oral administration whereas the parent drug is not. Prodrugs may also have improved solubility in pharmacological compositions over the parent drug.

[0057] Prodrugs have many useful properties. For example, they may be more soluble in water than the final drug, facilitating intravenous administration of the drug. Prodrugs may also have a higher level of oral bioavailability than the final drug. After administration, the prodrug is enzymatically or chemically cleaved to deliver the final drug into the blood or tissues.

[0058] Ester prodrugs of the compounds disclosed herein are specifically contemplated. For example, a prodrug would be a compound of the invention that is administered in the form of an ester (the "prodrug") to facilitate passage across a cell membrane where water solubility is disadvantageous, but is metabolically hydrolyzed once inside a cell where water solubility is advantageous.

[0059] Without intending to be limiting, the ester can be an alkyl ester, an aryl ester, or a heteroaryl ester. The term alkyl has the meaning commonly understood by those of ordinary skill in the art and refers to straight-chain, branched, or cyclic alkyl moieties. 1-6 Alkyl esters are particularly useful, where the alkyl portion of the ester has 1 to 6 carbon atoms and includes, but is not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, iso-butyl, t-butyl, pentyl isomers, hexyl isomers, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and combinations thereof.

[0060] Certain compounds can exist in tautomeric forms, and the present invention includes all such tautomeric forms of those compounds unless otherwise specified. Unless otherwise indicated, a structure depicted herein is intended to include all stereochemical forms of the structure, i.e., the R and L configurations for each asymmetric center. Thus, single stereochemical isomers of the compounds of the present invention as well as enantiomeric and diastereomeric mixtures are included within the scope of the present invention. Thus, the present invention encompasses each diastereomer or enantiomer substantially free of the other isomer (>90%, preferably >95% free of the other stereoisomer on a molar basis), as well as mixtures of such isomers in any ratio.

[0061] Optical isomers can be obtained by resolving the racemic mixtures according to conventional methods, for example. by formation of diastereomeric salts, by treatment with optically active acids or bases, or enzymatically. Examples of suitable acids are tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, ditoluoyltartaric acid, and camphorsulfonic acid, followed by separation of the mixture of diastereomers by crystallization, followed by liberation of the optically active bases from these salts. Another method for separating optical isomers includes the use of chiral chromatographic columns optimally selected to maximize the separation of the enantiomers. Yet another method includes the synthesis of covalent diastereomers by reacting the compounds of the invention with an optically pure activated acid or an optically pure isocyanate. The synthesized diastereomers can be separated by conventional means such as chromatography, distillation, crystallization, or sublimation, followed by hydrolysis to give enantiomerically pure compounds. The optically active compounds of the invention can also be obtained using active starting materials. These isomers can be in the form of free acids, free bases, esters, or salts.

[0062] The C5aR1 inhibitors of the present invention may be in amorphous or crystalline form, including any polymorphic form. Typically, C5aR1 inhibitors for use in accordance with the present invention will be administered in the form of a pharmaceutical composition.

[0063] Thus, a second aspect of the present invention relates to a pharmaceutical composition comprising a C5aR1 inhibitor (as defined above) and at least one pharma- ceutically acceptable excipient for use in the prevention or treatment of hypersensitivity reactions (HSR) to taxanes in an individual as described above.

[0064] Preferably, the pharmaceutical composition of the present invention is prepared into a suitable dosage form containing an effective amount of a C5aR1 inhibitor, a pharma- ceutically acceptable salt or a prodrug thereof, and at least one pharma- ceutically acceptable excipient.

[0065] Administration of the pharmaceutical compositions of the present invention to a patient may be in accordance with known methods and may include oral administration, parenteral administration, preferably parenteral administration selected from intravenous, intraperitoneal, intramuscular, intraarterial, subcutaneous, topical, buccal, or rectal (suppository) administration.

[0066] In this description and in the claims that follow, the expression "effective amount" refers to the administration of a drug compound sufficient to significantly achieve a desired clinical response. The dosage and treatment regimen of a C5aR1 inhibitor for use for a particular patient in accordance with the present invention will vary depending on a number of factors that are within the knowledge and expertise of one of ordinary skill in the art, including, for example, the inhibitory concentration (IC 50 ) and half-life, the formulation and route of administration used, the patient's age, weight, general health, sex, and diet.

[0067] As described herein, the pharmaceutical compositions of the present invention comprise a C5aR1 inhibitor together with a pharma- ceutically acceptable excipient, which as used herein includes any and all solvents, diluents or other vehicles, dispersing or suspending aids, surfactants, isotonicity agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, and the like, appropriate for the particular dosage form desired.

[0068] Some examples of substances that can function as pharma- ceutically acceptable excipients include, but are not limited to, sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; sterile water; Ringer's solution; buffered saline; dextrose solutions; maltodextrin solutions; ethyl alcohol; and phosphate buffers.

[0069] Furthermore, the composition of the present invention can be formulated into inhalable or injectable dosage forms such as solutions, suspensions, and emulsions by further adding diluents, dispersants, and surfactants.

[0070] Furthermore, the compositions of the present invention may be suitably formulated using any suitable method known in the art or by the methods disclosed in Remington's Pharmaceutical Sciences (latest edition), Mack Publishing Company, Easton, Pa.

[0071] The terms "pharmaceutical acceptable" and "physiologically acceptable" are intended to define, without limitation, any non-drug material that is suitable for preparing a pharmaceutical composition to be administered to an organism.

[0072] The dosage forms may also contain other conventional ingredients such as preservatives, stabilizers, surfactants, buffers, osmotic pressure regulating agents, emulsifiers, sweeteners, colorants, flavoring agents, and the like. The dosage forms of the pharmaceutical compositions of the present invention can be manufactured by techniques such as mixing, granulating, compressing, dissolving, sterilizing and the like, which are well known to pharmaceutical chemists.

[0073] In a further aspect, the present invention relates to a method for the prevention or treatment of a hypersensitivity reaction (HSR) to a taxane in an individual, the method comprising the step of administering to said individual a C5aR1 inhibitor as defined above.

[0074] Preferably, in the method according to the invention, said C5aR1 is administered in the form of a pharmaceutical composition as defined above. The invention is further described in the following examples, which do not limit the scope of the invention, which is defined in the claims. EXAMPLES

[0075] Experimental section Examples 1 to 3 BIAcore technology was used to analyze the interaction between paclitaxel and complement component 5a receptor 1 (C5aR1) (Example 1), complement component 5a receptor 2 (C5AR2) (Example 2) or complement component 5a (Example 3).

[0076] The BIAcore system utilizes the phenomenon of surface plasmon resonance (SPR) to monitor interactions between molecules in real time. Briefly, the SPR phenomenon occurs at a thin conductive film at the interface between media of different refractive index. In the BIAcore system, the media are the glass of the sensor chip and the sample solution, and the conductive film is a thin layer of gold on the sensor chip surface. BIAcore monitors the interaction between two molecules, one called a ligand that is bound to the sensor surface, and the other called an analyte that is free in solution.

[0077] Example 1 Binding of paclitaxel to C5aR1 a) Immobilization of C5aR1 on the sensor chip CM5 Anti-c-Myc antibody (9E10, sc-40, Santa Cruz Technology) was immobilized on a sensor chip CM5 (#29149604, Cytiva). Specifically, the antibody was diluted to a concentration of 50 μg / ml in 10 mM acetate buffer at pH 4.5 and injected over the entire surface of the sensor chip. Specifically, the dextran matrix on the sensor chip surface was first activated with a mixture of 1-ethyl-3-carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to form reactive succinimide esters. The anti-Myc antibody was then flowed over the surface, where the esters reacted spontaneously with the uncharged amino groups, covalently binding the ligand to the dextran. The surface was then inactivated with ethanolamine-HCl.

[0078] Covalent immobilization of anti-c-Myc antibodies by amine coupling chemistry resulted in stable binding of the ligand to the surface, measured at 2782 resonance units (RU). Once the antibody was immobilized, the receptor was captured on the sensor chip surface. This procedure allowed for directional immobilization of the ligand of interest. Specifically, C5AR1-cMyc (TP303784, OriGene) at a concentration of 20 μg / ml in running buffer (50 mM HEPES buffer, pH 7.0, containing 0.01% CHS, 0.1% CHAPS, and 0.33 mM lipid) was injected over the sensor chip prepared as described above at a flow rate of 5 μl / min.

[0079] As shown in Figure 1, this method resulted in stable binding of 77 RU of C5aR1-cMyc to the surface. b) Binding of C5a to immobilized C5aR1 To verify that the receptor was immobilized in the correct conformational state, binding of the receptor's natural ligand, C5a, was analyzed. Specifically, 5 μM C5a (A144, Complement Technologies) in running buffer was injected over the C5aR1 surface of the sensor chip prepared in Example 1a).

[0080] As shown in Figure 2, C5a was able to bind to the C5aR1 receptor, confirming the correct conformational state of the orientationally immobilized receptor. c) Binding of paclitaxel to immobilized C5aR1 To initially assess the direct binding of paclitaxel to C5aR1, the compound (paclitaxel) was injected over the C5aR1 surface of the sensor chip prepared in Example 1a) at a concentration of 300 μM in running buffer.

[0081] As shown in FIG. 3, paclitaxel binds to the receptor with 300 RU at the concentrations tested. To determine the affinity of the interaction, paclitaxel was diluted in running buffer at various concentrations (4.06–37.5 μM) and injected over the sensor chip for 4 min, followed by dissociation for 5 min in running buffer without paclitaxel, and the kinetic constant was evaluated as a function of time.

[0082] As shown in FIG. 4, binding of paclitaxel to the receptor was dose-dependent with a Kd value of 670 nM. The affinity value of the paclitaxel / Myc-C5aR1 interaction at equilibrium was also determined by measuring the steady-state binding levels by Scatchard analysis of the RU values ​​at equilibrium. The BIAcore provided an affinity constant of 525 nM as a function of analyte concentration in solution, as shown in Figure 5.

[0083] Example 2 Binding of paclitaxel to C5aR2 a) Immobilization of C5aR2 on the sensor chip CM5 Polyclonal anti-His antibody (ab137839, Abcam) was immobilized on a sensor chip CM5 (#29149604, Cytiva). Specifically, the antibody was diluted to a concentration of 50 μg / ml in 10 mM acetate buffer, pH 4.0, and injected over the surface of the sensor chip. Activation and deactivation of the sensor chip were performed as described in Example 1a).

[0084] Covalent immobilization of anti-His antibody by amine coupling chemistry resulted in stable binding of 2510 RU of ligand to the surface. Once the antibody was immobilized, the receptor was orientated and captured on the sensor chip surface. Specifically, C5AR2-His (CSB-CF868390HU, CUSABIO) at a concentration of 100 μg / ml was injected over the sensor chip at a flow rate of 5 microliters / min in running buffer.

[0085] As shown in FIG. 6, this method results in stable binding of 569 RU of C5aR2 to the surface. b) Binding of C5a to immobilized C5aR2 Again, to demonstrate that the receptor was immobilized in the correct conformational state, binding of the receptor's natural ligand, C5a, was analyzed: 5 μM C5a (A144, Complement Technologies) was injected over the C5AR2 surface of the sensor chip prepared in Example 2a).

[0086] As shown in Figure 7, C5a was able to bind to the C5aR2 receptor immobilized on the sensor chip, confirming that C5aR2 was immobilized in the correct conformation. c) Binding of paclitaxel to immobilized C5aR2 Paclitaxel was injected at various concentrations (10, 100 and 200 microM) in the running buffer onto the C5aR2 surface of the sensor chip prepared in Example 2a) and the amount of RU obtained was evaluated.

[0087] As shown in FIG. 8, paclitaxel was unable to bind to C5aR2 at any concentration tested. Example 3 Binding of paclitaxel to C5a a) Immobilization of C5a on the sensor chip CM5 C5a was immobilized on a sensor chip (#29149604, Cytiva). In detail, 20 μg / ml of recombinant protein C5a was diluted in 10 mM acetate buffer at pH 4.5 and injected over the surface of the sensor chip. Activation and deactivation of the sensor chip were performed as described in Example 1a).

[0088] Covalent immobilization of C5a by amine coupling chemistry resulted in stable binding of 1330 RU of C5a to the surface. b) Binding of paclitaxel to immobilized C5a Paclitaxel was injected at a concentration of 300 microM in running buffer onto the C5A surface of the sensor chip prepared in Example 2a) and the RUs can be detected by BIAcore.

[0089] As shown in FIG. 9, no RU was detectable by BIAcore, indicating that paclitaxel is unable to bind to C5a at the high concentrations tested. Example 4 RAW264.7 (ATCC TIB-71) mouse macrophages were cultured in DMEM medium (Euroclone, Milan, Italy) supplemented with 10% FBS (non-heat inactivated) (ATCC, USA) according to the manufacturer's instructions at 37 °C in a humidified 95% air-5% CO2 atmosphere, at a density of 1 × 104 cells / cm. 2 The seeds were sown in.

[0090] After 24 h, cells were treated with C5a (diluted in medium) (10 nM, R&D, USA), paclitaxel (diluted in medium) (10 nM, Sigma, USA) in the absence and presence of the C5a inhibitor DF3966A (10 μM), and anti-C5aR Ab (BioLegend, CA, USA; 50 nM) or Avacopan (10 μM) for 24 h, and then the medium was replaced with culture medium (5 ml) for 24 h and then harvested.

[0091] Conditioned media was centrifuged to remove particulate matter and assayed immediately. Sample volume was adjusted as suggested (500 μl). Reagents were prepared according to the manufacturer's protocol (R&D, USA). Briefly, membranes were incubated with Array Buffer 6 for 1 h on a rocking platform shaker. Samples were prepared by adding up to 1 ml of Array Buffer 4 to two separate tubes, then 15 microliters of reconstituted mouse cytokine detection antibody cocktail (>40 mouse cytokines) was added to each prepared sample and incubated for 1 h. Array Buffer 6 was then aspirated and replaced with the sample / antibody mix and incubated overnight at 4°C. The following day, membranes were washed three times and incubated with streptavidin-HRP (1:2000) for 30 min at room temperature on a rocking platform shaker. Membranes were washed again and 1 ml of prepared Chemi Reagent mix was sprayed onto each membrane. Multiple exposure times were acquired using a UVITEC digital analyzer (Alliance, Cambridge, UK).

[0092] Positive signals seen on the developed membrane can be identified by placing a transparent overlay template onto the array image and aligning it with pairs of reference spots in the three corners of each array (see Figure 10). The reference spots are included to indicate that the arrays were incubated with streptavidin-HRP during the assay procedure. Pixel density (average signal of duplicate pairs representing each cytokine) was analyzed by ImageJ and the average background was subtracted from each spot.

[0093] Figure 11 shows the proteomic profiling of cytokines released from RAW264.7 cells treated with paclitaxel (PAC) or C5a for 24 hours, which shows overlap of all cytokines analyzed except C5a, which is only released by RAW264.7 cells when stimulated with C5a but not paclitaxel. Furthermore, induction of cytokine release by C5a and paclitaxel is inhibited by DF3966A, C5aR Ab, and Avacopan, confirming overlap in activation of C5aR1 receptor by these two molecules.

[0094] Example 5 F11 hybridoma cells (ECACC 08062601), selected as an alternative model for DRG neurons, were cultured in DMEM medium (Euroclone, Milan, Italy) supplemented with 10% FBS (Sigma-Aldrich, St. Louis, CO, USA), 1% penicillin / streptomycin (Euroclone), and 1% glutamine (Euroclone) at 37°C in a humidified 95% air-5% CO2 atmosphere. Cells from passage 18 were used in the experiments, with a density of 1x104 cells / cm. 2 The seeds were sown in.

[0095] After 24 hours, cells were differentiated with rat NGF (rNGF) (Sigma). rNGF was dissolved in DMEM (without FBS) with 1% penicillin / streptomycin and 1% glutamine at a final concentration of 50 ng / ml. Medium was changed every 3 days until complete differentiation, which occurred after 7 days.

[0096] After neuronal differentiation, neurons were treated with paclitaxel (Sigma-Aldrich; final concentration 10 nM) or C5a (R&D Systems, Inc., Minnesota, USA; final concentration 50 nM) for 20 h.

[0097] RT-PCR The following protocol was used for gene expression analysis. Total RNA was extracted by Trizol reagent according to the manufacturer's instructions. Total RNA concentration was determined spectrophotometrically in RNAse-free water, and 1 μg aliquots of total RNA were reverse transcribed into cDNA using the ProtoScript First Strand cDNA Synthesis Kit (NEB). RT-PCR was performed on an ABI 7300HT Sequencing System (ABI) in a total volume of 20 ml containing EagleTaq Universal Master Mix (Roque), DEPC water, and 4 microliters of cDNA, and the following Prime Time qPCR Assay for IL-8 Mm04207460m1 (Applied Biosystem, USA). To normalize the expression of the target genes, the reference gene GADPH ID Mm04207460g1 (Applied Biosystem, USA) was used as an internal control (endogenous control). The protocol for RT-PCR was as follows: A preheating step of 3 min at 95°C, 40 cycles of 10 s at 95°C and 30 s at 60°C, and a final step of 10 s at 65°C. Relative expression levels were calculated for each sample using the ΔΔCt method for comparing relative fold expression differences after normalization to a reference gene as previously reported [Livak et al., 2001 DOI:10.1006 / meth.2001.1262]. Figure 12 shows the results of real-time PCR analysis of the relative mRNA expression of IL-8 in F11 cells treated for 20 h with 10 nM paclitaxel (PAC) or 50 nM C5a. In agreement with the results obtained with macrophages, both treatments increase the IL-8 expression level of differentiated F11. C5a increases the IL-8 expression level, sharing the exact molecular mechanism of paclitaxel.

[0098] The results obtained in these examples demonstrate that paclitaxel can bind to C5aR1 and activate the same C5a signaling pathway in RAW264 murine macrophages, and share the same molecular mechanism of C5a in F11 cells.

[0099] Example 6 Inhibition of C5aR1 prevents paclitaxel- or C5a-induced cytokine release in macrophages Using mouse (RAW264.7) and human (THP-1) macrophages, it was shown that C5aR1 mediates paclitaxel-induced HSR. Specifically, cytokines (>40 cytokines) released from RAW264.7 and human macrophages challenged with paclitaxel (Sigma-Aldrich; 10 nM) or C5a (R&D Systems, Inc. Minnesota, USA; 10 nM) for 24 h were profiled using the Proteome Profiler assay. In both treatments, the mouse and human proteome profiles resembled those of typical anaphylaxis, with overlap of all cytokines analyzed, except for C5a, which was released only by C5a-stimulated cells (Figures 13 and 15). The experimental assay is described in Example 4.

[0100] For human macrophages, Human XL Cytokine Array Kit #Ary022B was used, and for mouse macrophages, Mouse Cytokine Array Panel A #Ary006 was used (R&D systems, USA).

[0101] In RAW264.7 cells treated with paclitaxel and C5a, the levels of monocyte chemoattractant protein-1 (MCP-1) were significantly increased compared to the control group, as were the levels of macrophage inflammatory protein (MIP)-1α and MIP-1β, as well as MIP-2 and IL1Ra (Figures 13A-B).

[0102] The release of these cytokines was prevented by inhibition of C5aR1 mediated by Avacopan (Selleckchem, USA), anti-C5aR1 antibody (BioLegend, CA, USA) or DF3966A (Figure 13A-B). Analysis of human macrophages confirmed that the data obtained with mouse macrophages as a profile of cytokines released by C5a overlapped with the data obtained with paclitaxel stimulation, and also that under these experimental conditions, Avacopan (Selleckchem, USA), anti-C5aR1 antibody (BioLegend, CA, USA) or DF3966A effectively blocked C5a and paclitaxel-induced cytokine release (Figure 15A-B).

[0103] Example 7 Therefore, Balb / c mice were divided into four groups and treated with paclitaxel in Cremophor EL (20 mg / kg and 0.1 mL / Kg, iv), paclitaxel in DMSO / PEG / TWEEN (20 mg / kg and 0.1 mL / Kg, iv), Cremophor EL alone (0.1 mL / Kg, iv), or DMSO / PEG / TWEEN alone (0.1 mL / Kg, iv). Mice treated with paclitaxel in Cremophor EL or DMSO / PEG / TWEEN developed a more severe acute HSR, exhibiting greater vascular leakage (hyperpermeability) and plasma extravasation, compared with mice treated with both vehicles alone (Figures 17A and B), confirming that paclitaxel itself plays an important role in inducing the HSR.

[0104] Example 8 Inhibition of C5aR1 prevents paclitaxel-induced HSR in vivo Having identified paclitaxel as a key factor involved in the HSR-specific cytokine release induced by chemotherapy treatment, the potential efficacy of inhibition of C5aR1 as a preventive treatment was investigated.

[0105] Paclitaxel (Selleckchem, USA, cod. S1150) was dissolved in Cremophor EL (Polyoxyl 35 Castor Oil, Sigma-Aldrich, cod. 238470) and administered intravenously at 20 mg / kg (0.1 mL / Kg). DF3966A (30 mg / kg, os) or vehicle (saline) for DF3966A (PAC+DF in Crem and PAC+Veh DF in Cremophor, respectively) was administered 3 hours before paclitaxel. Meanwhile, the vehicle used for Avacopan (PEG-400 / Solutol-HS15 70:30, 5 mL / kg) was administered intravenously at 20 mg / kg (0.1 mL / Kg). 50 ) (PAC+Veh AVA in Cremophor) or Avacopan (30 mg / kg, os) (PAC+AVA in Crem) were administered 1 hour prior to paclitaxel according to their pharmacokinetic profiles. Cremophor alone (Cremophor) was used as control.

[0106] a) Assessment of vascular leakage To evaluate the cutaneous manifestations of HSR induced by paclitaxel injection, the ears were visualized as blue using Evans Blue (EB, 0.8% / 0.3 mL / iv) and the degree of vascular leakage in the ears was quantified. Thirty minutes after administration of paclitaxel, mice were administered EB. The degree of ear coloration was evaluated using a score of 0 to 5. "0" represents no visible blue areas, and "1 to 5" indicates the percentage of visible blue areas.

[0107] b) Plasma extravasation Approximately 40 minutes after paclitaxel administration, the animals were sacrificed and the ears were excised, minced, and stored in 2 mL of formaldehyde overnight at room temperature. The next day, the ears were centrifuged and the supernatant was removed and analyzed by spectrophotometer (620 nm OD).

[0108] c) ELISA Assay Intracardiac injection was performed and blood was collected in test tubes and left at room temperature. After 30 min, serum was separated from blood cells by centrifugation (2000×g for 10 min, 4° C.). Aliquots were prepared and stored at −20° C. The following markers were evaluated using ELISA kits: histamine and SC5b-9 (Mybiosource, San Diego, USA, Cat. Nos. MBS9361379, MBS725193).

[0109] d) Results Pharmacological inhibition of C5aR1 by both treatments effectively prevented the development of HSR and abolished the paclitaxel-induced increase in vascular leakage and plasma extravasation (Figure 18A and B, respectively). Furthermore, pretreatment with DF3966A or Avacopan significantly reduced the increase in serum concentrations of both histamine and complement 5b-9 (SC5b-9) in paclitaxel-treated mice (Figure 18C and D, respectively), indicating that inhibition of C5aR1 is an effective pharmacological countermeasure for the prevention of paclitaxel-induced HSR.

Claims

1. A pharmaceutical composition comprising a C5aR1 inhibitor for use in the prevention or treatment of hypersensitivity reactions (HSR) to taxanes in an individual.

2. The taxane is paclitaxel, Nab ＴＭ The pharmaceutical composition according to claim 1, selected from paclitaxel, docetaxel and cabazitaxel.

3. A pharmaceutical composition as described in claim 1 or 2, for use in preventing a hypersensitivity reaction to a taxane in the individual, wherein the inhibitor is administered one or more times prior to each administration of the taxane.

4. 4. The pharmaceutical composition of claim 3, wherein the C5aR1 inhibitor is administered one or more times 30 minutes to 24 hours prior to administration of a taxane to the individual.

5. 5. The pharmaceutical composition of claim 3 or 4, wherein the individual is a cancer patient with a previous episode of HSR to a taxane, a positive skin prick test result for sensitivity to the taxane, and / or a history of atopy.

6. The pharmaceutical composition of claim 3, wherein the C5aR1 inhibitor is administered in combination with one or more corticosteroids and / or antihistamines.

7. A pharmaceutical composition described in claim 1 or 2, for use in the acute treatment of a hypersensitivity reaction to a taxane in the individual, wherein the inhibitor is administered at the onset of symptoms of the hypersensitivity reaction.

8. The pharmaceutical composition according to any one of claims 1 to 7, wherein the C5aR1 inhibitor is selected from a C5aR1 competitive antagonist, an anti-C5aR1 antibody capable of blocking the C5a binding site on the receptor, and a C5aR1 non-competitive allosteric inhibitor.

9. The C5aR1 inhibitor is - (2R,3S)-2-[4-(cyclopentylamino)phenyl]-1-(2-fluoro-6-methylbenzoyl)-N-[4-methyl-3-(trifluoromethyl)phenyl]piperidine-3-carboxamide (Avacopan, Vynpenta®); - N-acetyl-L-phenylalanyl-L-ornithine-L-prolyl-3-cyclohexyl-D-alanyl-L-tryptophyl-L-arginine-N-5.2-C-1.6-lactam (PMX-53); - acetylated phenylalanine-[ornithine-proline-(D)cyclohexylalanine-tryptophyl-arginine]; - L-alanyl-L-seryl-glycyl-L-alanyl-L-prolyl-L-alanyl-L-prolyl-glycyl-L-prolyl-L-alanyl-glycyl-L-prolyl-L-alanyl-glycyl-L-prolyl-L-leucyl-L-arginyl-L-prolyl-L-methionyl-L-phenylalanine; - N,N-bis(1,3-benzodioxol-5-ylmethyl)-N-(1-butyl-2,4-diphenyl-1H-imidazol-5-ylmethyl)amine; - N-[2-(4-chlorophenyl)ethyl]-N-(1,4-dioxaspiro[4.5]dec-8-yl)-2-isobutylbenzamide; - N-[2-(4-chlorophenyl)ethyl]-N-(4-hydroxycyclohexyl)-1-benzothiophene-3-carboxamide; - N-[2-(4-chlorophenyl)ethyl]-N-(4-hydroxycyclohexyl)naphthalene-1-carboxamide; 2-(2-ethyl-6-methylphenyl)-4-methoxy-N-(5-methoxy-2-methylphenyl)-5,6,7,8-tetrahydroquinolin-5-amine; 2-(2,6-diethylphenyl)-N-ethyl-4-methoxy-N-(1-naphthyl)-5,6,7,8-tetrahydroquinolin-5-amine; - N-[2,6-dioxohexahydropyrimidin-4(S)-ylcarbonyl]-L-phenylalanyl-L-ornityl-L-prolyl-5-methyl-L-norleucyl-4-fluoro-L-phenylalanyl-L-phenylalaninamide (JPE-1375; JSM-1375); - N-(3-phenylpropionyl)-L-ornithine-L-prolyl-3-cyclohexyl-D-alanyl-L-tryptophyl-L-arginine N-5.1-C-1.5-lactam (PMX-205); - N,N'-bis(4-amino-2-methylquinolin-6-yl)urea (NSC12155); - N-[4-(dimethylamino)benzyl]-N-(4-isopropylphenyl)-7-methoxy-1,2,3,4-tetrahydronaphthalene-1-carboxamide hydrochloride (W54011); - L-phenylalanyl-L-ornithine-L-prolyl-3-cyclohexyl-D-alanyl-L-tryptophyl-L-arginine N-5.2-C-1.6-cyclic peptide; - (4aR,16aS)-6,18-dihydroxy-23(S)-[2(S)-hydroxy-2-[2(R)-hydroxy-6(R)-methyl-5(R)-[2(S)-methylbutyl]tetrahydro-2H-pyran-2-yl]propionamido]-22(S)-isopropyl-7(S),19(R)-dimethyldocosahydro-13H,22H-dipyridazino[6,1-f:6',1'-o][1,4,7,10,13,16]oxapentaazacyclononadecyne-5,7,11,17,20,24-hexanone (L-156602) The pharmaceutical composition according to any one of claims 1 to 8, which is a C5aR1 antagonist selected from the group consisting of:

10. The C5aR1 inhibitor is - Avdoralimab (IPH-5401) - MOR-044254; - NOX-D20; - anti-C5aR1ab-C5-SiRNA; - m20 / 70 mlgG2a. 1; - 3C5 - 7F3 The pharmaceutical composition according to any one of claims 1 to 8, which is an anti-C5aR1 antibody selected from the group consisting of:

11. The C5aR1 inhibitor is - (2R)-2-[3-(furan-2-carbonyl)phenyl]-N-[4-(trifluoromethyl)-1,3-thiazol-2-yl]propenamide (DF2427); and A compound having the general formula (I) or a pharma- ceutically acceptable salt thereof: 【Chemistry 1】 [In the formula, i) R is 2-thiazolyl or 2-oxazolyl, which is unsubstituted or substituted by a group selected from the methyl, tert-butyl or trifluoromethyl groups; - C(Ra)=N-W (W is linear or branched C １ -C ４ alkyl); - COOO、SOOO、SO ２ RO、PORO、RO ２ Rha Selected from In the above formula, Ra is - C １ -C ５ -Alkyl, C ３ -C ６ -Cycloalkyl, C ２ -C ５ -alkenyl, unsubstituted phenyl or halogen, C １ -C ４ -Alkyl, C １ -C ４ -Alkoxy, halo-C １ -C ４ -Alkoxy, hydroxy, C １ -C ４ - a substituted phenyl substituted with a group selected from acyloxy, phenoxy, cyano, nitro, amino; a heteroaryl group selected from pyridine, pyrimidine, pyrrole, thiophene, furan, indole, thiazole, oxazole, said heteroaryl being unsubstituted or halogen, C １ -C ４ -Alkyl, C １ -C ４ -Alkoxy, halo-C １ -C ４ -Alkoxy, hydroxy, C １ -C ４ -substituted with a group selected from acyloxy, phenoxy, cyano, nitro, amino; - α or β carboxyalkyl residues (linear or branched C １ -C ６ -Alkyl, C ３ -C ６ -Cycloalkyl, C ２ -C ６ -Alkenyl, C １ -C ６ -phenylalkyl, optionally substituted with additional carboxy (COOH) groups; an ω-aminoalkylamino group of formula (II): 【Chemistry 2】 In the formula (II), X is, - Linear or branched C １ -C ６ Alkylene, C ４ -C ６ Alkenylene, C ４ -C ６ Alkynylene (optionally, a) CO ２ R4 group, where R4 is hydrogen or a straight or branched C １ -C ６ Alkyl group or linear or branched C ２ -C ６ alkenylene group), or b) by a CONHR group, where R5 is hydrogen, a straight or branched C ２ -C ６ optionally substituted by alkyl or an OR group, where R is as defined above; - (CH ２ ) ｍ -B-(CH ２ ) ｎ A group (CO as defined above) ２ R4 or CONHR5 groups, where a) B is an oxygen or sulfur atom or a nitrogen atom (C １ -C ４ m is an integer from 0 to 3 and n is an integer from 2 to 3; or b) B is a CO, SO or CONH group, m is an integer from 1 to 3 and n is an integer from 2 to 3. and R2 and R3 are independently hydrogen, straight chain or branched C １ -C ６ Alkyl (which may contain an oxygen or sulfur atom), C ３ -C ７ Cycloalkyl, C ３ -C ６ Alkenyl, C ３ -C ６ -Alkynyl, aryl-C １ -C ３ -Alkyl, hydroxy-C ２ -C ３ - is an alkyl group; or R2 and R3, together with the N atom to which they are attached, form a group of the formula (III): 【Chemistry 3】 (In the above formula, Y is, - Single bond, CH ２ , O, S, or N-R6 group, where R6 is hydrogen, C １ -C ４ Alkyl, C １ -C ４ acyl, unsubstituted phenyl or halogen, C １ -C ４ -Alkyl, C １ -C ４ -Alkoxy, hydroxy, C １ -C ４ -representing a substituted phenyl substituted with a group selected from acyloxy, phenoxy, cyano, nitro, and amino); and p represents an integer from 0 to 3; - Formula SO ２ Residue of R7, where R7 is C １ -C ６ -Alkyl, C ３ -C ６ -Cycloalkyl, C ２ -C ６ -alkenyl, aryl and heteroaryl; (represents or forming a 3- to 7-membered nitrogen heterocyclic ring of or In the formula (II), X, together with the nitrogen atom to which it is attached and the R2 group, forms a nitrogen-containing 3- to 7-membered heterocyclic, monocyclic or polycyclic ring; R3 is hydrogen, C １ -C ４ Alkyl, C １ -C ４ acyl, unsubstituted phenyl or halogen, C １ -C ４ -Alkyl, C １ -C ４ -Alkoxy, hydroxy, C １ -C ４ -represents a substituted phenyl substituted with a group selected from acyloxy, phenoxy, cyano, nitro, and amino. Selected from; ii) R1 is a straight or branched C １ -C ５ Alkyl, C ３ -C ５ is cycloalkyl; iii) Ar is - unsubstituted phenyl or halogen, C １ -C ４ -Alkyl, C １ -C ４ -Alkoxy, hydroxy, C １ -C ４ -Acyloxy, phenoxy, cyano, nitro, amino, C １ -C ４ -Acylamino, halo-C １ -C ３ -Alkyl, halo-C １ -C ３ -alkoxy, benzoyl, heteroarylcarbonyl, heteroaryl, linear or branched C １ -C ８ -Alkanesulfonates, linear or branched C １ -C ８ -Alkanesulfonamides, linear or branched C １ -C ８ a substituted phenyl group substituted with one or more groups independently selected from alkylsulfonylmethyl; or - a heteroaryl ring selected from pyridine, pyrrole, thiophene, furan, and indole; is] is a non-competitive allosteric inhibitor selected from Among the above compounds, particularly preferred are R is, 2-thiazolyl or 2-oxazolyl, which is unsubstituted or substituted by a group selected from the methyl, tert-butyl or trifluoromethyl groups; - C(Ra)=N-W (W is linear or branched C １ -C ４ alkyl); - CORa, SORa or SO ２ Ra (wherein Ra is as defined above) Selected from; and Ar is 3'-benzoylphenyl, 3'-(4-chloro-benzoyl)-phenyl, 3'-(4-methyl-benzoyl)-phenyl, 3'-acetyl-phenyl, 3'-propionyl-phenyl, 3'-isobutanoyl-phenyl, 4'-isobutyl-phenyl, 4'-trifluoromethanesulfonyloxy-phenyl, 4'-benzenesulfonyloxy-phenyl, 4'-trifluoromethanesulfonylamino-phenyl, 4'-benzenesulfonylamino-phenyl, 4'-benzenesulfonylmethyl-phenyl, 4'-acetoxyphenyl, 4'-propionyloxy-phenyl, 4'-benzoyloxy-phenyl, 4'-acetylamino-phenyl, 4'-propionylamino-phenyl, 4'-phenyloxy ... '-Benzoylamino-phenyl, 3'-(furan-2-carbonyl)-phenyl, 3'-(benzofuran-2-carbonyl)-phenyl, 3'-(thiophene-2-carbonyl)-phenyl, 3'-(pyridine-2-carbonyl)-phenyl, 3'-(thiazole-2-carbonyl)-phenyl, 3'-(oxazole-2-carbonyl)-phenyl, 3'-(2-furyl)-phenyl, 3'-(2-oxazolyl)-phenyl, 3'-(3-isoxazolyl)-phenyl, 3'-(2-benzoxazolyl)-phenyl, 3'-(3-benzoisoxazolyl)-phenyl, 3'-(2-thiazolyl)-phenyl, 3'-(2-pyridyl)-phenyl, 3'-(2-thiophenyl)-phenyl Selected from; or Ar is a heteroaryl ring selected from pyridine, pyrrole, thiophene, furan or indole; The pharmaceutical composition according to any one of claims 1 to 8, which is the compound of formula (I) or a pharma- ceutically acceptable salt thereof.

12. The C5aR1 inhibitor is - (2R)-2-[3-(furan-2-carbonyl)phenyl]-N-[4-(trifluoromethyl)-1,3-thiazol-2-yl]propenamide (DF2427); 4-{(1R)-1-[(phenylsulfonyl)amino]ethyl}phenyl trifluoromethanesulfonate; - N-[(1R)-1-(3-benzoylphenyl)ethyl]benzenesulfonamide; 4-{(1R)-1-[(pyridin-3-ylsulfonyl)amino]ethyl}phenyl trifluoromethanesulfonate; - N-[(1R)-1-(3-benzoylphenyl)ethyl]methanesulfonamide; - N-{(1R)-1-[3-(2-furoyl)phenyl]ethyl}thiophene-2-sulfonamide; - N-{(1R)-1-[3-(2-furoyl)phenyl]ethyl}methanesulfonamide; 4-{(1R)-1-[(thien-2-ylsulfonyl)amino]ethyl}phenyl trifluoromethanesulfonate; - N-[(1R)-1-(3-benzoylphenyl)ethyl]thiophene-2-sulfonamide; - N-[(1R)-1-(3-benzoylphenyl)ethyl]-3-pyrrolidin-1-ylpropane-1-sulfonamide; - methyl 5-({[(1R)-1-(3-benzoylphenyl)ethyl]amino}sulfonyl)-2-furoate; 5-({[(1R)-1-(3-benzoylphenyl)ethyl]amino}sulfonyl)-2-furoic acid; 4-{(1R)-2-methyl-1-[(methylsulfonyl)amino]propyl}phenyl trifluoromethanesulfonate; - N-((1R)-1-{4-[1-methyl-1-(phenylsulfonyl)ethyl]phenyl}ethyl)methanesulfonamide; 4-[(1R)-1-(isobutyrylamino)ethyl]phenyl trifluoromethanesulfonate; 4-{[(1R)-1-(pyridin-3-ylcarbonyl)amino]ethyl]}phenyl trifluoromethanesulfonate; - N-[(1R)-1-(3-benzoylphenyl)ethyl]benzamide; - N-[(1R)-1-(3-benzoylphenyl)ethyl]-2-furamide; - N-[(1R)-1-(3-benzoylphenyl)ethyl]cyclobutanecarboxamide; - N-[(1R)-1-(4-trifluoromethanesulfonyloxy)phenylethyl]-4-piperidin-1-yl butanamide (DF2593Y); 4-{(1R)-1-[(4-pyrrolidin-1-ylbutanoyl)amino]ethyl]}phenyl trifluoromethanesulfonate; - 3-{(1R)-1-[4-(4-trifluoromethyl-1,3-thiazol-2-yl)amino]ethyl}phenyl)(phenyl)methanone; - R(-)-2-[(4'-trifluoromethanesulfonyloxy)phenyl]-N-[3-(N'-pyrrolidinyl)propyl]propionamide (DF2297X) or its chloride salt (DF2297A); and 5-[(1R)-1-(4-{[4-(trifluoromethyl)-1,3-thiazol-2-yl]amino}phenyl)ethyl]tetrazol-2-ide (DF3966Y) or its sodium salt (DF3966A) The pharmaceutical composition according to any one of claims 1 to 8 and 11, selected from

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