Selective glucocorticoid receptor modifiers for the treatment of impaired cutaneous wound healing

Abstract

The present invention relates to selective glucocorticoid receptor modulators (SEGRMs) or pharmaceutically acceptable salts thereof for treating impaired skin wound healing in a subject, in vitro methods for identifying subjects with impaired skin wound healing responsive to treatment with selective glucocorticoid receptor modulators (SEGRMs) or pharmaceutically acceptable salts thereof, and kits and kits related thereto.

Description

[0001] The present invention relates to selective glucocorticoid receptor modulators (SEGRMs) or pharmaceutically acceptable salts thereof for treating impaired skin wound healing in a subject, in vitro methods for identifying subjects with impaired skin wound healing who are responsive to treatment with selective glucocorticoid receptor modulators (SEGRMs) or pharmaceutically acceptable salts thereof, and kits and kits-of-parts related thereto.

[0002] Chronic wounds are a major health problem worldwide, affecting 6.5 million patients in the United States alone, and the number of patients affected is expected to increase due to an aging population and an increasing incidence of metabolic diseases [Sen CK et al (2009) Wound Repair Regen 17:763–771; Gould L et al (2015) Wound Repair Regen 23:1-13].

[0003] Chronic wounds have a multifactorial etiology and depend on different variables: a) underlying disease, such as diabetes, arterial or venous insufficiency; b) stress; c) age and nutritional status and d) the microbial environment [Gould L et al (2015) Wound Repair Regen 23:1-13].

[0004] Chronic wounds are generally understood as wounds that do not heal within 2 months. They are a major health problem worldwide. In developed countries including the United States and the European Union, it is estimated that 1% to 2% of the total population will experience a chronic wound in their lifetime [Gottrup F (2004) Am J Surg 187:38S-43S].

[0005] The main chronic wound indications are venous ulcers, pressure ulcers and diabetic foot ulcers. Venous ulcers are pathologically altered tissue defects in the lower leg based on chronic venous insufficiency, often accompanied by deep vein thrombosis. Pressure ulcers are the result of severe tissue hypoxia in immobilized patients. Diabetic foot ulcers can affect up to 25% of diabetic patients in their lifetime and often lead to lower limb amputation. The standard of care recommended by the German Society of Dermatology for all these wounds [Dissemond J et al (2014) JDDG 1610-0379 / 2014 / 1207:541-554] includes wound dressings, surgical and biological (maggot) debridement, infection control and negative pressure therapy. (PDGF: Platelet-derived growth factor) was the only registered drug treatment for a long time, but its therapeutic effect was minimal, as was the case with successful cell therapy. Recombinant human EGF (rhEGF) is registered in many countries as For the treatment of ulcers of diabetic foot ulcer syndrome. In addition, Trafermin (trade name: ), also known as recombinant human basic fibroblast growth factor (rhbFGF), is a recombinant form of human basic fibroblast growth factor (bFGF) and is sold in Japan as a topical spray for the treatment of skin ulcers.

[0006] Regardless of the treatment used, one-third of chronic wounds will recur.

[0007] Although glucocorticoids have anti-inflammatory effects in other conditions, they cannot be used topically because one of their side effects is actually delayed wound healing [Hengge UR (2006) J Am Acad Dermatol 54:1-15]. Therefore, as a matter of current art dogma, topical glucocorticoids will affect wound healing [Wicke C et al (2000) Arch Surg 135:1265-1270; Anderson K et al (2014) J Am Coll Clin Wound Spec 4:84–91]. In addition, nonsteroidal anti-inflammatory drugs (such as ibuprofen) are only effective in improving wound pain [Dissemond J et al (2014), supra].

[0008] Therefore, there is a continuing and strong medical need for reliable and effective therapies for treating impaired skin wound healing in patients.

[0009] As shown in the examples and corresponding figures, it was surprisingly found in the present application that selective glucocorticoid receptor modulators (SEGRMs) exhibited significant enhancement of fibroblast proliferation (2D) and fibroblast-derived matrix formation (3D) in a human-related ex vivo wound healing model using wound exudates from chronic wound patients, increased collagen 1 and 3 expression and inhibited IL-1β secretion. For example, the effect of mapracorat (as an exemplary SEGRM) on fibroblast proliferation was studied in the presence of >80 human wound exudates, resulting in an increase of >120%.

[0010] Furthermore, in a porcine model of delayed wound healing, mapracorat was found to reduce wound scores from day 6 to day 12 in wounds treated with wound exudate from chronic human wounds and / or the TLR 7 / 8 agonist R848 (resiquimod) as an inducer of delayed wound healing ( Fig.13 ).

[0011] Furthermore, in the Examples, BI-653048, but not its inactive analog BI-3047, was found to enhance fibroblast proliferation in a dose-dependent manner in the presence of different aggressive wound exudates ( Figure 1 ) and inhibited IL-1β secretion in these cultures ( Figure 2 In the presence of wound exudate, chemically distinct SEGRMs (mapracorat, ZK216348, and HY14234) had similar effects on proliferation and IL-1β secretion or mRNA expression ( Figure 5 , 8 and 9-11). Although qualitatively similar, the magnitude of the effect varies between SEGRM compounds and individual wound exudates. Thus, in a preferred embodiment, a personalized medicine approach can be performed by pre-testing patient exudates in vitro for different compounds using the methods of the invention described herein. In a fibroblast proliferation assay in wound exudate (rather than culture medium), BI-653048 and mapracorat increased the expression of collagen 1 and collagen 3 mRNA ( Figure 3 and 6 -8). This effect was reversed by the glucocorticoid receptor antagonist mifepristone ( Figure 4 ).

[0012] Furthermore, positive effects were observed in 3D fibroblast cultures of wound exudate from patients with chronic, non-healing wounds in the presence of multiple structurally distinct SEGRMs, including BI-653048 (but not its inactive analog BI-3047), mapracorat, ZK216348, AZD7594, and HY14234 ( Fig.12 ). Fibroblast proliferation assay (2D) and fibroblast-derived matrix formation assay (3D) are human-relevant ex vivo assays for wound healing.

[0013] Thus, in one embodiment, the present invention is directed to a selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof for use in treating impaired cutaneous wound healing in a subject.

[0014] The human glucocorticoid receptor, also known as "GR", "GC" or "GCR", also known as NR3C1 (nuclear receptor subfamily 3, group C, member 1), is a receptor to which Cortisol and other glucocorticoids bind. In a preferred embodiment, the glucocorticoid receptor is a human glucocorticoid receptor.

[0015] The GR is expressed in nearly every cell in the body and regulates genes that control development, metabolism, and immune responses. Because the receptor gene is expressed in multiple forms, it has many different (pleiotropic) effects in different parts of the body.

[0016] When the GR binds to glucocorticoids, its primary mechanism of action is the regulation of gene transcription. Unbound receptors are found in the cytoplasm of the cell. After the receptor binds to a glucocorticoid, the receptor-glucocorticoid complex can take different paths. The activated GR complex upregulates the expression of anti-inflammatory proteins in the nucleus or inhibits the expression of pro-inflammatory proteins in the cytoplasm (by preventing the translocation of other transcription factors from the cytoplasm into the nucleus). Alternatively, inhibition can be achieved by the receptor binding to DNA at the same site where another transcription factor would bind, thereby eliminating the effects of the other transcription factor.

[0017] In humans, the GR protein is encoded by the NR3C1 gene located on chromosome 5 (5q31). Various alternatively spliced ​​isoforms exist.

[0018] "Selective glucocorticoid receptor modulators" or "SEGRMs" are well-known and established classes of compounds. Selective glucocorticoid receptor modulators are preferably non-steroidal selective glucocorticoid receptor modulators (SEGRMs). In earlier prior art documents, selective glucocorticoid receptor modulators were also referred to as "selective glucocorticoid receptor agonists" or "SEGRAs" or "dissociated glucocorticoid receptor agonists" or "DIGRAs". Further, in another preferred embodiment, SEGRMs are also referred to as "SGRMs", "GR agonists" or "SEDIGRAMs" in some prior art documents. Therefore, in a preferred embodiment, "selective glucocorticoid receptor activators" or "SEGRAs" are well-known and established classes of compounds. Selective glucocorticoid receptor activators are preferably non-steroidal selective glucocorticoid receptor activators (SEGRAs). In another preferred embodiment, SEGRMs or SEGRAs are also referred to as "selective glucocorticoid receptor activators and modulators" or "SEGRAMs" in the prior art. Therefore, in a preferred embodiment, the terms "SEGRM" and "SEGRA" are synonyms. Preferably, the synonymous meaning is reflected by the term "SEGRAM" in some prior arts.

[0019] Glucocorticoids are known to exhibit anti-inflammatory properties. However, as mentioned above, glucocorticoids also exhibit serious side effects, such as skin atrophy, etc. As mentioned above, topical glucocorticoids cannot be used for other common types of skin wounds such as diabetic ulcers and delayed wound healing, because one of their side effects is actually delayed wound healing.

[0020] SEGRMs achieve their selectivity by triggering only a subset of the glucocorticoid receptor's mechanisms of action. In particular, SEGRMs bind specifically to the glucocorticoid receptor (GC) but only trigger a subset of the GC's mechanisms of action.

[0021] Both non-selective glucocorticoids and SEGRMs act by binding to and activating the glucocorticoid receptor (GR). In contrast to glucocorticoids, which activate the GR via at least two signal transduction pathways, generally referred to as "transactivation" and "transinhibition," SEGRMs activate the GR in such a way that they act only or primarily via one of these two major possible pathways, preferably via transinhibition.

[0022] In the absence of glucocorticoids, GR exists in the cytoplasm in an inactive state complexed with heat shock proteins (HSPs) and immunophilins. Binding of glucocorticoids to GR activates the receptor by causing conformational changes in GR and dissociation of bound HSPs. The activated GR can then regulate gene expression through one of two pathways: "transactivation" and "transrepression":

[0023] Transactivation: The direct pathway is called transactivation, whereby the activated GR dimerizes, translocates into the nucleus and binds to a specific DNA sequence called the glucocorticoid response element (GRE). The GR / DNA complex recruits other proteins that transcribe the downstream DNA into mRNA and ultimately translate it into protein. Examples of glucocorticoid-responsive genes include genes encoding tyrosine aminotransferase (TAT), annexin A1, T22D3, angiotensin-converting enzyme, neutral endopeptidase, dual-specificity phosphatase 1, interferon regulatory factor 1, and other anti-inflammatory proteins.

[0024] Transrepression: The second indirect pathway is called transrepression, in which the activated monomeric GR binds to other transcription factors such as NF-κB and AP-1 and prevents them from upregulating the expression of their target genes. These target genes encode proteins such as cyclooxygenase, NO synthase, phospholipase A2, tumor necrosis factor, transforming growth factor β, ICAM-1, and many other proinflammatory proteins.

[0025] Therefore, the anti-inflammatory effects of glucocorticoids result from both transactivation and transrepression.

[0026] Therefore, SEGRM is a compound that is more potent in trans-inhibition than in trans-activation.

[0027] Assays for determining transinhibition are known in the art and include determining LPS-induced secretion of the cytokines IL-12 or TNFα in PBMC cells, such as, for example et al. (2004; PNAS, 101: 227-232); or inhibition of collagenase promoter activity in stimulated human primary cells, inhibition of cytokine (e.g., IL-12 or IFNγ) secretion; or inhibition of lymphocyte proliferation in mixed lymphocyte reactions, such as et al. (2009; Br. J. Pharmacol., 158: 1088-1103).

[0028] Assays for determining transactivation are known in the art and include determining the induction of tyrosine aminotransferase (TAT) activity in hepatocytes, such as, for example et al. (2004; PNAS, 101: 227-232); or the induction of MMTV promoter activity or the induction of TAT activity, such as et al. (2009; Br. J. Pharmacol., 158: 1088-1103).

[0029] Preferably, the assays for determining transactivation and transrepression can be as follows et al (2009):

[0030] Preferably, the inhibition of collagenase promoter activity can be as follows et al (2009): HeLa cells stably transfected with a luciferase reporter gene linked to a collagenase promoter were cultured in a nutrient solution supplemented with 3% charcoal-absorbed fetal calf serum (FCS), 50 units mL -1 Penicillin and 50 mg mL -1 Streptomycin, 4mmol·L -1 L-glutamine and 300 μg mL -1 The cells were cultured in Dulbecco's modified Eagle's medium containing geneticin for 24 hours. The cells were then seeded into 96-well culture plates (1x10 4 After 24 hours, cells were incubated with or without increasing concentrations of reference or test compounds (1 pmol·L -1 To 1mmol·L -1 ) of inflammatory stimulants [10 ng·mL -1 12-o-tetradecanoylphorbol 13-acetate (TPA)] were incubated. As negative control (unstimulated cells), cells were incubated with 0.1% dimethyl sulfoxide (DMSO), and as positive control (stimulated cells), cells were incubated with 10 mg mL -1 TPA plus 0.1% DMSO was incubated and luciferase assay was performed 18 hours later.

[0031] Preferably, the inhibition of the secretion of cytokines IFN-γ and IL-12p40 in stimulated human primary cells can be as follows et al (2009): using 10 ng mL -1The effects of compounds on monocyte secretion of IL-12p40 were examined after lipopolysaccharide-stimulated peripheral blood mononuclear cells (PBMCs) from healthy donors (Escherichia coli serotype 0127:B8; Sigma). -1 The effect of mitogen-activated protein glutamate (PLG) on interferon (IFN)-γ secretion was detected after PBMCs were stimulated with PLG. The cells were incubated for 24 h (37°C, 5% CO 2 ), the cytokine concentrations in the supernatants of the treated cells were determined using specific ELISA kits (IFN-γ and IL-12p40 ELISA (R&D Systems)).

[0032] Preferably, the induction of MMTV promoter activity can be as follows et al (2009): The MMTV promoter was linked to a luciferase reporter gene and HeLa cells were stably transfected with this construct. The cells were grown in a humidified medium supplemented with 50 units of penicillin and 300 μg mL -1 To study the transactivation activity of GR ligands, cells were cultured in medium supplemented with 3% charcoal-absorbed FCS for 24 h. Cells were then seeded into 96-well plates at 1 x 10 cells per well. 4 cells. After 24 hours, the cells were incubated with increasing concentrations of reference (dexamethasone) or test compounds. As a negative control (unstimulated cells), cells were treated with 0.1% DMSO. Cells were incubated with compounds for 18 hours, and then luciferase activity was determined as a measure of GR activity.

[0033] Preferably, the induction of TAT activity can be as follows Determination as described in et al (2009): The induction of TAT by the test compounds was confirmed in vitro using the human hepatoma cell line HepG2. HepG2 cells were cultured in the presence of 2 mmol·L -1 The cells were cultured in minimal essential medium containing glutamine, 10% heat-inactivated FCS and 1% non-essential amino acids. To detect the induction of TAT by the test compounds, cells were seeded in 96-well plates at 1x10 5 After 24 hours, the cells were incubated with test medium containing increasing concentrations of test and reference compounds. After 24 hours of cell lysis, TAT activity was measured by the absorption at 340 nm of the aromatic p-hydroxybenzaldehyde after conversion of p-hydroxyphenylpyruvate.

[0034] Therefore, in another preferred embodiment of the present invention, the selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof is a non-steroidal selective glucocorticoid receptor modulator (SEGRM). The selective glucocorticoid receptor modulator is combined with the glucocorticoid receptor (GC). In a more preferred embodiment, the SEGRM achieves their selectivity by only triggering a subset of the mechanism of action with an affinity (KD) of less than 100nM, 70nM, 60nM, 50nM, 40nM, 30nM, 20nM or 10nM. Affinity (KD) can be determined by methods known in the art, for example, by measuring using surface plasmon resonance (SPR), for example, using a Biacore device. Preferably, affinity is determined at about 20°C or 25°C.

[0035] In another preferred embodiment of the present invention, the non-steroidal selective glucocorticoid receptor modulator (SEGRM) triggers trans-inhibition when binding to the glucocorticoid receptor (GC) in cells and exhibits less trans-activation when binding to the glucocorticoid receptor (GC) in cells compared to dexamethasone.

[0036] In a preferred embodiment, the IC of the transinhibitory activity of the SEGRM for a particular use of the present invention is 50 The IC value of dexamethasone for transinhibitory activity 50 In a more preferred embodiment, the transinhibitory activity of the SEGRM has an IC value of at most 100 times, 50 times, 30 times, at most 10 times, at most 5 times, or at most 2 times higher. 50 The IC value is similar to that of dexamethasone for transinhibitory activity 50 Values, as found, for example, for mapracorat. For example, the IC of the transinhibitory activity of dexamethasone 50 The value can also be lower than the IC of dexamethasone's transinhibitory activity. 50 Preferably these values ​​are determined in one of the above-mentioned transinhibition assays.

[0037] In a preferred embodiment, the potency of the transinhibitory activity of the SEGRM for a particular use of the invention is at least 30%, 40%, 50%, 60%, 70%, 80% or 90% of the potency of the transinhibitory activity of dexamethasone. These values ​​are preferably determined in one of the above-mentioned transinhibition assays.

[0038] In a preferred embodiment, the transactivation activity of the SEGRM of the present invention is 50 EC value of transactivation activity of dexamethasone 50 The values ​​are preferably at least 20-fold, 40-fold, 50-fold or 100-fold higher. These values ​​are preferably determined in one of the above-mentioned transactivation assays.

[0039] As mentioned above, SEGRMs exert their effects by binding to and activating glucocorticoid receptors (GRs).

[0040] Thus, in a preferred embodiment, the SEGRM for a particular use according to the invention is an activator of the glucocorticoid receptor (GR).

[0041] Thus, in a preferred embodiment, the SEGRM for specific use according to the present invention is a glucocorticoid receptor (GR) agonist.

[0042] Preferably, an activator of a glucocorticoid receptor (GR) or a glucocorticoid receptor (GR) agonist is understood to be a compound that activates the GR by transactivation and / or transinhibition of the GR. Transactivation and transinhibition can be determined using the above-mentioned assays.

[0043] As described above, the SEGRMs for specific uses according to the invention are compounds that are stronger trans-inhibitory than trans-activating. Thus, in preferred embodiments, the SEGRMs for specific uses according to the invention activate GR by trans-inhibition and optionally trans-activation of GR. In more preferred embodiments, the SEGRMs for specific uses according to the invention activate GR by trans-inhibition and trans-activation of GR, wherein the trans-inhibition of the SEGRM is stronger than the trans-activation. Assays for determining trans-activation and trans-inhibition and for determining whether the trans-inhibition of the SEGRM is stronger than the trans-activation are as described above.

[0044] Preferably, a SEGRM that is an activator of the glucocorticoid receptor (GR) or a glucocorticoid receptor (GR) agonist is capable of being positively detected in an in vitro translocation assay as described in Example 1.10. The assay represents an additional positive detection of a SEGRM that is an activator of the glucocorticoid receptor (GR) or a glucocorticoid receptor (GR) agonist. In particular, the assay cannot distinguish between active glucocorticoids and active SEGRMs, because for the assay in Example 1.10, glucocorticoids at a concentration of 10 nM at 37° C. also effect translocation of the glucocorticoid receptor from the cytoplasm to the nucleus in primary human fibroblasts in in vitro culture.

[0045] Therefore, in a preferred embodiment, SEGRM as an activator of glucocorticoid receptor (GR) or a glucocorticoid receptor (GR) agonist for the specific use of the present invention achieves translocation of glucocorticoid receptor from the cytoplasm to the nucleus in primary human fibroblasts in vitro culture at 37°C with a SEGRM concentration of 10nM or 100nM. The detection is as described in detail in Example 1.10. In particular, the cultured cells are starved overnight, incubated with the compound at 37°C for 45 minutes, and fixed with 4% paraformaldehyde at room temperature for 10 minutes, and then permeabilized with 1% BSA in PBS with 0.5% Triton X100 at room temperature for 10 minutes. Subsequently, the cells are stained with mouse anti-glucocorticoid receptor monoclonal antibodies. Detection can be performed by immunofluorescence microscopy using secondary antibodies labeled with fluorescent markers.

[0046] In another preferred embodiment of the present invention, the non-steroidal selective glucocorticoid receptor modulator (SEGRM) specifically binds to the glucocorticoid receptor (GC) with an affinity (KD) of less than 100 nM.

[0047] In another preferred embodiment of the present invention, the nonsteroidal selective glucocorticoid receptor modulator (SEGRM) specifically binds to the glucocorticoid receptor (GC). If the competitive factor of the SEGRM for the glucocorticoid receptor (GC) (defined as the IC of the SEGRM test compound) is 50 / IC of reference compound 50 ) is less than 20, 10, 5, 4 or 2, and the competition factor for the progesterone receptor (PR), androgen receptor (AR) and mineralocorticoid receptor (MR) is at least 5, 10, 15, 20, 30, 40 or 50, then the SEGRM is understood to specifically bind to the glucocorticoid receptor (GC). et al. (2009; Br. J. Pharmacol., 158: 1088-1103). et al., a suitable reference compound for GC is dexamethasone, a suitable reference compound for PR is progesterone, a suitable reference compound for AR is metribolone, and a suitable reference compound for MR is aldosterone.

[0048] Preferably, the competitive factor can be as follows et al (2009) as described:

[0049] IC for determination of receptor binding 50 The value can be as follows et al (2009) as described: Extracts from Sf9 cells infected with recombinant baculovirus encoding human GR, progesterone receptor (PR), androgen receptor (AR), or mineralocorticoid receptor (MR) were used for receptor binding assays as described in Alexander et al. (2004). All receptor nomenclature follows the “Receptor and Channel Guide” (Alexander et al., 2008). For GR, PR, AR, and MR binding assays, [1,2,4,6,7-3H]dexamethasone (approximately 3.18 GBq·mmol -1 )(~20nmol·L -1 ), [1,2,6,7-3H(N)]progesterone (about 3.7 GBq·mmol-1), [17a-methyl3H]estratrienolone (about 3.18 GBq·mmol-1) or D[1,2,6,7-3H(N)]aldosterone (about 2.81 GBq·mmol-1) were respectively incubated with SF9 cytoplasm (100-500 μg protein), test compounds and binding buffer (10 mmol·L -1 Tris / HCL pH 7.4, 1.5mmol·L-1 EDTA, 10% glycerol) were mixed in a total volume of 50μL and incubated at room temperature for 1 hour. After incubation, 50μL of cold charcoal suspension was added for 5 minutes and the mixture was then transferred to a microtiter filter plate. The mixture was filtered into Picoplates (Canberra Packard) and mixed with 200μL Microszint-40 (Canberra Packard). Bound radioactivity was determined using a Packard Top Count microplate reader. Specific binding was defined as the concentration of 10μmol·L -1 The difference between the binding of [1,2,4,6,7-3H]dexamethasone, [1,2,6,7-3H(N)]progesterone, [17a-methyl-3H]estratrienolone and D[1,2,6,7-3H(N)]aldosterone in the presence and absence of unlabeled dexamethasone, progesterone, metribolone or aldosterone. The concentration of the test compound that produces 50% specific binding inhibition (IC50) is determined by Hill analysis of the binding curve. The competition factor (CF) is defined as IC50 of the test compound / IC50 of the reference compound and can be determined accordingly. By definition, the CF of the reference compound is 1.0. The reference compound for GC is preferably dexamethasone, the suitable reference compound for PR is progesterone, the suitable reference compound for AR is metribolone, and the suitable reference compound for MR is aldosterone.

[0050] In a preferred embodiment, the selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof for a specific use of the present invention is a selective glucocorticoid receptor modulator (SEGRM) characterized in that:

[0051] - SEGRM specifically binds to the glucocorticoid receptor (GC) with an affinity (KD) of less than 100 nM, and / or

[0052] - the competition factor of the SEGRM is i) less than 20 for the glucocorticoid receptor (GC), and ii) at least 5 for the progesterone receptor (PR), androgen receptor (AR) and mineralocorticoid receptor (MR), wherein the competition factor is defined as the IC50 value of the SEGRM / the IC50 value of the reference compound, and wherein the reference compound for GC is dexamethasone, the reference compound for PR is progesterone, the reference compound for AR is metribolone, and the reference compound for MR is aldosterone, and / or

[0053] - The EC50 value of the transactivation activity of SEGRM after binding to the glucocorticoid receptor (GC) in a cell is at least 20 times higher than the EC50 value of the transactivation activity of dexamethasone, and / or the IC50 value of the transinhibitory activity of SEGRM after binding to the glucocorticoid receptor (GC) in a cell is up to 100 times higher than the IC50 value of the transinhibitory activity of dexamethasone.

[0054] Therefore, in a preferred embodiment, the selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof for a particular use of the present invention is a non-steroidal selective glucocorticoid receptor modulator (SEGRM), and:

[0055] - SEGRM specifically binds to the glucocorticoid receptor (GC) with an affinity (KD) of less than 100 nM, and / or

[0056] - the competition factor of the SEGRM is i) less than 20 for the glucocorticoid receptor (GC), and ii) at least 5 for the progesterone receptor (PR), androgen receptor (AR) and mineralocorticoid receptor (MR), wherein the competition factor is defined as the IC50 value of the SEGRM / the IC50 value of the reference compound, and wherein the reference compound for GC is dexamethasone, the reference compound for PR is progesterone, the reference compound for AR is metribolone, and the reference compound for MR is aldosterone, and / or

[0057] - The EC50 value of the transactivation activity of SEGRM after binding to the glucocorticoid receptor (GC) in a cell is at least 20 times higher than the EC50 value of the transactivation activity of dexamethasone, and / or the IC50 value of the transinhibitory activity of SEGRM after binding to the glucocorticoid receptor (GC) in a cell is up to 100 times higher than the IC50 value of the transinhibitory activity of dexamethasone.

[0058] In yet another preferred embodiment, the selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof specifically binds to the glucocorticoid receptor (GC) with an affinity (KD) of less than 100 nM, and / or is a non-steroidal SEGRM.

[0059] Preferably, for SEGRMs that are more trans-inhibitory than trans-activating as defined above, the activity of the SEGRM can be positively tested in addition in an in vitro assay as described in Examples 1.10, 1.11 and / or 1.12. The assays represent additional positive assays for SEGRMs that are more trans-inhibitory than trans-activating, and these assays can be used to distinguish between active and inactive stereoisomers of the same structure, such as BI-3047. In particular, the assays cannot distinguish between active glucocorticoids and active SEGRMs, because in the case of the translocation assay in Example 1.10, glucocorticoids at a concentration of 10 nM at 37°C also achieve translocation of the glucocorticoid receptor from the cytoplasm to the nucleus in primary human fibroblasts in in vitro culture.

[0060] Therefore, in a preferred embodiment, the SEGRM for the specific use of the present invention achieves the translocation of the glucocorticoid receptor from the cytoplasm to the nucleus in primary human fibroblasts in vitro culture at 37°C with a SEGRM concentration of 10nM or 100nM. The detection is as described in detail in Example 1.10. In particular, the cultured cells are starved overnight, incubated with the compound at 37°C for 45 minutes, and fixed with 4% paraformaldehyde at room temperature for 10 minutes, and then permeabilized with 1% BSA in PBS with 0.5% Triton X100 at room temperature for 10 minutes. Subsequently, the cells are stained with mouse anti-glucocorticoid receptor monoclonal antibodies. Detection can be performed by immunofluorescence microscopy using secondary antibodies labeled with fluorescent markers.

[0061] Thus, in another preferred embodiment, the SEGRM for the specific use of the invention reduces the spontaneous IL-8 secretion of human monocytes in vitro by at least 20%, 30% or 50% at a concentration of 100 nM at 37° C. compared to control cells incubated without the SEGRM compound. The assay is as described in detail in Example 1.11.

[0062] Thus, in another preferred embodiment, the SEGRM for the specific use of the present invention reduces the IL-8 secretion of U937 cells stimulated in vitro by LPS at a concentration of 100 nM at 37° C. by at least 20%, 30% or 50% compared to control cells incubated without the SEGRM compound. The assay is as described in detail in Example 1.12.

[0063] Suitable SEGRMs for particular uses of the present invention are known in the art and include mapracorat (also known as ZK-245186) and related 5-substituted quinolone and isoquinoline derivative compounds 5-{[(1S,2S)-1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(methoxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one, BI653048, HY14234, LGD-5552, MK-5932, Org 214007-0, Compound A, AL-438, ZK-216348, PF-802, Fosdagrocorat and Compound 10. Other suitable SEGRMs for the specific purposes of the present invention are known in the art, including [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl] 4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[[(2R)-tetrahydrofuran-2-carbonyl]amino]propoxy]benzoate, 5-{(1S,2R)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-([methylsulfonyl]methyl)propyl]amino}-1H-quinolin-2-one, AZD-7594, AZD-5423, AZD-2906, JPT-117968 and SEGRM compounds disclosed in WO 2018 / 236749 A2, WO SEGRM disclosed in 2018 / 049255 A1 and WO 2018 / 183947 A1; WO 2017 / 046096, WO 2009 / 023471 A2, WO 2008 / 027796 A2, WO 2008 / 033655 A2, WO 2008 / 005686A2, WO 2008 / 021728 A2, WO 2008 / 021729 A2, WO 2008 / 154129 A1, WO 2009 / 042377 A1, WO2010 / 123769 A1, WO 2012 / 170175 BI-54903, BI-607812, GW870086X, PF-00251802, BOL-303242-X (which is a synonym for mapracorate) and other DIGRA compounds and their preparations disclosed in A1; NO donor DIGRA compounds disclosed in WO2013 / 126156 A1; Badarau E.et al.Cortivazol, fluorocortivazol, and spirocyclic analogs of fluorocortivazol disclosed in (European Journal of Medicinal Chemistry, 2019, 161: 354-363); WO 2003 / 086294 A2, WO 2004026248 A2, WO 2004 / 066920 A2, WO 2004 / 075840 A2, WO 2004 / 093805 A2, WO 2009 / 108525 A2, WO2009 / 111214 A1, WO 2010 / 138421 A1, WO 2010 / 141247 A1, WO 2011 / 031574 A1, WO 2011 / 053567 SEGRM compounds disclosed in A1; compounds disclosed in WO 2005 / 082909 A1, WO 2006 / 019716 A1 and WO 2009 / 103007 A2, WO 00 / 66522 A1 and RU24858. .

[0064] Fosdagrocorat, also known as dagrocorat 2-(dihydrogen phosphate), has the following structure:

[0065]

[0066] HY14234 has the following structure:

[0067]

[0068] LGD-5552 has the following structure:

[0069]

[0070] MK-5932 has the following structure:

[0071]

[0072] Org 214007-0 has the following structure:

[0073] The chloride salt of compound A has the following structure:

[0074] AL-438 has the following structure:

[0075]

[0076] PF-802 has the following structure:

[0077]

[0078] Compound 10 has the following structure:

[0079]

[0080] The IUPAC name of AZD-7594 is 3-(5-((1R,2S)-2-(2,2-difluoropropanylamino)-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)propoxy)-1H-indol-1-yl)-N-(tetrahydrofuran-3-yl)benzamide, which has the following structure:

[0081]

[0082] The IUPAC name of AZD-5423 is 2,2,2-trifluoro-N-((1R,2S)-1-((1-(4-fluorophenyl)-1H-indol-5-yl)oxy)-1-(3-methoxyphenyl)-2-propanyl)acetamide, which has the following structure:

[0083]

[0084] The IUPAC name of AZD-2906 is N-((1R,2S)-1-((1-(4-fluorophenyl)-1H-indol-5-yl)oxy)-1-(6-methoxypyridin-3-yl)propan-2-yl)cyclopropanamide, which has the following structure:

[0085]

[0086] JPT-117968 has the following structure:

[0087]

[0088] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[[(2R)-tetrahydrofuran-2-carbonyl]amino]propoxy]benzoate has the following structure:

[0089]

[0090] WO 2008 / 076048 A1 discloses a compound of formula (I) or a pharmaceutically acceptable salt thereof:

[0091]

[0092] in:

[0093] A is C1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 Cyanoalkyl, cyano, C 1-6 Nitroalkyl, nitro, C 1-6 Alkyl S(O) n , C 1-6 Alkoxy, C 3-7 Cycloalkyl C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-7 Heterocycloalkyl, C 3-7 Heterocycloalkyl C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkyl-C 1-6 Thioalkyl, C 1-6 Thioalkyl, C 1-6 Alkyl OC 1-6 Alkyl, C 1-6 Alkyl OC 1-6 Alkyl O, C 1-6 Alkyl C(O)C 1-6 Alkyl, C 1-6 Alkyl C(O), C 1-6 Alkyl C(O)OC 1-6 Alkyl, C 1-6 Alkyl C(O)O, C 1-6 Alkyl OC(O)C 1-6 Alkyl, C 1-6 Alkyl OC(O), HOC(O), NR 5 R 6 C 1-6 Alkyl, NR 5 R 6 NR 5 R 6 C(O)C 1-6 Alkyl, NR 5 R 6 C(O),NR 5 R 6 OC(O)C 1-6 Alkyl, NR 5 R 6 OC(O),R 7 NH, C 5-10 Aryl, C 1-3 Alkyl, C 5-10 Aryl, C 5-10 Heteroaryl, C 1-3 Alkyl or C 5-10 wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl may be optionally substituted with one or more substituents independently selected from the group consisting of halogen, cyano, hydroxyl, C1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Alkyl OC(O), C 1-4 Alkyl OC 1-4 Alkyl, C 1-4 Alkyl S(O) 2 and C 1-4 Haloalkyl O, and R x is hydrogen, or

[0094] A and R x together form a 5- to 6-membered nitrogen heterocyclic ring optionally having one or more additional heteroatoms independently selected from O, N and S;

[0095] R 1 and R 1a are independently selected from hydrogen, C 1-4 Alkyl, C 1-4 Hydroxyalkyl, C 1-4 Alkyl OC 1-4 Alkyl, C 1-4 Alkyl C 1-4 Thioalkyl and C 1-4 A haloalkyl group, or R 1 and R 1a Together for the oxo;

[0096] R 2 is hydrogen or C 1-4 alkyl;

[0097] R 3 C 5-10 Aryl, C 5-10 Aryl C 1-4 Alkyl, C 5-10 Aryl O, C 5-10 Aryl OC 1-4 Alkyl or C 5-10 Heteroaryl, these groups may be optionally substituted by one or more substituents independently selected from B;

[0098] B is C 1-3 Hydroxyalkyl, hydroxy, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkyl C 1-4 Thioalkyl, C 1-4 Thioalkyl, C 3-6 Cycloalkyl C 1-4 Thioalkyl, C 3-6 Cycloalkyl S, C 1-3 Alkyl S(O) n C 1-4 Alkyl, C 1-3 Alkyl S(O)n , C 1-4 Haloalkyl, C 1-4 HaloalkylO, halogen, nitro, cyano, C 1-4 Alkyl OC 1-4 Alkyl OC 1-4 Alkyl, C 1-4 Alkyl C(O)C 1-4 Alkyl, C 1-4 Alkyl C(O), C 1-4 Alkyl C(O)OC 1-4 Alkyl, C 1-4 Alkyl C(O)O, C 1-4 Alkyl OC(O)C 1-4 Alkyl, C 1-4 Alkyl OC(O), NR 8 R 9 C 1-4 Alkyl, NR 8 R 9 NR 8 R 9 C(O)C 1-4 Alkyl, NR 8 R 9 C(O),NR 8 R 9 OC(O)C 1-4 Alkyl, NR 8 R 9 OC(O),NR 8 R 9 C(O)OC 1-4 Alkyl, NR 8 R 9 C(O)O、R 9 C(O)R 8 NC 1-4 Alkyl, R 9 C(O)R 8 NH, C 1-4 Alkyl NH, C 1-4 Alkyl OC(O)NH, C 1-4 Alkyl C(O)OC 1-4 Alkyl NH, C 1-4 Alkyl C(O)C 1-4 Alkyl NH, C 1-4 Alkyl C(O)NH, NR 8 R 9 S(O) n C 1-4 Alkyl or NR 8 R 9 S(O) n ; n is 1 or 2;

[0099] R4 For hydrogen, hydroxyl, halogen, C 1-4 Alkyl or;

[0100] W is hydrogen, or phenyl, C 1-4 Alkyl, C 3-7 cycloalkyl, thienyl, isoxazolyl, pyrazolyl, pyridinyl, pyridazinyl or pyrimidinyl, all of which are optionally substituted by one or more substituents independently selected from the group consisting of: 1-3 Hydroxyalkyl, hydroxy, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkyl C 1-4 Thioalkyl, C 1-4 Thioalkyl, C 3-6 Cycloalkyl C 1-4 Thioalkyl, C 3-6 Cycloalkyl S, C 3-6 Cycloalkyl, C 3-6 Cycloalkyl C 1-4 Alkyl, C 3-6 Heterocycloalkyl, C 3-6 Heterocycloalkyl C 1-4 Alkyl, C 1-4 Alkyl S(O) n C 1-4 Alkyl, C 1-4 Alkyl S(O) n , C 1-4 Haloalkyl, C 1-4 HaloalkylO, halogen, nitro, cyano, C 1-4 Alkyl OC 1-4 Alkyl, C 1-4 Alkyl OC 1-4 Alkyl OC 1-4 Alkyl, C 1-4 Alkyl C(O)C 1-4 Alkyl, C 1-4 Alkyl C(O), C 1-4 Alkyl C(O)OC 1-4 Alkyl, C 1-4 Alkyl C(O)O, C 1-4 Alkyl OC(O)C 1-4 Alkyl, C 1-4 Alkyl OC(O), NR 10 R 11 NR 10 R n C(O)C 1-4 Alkyl, NR 10 R 11 C(O),NR 10 R 11 C(O)O、NR 10 R11 C 1-4 Alkyl, NR 10 R 11 OC(O),R 11 C(O)R 10 NC 1-4 Alkyl, R 11 C(O)R 10 NH, C 1-4 Alkyl OC(O)C 1-4 Alkyl NH, C 1-4 Alkyl OC(O)NH, C 1-4 Alkyl C(O)OC 1-4 Alkyl NH, C 1-4 Alkyl C(O)C 1-4 Alkyl NH, C 1-4 Alkyl C(O)NH, NR 10 R 11 S(O) n C 1-4 Alkyl or NR 10 R 11 S(O) n ;

[0101] X is CH 2 , O, S, S(O) n , NH or NC 1-4 alkyl;

[0102] Y is hydrogen, halogen, C 1-4 Thioalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkyl O, nitro, cyano, hydroxyl, R 12 C(O), R 12 OC(O),R 12 C(O)O、C 1-6 Alkyl S(O) n , R 12 R 13 NS(O) n , benzyloxy, imidazolyl, C 1-4 Alkyl NHC(O), NR 12 R 13 C(O), C 1-4 Alkyl C(O)NH or NR 12 R 13 ;

[0103] Z is O or S;

[0104] R 5 , R 6 , R 8 , R 9 , R 10, R 11 , R 12 and R 13 are independently selected from hydrogen, C 1-6 Alkyl C(O), NHR 7 C(O) and C 1-6 Alkyl; and R 7 For hydrogen, C 1-6 Alkyl, C 1-6 Alkyl C(O)OC 1-3 Alkyl, C 1-6 Alkyl C(O)O, C 1-6 Alkyl OC(O)C 1-3 Alkyl, C 1-6 Alkyl OC(O), C 1-6 Alkyl C(O), C 5-10 Heteroaryl C 1-3 Alkyl, C 5-10 Heteroaryl, C 5-10 Aryl C 1-3 Alkyl, C 5-10 Aryl, C 3-6 Cycloalkyl C 1-3 Alkyl or C 3-6 Cycloalkyl.

[0105] Badarau E.et al. (European Journal of Medicinal Chemistry, 2019, 161: 354-363) discloses spirocyclic analogs of fluorinated cortivaazole that can be used according to the present invention. Exemplary compounds and their syntheses that can be used according to the present invention are:

[0106] - Compounds of formula 8a-e and 9a-e:

[0107]

[0108] in:

[0109] In compounds 8a and 8b: n = 1, R = phenyl,

[0110] In compounds 8b and 9b: n = 2, R = dibenzyl,

[0111] In compounds 8c and 9c: n = 3, R = phenyl,

[0112] In compounds 8d and 9d: n = 3, R = phenyl,

[0113] In compounds 8e and 9e: n=3, R=benzyl.

[0114] - Compounds of formula 10a-c:

[0115] as well as

[0116] - Compounds of formula 11a-b, 12a-b, 13a-b and 14a:

[0117]

[0118] in:

[0119] In compounds 11a, 12a, 13a, 14a: R = benzyl

[0120] In compounds 11b, 12b, 13b: R = propargyl.

[0121] BI-54903 is also known as tiotropium bromide, particularly its bromide salt.

[0122] BI-607812 has the following structure:

[0123]

[0124] GW870086X has the following structure:

[0125]

[0126] PF-00251802 is an active metabolite of the active metabolite of fosdagrocorat. PF-00251802 is also known as dagrocorat and has the following structure:

[0127]

[0128] Cortivazole has the following structure:

[0129]

[0130] Fluorinated cortivaazole is known in the art and disclosed, for example, in Badarau E. (2019; see above).

[0131] WO 2008 / 021728 A2, WO 2008 / 021729 A2, WO 2008 / 005686 A2, WO 2008 / 027796 A2, WO 2009 / 042377 A1, WO 2010 / 123769 A1, WO 2009 / 023471 A2, WO 2012 / 170175 A1 and WO 2008 / 033655 A2 disclose DIGRA compounds of formula (I):

[0132]

[0133] wherein A and Q are independently selected from unsubstituted and substituted aryl and heteroaryl, unsubstituted and substituted cycloalkyl and heterocycloalkyl, unsubstituted and substituted cycloalkenyl and heterocycloalkenyl, unsubstituted and substituted cycloalkynyl and heterocycloalkynyl, and unsubstituted and substituted heterocyclyl; R 1 and R 2 are independently selected from hydrogen, unsubstituted C 1 -C 15 (Alternatively, C 1 -C 10 or C 1 -C 5 or C 1 -C 3 ) Straight or branched chain alkyl, substituted C 1 -C 15 (Alternatively, C 1 -C 10 or C 1 -C 5 or C 1 -C 3 ) Straight or branched alkyl, unsubstituted C 3 -C 15 Cycloalkyl, and substituted C 3 -C 15 (Alternatively, C 3 -C 6 or C 3 -C 5 )cycloalkyl; R 3 is selected from hydrogen, unsubstituted C 1 -C 15 (Alternatively, C 1 -C 10 or C 1 -C 5 or C 1 -C 3 ) Straight or branched chain alkyl, substituted C 1 -C 15 (Alternatively, C 1 -C 10 or C 1 -C 5 or C 1 -C 3 ) Straight or branched alkyl, unsubstituted C 3 -C 15 (Alternatively, C 3 -C 6 or C 3 -C 5 ) Cycloalkyl and heterocycloalkyl, substituted C 3 -C 15 (Alternatively, C 3 -C 6 or C3 -C 5 ) cycloalkyl and heterocycloalkyl, aryl, heteroaryl and heterocyclic groups; B contains a carbonyl, amino, divalent hydrocarbon or heterohydrocarbon group; E is hydroxy or amino; and D is absent or contains a carbonyl, -NH- or -NR'-, wherein R' contains an unsubstituted or substituted C 1 -C 15 (Alternatively, C 1 -C 10 or C 1 -C 5 or C 1 -C 3 ) a straight or branched chain alkyl group; and wherein R 1 and R 2 can together form unsubstituted or substituted C 3 -C 15 Cycloalkyl.

[0134] In one embodiment, B may contain one or more unsaturated carbon-carbon bonds.

[0135] In another embodiment, B can comprise an alkylenecarbonyl, alkyleneoxycarbonyl, alkylenecarbonyloxy, alkyleneoxycarbonylamino, alkyleneamino, alkenylenecarbonyl, alkenyleneoxycarbonyl, alkenylenecarbonyloxy, alkenyleneoxycarbonylamino, alkenyleneamino, alkynylenecarbonyl, alkynyleneoxycarbonyl, alkynylenecarbonyloxy, alkynyleneoxycarbonylamino, alkynyleneamino, arylcarbonyloxy, aryloxycarbonyl, or urea group.

[0136] In yet another embodiment, A and Q are independently selected from the group consisting of 1 -C 10 Alkoxy (alternatively, C 1 -C 5 Alkoxy or C 1 -C 3 alkyloxy) substituted aryl and heteroaryl; R 1 , R 2 and R 3 independently selected from unsubstituted and substituted C 1 -C 5 Alkyl (preferably, C 1 -C 3 alkyl); B is C 1 -C 5 Alkylene (alternatively, C 1 -C 3 alkyl); D is -NH- or -NR'- group, wherein R' is C 1 -C 5 Alkyl (preferably, C 1 -C 3alkyl); and E is hydroxy.

[0137] In yet another embodiment, A comprises a dihydrobenzofuranyl group substituted with a halogen atom; Q comprises a 1 -C 10 Alkyl substituted quinolyl or isoquinolyl; R 1 and R 2 independently selected from unsubstituted and substituted C 1 -C 5 Alkyl (preferably, C 1 -C 3 alkyl); B is C 1 -C 3 alkylene; D is a -NH- group; E is a hydroxyl group; and R 3 Contains fully halogenated C 1 -C 10 Alkyl (preferably fully halogenated C 1 -C 5 More preferably, fully halogenated C 1 -C 3 alkyl).

[0138] In yet another embodiment, A comprises a dihydrobenzofuranyl group substituted with a fluorine atom; Q comprises a quinolyl or isoquinolyl group substituted with a methyl group; R 1 and R 2 independently selected from unsubstituted and substituted C 1 -C 5 Alkyl; B is C 1 -C 3 alkylene; D is a -NH- group; E is a hydroxyl group; and R comprises a trifluoromethyl group.

[0139] Furthermore, disclosed is a DIGRA compound of formula (IV) which is mapracorat:

[0140]

[0141] WO 2013 / 126156 A1 discloses NO donor DIGRA compounds.

[0142] BI653048 is the compound (R)-2-(4-((5-(ethylsulfonyl)-1H-pyrrolo[2,3-c]pyridin-2-yl)methyl)-5,5,5-trifluoro-4-hydroxy-2-methylpentan-2-yl)-5-fluorobenzamide.

[0143] Its phosphate has the following structure:

[0144]

[0145] WO 2009 / 149139 A1 and EP 2 300 472 disclose BI653048 and related compounds as well as their synthesis.

[0146] WO 2017 / 046096 discloses the following SEGRM compounds and their synthesis. The SEGRM compounds in WO 2017 / 046096 are particularly preferably used for the purposes of the present invention:

[0147] A compound of the following formula (I) or a pharmaceutically acceptable salt, hydrate or solvate thereof:

[0148]

[0149] in:

[0150] R 1 is selected from 5- and 6-membered heteroaryl, (C 1 -C 6 ) alkyl, (C 3 -C 6 )cycloalkyl, (4-6) membered heterocycloalkyl and phenyl, wherein the 5 and 6 membered heteroaryl, (C 1 -C 6 ) alkyl, (C 3 -C 6 )cycloalkyl, (4-6) membered heterocycloalkyl and phenyl are optionally substituted by one or more substituents independently selected from the following: (C 1 -C 4 ) alkyl, (C 1 -C 4 ) alkoxy, halogen, hydroxy and cyano;

[0151] R 2 Selected from (C 1 -C 3 ) alkyl and halogen (C 1 -C 3 )alkyl;

[0152] R 3 is selected from phenyl, 5-membered heteroaryl and 6-membered heteroaryl, wherein the phenyl, 5-membered heteroaryl and 6-membered heteroaryl are optionally substituted by one or more independently selected from R 5 Substituents are substituted;

[0153] R 4 is selected from hydrogen, halogen, (C 1 -C 4 ) alkyl and halogenated (C 1 -C 4 )alkyl;

[0154] R 5 Selected from halogen, cyano, (C1 -C 6 ) alkyl, (C 3 -C 6 )cycloalkyl, (C 1 -C 6 ) alkoxy, halogenated (C 1 -C 6 ) alkyl, halogenated (C 1 -C 6 ) alkoxy, hydroxyl (C 1 -C 6 )alkyl, phenyl, 5-membered heteroaryl, 6-membered heteroaryl and -S(O) 2 R a , where R a Indicates (C 1 -C 4 )alkyl;

[0155] X 1 Selected from CH, C(R b ) and N, where R b Represents halogen, (C 1 -C 4 ) alkyl or halogenated (C 1 -C 4 )alkyl;

[0156] X 2 is selected from CH and N;

[0157] Y is selected from -NH- and -O-;

[0158] m is 0 or 1; n is 0 or 1;

[0159] L represents a bond, -O-, -NH- or -N(R c )-, where R c Indicates (C 1 -C 4 )alkyl.

[0160] Preferably, the SEGRM compound used for the specific use of the present invention is selected from the following compounds or pharmaceutically acceptable salts, hydrates or solvates thereof:

[0161] N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-5-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]pyridine-2-carboxamide,

[0162] 5-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]-N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]pyridine-2-carboxamide,

[0163] 5-[(1R,2S)-1-(4-cyclopropylphenyl)-2-(2,2-difluoropropionylamino)propoxy]-N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]pyridine-2-carboxamide,

[0164] N-[(1S,2R)-2-(4-cyclopropylphenyl)-1-methyl-2-[[6-[[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]carbamoyl]-3-pyridinyl]oxy]ethyl]isothiazole-3-carboxamide,

[0165] N-[(1S,2R)-2-(4-cyclopropylphenyl)-1-methyl-2-[[6-[[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]carbamoyl]-3-pyridinyl]oxy]ethyl]isothiazole-5-carboxamide,

[0166] N-[(1S,2R)-2-(4-cyclopropylphenyl)-1-methyl-2-[[6-[[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]carbamoyl]-3-pyridinyl]oxy]ethyl]oxazole-2-carboxamide,

[0167] N-[(1S,2R)-2-(4-cyclopropylphenyl)-1-methyl-2-[[6-[[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]carbamoyl]-3-pyridinyl]oxy]ethyl]thiazole-4-carboxamide,

[0168] N-[(1S,2R)-2-(4-cyclopropylphenyl)-1-methyl-2-[[6-[[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]carbamoyl]-3-pyridinyl]oxy]ethyl]-3-methyl-isoxazole-5-carboxamide,

[0169] N-[(1S,2R)-2-(4-cyclopropylphenyl)-1-methyl-2-[[6-[[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]carbamoyl]-3-pyridinyl]oxy]ethyl]oxazole-5-carboxamide,

[0170] 5-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[[(2S)-2-hydroxybutyryl]amino]propoxy]-N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]pyridine-2-carboxamide,

[0171] 5-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[[(2R)-2-hydroxypropionyl]amino]propoxy]-N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]pyridine-2-carboxamide,

[0172] N-[(1S,2R)-2-(4-cyclopropylphenyl)-1-methyl-2-[[6-[[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]carbamoyl]-3-pyridinyl]oxy]ethyl]isoxazole-5-carboxamide,

[0173] 5-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[[(2R)-2-hydroxybutyryl]amino]propoxy]-N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]pyridine-2-carboxamide,

[0174] 5-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(2-methoxyacetyl)amino]propoxy]-N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]pyridine-2-carboxamide,

[0175] 5-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(2-hydroxy-2-methyl-propionyl)amino]propoxy]-N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]pyridine-2-carboxamide,

[0176] 5-[(1R,2S)-1-(4-cyclopropylphenyl)-2-(3-hydroxypropionylamino)propoxy]-N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]pyridine-2-carboxamide,

[0177] 5-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(1-hydroxycyclobutanecarbonyl)amino]propoxy]-N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]pyridine-2-carboxamide,

[0178] 5-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(1-hydroxycyclopropanecarbonyl)amino]propoxy]-N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]pyridine-2-carboxamide,

[0179] N-[(1S,2R)-2-(4-cyclopropylphenyl)-1-methyl-2-[[6-[[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]carbamoyl]-3-pyridinyl]oxy]ethyl]oxazole-4-carboxamide,

[0180] 5-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[[(2S)-tetrahydrofuran-2-carbonyl]amino]propoxy]-N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]pyridine-2-carboxamide,

[0181] 5-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(2-hydroxyacetyl)amino]propoxy]-N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]pyridine-2-carboxamide,

[0182] 5-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[[(2R)-tetrahydrofuran-2-carbonyl]amino]propoxy]-N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]pyridine-2-carboxamide,

[0183] N-[(1S,2R)-2-(4-cyclopropylphenyl)-1-methyl-2-[[6-[[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]carbamoyl]-3-pyridinyl]oxy]ethyl]isoxazole-3-carboxamide,

[0184] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]benzoate,

[0185] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-(1,2,5-thiadiazole-3-carbonylamino)propoxy]benzoate,

[0186] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(5-methylthiazole-2-carbonyl)amino]propoxy]benzoate,

[0187] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-(thiazole-5-carbonylamino)propoxy]benzoate,

[0188] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(4-methyl-1,2,5-oxadiazole-3-carbonyl)amino]propoxy]benzoate,

[0189] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(5-methylthiazole-4-carbonyl)amino]propoxy]benzoate,

[0190] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(4-methylthiazole-5-carbonyl)amino]propoxy]benzoate,

[0191] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(5-methyl-1,3,4-oxadiazole-2-carbonyl)amino]propoxy]benzoate,

[0192] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(4-methylthiadiazole-5-carbonyl)amino]propoxy]benzoate,

[0193] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(4-methyloxazole-5-carbonyl)amino]propoxy]benzoate,

[0194] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(2-methoxyacetyl)amino]propoxy]benzoate,

[0195] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(2-methylpyrazole-3-carbonyl)amino]propoxy]benzoate,

[0196] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(2-methylthiazole-5-carbonyl)amino]propoxy]benzoate,

[0197] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[[(2R)-tetrahydrofuran-2-carbonyl]amino]propoxy]benzoate,

[0198] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(3-methyltriazole-4-carbonyl)amino]propoxy]benzoate,

[0199] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-(1,2,4-oxadiazole-3-carbonylamino)propoxy]benzoate,

[0200] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(1-methylimidazole-2-carbonyl)amino]propoxy]benzoate,

[0201] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(3-methylisoxazole-5-carbonyl)amino]propoxy]benzoate,

[0202] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(5-methyl-1,2,4-oxadiazole-3-carbonyl)amino]propoxy]benzoate,

[0203] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(1-methylimidazole-4-carbonyl)amino]propoxy]benzoate,

[0204] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-(isothiazole-3-carbonylamino)propoxy]benzoate,

[0205] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(3-methylisoxazole-4-carbonyl)amino]propoxy]benzoate,

[0206] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-(thiazole-2-carbonylamino)propoxy]benzoate,

[0207] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-(isothiazole-5-carbonylamino)propoxy]benzoate,

[0208] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(5-methylisothiazole-4-carbonyl)amino]propoxy]benzoate,

[0209] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-(oxazole-2-carbonylamino)propoxy]benzoate,

[0210] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-(oxazole-5-carbonylamino)propoxy]benzoate,

[0211] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-(isoxazole-3-carbonylamino)propoxy]benzoate,

[0212] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-(thiadiazole-4-carbonylamino)propoxy]benzoate,

[0213] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(5-methylisoxazole-3-carbonyl)amino]propoxy]benzoate,

[0214] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-(isoxazole-5-carbonylamino)propoxy]benzoate,

[0215] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-(thiazole-4-carbonylamino)propoxy]benzoate,

[0216] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(3-methyl-1,2,4-oxadiazole-5-carbonyl)amino]propoxy]benzoate,

[0217] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[(1-methylpyrazole-3-carbonyl)amino]propoxy]benzoate,

[0218] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-(2,2-difluoropropionylamino)propoxy]benzoate,

[0219] N-[(3R)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-5-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]pyridine-2-carboxamide,

[0220] N-[(3R)-1-[(3S)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-5-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]pyridine-2-carboxamide,

[0221] N-[(3R)-1-[(2S)-5-oxotetrahydrofuran-2-carbonyl]-3-piperidinyl]-5-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]pyridine-2-carboxamide,

[0222] N-[(3R)-1-[(2R)-5-oxotetrahydrofuran-2-carbonyl]-3-piperidinyl]-5-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]pyridine-2-carboxamide,

[0223] N-[(3S)-1-[(3S)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-5-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]pyridine-2-carboxamide,

[0224] N-[(3S)-1-[(2S)-5-oxotetrahydrofuran-2-carbonyl]-3-piperidinyl]-5-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]pyridine-2-carboxamide,

[0225] N-[(3R)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]pyrrolidin-3-yl]-5-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]pyridine-2-carboxamide,

[0226] 5-[(1R,2S)-1-(4-ethylphenyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]-N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]pyridine-2-carboxamide,

[0227] 5-[(1R,2S)-1-(4-bromophenyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]-N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]pyridine-2-carboxamide,

[0228] N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-5-[(1R,2S)-1-(4-phenylphenyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]pyridine-2-carboxamide,

[0229] N-[(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]benzamide,

[0230] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]4-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]benzoate,

[0231] N-[(3S)-1-[[(3S)-5-oxotetrahydrofuran-3-yl]carbamoyl]-3-piperidinyl]-5-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]pyridine-2-carboxamide,

[0232] N-[(3S)-1-[[(3R)-5-oxotetrahydrofuran-3-yl]carbamoyl]-3-piperidinyl]-5-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]pyridine-2-carboxamide,

[0233] [(3S)-2-oxotetrahydrofuran-3-yl](3S)-3-[[5-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]pyridine-2-carbonyl]amino]piperidine-1-carboxylate,

[0234] [(3R)-2-oxotetrahydrofuran-3-yl](3S)-3-[[5-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]pyridine-2-carbonyl]amino]piperidine-1-carboxylate,

[0235] [(3S)-5-oxotetrahydrofuran-3-yl](3S)-3-[[5-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]pyridine-2-carbonyl]amino]piperidine-1-carboxylate, or

[0236] [(34)-5-Oxotetrahydrofuran-3-yl](3S)-3-[[5-[(1R,2S)-1-(p-tolyl)-2-[(2,2,2-trifluoroacetyl)amino]propoxy]pyridine-2-carbonyl]amino]piperidine-1-carboxylate.

[0237] In a more preferred embodiment, [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]-4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[[(2R)-tetrahydrofuran-2-carbonyl)amino]propoxy]benzoate or a pharmaceutically acceptable salt thereof can be used in the present invention. The compound corresponds to "Compound 37" in WO 2017 / 046096, which has the following structure:

[0238]

[0239] WO 2009 / 065503 discloses the following three SEGRM compounds and their synthesis.

[0240] 5-{[(1S,2S)-1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(methoxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one has the following structure:

[0241]

[0242] This compound is known as Example 5 from WO 2009 / 065503 A (without stereochemistry as racemate) and can be obtained from the racemic form by chiral HPLC on Chiralpak IC with 5 μm eluent (hexane / ethanol (4:1)).

[0243] In an even more preferred embodiment, 5-{[(1S,2S)-1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(methoxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one or a pharmaceutically acceptable salt thereof can be used in the present invention.

[0244] 5-{(1S,2S)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(hydroxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one has the following structure:

[0245]

[0246] This compound is known as Example 7 from WO 2009 / 065503 A (without stereochemistry as racemate) and can be obtained from the racemic form by chiral HPLC on Chiralpak IC with 5 μm eluent (hexane / ethanol (4:1)).

[0247] In a more preferred embodiment, 5-{(1S,2S)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(hydroxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one or a pharmaceutically acceptable salt thereof can be used in the present invention.

[0248] 5-{(1S,2R)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-([methylsulfonyl]methyl)propyl]amino}-1H-quinolin-2-one has the following structure:

[0249]

[0250] This compound is known as Example 3 from WO 2009 / 065503 A (without stereochemistry as racemate) and can be obtained from the racemic form by chiral HPLC on Chiralpak IC with 5 μm eluent (hexane / ethanol (4:1)).

[0251] In a more preferred embodiment, 5-{(1S,2R)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-([methylsulfonyl]methyl)propyl]amino}-1H-quinolin-2-one or a pharmaceutically acceptable salt thereof can be used in the present invention.

[0252] These SEGRM compounds and their synthesis are disclosed in WO 2009 / 065503.

[0253] Therefore, in one embodiment, preferred according to the invention is the use of a compound of formula (I) and a salt, solvate or salt of a solvate thereof:

[0254]

[0255] in:

[0256] R 1 and R 2 independently represent a hydrogen atom, a hydroxyl group, a halogen atom, an optionally substituted (C 1 -C 10 )-alkyl, optionally substituted (C 1 -C 10 )-alkoxy, (C 1 -C 10 )-alkylthio, (C 1 -C 5 )-perfluoroalkyl, cyano, nitro, or, R 1 and R 2 Together they represent a group selected from: -O-(CH2 ) p -O-, -O-(CH 2 ) p -CH 2 -, -O-CH=CH-, -(CH 2 ) p+2 -、-NH-(CH 2 ) p+1 、-N(C 1 -C 3 -alkyl)-(CH 2 ) p+1 and -NH-N=CH-, wherein p=1 or 2, and the terminal oxygen atom and / or carbon atom and / or nitrogen atom is attached to the directly adjacent ring carbon atom, or NR 6 R 7 , where R 6 and R 7 Independently represent hydrogen, C 1 -C 5 -alkyl or (CO)-(C 1 -C 5 )-alkyl,

[0257] R 3 represents a hydrogen atom, a hydroxyl group, a halogen atom, a cyano group, an optionally substituted (C 1 -C 10 )-alkyl, (C 1 -C 10 )-alkoxy, (C 1 -C 10 )-alkylthio or (C 1 -C 5 )-perfluoroalkyl,

[0258] R 4 represents hydrogen, halogen, hydroxyl, (C 1 -C 5 )-alkyl, (C 1 -C 5 )-alkoxy, (C 1 -C 5 )-alkylthio, (C 1 -C 5 )-perfluoroalkyl, cyano, nitro, NR 6 R 7 、COOR 9 、(CO)NR 6 R 7 or (C 1 -C 5 -alkylene)-O-(CO)-(C 1 -C 5 )alkyl,

[0259] R 5 Indicates except -CH(CH 3 ) 2 or -C(CH 3 )=CH 2 The other group is selected from the following groups: -(C 1 -C 10 )-alkyl, -(C 2 -C 10 )-alkenyl, -(C 2 -C 10 )-alkynyl, (C 3 -C 7 )cycloalkyl-(C 1 -C 8 ) alkyl, (C 3 -C 7 )cycloalkyl-(C 1 -C 8 ) alkenyl, (C 3 -C 7 )cycloalkyl-(C 2 -C 8 ) alkynyl, heterocyclic-(C 1 -C 8 )alkyl, heterocyclic-(C 1 -C 8 )alkenyl, heterocyclic-(C 2 -C 8 ) alkynyl, -R 8 , R 8 -(C 1 -C 8 ) alkyl, R 8 -(C 2 -C 8 ) alkenyl, R 8 -(C 2 -C 8 )alkynyl, -S-(C 1 -C 10 )-alkyl, -SO 2 -(C 1 -C 10 )-alkyl, -SR 8 、-SO 2 -R 8 、-CN、-Hal、-O-(C 1 -C 10 )-alkyl, wherein R 6 and R 7 -NR having the meaning defined above 6 R 7 、-OR 8 、-OH,

[0260] R 8 It can be optionally replaced by 1-3 hydroxyl groups, halogen groups, C 1 -C 5 Alkyl, C 1 -C 5 -Alkoxy, cyano, CF 3 , nitro, COO(C 1 -C 5 -alkyl) or C(O)OCH 2 -phenyl substituted aryl, or may contain 1-3 heteroatoms of heteroaryl, the heteroatoms may be optionally replaced by 1-3 alkyl, hydroxy, halogen, cyano or C 1 -C 5 -Alkoxy substitution.

[0261] More preferred is the use of a compound of formula (I) and a salt, solvate or salt of a solvate thereof, wherein:

[0262] R 1 and R 2 independently represent a hydrogen atom, a hydroxyl group, a halogen atom, an optionally substituted (C 1 -C 10 )-alkyl, optionally substituted (C 1 -C 10 )-alkoxy, (C 1 -C 10 )-alkylthio, (C 1 -C 5 )-perfluoroalkyl, cyano, nitro, or, R 1 and R 2 Together they represent a group selected from: -O-(CH 2 ) p -O-, -O-(CH 2 ) p -CH 2 -, -O-CH=CH-, -(CH 2 ) p+2 -、-NH-(CH 2 ) p+1 、-N(C 1 -C 3 -alkyl)-(CH 2 ) p+1 and -NH-N=CH-, wherein p=1 or 2, and the terminal oxygen atom and / or carbon atom and / or nitrogen atom is attached to the directly adjacent ring carbon atom, or NR 6 R 7 , where R 6 and R 7 Independently represent hydrogen, C 1-C 5 -alkyl or (CO)-(C 1 -C 5 )-alkyl,

[0263] R 3 represents a hydrogen atom, a hydroxyl group, a halogen atom, a cyano group, an optionally substituted (C 1 -C 10 )-alkyl, (C 1 -C 10 )-alkoxy, (C 1 -C 10 )-alkylthio or (C 1 -C 5 )-perfluoroalkyl,

[0264] R 4 represents a hydrogen atom, a hydroxyl group, or a halogen atom,

[0265] R 5 Indicates except -CH(CH 3 ) 2 or -C(CH 3 )=CH 2 The other group is selected from the following groups: -(C 1 -C 10 )-alkyl, -(C 2 -C 10 )-alkenyl, -(C 2 -C 10 )-alkynyl, (C 3 -C 7 )cycloalkyl-(C 1 -C 8 ) alkyl, (C 3 -C 7 )cycloalkyl-(C 2 -C 8 ) alkenyl, (C 3 -C 7 )cycloalkyl-(C 2 -C 8 ) alkynyl, heterocyclic-(C 1 -C 8 )alkyl, heterocyclic-(C 2 -C 8 )alkenyl, heterocyclic-(C 2 -C 8 ) alkynyl, -R 8 , R 8 -(C 1 -C 8 ) alkyl, R 8 -(C 2 -C 8 ) alkenyl, R8 -(C 2 -C 8 )alkynyl, -S-(C 1 -C 10 )-alkyl, -SR 8 、-SO 2 -R 8 、-SO 2 -(C 1 -C 10 )-alkyl-CN, -Hal, -O-(C 1 -C 10 )-alkyl, wherein R 6 and R 7 -NR having the meaning defined above 6 R 7 、-OR 8 、-OH,

[0266] R 8 It may be optionally replaced by 1-3 alkyl, hydroxy, halogen, cyano or C 1 -C 5 -alkoxy substituted aryl, or may contain 1-3 heteroatoms of heteroaryl, the heteroatoms may be optionally replaced by 1-3 alkyl, hydroxy, halogen, cyano or C 1 -C 5 -alkoxy substituted, n represents an integer selected from 1, 2, 3, 4, 5.

[0267] More preferred is the use of a compound of formula (I) and its salt, solvate or salt of a solvate, wherein R 1 and R 2 independently represent a hydrogen atom, a hydroxyl group, a halogen atom, an optionally substituted (C 1 -C 10 )-alkyl, optionally substituted (C 1 -C 10 )-alkoxy, (C 1 -C 5 )-perfluoroalkyl, cyano, or NR 6 R 7 , where R 6 and R 7 Independently represent hydrogen, C 1 -C 5 -alkyl or (CO)-(C 1 -C 5 )-alkyl, R 3 represents a hydrogen atom, a hydroxyl group, a halogen atom, a cyano group, an optionally substituted (C 1 -C 10 )-alkyl, (C 1 -C10 )-alkoxy or (C 1 -C 5 )-perfluoroalkyl, R 4 Represents hydrogen, C 1 -C 3 -alkyl, C 1 -C 3 -alkoxy, hydroxy, halogen, R 5 Indicates except -CH(CH 3 ) 2 or -C(CH 3 )=CH 2 The other group is selected from the following groups: -(C 1 -C 10 )-alkyl, -(C 2 -C 10 )-alkenyl, -(C 2 -C 10 )-alkynyl, -(C 3 -C 7 )cycloalkyl-(C 1 -C 8 )alkyl, -(C 3 -C 7 )cycloalkyl-(C 2 -C 8 ) alkenyl, -S-(C 1 -C 10 )-alkyl, -SO 2 -(C 1 -C 10 )-alkyl, -CN, -Hal, -O-(C 1 -C 10 )-alkyl, wherein R 6 and R 7 -NR having the meaning defined above 6 R 7 、-OH.

[0268] Another aspect of the present invention is a compound of the general formula I according to claim 1 and a salt, a solvate or a salt of a solvate thereof, wherein R 1 , R 2 and R 3 are independently hydrogen, fluorine, chlorine, bromine, cyano, methoxy, ethoxy, hydroxyl, R 4 It is hydrogen, C 1 -C 3 -alkyl, halogen, R 5 Hydroxyl, chlorine, -S-CH 3 、-S-CH 2 -CH 3 、-S-CH 2-CH 2 -CH 3 、-O-CH 3 or -O-CH 2 -CH 3 、-O-CH 2 -CH 2 -CH 3 、-N-(CH 3 ) 2 、-N-(CH 2 -CH 3 ) 2 .

[0269] Another aspect of the present invention is a compound of the general formula I according to claim 1 and a salt, a solvate or a salt of a solvate thereof, wherein R 1 , R 2 and R 3 are independently hydrogen, fluorine, chlorine, bromine, cyano, methoxy, ethoxy, hydroxyl, R 4 It is hydrogen, C 1 -C 3 -alkyl, halogen, R 5 Hydroxyl, chlorine, -S-CH 3 、-S-CH 2 -CH 3 、-S-CH 2 -CH 2 -CH 3 、-O-CH 3 、-O-CH 2 -CH 3 、-O-CH 2 -CH 2 -CH 3 or N-(CH 3 ) 2 .

[0270] More preferred is the use of a compound of formula (I) and a salt, solvate or salt of a solvate thereof, wherein R 1 and R 2 and R 3 are independently hydrogen, fluorine, chlorine, bromine, cyano, methoxy, ethoxy, hydroxyl, R 4 It is hydrogen, CrC 3 -alkyl, halogen, R 5 Hydroxyl, chlorine, -S-CH 3 、-S-CH 2 -CH 3 、-S-CH 2 -CH 2 -CH 3 、-O-CH 3 or -O-CH2 -CH 3 、-O-CH 2 -CH 2 -CH 3 .

[0271] More preferred is the use of a compound of formula (I) and a salt, solvate or salt of a solvate thereof, wherein R 1 and R 2 are independently hydrogen, fluorine, chlorine, methoxy, hydroxyl, R 3 is hydrogen, fluorine, chlorine or methoxy, R 4 is hydrogen or fluorine, R 5 It is a hydroxyl group, a chlorine atom, -S-CH 3 、-S-CH 2 -CH 3 、-O-CH 3 、-O-CH 2 -CH 3 or N-(CH 3 ) 2 .

[0272] Another aspect of the present invention is a compound of the general formula I according to claim 1 and a salt, a solvate or a salt of a solvate thereof, wherein R 1 and R 2 are independently hydrogen, fluorine, chlorine, methoxy, R 3 is hydrogen, fluorine, chlorine or methoxy, R 4 is hydrogen or fluorine, R 5 It is a hydroxyl group, a chlorine atom, -S-CH 3 、-S-CH 2 -CH 3 、-O-CH 3 or -O-CH 2 -CH 3 .

[0273] In a more preferred embodiment, the compounds for particular use and their salts, solvates or salts or solvates are in enantiomerically pure form.

[0274] In a more preferred embodiment, the compound for specific use is selected from the following list and their salts, solvates or salts of solvates:

[0275] 5-{[1-(2-Fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-([methylthio]methyl)propyl]amino}-1H-quinolin-2-one

[0276] 5-{[2-([Ethylthio]methyl)-1-(2-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-1H-quinolin-2-one

[0277] 5-{[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-([methylthio]methyl)propyl]amino}-1H-quinolin-2-one

[0278] 5-{[1-(2-chloro-3-fluoro-4-methoxyphenyl)-2-([ethylthio]methyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-7-fluoro-1H-quinolin-2-one

[0279] 5-{[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(methoxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one

[0280] 5-{[1-(2-chloro-3-fluoro-4-methoxyphenyl)-2-(ethoxymethyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-7-fluoro-1H-quinolin-2-one

[0281] 5-{[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(hydroxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one

[0282] 5-{[1-(5-chloro-3-fluoro-2-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(hydroxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one

[0283] 5-{[1-(5-chloro-3-fluoro-2-methoxyphenyl)-2-(chloromethyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-7-fluoro-1H-quinolin-2-one

[0284] 5-{[3,3,3-Trifluoro-2-hydroxy-2-([methoxymethyl)-1-phenylpropyl]amino}-1H-quinolin-1-one

[0285] 5-{[1-(2-chloro-3-fluoro-4-methoxyphenyl)-2-(diaminomethyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-7-fluoro-1H-quinolin-2-one

[0286] 5-{[1-(4-chloro-3-fluoro-2-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(methoxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one

[0287] 5-{[1-(2-chloro-3-fluoro-4-methoxyphenyl)-2-(ethoxymethyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-7-fluoro-1H-quinolin-2-one

[0288] 5-{[1-(2-Chloro-3-fluoro-4-hydroxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(hydroxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one.

[0289] More preferred are compounds selected from the following (especially enantiomerically pure compounds) and their salts, solvates or salts of solvates:

[0290] 5-{[1-(2-Fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-([methylthio]methyl)propyl]amino}-1H-quinolin-2-one

[0291] 5-{[2-([Ethylthio]methyl)-1-(2-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-1H-quinolin-2-one

[0292] 5-{[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-([methylthio]methyl)propyl]amino}-1H-quinolin-2-one

[0293] 5-{[1-(2-chloro-3-fluoro-4-methoxyphenyl)-2-([ethylthio]methyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-7-fluoro-1H-quinolin-2-one

[0294] 5-{[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(methoxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one

[0295] 5-{[1-(2-chloro-3-fluoro-4-methoxyphenyl)-2-(ethoxymethyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-7-fluoro-1H-quinolin-2-one

[0296] 5-{[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(hydroxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one

[0297] 5-{[1-(5-chloro-3-fluoro-2-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(hydroxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one

[0298] 5-{[1-(5-chloro-3-fluoro-2-methoxyphenyl)-2-(chloromethyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-7-fluoro-1H-quinolin-2-one

[0299] 5-{[3,3,3-Trifluoro-2-hydroxy-2-([methoxymethyl)-1-phenylpropyl]amino}-1H-quinolin-1-one.

[0300] More preferred is the use of compounds selected from the following list (especially in enantiomerically pure form) and their salts, solvates or salts of solvates:

[0301] 5-{(1S,2R)[1-(2-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-([methylthio]methyl)propyl]amino}-1H-quinolin-2-one

[0302] 5-{(1S,2R)[2-([ethylthio]methyl)-1-(2-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-1H-quinolin-2-one

[0303] 5-{(1S,2R)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-([methylthio]methyl)propyl]amino}-1H-quinolin-2-one

[0304] 5-{(1S,2R)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-2-([ethylthio]methyl)-trifluoro-2-hydroxypropyl]amino}-7-fluoro-1H-quinolin-2-one

[0305] 5-{(1S,2S)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(methoxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one

[0306] 5-{(1S,2S)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-2-(ethoxymethyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-7-fluoro-1H-quinolin-2-one

[0307] 5-{(1S,2S)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(hydroxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one

[0308] 5-{(1S,2S)[1-(5-chloro-3-fluoro-2-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(hydroxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one

[0309] 5-{(1S,2R)[1-(5-chloro-3-fluoro-2-methoxyphenyl)-2-(chloromethyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-7-fluoro-1H-quinolin-2-one

[0310] 5-{(1S,2S)[3,3,3-trifluoro-2-hydroxy-2-([methoxymethyl)-1-phenylpropyl]amino}-1H-quinolin-1-one

[0311] 5-{(1S,2R)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-2-(diaminomethyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-7-fluoro-1H-quinolin-2-one

[0312] 5-{(1S,2S)[1-(2-chloro-3-fluoro-4-hydroxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(hydroxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one.

[0313] More preferred is the use of compounds selected from the group consisting of the following (especially in enantiomerically pure form) and their salts, solvates or salts of solvates:

[0314] 5-{(1S,2R)[1-(2-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-([methylthio]methyl)propyl]amino}-1H-quinolin-2-one

[0315] 5-{(1S,2R)[2-([ethylthio]methyl)-1-(2-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-1H-quinolin-2-one

[0316] 5-{(1S,2R)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-([methylthio]methyl)propyl]amino}-1H-quinolin-2-one

[0317] 5-{(1S,2R)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-2-([ethylthio]methyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-7-fluoro-1H-quinolin-2-one

[0318] 5-{(1S,2S)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(methoxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one

[0319] 5-{(1S,2S)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-2-(ethoxymethyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-7-fluoro-1H-quinolin-2-one

[0320] 5-{(1S,2S)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(hydroxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one

[0321] 5-{(1S,2S)[1-(5-chloro-3-fluoro-2-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(hydroxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one

[0322] 5-{(1S,2R)[1-(5-chloro-3-fluoro-2-methoxyphenyl)-2-(chloromethyl)-3,3,3-trifluoro-2-hydroxypropyl]amino}-7-fluoro-1H-quinolin-2-one

[0323] 5-{(1S,2S)[3,3,3-Trifluoro-2-hydroxy-2-([methoxymethyl)-1-phenylpropyl]amino}-1H-quinolin-1-one.

[0324] In yet another embodiment, preferred according to the present invention is the use of a compound of formula (I) or a tautomer, optical isomer, prodrug, cocrystal or pharmaceutically acceptable salt thereof:

[0325]

[0326] in:

[0327] R 1 is aryl or heteroaryl, each optionally independently substituted by one, two or three substituents selected from the following: 1 -C 5 Alkyl, aminocarbonyl, C 1 -C 5 Alkylaminocarbonyl, C 1 -C 5 Dialkylaminocarbonyl, aminosulfonyl, C 1 -C 5 Alkylaminosulfonyl, C 1 -C 5 dialkylaminosulfonyl, halogen, hydroxyl, cyano and C 1 -C 5 Alkylthio, wherein the sulfur atom is optionally oxidized to a sulfoxide or sulfone;

[0328] R 2 C 1 -C 5Alkylthio, wherein the sulfur atom is optionally oxidized to a sulfoxide or sulfone, optionally substituted independently with one, two or three substituents selected from halogen, hydroxy, oxo, cyano, alkoxyalkyl and aminocarbonyl;

[0329] X is CH or N; and

[0330] Y is CH or N,

[0331] Where X and Y are not both CH.

[0332] Preferred is the use of a compound of formula (I) or a tautomer, prodrug, cocrystal or pharmaceutically acceptable salt thereof, wherein:

[0333] R 1 is aryl or heteroaryl, each optionally substituted independently by one, two or three substituents selected from the following: 1 -C 5 Alkyl, aminocarbonyl, C 1 -C 5 Alkylaminocarbonyl, C 1 -C 5 Dialkylaminocarbonyl, aminosulfonyl, C 1 -C 5 Alkylaminosulfonyl, C 1 -C 5 dialkylaminosulfonyl, halogen, hydroxyl, cyano and C 1 -C 5 Alkylthio, wherein the sulfur atom is optionally oxidized to a sulfoxide or sulfone;

[0334] R 2 C 1 -C 5 Alkylthio, wherein the sulfur atom is optionally oxidized to a sulfoxide or sulfone, each optionally substituted independently with one to three substituents selected from halogen, hydroxy, oxo, cyano, alkoxyalkyl and aminocarbonyl;

[0335] X is CH; and

[0336] Y is N.

[0337] Preferred is the use of a compound of formula (I) or a tautomer, prodrug, cocrystal or pharmaceutically acceptable salt thereof, wherein:

[0338] R 1 is aryl, optionally substituted by one, two or three substituents independently selected from the following: 1 , C 2 or C 3 Alkyl, aminocarbonyl, halogen, and C 1 , C 2 or C 3Alkylthio, wherein the sulfur atom is optionally oxidized to a sulfoxide or sulfone;

[0339] R 2 C 1 , C 2 or C 3 Alkylthio, wherein the sulfur atom is optionally oxidized to a sulfoxide or sulfone, each optionally substituted independently with one to three substituents selected from halogen, hydroxy, oxo, cyano, alkoxyalkyl and aminocarbonyl;

[0340] X is CH; and

[0341] Y is N.

[0342] Preferred is the use of a compound of formula (I) or a tautomer, prodrug, cocrystal or salt thereof, wherein:

[0343] R 1 is phenyl, optionally substituted by one or two substituents independently selected from the group consisting of aminocarbonyl, methyl, fluorine, chlorine, bromine and C 1 or C 2 Alkylthio, wherein the sulfur atom is optionally oxidized to a sulfoxide or sulfone;

[0344] R 2 C 1 , C 2 or C 3 Alkylthio, wherein the sulfur atom is optionally oxidized to a sulfoxide or sulfone;

[0345] X is CH; and

[0346] Y is N.

[0347] More preferred is the use of a compound of formula (I) or a tautomer, prodrug, cocrystal or pharmaceutically acceptable salt thereof, wherein:

[0348] R 1 is phenyl, optionally substituted by one or two substituents independently selected from the group consisting of aminocarbonyl, methyl, fluorine, chlorine, bromine and C 1 or C 2 Alkylthio, wherein the sulfur atom is optionally oxidized to a sulfoxide or sulfone;

[0349] R 2 C 1 or C 2 Alkylthio, wherein the sulfur atom is optionally oxidized to a sulfoxide or sulfone;

[0350] X is CH; and

[0351] Y is N.

[0352] WO 2018 / 046678 A1 discloses suitable topical preparations of these compounds for specific uses of the present invention. WO 2018 / 046685 A1 discloses the use of these compounds in topical treatment of T cell-mediated inflammatory skin diseases such as psoriasis, atopic dermatitis, and allergic contact dermatitis.

[0353] BI-3047 has the following structure and is an inactive analog of BI-653048:

[0354]

[0355] BI-3047 is not a SEGRM.

[0356] In particular, it was found in the Examples that BI-653048 was able to rescue primary human fibroblasts in monolayer (2D) culture from the growth inhibitory effects of wound exudate from non-healing wounds, such as for WE-1 to WE-3. Figure 1 A-1C. This effect was dose-dependent, with a maximum at 10 μM. In contrast, BI-3047 (an inactive analog of BI-653048) had no proliferation-promoting effect at all tested concentrations. In the absence of WE, both compounds showed similar fibroblast growth inhibition ( Figure 1 D).

[0357] Furthermore, BI-653048, but not BI-3047, reduced IL-1β secretion in fibroblast cultures at the same concentrations that enhanced proliferation in the presence of WE ( Figure 2 ).

[0358] Collagen 1 and 3, produced by fibroblasts, are important for the wound healing process. Collagen 1 and 3 mRNA expression is reduced by aggressive wound exudate (e.g., compare Figure 3 A and 3C and targeting collagen 3 Figure 3 WE and culture medium in B and 3D). Figure 3 BI-653048, but not BI-3047, induced mRNA expression of collagens 1 and 3 in the presence of wound exudate, as shown in B. For both compounds, there was no significant effect on collagen 1 mRNA in the presence of medium, and collagen 3 mRNA was reduced in the absence of medium.

[0359] In a fibroblast-derived matrix formation assay (3D fibroblast culture), BI-653048 effectively induced matrix formation in the presence of wound exudate, as Figure 4This effect could be completely abolished by the glucocorticoid receptor antagonist mifepristone, indicating that the substrate-promoting activity of SEGRM is mediated by the glucocorticoid receptor.

[0360] WO2006 / 050998A1 discloses Mapracorat and related 5-substituted quinolone and isoquinoline derivative compounds (including ZK-216448) and their synthesis. In particular, compounds of the following formula (IIa) or (IIb) and their racemates or independently existing stereoisomers and optionally their pharmaceutically acceptable salts or their prodrugs can be used:

[0361]

[0362] in

[0363] R1 and R2 independently of each other may be a hydrogen atom, a C1-3 alkyl group, a halogen atom, a cyano group, a C1-3 alkoxy group or a hydroxyl group.

[0364] The designation halogen atom or halogen denotes a fluorine, chlorine, bromine or iodine atom. Preference is given to fluorine, chlorine or bromine atoms.

[0365] The C1-C3 alkyl group and the C1-C5 alkyl group may be linear or branched and represent methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl or n-pentyl, 2,2-dimethylpropyl, 2-methylbutyl or 3-methylbutyl.

[0366] Preferred is methyl or ethyl.

[0367] The radicals R1 and R2 preferably represent hydrogen, C1-3 alkyl, halogen or hydroxy. Particular preference is given to hydrogen, methyl, chlorine and hydroxy.

[0368] Therefore, preference is given to the use of 5-substituted quinolone and isoquinoline derivative compounds of the general formulae IIa and IIb, in which R1 and R2 independently of one another preferably represent hydrogen, C1-3 alkyl, halogen or hydroxy.

[0369] Particularly preferred is the use of 5-substituted quinolone and isoquinoline derivative compounds of the formula I and IIb, in which R1 and R2 independently of one another preferably represent hydrogen, methyl, chlorine or hydroxyl.

[0370] More preferred is the use of 5-substituted quinolone and isoquinoline derivative compounds of the general formula IIa.

[0371] Due to the presence of asymmetric centers, the 5-substituted quinolone and isoquinoline derivative compounds of the general formula (IIa) and (IIb) for the specific use of the present invention can exist in different stereoisomers. Both the racemates and the individual stereoisomers belong to the subject matter of the present invention.

[0372] In particular, the independently existing stereoisomers of Examples 1, 2, 3, 4, 5, 11 and 12 of WO 2006 / 050998 A1, ie the (+)-enantiomer and the (-)-enantiomer, are particularly preferred for the specific use according to the present invention.

[0373] The compounds for the specific use according to the invention, if they contain a hydroxyl group in the alpha position to the quinolyl or isoquinolyl nitrogen atom, can be distinguished by the presence of keto-enol tautomerism. According to the invention, even if, for example, only one of the two tautomeric forms is mentioned in the experimental part, both forms are part of the subject matter of the invention.

[0374] In particular, the following SEGRMs may be used according to the invention:

[0375] 5-[4-(5-Fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2-methylquinoline,

[0376] 5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-1-methylisoquinoline),

[0377] 5-[4-(5-Fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]isoquinol-1(2H)-one,

[0378] 5-[4-(5-Fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2,6-dimethylquinoline,

[0379] 5-[4-(5-Fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-6-chloro-2-methylquinoline,

[0380] 5-[4-(5-Fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]isoquinoline,

[0381] 5-[4-(5-Fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]quinoline,

[0382] 5-[4-(2,3-dihydro-5-fluoro-7-benzofuranyl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]quinolin-2[1H]-one,

[0383] 6-Fluoro-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2-methylquinoline,

[0384] 8-Fluoro-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2-methylquinoline,

[0385] 5-[4-(5-Fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2-methylisoquinol-1(2H)-one, and their individual enantiomers:

[0386] 2(R)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2-methylquinoline),

[0387] 2(R)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-1-methylisoquinoline,

[0388] 2(R)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]isoquinol-1(2H)-one,

[0389] 2(R)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2,6-dimethylquinoline,

[0390] 2(R)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-6-chloro-2-methylquinoline,

[0391] 2(R)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]isoquinoline,

[0392] 2(R)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]quinoline,

[0393] 2(R)-5-[4-(2,3-dihydro-5-fluoro-7-benzofuranyl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]quinolin-2[1H]-one,

[0394] 2(R)-6-Fluoro-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2-methylquinoline,

[0395] 2(R)-8-Fluoro-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2-methylquinoline,

[0396] 2(R)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2-methylquinolin-1(2H)-one,

[0397] 2(S)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2-methylquinoline),

[0398] 2(S)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-1-methylisoquinoline),

[0399] 2(S)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]isoquinol-1(2H)-one,

[0400] 2(S)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2,6-dimethylquinoline,

[0401] 2(S)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-6-chloro-2-methylquinoline,

[0402] 2(S)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]isoquinoline,

[0403] 2(S)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]quinoline,

[0404] 2(S)-5-[4-(2,3-dihydro-5-fluoro-7-benzofuranyl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]quinolin-2[1H]-one,

[0405] 2(S)-6-Fluoro-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2-methylquinoline,

[0406] 2(S)-8-Fluoro-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2-methylquinoline,

[0407] 2(S)-5-[4-(5-Fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2-methylisoquinol-1(2H)-one.

[0408] Particularly preferred are 5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2-methylquinoline and its individual enantiomers 2-(R)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2-methylquinoline and 2-(S)-5-[4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-2-hydroxy-4-methyl-2-trifluoromethyl-pentylamino]-2-methylquinoline.

[0409] In an even more preferred embodiment, mapracorat or a pharmaceutically acceptable salt thereof may be used according to the present invention. Mapracorat is the INN name of the compound having the following formula (I):

[0410]

[0411] ZK-216348 has the following structure (III)

[0412]

[0413] In particular, it was found in the Examples that mapracorat enhanced the proliferation of fibroblasts in 2D culture in the presence of wound exudate in the same manner as BI-653048 ( Figure 5 ).

[0414] In the presence of wound exudate (but not culture medium), gradient concentrations of mapracorat enhanced the mRNA expression of collagen 1 and collagen 3 in 2D fibroblast cultures ( Figure 6 and Figure 7 ). In contrast, mapracorat dose-dependently inhibited IL-1β mRNA expression in these cultures, as shown in Figure 8As shown in A (IL-1β mRNA level in the culture medium was below the detection limit, Figure 8 B).

[0415] Furthermore, in a porcine model of delayed wound healing, mapracorat was found to reduce wound scores from day 6 to day 12 in wounds treated with wound exudate from chronic human wounds and the TLR7 / 8 agonist R848 as an inducer of delayed wound healing. Mapracorat did not have any negative effect on the healing of control wounds in the presence of human serum.

[0416] In particular, it was found in the Examples that ZK-216348 and HY14234 were able to rescue fibroblasts in 2D culture from wound exudate-induced proliferation inhibition ( Fig. 9 A, 10A and 11A). ZK216348 was less efficient. In the presence of three different wound exudates, both ZK-216348 and HY14234 inhibited wound exudate-induced Il-1β secretion in fibroblast culture ( Fig. 9 B, 10B and 11B).

[0417] In a particularly preferred embodiment, the SEGRM for a particular use of the present invention is selected from:

[0418] (i) a compound of the following formula (IIa) or (IIb) and its racemate or independently existing stereoisomer and optionally their pharmaceutically acceptable salts or their prodrugs:

[0419]

[0420] Where R 1 and R 2 can be independently a hydrogen atom, a C 1-3 Alkyl, halogen atom, cyano, C 1-3 Alkoxy or hydroxyl;

[0421] (ii) compound (R)-2-(4-((5-(ethylsulfonyl)-1H-pyrrolo[2,3-c]pyridin-2-yl)methyl)-5,5,5-trifluoro-4-hydroxy-2-methylpentan-2-yl)-5-fluorobenzamide, or a pharmaceutically acceptable salt thereof;

[0422] (iii) a compound of the formula, or a pharmaceutically acceptable salt thereof,

[0423]

[0424] (iv) a compound of the formula, or a pharmaceutically acceptable salt thereof,

[0425]

[0426] (v) a compound of the following formula (I) or a pharmaceutically acceptable salt, hydrate or solvate thereof,

[0427]

[0428] in:

[0429] R 1 is selected from 5- and 6-membered heteroaryl, (C 1 -C 6 ) alkyl, (C 3 -C 6 )cycloalkyl, (4-6) membered heterocycloalkyl and phenyl, wherein the 5 and 6 membered heteroaryl, (C 1 -C 6 ) alkyl, (C 3 -C 6 )cycloalkyl, (4-6) membered heterocycloalkyl and phenyl are optionally substituted by one or more substituents independently selected from the following: (C 1 -C 4 ) alkyl, (C 1 -C 4 ) alkoxy, halogen, hydroxy and cyano;

[0430] R 2 Selected from (C 1 -C 3 ) alkyl and halogen (C 1 -C 3 )alkyl;

[0431] R 3 is selected from phenyl, 5-membered heteroaryl and 6-membered heteroaryl, wherein the phenyl, 5-membered heteroaryl and 6-membered heteroaryl are optionally substituted by one or more independently selected from R 5 Substituents are substituted;

[0432] R 4 is selected from hydrogen, halogen, (C 1 -C 4 ) alkyl and halogenated (C 1 -C 4 )alkyl;

[0433] R 5 Selected from halogen, cyano, (C 1 -C 6 ) alkyl, (C 3 -C 6 )cycloalkyl, (C 1 -C 6 ) alkoxy, halogenated (C 1 -C 6 ) alkyl, halogenated (C 1 -C6 ) alkoxy, hydroxyl (C 1 -C 6 )alkyl, phenyl, 5-membered heteroaryl, 6-membered heteroaryl and -S(O) 2 R a , where R a Indicates (C 1 -C 4 )alkyl;

[0434] X 1 Selected from CH, C(R b ) and N, where R b Represents halogen, (C 1 -C 4 ) alkyl or halogenated (C 1 -C 4 )alkyl;

[0435] X 2 is selected from CH and N;

[0436] Y is selected from -NH- and -O-;

[0437] m is 0 or 1; n is 0 or 1;

[0438] L represents a bond, -O-, -NH- or -N(R c )-, where R c Indicates (C 1 -C 4 )alkyl;

[0439] as well as

[0440] (vi) a compound of the following formula (III) and a salt, solvate or salt of a solvate thereof,

[0441] in:

[0442] R 1 and R 2 independently represent a hydrogen atom, a hydroxyl group, a halogen atom, an optionally substituted (C 1 -C 10 )-alkyl, optionally substituted (C 1 -C 10 )-alkoxy, (C 1 -C 10 )-alkylthio, (C 1 -C 5 )-perfluoroalkyl, cyano, nitro, or, R 1 and R 2 Together they represent a group selected from: -O-(CH 2 ) p -O-, -O-(CH2 ) p -CH 2 -, -O-CH=CH-, -(CH 2 ) p+2 -、-NH-(CH 2 ) p+1 、-N(C 1 -C 3 -alkyl)-(CH 2 ) p+1 and -NH-N=CH-, wherein p=1 or 2, and the terminal oxygen atom and / or carbon atom and / or nitrogen atom is attached to the directly adjacent ring carbon atom, or NR 6 R 7 , where R 6 and R 7 Independently represent hydrogen, C 1 -C 5 -alkyl or (CO)-(C 1 -C 5 )-alkyl,

[0443] R 3 represents a hydrogen atom, a hydroxyl group, a halogen atom, a cyano group, an optionally substituted (C 1 -C 10 )-alkyl, (C 1 -C 10 )-alkoxy, (C 1 -C 10 )-alkylthio or (C 1 -C 5 )-perfluoroalkyl,

[0444] R 4 represents hydrogen, halogen, hydroxyl, (C 1 -C 5 )-alkyl, (C 1 -C 5 )-alkoxy, (C 1 -C 5 )-alkylthio, (C 1 -C 5 )-perfluoroalkyl, cyano, nitro, NR 6 R 7 、COOR 9 、(CO)NR 6 R 7 or (C 1 -C 5 -alkylene)-O-(CO)-(C 1 -C 5 )alkyl,

[0445] R 5 Indicates except -CH(CH3 ) 2 or -C(CH 3 )=CH 2 The other group is selected from the following groups: -(C 1 -C 10 )-alkyl, -(C 2 -C 10 )-alkenyl, -(C 2 -C 10 )-alkynyl, (C 3 -C 7 )cycloalkyl-(C 1 -C 8 ) alkyl, (C 3 -C 7 )cycloalkyl-(C 1 -C 8 ) alkenyl, (C 3 -C 7 )cycloalkyl-(C 2 -C 8 ) alkynyl, heterocyclic-(C 1 -C 8 )alkyl, heterocyclic-(C 1 -C 8 )alkenyl, heterocyclic-(C 2 -C 8 ) alkynyl, -R 8 , R 8 -(C 1 -C 8 ) alkyl, R 8 -(C 2 -C 8 ) alkenyl, R 8 -(C 2 -C 8 )alkynyl, -S-(C 1 -C 10 )-alkyl, -SO 2 -(C 1 -C 10 )-alkyl, -SR 8 、-SO 2 -R 8 、-CN、-Hal、-O-(C 1 -C 10 )-alkyl, wherein R 6 and R 7 -NR having the meaning defined above 6 R 7 、-OR 8 、-OH,

[0446] R 8It can be optionally replaced by 1-3 hydroxyl groups, halogen groups, C 1 -C 5 Alkyl, C 1 -C 5 -Alkoxy, cyano, CF 3 , nitro, COO(C 1 -C 5 -alkyl) or C(O)OCH 2 -phenyl substituted aryl, or may contain 1-3 heteroatoms of heteroaryl, the heteroatoms may be optionally replaced by 1-3 alkyl, hydroxy, halogen, cyano or C 1 -C 5 -Alkoxy substitution.

[0447] In a more preferred embodiment, the selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof for a specific use of the present invention is selected from the following compounds or a pharmaceutically acceptable salt thereof:

[0448]

[0449] (R)-2-(4-((5-(ethylsulfonyl)-1H-pyrrolo[2,3-c]pyridin-2-yl)methyl)-5,5,5-trifluoro-4-hydroxy-2-methylpentan-2-yl)-5-fluorobenzamide;

[0450]

[0451] [(3S)-1-[(3R)-5-oxotetrahydrofuran-3-carbonyl]-3-piperidinyl]4-[(1R,2S)-1-(4-cyclopropylphenyl)-2-[[(2R)-tetrahydrofuran-2-carbonyl]amino]propoxy]benzoate;

[0452] 5-{[(1S,2S)-1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(methoxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one;

[0453] 5-{(1S,2S)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(hydroxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one,

[0454] and 5-{(1S,2S)[1-(2-chloro-3-fluoro-4-methoxyphenyl)-3,3,3-trifluoro-2-hydroxy-2-(hydroxymethyl)propyl]amino}-7-fluoro-1H-quinolin-2-one.

[0455] Specifically, several SEGRMs were tested in 3D fibroblast cultures with wound exudate from patient #92. Fig.12 AZD7594 was the most active SEGRM tested in 3D culture, followed by mapracorat and BI653048. This is consistent with their potency on glucocorticoid receptor activation (EC 50 The values ​​of HY-14234 and HY-14234 were consistent (0.9 nM, 1.9 nM and 55 nM, respectively). In addition, HY-14234 was found to be active. The inactive analog BI3047 did not induce matrix formation.

[0456] The term "pharmaceutically acceptable" is used to indicate that the modified noun is suitable for use as a pharmaceutical product or as a part of a pharmaceutical product. Pharmaceutically acceptable salts include salts that are commonly used to form alkali metal salts and to form addition salts of free acids or free bases. Generally, these salts can be prepared by conventional methods, for example, by reacting a suitable acid or base with the compounds used in the present invention.

[0457] Pharmaceutically acceptable acid addition salts can be prepared from inorganic or organic acids. Examples of suitable inorganic acids generally include hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, sulfuric acid and phosphoric acid. Suitable organic acids generally include aliphatic acids, cycloaliphatic acids, aromatic acids, araliphatic acids, heterocycles, carboxylic acids and sulfonic acids such as organic acids. Generally, specific examples of suitable organic acids include acetic acid, trifluoroacetic acid, formic acid, propionic acid, succinic acid, glycolic acid, gluconic acid, digluconic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, glucuronic acid, maleic acid, fumaric acid, pyruvic acid, aspartic acid, glutamic acid, benzoic acid, anthranilic acid, methanesulfonic acid, stearic acid, salicylic acid, p-hydroxybenzoic acid, phenylacetic acid, mandelic acid, pamoic acid (pamoic acid), ethanesulfonic acid, benzenesulfonic acid, Pantothenic acid, 2-hydroxyethanesulfonic acid, sulfanilic acid, cyclohexylaminosulfonic acid, alginic acid, beta-hydroxybutyric acid, mucic acid, galacturonic acid, adipic acid, alginic acid, bisulfuric acid, butyric acid, camphoric acid, camphorsulfonic acid, cyclopentanepropionic acid, dodecylsulfuric acid, saccharoheptanoic acid, glycerophosphoric acid, heptanoic acid, hexanoic acid, nicotinic acid, oxalic acid, palmoate, pectic acid, 2-naphthalenesulfonic acid, 3-phenylpropionic acid, picric acid, trimethylacetic acid, thiocyanic acid, toluenesulfonic acid and undecanoic acid.

[0458] Pharmaceutically acceptable base addition salts include, for example, metal salts and organic salts. Preferred metal salts include alkali metal (Group Ia) salts, alkaline earth metal (Group IIa) salts, and other physiologically acceptable metal salts. Such salts can be made from aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc. Preferred organic salts can be made from amines, such as tromethamine, diethylamine, N, N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine), and procaine. Basic nitrogen-containing groups can be quaternized using reagents such as lower alkyl (C1-C6) halides (e.g., methyl, ethyl, propyl and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, dibutyl and diamyl sulfates), long chain halides (e.g., decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides), aryl alkyl halides (e.g., benzyl and phenethyl bromides), and the like.

[0459] Preferred physiologically acceptable salts of Mapracorat include acid addition salts of inorganic acids, carboxylic acids and sulfonic acids, such as salts of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, naphthalene disulfonic acid, acetic acid, trifluoroacetic acid, propionic acid, lactic acid, tartaric acid, malic acid, citric acid, fumaric acid, maleic acid and benzoic acid. Physiologically acceptable salts of Mapracorat also include salts of conventional bases, such as exemplarily and preferably, alkali metal salts (such as sodium and potassium salts), alkaline earth metal salts (such as calcium and magnesium salts), and ammonium salts derived from ammonia or organic amines having 1 to 16 carbon atoms, such as exemplarily and preferably, ethylamine, diethylamine, triethylamine, ethyldiisopropylamine, monoethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, dimethylaminoethanol, procaine, dibenzylamine, N-methylmorpholine, arginine, lysine, ethylenediamine and N-methylpiperidine.

[0460] Terminology C 2 -C 8-Alkenyl is a straight or branched, substituted or unsubstituted chain, including isomers with double bonds in E- or Z-configuration, such as vinyl, prop-1-enyl, prop-2-enyl (allyl), but-1-en-1-yl, but-1-en-2-yl, but-2-en-1-yl, but-2-en-2-yl, 2-methyl-prop-2-en-1-yl, 2-methyl-prop-1-en-1-yl, but-1-en-3-yl, but-3-en-1-yl. If the alkenyl group is located between two other groups, the term alkenyl refers to alkenylene, such as ethenylene, prop-1-enylene, prop-2-enylene (allylene), but-1-en-1-ylene, but-1-en-2-ylene, but-2-en-1-ylene, but-2-en-2-ylene, 2-methyl-prop-2-en-1-ylene, 2-methyl-prop-1-en-1-ylene, but-1-en-3-ylene, but-3-en-1-ylene.

[0461] Terminology C 2 -C 8 -Alkynyl represents a straight chain or branched chain, such as -C≡CH, -CH 2 -C≡CH, -C≡C-CH 3 、-CH(CH 3 )-C≡CH, -C≡C-CH 2 (CH 3 )、-C(CH 3 ) 2 -C≡CH, -C≡C-CH(CH 3 ) 2 、-CH(CH 3 )-C≡C-CH 3 、-CH 2 -C≡C-CH 2 (CH 3 ), or, if the alkynyl group is between two other groups, the term alkynyl refers to an alkynylene group, such as -C≡C-, -CH 2 -C≡C-, -C≡C-CH 2 -、-CH(CH 3 )-C≡C-, -C≡C-CH(CH 3 )-、-C(CH 3 ) 2 -C≡C-, -C≡CC-(CH 3 ) 2 -、-CH(CH 3 )-C≡C-CH 2 -、-CH 2 -C≡C-CH(CH 3 )-.

[0462] Terminology C3 -C 7 -Cycloalkyl represents a substituted or unsubstituted group selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl. Possible substituents may be selected from hydroxyl, halogen, (C 1 -C 5 )-alkyl, (C 1 -C 5 )-alkoxy, NR 4 R 5 、COO(C 1 -C 5 )-alkyl, CHO, cyano.

[0463] Terminology C 3 -C 7 -cycloalkyl-(C 1 -C 10 )-alkyl- represents, for example, -(CH 2 )-cycloalkyl, -(C 2 H 4 )-cycloalkyl, -(C 3 H 6 )-cycloalkyl, -(C 4 H 8 )-cycloalkyl, -(C 5 H 10 )-cycloalkyl, wherein the cycloalkyl represents, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl.

[0464] Terminology C 3 -C 7 -cycloalkyl-(C 2 -C 8 )-alkenyl represents, for example, -(CH=CH)-cycloalkyl, -[C(CH 3 )=CH]-cycloalkyl, -[CH=C(CH 3 )]-cycloalkyl, -(CH=CH-CH 2 )-cycloalkyl, -(CH 2 -(CH=CH)-cycloalkyl, -(CH=CH-CH 2 -CH 2 )-cycloalkyl, -(CH 2 -CH=CH-CH 2 )-cycloalkyl, -(CH 2 -CH 2 -CH=CH)-cycloalkyl, -(C(CH 3 )=CH-CH 2 )-cycloalkyl, -(CH=C(CH 3 )-CH 2 )-cycloalkyl, wherein the term cycloalkyl is as defined above.

[0465] The term heterocyclyl denotes, for example, piperidinyl-, morpholinyl-, thiomorpholinyl-, piperazinyl-, tetrahydrofuranyl-, tetrahydrothienyl-, imidazolidinyl- or pyrrolidinyl-, wherein the heterocyclyl can be attached via any possible ring atom. The heterocyclyl can be C 1 -C 5 -alkyl (optionally substituted), hydroxy-, C 1 -C 5 -alkoxy-、NR 4 R 5 -, halogen, cyano-, COOR 8 -, CHO-. If possible, these substituents may also be attached to a free nitrogen atom (if any). N-oxides are also included in this definition.

[0466] The term heterocyclyl-(C 1 -C 10 )-Alkyl- represents an alkylene group as defined above attached to the heterocyclyl group which has also been defined above.

[0467] The term heterocyclyl-(C 2 -C 8 )-Alkenyl- represents an alkenylene group as defined above attached to a heterocyclyl group which has also been defined above.

[0468] In the sense of the present invention, the term aryl means an aromatic or partially aromatic carbocyclic ring having 6 to 14 carbon atoms, for example phenyl, which may also have a fused second or third ring, such as naphthyl or anthracenyl. Other examples are phenyl, naphthyl, tetralinyl, anthracenyl, benzoxazinone, indolinone, indanyl and indenyl. The aryl group may be replaced by one or more substituents (e.g. 1-3 substituents, such as hydroxyl, halogen, C 1 -C 5 -alkyl, C 1 -C 5 -Alkoxy, cyano, CF 3 , nitro, COO(C 1 -C 5 -alkyl or benzyl), or heteroaryl) to form a stable molecule, preferably 1-3 C 1 -C 5 -alkyl, hydroxy, halogen, cyano or C 1 -C 5 -Alkoxy substitution.

[0469] The term heteroaryl denotes an aromatic ring system having 1 to 3 heteroatoms selected from nitrogen, oxygen or sulfur, the maximum number of heteroatoms being three for a five-membered ring, wherein only two oxygen or sulfur atoms are permitted provided that the two atoms are not directly bonded to each other.

[0470] Possible heteroaryl rings are, for example, thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, benzofuranyl, benzothiophenyl, benzothiazole, benzoxazolyl, benzimidazolyl, indazolyl, indolyl, isoindolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, azaindolizinyl, benzopyridinyl, benzopyridazinyl, benzopyrimidinyl, benzopyrazinyl, benzotriazinyl, quinolyl, isoquinolyl, phthalyl, thiophthalidyl, indolonyl, dihydroindolinonyl, isoindolinonyl, dihydroisoindolonyl, benzofuranyl or benzimidazolyl.

[0471] The SEGRM for the specific use of the present invention is administered to a subject in a therapeutically effective amount. For systemic application, the corresponding SEGRM dosage is in the range of about 10 to 1000 mg / day, depending on the corresponding SEGRM. Topical preparations, such as skin or intradermal preparations of SEGRM, such as creams, gels, lotions, ointments, liposomes or nanoparticle preparations, etc., can be administered at a concentration of about 0.00001 to 10% (w / v), about 0.00001 to 6% (w / v) or about 0.00001 to 1% (w / v) (e.g., 0.0001 to 0.1% (w / v) or about 0.001 to 1% (w / v)). Intradermal preparations can be administered, for example, using microneedles.

[0472] For example, topical formulations of mapracorat may preferably contain mapracorat in a concentration of >0.01 wt % to <10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 wt %, in particular >0.01 wt % to <5, 4, 3, 2 or 1 wt %, or >0.05 wt % to 0.15 wt %, or >0.02 wt % to <0.5 wt %, or any combination range of these values.

[0473] For example, according to the present invention, the topical preparation for the compound of WO2009 / 065503 disclosed in WO 2018 / 046678 can be used. The preparation relates to an anhydrous multiphase gel system comprising an outer lipid matrix and an inner phase gelled by a polymer, characterized in that the multiphase gel system comprises:

[0474] (a) an outer lipid matrix; and

[0475] (b) comprising an internal phase gelled by at least one polymer selected from cellulose derivatives, acrylate polymers and their derivatives or mixtures thereof,

[0476] And the active ingredient is selected from the compounds of general formula (I)

[0477]

[0478] wherein the compound is as defined above.

[0479] In particular, such external preparations are preferably used for the specific use of the present invention, wherein the active ingredient is selected from the following three compounds:

[0480]

[0481] Preferably, the lipid phase comprises a skin-compatible lipid selected from the group consisting of petrolatum, paraffin, microcrystalline wax, squalene, cetyl stearyl octanoate, ethyl oleate, tricaprylic / capric glyceryl, myristylmyristate, propylene glycol dicaprate, cetyl esters, isopropyl myristate, isopropyl palmitate, mono-, di- and triglycerides, ethoxylated glycerides, polyethylene glycol esters, sorbitan esters, stearin, dibutyl adipate, ethyl linoleate, crodamols, isocetyl stearate, cetyl palmitate, cetyl alcohol, oleyl alcohol, stearyl alcohol, dicaprylyl alcohol, oleic acid, waxes (particularly yojoba wax and beeswax), cholesterol, polyethylene glycol, lanolin, lanolin alcohol, silicone oils and mixtures thereof. Preferably, the lipid phase constitutes a proportion of 60 to 95% by weight, more preferably 65 to 92% by weight, more preferably 70 to 90% by weight of the multiphase gel system.

[0482] Preferably, the lipid phase contains: 50 to 90 wt.% petrolatum, more preferably 55 to 85 wt.%, more preferably 58 to 80 wt.%; 0 to 20 wt.% paraffin oil, more preferably 1 to 15 wt.%, more preferably 5 to 13 wt.%; 0 to 8 wt.% beeswax, more preferably 1 to 6 wt.%, more preferably 1 to 4 wt.%; 0 to 20 wt.% hard paraffin, more preferably 1 to 15 wt.%, more preferably 5 to 13 wt.%; and 0 to 8 wt.% cyclomethicone, more preferably 1 to 6 wt.%, more preferably 1 to 4 wt.%, weight based on the multiphase gel system. Preferably, the inner gel phase comprises at least one polymer selected from the group consisting of a cellulose derivative, an acrylate polymer or a derivative thereof or a mixture thereof. Preferably, the inner gel phase comprises a proportion of 5 to 40% by weight of the multiphase gel system, more preferably 8 to 35% by weight, more preferably 10 to 30% by weight. Preferably, a combination of a cross-linked acrylate polymer and hydroxypropyl cellulose is used in the gelled inner phase. Preferably, the inner gel phase comprises a mixture of a polyol and a carbonic acid diester. Preferably, the inner gel phase comprises a mixture of propylene glycol and propylene carbonate. Preferably, the inner gel phase comprises 0.05 to 0.5% by weight of an acrylate polymer, more preferably 0.075 to 0.4% by weight, more preferably 0.1 to 0.3% by weight; 0.01-0.1 wt.% cellulose derivative (especially hydroxypropyl cellulose), more preferably 0.01 to 0.08 wt.%, more preferably 0.01 to 0.06 wt.%; 1 to 15 wt.% carbonic acid diester (especially propylene carbonate), more preferably 2 to 13 wt.%, more preferably 3 to 10 wt.%; and 0.5 to 20 wt.% polyol (especially propylene glycol), more preferably 0.75 to 17 wt.%, more preferably 1 to 15 wt.%; the weights are always based on the multiphase gel system.

[0483] The present invention can be used to treat or prevent different types of skin wounds showing impaired skin wound healing. Different types of skin wounds showing impaired skin wound healing that can be treated according to the present invention include wounds of diabetic patients, skin wounds infected by at least one microorganism, ischemic wounds, wounds of patients suffering from insufficient blood supply or venous stasis, ulcers such as diabetic ulcers, venous ulcers, arterial ulcers (such as lower extremity arterial ulcers), mixed ulcers or pressure ulcers, neuropathic wounds, lower extremity ulcers, surgical wounds, burns, fissures, tumor ulcers, bullous dermatoses (such as epidermolysis bullosa) and rare ulcers. Microorganisms that infect skin wounds are known in the art, including bacteria and fungi, such as Corynebacterium, Staphylococcus, Streptococcus and yeast (such as Candida).

[0484] Therefore, in a preferred embodiment of the present invention, the skin wound is selected from the following wounds: wounds of diabetic patients, skin wounds infected by at least one microorganism, ischemic wounds, wounds of patients with insufficient blood supply or venous stasis, ulcers such as diabetic ulcers, venous ulcers, arterial ulcers (such as arterial ulcers of the lower limbs), mixed ulcers or pressure ulcers, neuropathic wounds, lower limb ulcers, surgical wounds, burns, fissures, tumor ulcers, bullous skin diseases (such as epidermolysis bullosa) and rare ulcers.

[0485] The subject or individual may be an otherwise healthy individual or may exhibit other diseases and / or co-morbidities, and / or be treated with medications for other diseases and / or co-morbidities. In a preferred embodiment, the subject or individual exhibits other diseases and / or co-morbidities, and / or is treated with medications for other diseases and / or co-morbidities, in addition to impaired skin wound healing.

[0486] Thus, in a preferred embodiment, the subject suffers from at least one comorbidity associated with impaired skin wound healing. Such comorbidities are, for example, diabetes, immune suppression after transplantation of a graft, and graft versus host disease (GvHD). Other comorbidities include obesity, elevated blood pressure, venous stasis, or peripheral arterial occlusion. Other comorbidities are diseases treatable with glucocorticoids.

[0487] Comorbidity is understood as the presence of one or more other diseases or conditions that co-occur with a given disease.

[0488] Therefore, in another preferred embodiment of the present invention, the subject has undergone transplantation of a transplant, and / or obtained immunosuppressive therapy, and / or treatment with at least one immunosuppressive drug. For example, immunosuppressive therapy is by administering a glucocorticoid and / or a calcineurin inhibitor. Therefore, the immunosuppressive drug can be selected from a glucocorticoid and a calcineurin inhibitor. Suitable calcineurin inhibitors are known in the art and include tacrolimus, pimecrolimus and cyclosporin A. Suitable glucocorticoids are known in the art and include Cortisol, cortisone acetate, prednisone, prednisolone, methylprednisolone, chloroprednisone, cloprednol, difluprednate, fludrocortisone acetate, fluocinolone, fluperolone, fluprednisolone, loteprednol, prednicarbate, tixocortol, triamcinolone, triamcinolone acetonide, tixocortol ... acetonide, dexamethasone, betamethasone, beclometasone, deoxycorticosterone acetate, alclometasone, clobetasol, clobetasone, clocortolone, desoximetasone, diflorasone, difluocortolone, fluclorolone, flumetasone, fluocortin, fluocortolone, fluprednidene, fluticasone, fluticasone furoate, halometasone, meprednisone, mometasone, mometasone furoatefuroate, paramethasone, prednylidene, rimexolone, ulobetasol, amcinonide, budesonide, ciclesonide, deflazacort, desonide, formocortal, fluclorolone acetonide, fludroxycortide, flunisolide, fluocinolone acetonide, fluocinonide, halcinonide, hydroxymethylprogesterone and medroxyprogesterone or a pharmaceutically acceptable salt thereof.

[0489] The present invention relates to a SEGRM or a pharmaceutically acceptable salt thereof for use in preventing and / or treating impaired skin wound healing in a subject.

[0490] "Skin wound" is understood to be an injury to the skin of a living individual, such as a cut, tear, burn or break. Preferably, a skin wound is understood to be an open injury to the skin of a living individual. The skin can be located in any area of ​​the individual, such as the head, arms, legs, chest or back. In addition, an individual can have one, two, three, four or more skin wounds. In addition, the area of ​​the skin wounds can vary. In a preferred embodiment, the skin wound forms a wound exudate. In another preferred embodiment, the skin wound forms a wound biofilm.

[0491] "Impaired skin wound healing" means that a skin wound does not heal at the expected rate. In a preferred embodiment, impaired skin wound healing is a non-healing skin wound or a chronic skin wound. Non-healing skin wounds are preferably understood as skin wounds that do not close within 2 months under standard treatment, preferably within 3 months or more under standard treatment. Preferably, non-healing skin wounds are characterized by lack of wound closure, increase in area and / or depth of the wound, necrosis and / or infection of the skin wound, and / or lack of granulation.

[0492] As used herein, "healed skin wounds" are understood to be skin wounds that heal at an expected rate, in particular skin wounds that close within 2 months under standard treatment. Preferably, healed skin wounds are characterized by sustained wound closure, granulation, absence of necrosis and / or absence of infection.

[0493] An "ulcer" is understood to be a sore on the skin, accompanied by a breakdown of the tissue. An ulcer can lead to a complete loss of the epidermis and often part of the dermis and even subcutaneous fat.

[0494] A "subject" or "individual" is an animal, preferably the individual is a vertebrate, particularly a mammal, more preferably a human.

[0495] In another preferred embodiment of the present invention, the subject suffers from diabetes and / or has at least one diabetic ulcer.

[0496] The subject's skin wound may have been treated, such as with standard therapies for treating wound healing, or the skin wound may have been left untreated.

[0497] "Standard therapy" is understood to be the treatment methods usually recommended by doctors for skin wounds, in particular one or more methods selected from the group consisting of: wound dressings, surgical and biological (maggot) debridement, infection control, negative pressure therapy, and biological or cellular therapy.

[0498] Thus, in a preferred embodiment, the skin wound of the subject may be untreated or treated with standard therapy for treating wound healing or treated with one or more of the following therapies for treating wound healing: compression, wound dressing, surgical debridement, biological debridement, infection control, antibiotic therapy, negative pressure therapy, proteins (particularly protein growth factors), antibodies, peptides, sugars, cells or cell components, artificial skin, human blood-derived products, gene therapy or genetic engineering wound bed modification, drugs, herbs or plant extracts. In a preferred embodiment, the skin wound of the subject may be untreated or treated with standard therapy for treating wound healing, wherein the standard therapy does not include treatment with protein growth factors. In another preferred embodiment, the skin wound of the subject may be untreated or treated with standard therapy for treating wound healing, wherein the standard therapy includes treatment with protein growth factors.

[0499] Therefore, in another preferred embodiment of the present invention, the subject suffers from diabetes and / or has at least one diabetic ulcer, and / or the subject (i) has undergone a transplantation of a transplant and, optionally, suffers from diabetes, and / or (ii) has received immunosuppressive therapy and, optionally, suffers from diabetes.

[0500] Furthermore, it was surprisingly found in the Examples that assays based on fibroblast proliferation and assays based on fibroblast-derived matrix formation can identify subjects with impaired skin wound healing who are responsive to treatment and / or prevention with SEGRM. Furthermore, the secretion of the IL-1 cytokine IL-1β was found to be a sensitive predictive marker. Therefore, in a preferred embodiment, the determination of IL-1 is used in combination with an assay based on fibroblast proliferation and / or an assay based on fibroblast-derived matrix formation.

[0501] The test can be used to successfully stratify and identify subjects with impaired skin wound healing. Thus, the test can be used in a personalized medicine approach.

[0502] Therefore, in a further preferred embodiment of the present invention, the subject is identified as responsive to treatment for impaired skin wound healing by performing steps i) and / or ii):

[0503] i) measuring the proliferation of fibroblasts and, optionally, the amount of at least one IL-1 cytokine marker in the supernatant of the fibroblasts in the presence of,

[0504] (1) a wound exudate sample or a wound biofilm sample obtained from a skin wound of the subject, and

[0505] (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof;

[0506] ii) Measuring fibroblast-derived matrix formation by fibroblasts in the presence of:

[0507] (1) a wound exudate sample or a wound biofilm sample obtained from a skin wound of the subject, and

[0508] (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof.

[0509] In a more preferred embodiment, a subject is identified as responsive to treatment with a selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof for impaired skin wound healing if the value of fibroblast proliferation measured in step i) and / or the value of fibroblast-derived matrix formation of the fibroblasts measured in step ii) is at least 20% higher than a control value established in the absence of (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof, and, optionally, if the value of the amount of at least one IL-1 cytokine marker in the supernatant of the fibroblasts obtained in step i) is lower than a control value established in the absence of (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof.

[0510] In a preferred embodiment of the present invention, the sample is a wound exudate sample. In another preferred embodiment, the sample is a wound biofilm sample. In a more preferred embodiment, the sample is a wound exudate sample.

[0511] The SEGRM administered to the subject where the subject is determined to be responsive can be the same SEGRM or a different SEGRM, preferably the same SEGRM.

[0512] Measuring the proliferation of fibroblasts in the presence of a wound exudate sample or a wound biofilm sample obtained from the skin wound and the SEGRM of (2) can be performed as shown in the examples. The assay is also referred to in the present application as a "HDF proliferation", "human dermal fibroblast proliferation", "fibroblast proliferation" or "2D fibroblast proliferation" assay. For the assay, fibroblasts are used, which can be primary fibroblasts (e.g., primary mammalian dermal fibroblasts) or cells of a fibroblast line, preferably primary fibroblasts. Methods for culturing fibroblasts are known in the art and are described, for example, in the examples. For example, cells can be cultured using DMEM medium containing FCS.

[0513] In a further preferred embodiment, the cells are incubated on a solid support so that the cells adhere to the support, such as described in the examples using a porous plate. In addition, the cells are contacted with a wound exudate sample or a wound biofilm sample (optionally diluted (e.g., diluted with a culture medium or saline solution)) and the SEGRM of (2). The contact can be performed before or after cell adhesion occurs. For example, the contact can be achieved by adding an optionally diluted liquid wound exudate sample or wound biofilm sample and the SEGRM of (2) to the cells before adhesion (e.g., when the cells are inoculated) or after adhesion. The contact can be achieved by, for example, pipetting and optionally gentle mixing. The cells are incubated for an appropriate time, such as 6 hours to 300 hours, more preferably 12 hours to 200 hours, and even more preferably 24 hours to 120 hours. In the examples, 72 hours were successfully used. For negative control samples, in addition to wound exudate or wound biofilm, the corresponding liquid in which the SEGRM of (2) is not present can be added, or only wound exudate or wound biofilm can be added. Subsequently, for example, by fixing cells and determining total protein content, determine the amount of fibroblasts, preferably cell number, including the formation of extracellular matrix. Cells can for example be fixed using paraformaldehyde. In addition, suitable dyes (such as sulforhodamine B) can be used to determine the amount of fibroblasts, preferably cell number, including the formation of extracellular matrix. Then the dyed cells including the extracellular matrix formed can be quantitatively measured according to the dye, for example, by determining the absorbance or fluorescence at a suitable wavelength. Preferably, these steps are carried out in 2D cell culture, which allows adherent culture cells on a solid support. Preferably, the sample is a wound exudate sample.

[0514] Therefore, in another preferred embodiment, the method steps include the following steps:

[0515] (i) culturing fibroblasts,

[0516] (ii) culturing the cells on a solid support so that the cells adhere to the support,

[0517] (iii) contacting the cells with (1) a wound exudate sample or a wound biofilm sample (optionally diluted) and (2) the SEGRM, wherein the contacting may be performed before or after cell adhesion occurs, and wherein the contacting of (1) and (2) may be performed simultaneously or sequentially, and

[0518] (iv) determining the amount of fibroblasts, preferably the number of cells, including the formation of extracellular matrix, e.g. by fixing the cells and determining the total protein content,

[0519] Preferably, wherein the method is performed in 2D cell culture.

[0520] In a preferred embodiment of the present invention, the sample is a wound exudate sample. In another preferred embodiment, the sample is a wound biofilm sample. In a more preferred embodiment, the sample is a wound exudate sample.

[0521] Optionally, the amount of at least one IL-1 cytokine marker is measured in the supernatant of the fibroblasts in i).

[0522] In the examples it was found that IL-1 cytokine (particularly IL-1β) is a sensitive marker in the context of the present invention.

[0523] "IL-1 cytokine" is understood to include IL-1 alpha and IL-1 beta. In a preferred embodiment, the IL-1 cytokine is IL-1 beta.

[0524] The amount of pro-inflammatory IL-1 cytokine secreted by fibroblasts was found to have a particular predictive effect on identifying healed skin wounds or unhealed skin wounds and monitoring wound healing. In particular, it was found that the secretion of these cytokines was higher in the presence of WE from unhealed wounds than in the presence of WE from healed wounds. The cytokines IL1α and IL-1β are proteins well known to the skilled person, preferably human proteins. IL-1α (also known as interleukin-1α or IL-1alpha) and IL-1β (also known as interleukin-1β or IL-1beta) can be determined by methods known in the art, for example by using immunological detection, even more preferably by using ELISA detection as described in the "Examples", or by determining IL-1 cytokine mRNA as described in the "Examples". IL-1α and IL-1β are known to be pro-inflammatory cytokines.

[0525] Thus, in a preferred embodiment, a subject is identified as responsive to treatment for impaired skin wound healing by performing step i) measuring the proliferation of fibroblasts and optionally the amount of at least one IL-1 cytokine marker in the supernatant of the fibroblasts.

[0526] Preferably, the measuring of the amount of at least one IL-1 cytokine marker in the supernatant of fibroblasts in i) comprises the following steps:

[0527] (i) culturing fibroblasts,

[0528] (ii) culturing the cells on a solid support so that the cells adhere to the support,

[0529] (iii) contacting the cells with (1) a wound exudate sample or a wound biofilm sample (optionally diluted) and (2) the SEGRM, wherein the contacting may be performed before or after cell adhesion occurs, and wherein the contacting of (1) and (2) may be performed simultaneously or sequentially, and

[0530] (iv) determining the amount of at least one IL-1 cytokine marker in the cell culture supernatant,

[0531] Preferably, wherein the at least one IL-1 cytokine marker is determined by using an immunological assay, more preferably by using an ELISA assay, and / or by determining IL-1 cytokine mRNA, wherein the method is performed in 2D cell culture.

[0532] In a preferred embodiment of the present invention, the sample is a wound exudate sample. In another preferred embodiment, the sample is a wound biofilm sample. In a more preferred embodiment, the sample is a wound exudate sample.

[0533] The culturing of the cells is preferably carried out at about 20°C to 40°C, more preferably 25°C to 38°C, even more preferably about 37°C.

[0534] Measuring fibroblast-derived matrix formation of fibroblasts in the presence of a wound exudate sample or a wound biofilm sample obtained from a skin wound can be performed as shown in the examples. Detection in the present application also refers to "ECM formation", "fibroblast-derived matrix" or "3D fibroblast-derived matrix" detection. For the detection, fibroblasts are used, which can be primary fibroblasts (e.g., primary mammalian dermal fibroblasts) or cells of a fibroblast line, preferably primary fibroblasts. In an embodiment, fibroblasts are seeded on a support, which is preferably pre-coated with an adhesion enhancer, such as gelatin. For example, the coating can be achieved by incubating the support with a solution or suspension containing an adhesion enhancer (e.g., gelatin). In an embodiment, a 0.2% gelatin solution was successfully used. Preferably, the cells are cultured until fusion is reached. Subsequently, the cells are contacted with (i) a matrix promoting supplement, (ii) a wound exudate sample or a wound biofilm sample (optionally diluted) and (iii) the SEGRM of (2), wherein (i), (ii) and (iii) can be performed simultaneously or sequentially. For example, a matrix promoting supplement (preferably selected from a solution comprising vitamin C or a physiologically acceptable salt thereof (e.g., a sodium salt), or 2-phospho-L-ascorbic acid or a physiologically acceptable salt thereof, and a combination of EGF and insulin) is added to the cells, for example, by pipetting and optionally gentle mixing. The wound exudate sample or wound biofilm sample (optionally diluted) can be contacted simultaneously or sequentially, and the SEGRM of (2) is added simultaneously or sequentially. For example, an optionally diluted wound exudate sample or wound biofilm sample can be mixed with a matrix promoting supplement, and the mixture can be added to the cells and then the SEGRM of (2) is added. Alternatively, optionally diluted wound exudate samples or wound biofilm samples can be added separately but simultaneously or separately, but after or before the SEGRM of matrix promotion supplement and / or (2). In the case of subsequent non-simultaneous contact, components (i), (ii) and (iii) are preferably contacted within 1 hour. The cells are then incubated, preferably for 12 hours to 20 days, wherein the culture medium is optionally replaced at least once with fresh culture medium supplemented with optionally diluted wound exudate or wound biofilm and matrix promotion supplement. In this embodiment, the culture medium is replaced once after 4 days of incubation. Due to the formation of a 3-dimensional fibroblast-derived matrix, the solid support preferably includes at least one cavity that allows filling space and therefore allows 3D cell culture. Subsequently, the amount of fibroblast-derived matrix is ​​determined, for example, by fixing cells and determining total protein content. Cells can be fixed, for example, using paraformaldehyde. In addition, the amount of fibroblasts, preferably cell number, including the formation of extracellular matrix can be determined using a suitable dye (such as sulforhodamine B).The stained cells including the formation of the extracellular matrix can then be quantified based on the dye, for example by determining absorbance or fluorescence at a suitable wavelength. For negative control samples, in addition to wound exudate or wound biofilm, the corresponding liquid of the SEGRM in the absence of (2) can be added, or only wound exudate or wound biofilm can be added. Preferably, the sample is a wound exudate sample.

[0535] Therefore, the method steps preferably include the following steps:

[0536] (i) seeding fibroblasts on a support, which is preferably pre-coated with an adhesion enhancer, such as gelatin,

[0537] (ii) culturing the cells on a support, preferably until confluence is achieved,

[0538] (iii) contacting the cells with (i) a matrix promoting supplement, (ii) a wound exudate sample or a wound biofilm sample (optionally diluted), and (iii) the SEGRM of (2), wherein (i) and (ii) may be performed simultaneously or sequentially,

[0539] (iv) determining the amount of fibroblast-derived matrix, for example by fixing the cells and determining the total protein content,

[0540] Preferably wherein the method is performed in 3D cell culture.

[0541] In a preferred embodiment of the present invention, the sample is a wound exudate sample. In another preferred embodiment, the sample is a wound biofilm sample. In a more preferred embodiment, the sample is a wound exudate sample.

[0542] "Fibroblast-derived matrix" or "FDM" is understood to be an extracellular matrix (ECM) formed by living fibroblasts in an environment conducive to matrix formation (e.g., in the presence of a matrix-promoting supplement). FDM can be obtained as described in the Examples. In particular, FDM can be obtained by the following steps: (i) seeding fibroblasts on a support pre-coated with an adhesion enhancer (e.g., gelatin), (ii) culturing cells on the support, preferably until fusion is achieved, and (iii) contacting the cells with a matrix-promoting supplement (e.g., vitamin C or a physiologically acceptable salt thereof, or 2-phospho-L-ascorbic acid or a physiologically acceptable salt thereof, or a combination of EGF and insulin).

[0543] "Matrix promotion supplement" is understood to be a compound or composition that promotes living fibroblasts to form a fibroblast-derived matrix in in vitro cell culture. Suitable matrix promotion supplements are vitamin C or its physiologically acceptable salt (e.g., sodium salt) or 2-phosphoric acid-L-ascorbic acid or its physiologically acceptable salt and a combination of EGF and insulin and a composition (e.g., solution or suspension) comprising these compounds. The combination of EGF and insulin can be provided to cell culture separately (e.g., as a separate solution comprising EGF or insulin respectively) or together (e.g., as a solution comprising EGF and insulin).

[0544] An "adhesion enhancer" is an agent that enhances cell adhesion to a solid support (eg, a plastic support) but does not substantially interfere with cell viability. In a preferred embodiment, the adhesion enhancer is gelatin or fibronectin, more preferably gelatin.

[0545] "2D cell culture" is understood as a cell culture, wherein the cells are cultured on a planar or substantially planar surface. In a preferred embodiment, the 2D cell culture is a culture of adherent cells.

[0546] "3D cell culture" is understood as a cell culture in which cells are cultured on a non-planar or substantially non-planar surface. In a preferred embodiment, the 3D cell culture is a culture of adherent cells and / or a culture of cells within a matrix (eg, ECM, in particular FDM).

[0547] "Support" or "solid support" is preferably selected from a chip, an array (e.g., a microarray or nanoarray), a plate (e.g., a multiwell plate) or a culture dish. For cell culture applications, the solid support is preferably suitable for culturing cells, e.g., the support may be a plastic support.

[0548] "Wound exudate" is understood to be the extracellular fluid located in and above a skin wound. Wound exudate is also referred to as a "liquid biopsy".

[0549] "Wound biofilm" is understood to be a substance produced by microorganisms that can form colonies and infect skin wounds. Typically, wound biofilms are sticky or colloidal substances. Wound biofilms contain microbial species selected from bacteria, fungi, yeasts, algae and other microorganisms, as well as cell debris. Wound biofilms are formed when certain types of microorganisms adhere to the surface of skin wounds by secreting sticky or colloidal substances. For example, wound biofilm samples can be obtained by surgical sharp debridement or by wiping the wound surface with a swab (e.g., a cotton swab or a nylon flocked swab) or a wound dressing material.

[0550] "Wound exudate sample" or "WE" is understood to be a wound exudate sample obtained from an individual skin wound. The method for obtaining a wound exudate sample is known in the art. For example, a wound exudate sample can be obtained by a physical or chemical method, in particular by applying negative pressure to a skin wound, for example, by using a negative pressure drainage device, using a method for capillary force, collecting wound exudate in a film dressing or a membrane, collecting wound exudate in a syringe, applying an absorbent material (absorbent beads or filter) or by using a swab, such as a cotton swab or a nylon flocked swab, in particular wherein the film dressing or the membrane is a cellulose layer and / or wherein the absorbent material is a cellulose layer. A preferred suitable cellulose layer is a nanocellulose layer. The volume of the wound exudate sample can vary, and can be in the range of 1nl to 1l, 10nl to 1l or 100nl to 1l, for example 1 μl to 1l, 1ml to 1l or 10ml to 1l. For example, the wound exudate samples studied in the examples have a volume of up to 400 ml and typically have a volume of 0.1 to 100 ml, in particular 1 to 50 ml. The wound exudate sample can be used in the method of the invention directly after obtaining the sample or can be stored before being used in the method of the invention, in particular stored at <4°C, <0°C or <10°C, for example about -20°C or -80°C.

[0551] "Wound biofilm sample" or "WB" is understood to be a wound biofilm sample obtained from an individual's skin wound. Methods for obtaining wound biofilm samples are known in the art. For example, wound biofilm samples can be obtained by surgical sharp debridement or by wiping the wound surface with a swab (e.g., cotton swab or nylon flocked swab) or wound dressing material. The volume of the wound biofilm sample can vary, and can be in the range of 1nl to 1l, 10nl to 1l or 100nl to 1l, for example, 1μl to 10ml, 1μl to 1ml or 10μl to 1ml. The wet weight of the wound biofilm can vary, and can be in the range of 10μg to 10g, 100μg to 10g, for example, 1mg to 10g, 10mg to 10g, 100mg to 10g or 1g to 10g. The wound biofilm sample can be directly used in the method of the present invention after obtaining the sample or can be stored before being used in the method of the present invention, particularly stored at <4°C, <0°C or <10°C. The wound biofilm sample can be extracted with a suitable liquid, such as a cell culture medium or a buffer, particularly 5 to 10 times the weight of the sample.

[0552] Surprisingly, it has been found that the above-described assays involving measuring proliferation of fibroblasts and measuring fibroblast-derived matrix formation by fibroblasts can reliably identify subjects responsive to treatment and / or prevention of impaired skin wound healing according to any of the above-described embodiments of the present invention.

[0553] Furthermore, the inventors have found that the accuracy of identifying a responding subject is improved when the proliferation of fibroblasts and optionally the amount of at least one IL-1 cytokine is measured. Thus, in a more preferred embodiment, a subject is identified as responding to a treatment for impaired skin wound healing if the value of the fibroblast proliferation measured in step i) and the value of the fibroblast-derived matrix formation of the fibroblasts measured in step ii) are at least 20% higher than a control value established in the absence of the SEGRM of (2), and, optionally, if the value of the amount of at least one IL-1 cytokine marker in the supernatant of the fibroblasts obtained in step i) is lower than a control value established in the absence of at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2).

[0554] Thus, in a more preferred embodiment, a subject is identified as responsive to treatment for impaired skin wound healing if the value of fibroblast proliferation measured in step i) and / or the value of fibroblast-derived matrix formation of the fibroblasts measured in step ii) is at least 30%, 40%, 50%, 60%, 70%, 80%, 100% or more higher than a control value established in the absence of the SEGRM of (2), and, optionally, if the value of the amount of at least one IL-1 cytokine marker in the supernatant of the fibroblasts obtained in step i) is lower than a control value established in the absence of at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2).

[0555] Thus, in a more preferred embodiment, a subject is identified as responsive to treatment for impaired skin wound healing if the value of fibroblast proliferation measured in step i) and / or the value of fibroblast-derived matrix formation of the fibroblasts measured in step ii) is at least 30%, 40%, 50%, 60%, 70%, 80%, 100% or more higher than a control value established in the absence of the SEGRM of (2), and, optionally, if the value of the amount of at least one IL-1 cytokine marker in the supernatant of the fibroblasts obtained in step i) is at least 5%, 10%, 15%, 20%, 30%, 40%, 50% or more lower than a control value established in the absence of at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2).

[0556] Control values ​​may be determined in parallel or may be established independently, preferably in parallel.

[0557] In addition, accuracy and reliability can be further improved by including one or more additional tests that determine macrophage M1 and M2 markers and / or cytokine markers IL1α, IL1β and / or TNFα in macrophage / fibroblast co-cultures in the presence of wound exudate or wound biofilm obtained from the respective subject. These M1 and M2 markers can be cell surface protein markers, protein markers in macrophage supernatants, or marker mRNA in macrophages.

[0558] Macrophages are professional phagocytes and antigen presenting cells (APCs) in tissue situ, which are different from circulating peripheral blood mononuclear cells. Technicians divide activated macrophages of different phenotypes into M1-macrophages and M2 macrophages. M1-macrophages are activated macrophages, which contain immune effector cells with acute inflammatory phenotypes. They are highly aggressive to bacteria and produce a large amount of cytokines. M2-macrophages are alternately activated and anti-inflammatory.

[0559] "M2 marker" is understood to be a protein marker specific for M2 macrophages. Preferably, the marker is secreted by macrophages. Suitable M2 markers are known in the art and are preferably selected from CCL22 and CCL18. The marker can be determined by methods known in the art, for example by using immunological detection, even more preferably by using ELISA detection.

[0560] "M1 marker" is understood to be a protein marker specific for M1 macrophages. Preferably, the marker is secreted by macrophages. Suitable M1 markers are known in the art and are preferably selected from CXCL10 and IL-23p19. The marker can be determined by methods known in the art, for example by using immunological detection, even more preferably by using ELISA detection.

[0561] "M1 cell surface marker" is understood to be a protein marker expressed on the surface of macrophages and specific for M1 macrophages. Suitable M1 cell surface markers are known in the art and are preferably selected from CD38, CD64 and CD197. The amount and / or frequency distribution of cell surface markers can be determined by immunological detection and / or fluorescence detection, in particular by FACS analysis, whereby the frequency distribution is usually determined.

[0562] "M2 cell surface marker" is understood to be a protein marker expressed on the surface of macrophages and specific for M2 macrophages. Suitable M2 cell surface markers are known in the art and are preferably selected from the group consisting of CD200 receptor (CD200R), CD206 and CD209. The amount and / or frequency distribution of cell surface markers can be determined by immunological detection and / or fluorescence detection, in particular by FACS analysis, whereby the frequency distribution is usually determined.

[0563] "M2 marker mRNA" is understood to be an mRNA expressed by macrophages and specific for M2 macrophages. Suitable M2 marker mRNAs are known in the art and are preferably selected from CD200 receptor (CD200R), CD206, CD209, CCL22 and CCL18. The marker mRNA can be determined by methods known in the art. Preferably, the amount can be determined by contacting a probe that specifically binds to the marker mRNA (wherein the probe is optionally labeled) with macrophage RNA under conditions that are conducive to hybridization, and detecting the hybridized probe. For example, the mRNA can be reverse transcribed into cDNA before detection.

[0564] "M1 marker mRNA" is understood to be an mRNA expressed by macrophages and specific for M1 macrophages. Suitable M1 marker mRNAs are known in the art and are preferably selected from CD38, CD64, CD197, CXCL10 and IL-23p19. Preferably, the amount can be determined by contacting a probe that specifically binds to the marker mRNA (wherein the probe is optionally labeled) with the macrophage RNA under conditions that are conducive to hybridization, and detecting the hybridized probe. For example, the mRNA can be reverse transcribed into cDNA before detection.

[0565] The ratio of the M1 / M2 markers in combination with one or more of the cell tests described above related to measuring fibroblast proliferation, measuring fibroblast-derived matrix (FDM) formation by fibroblasts, and measuring proliferation of keratinocytes indicates the responsiveness of the subject. In particular, an elevated ratio of the M1 / M2 markers, M1 / M2 cell surface markers, or M1 / M2 marker mRNAs indicates that the subject is non-responsive, while a low ratio of the M1 / M2 markers, M1 / M2 cell surface markers, or M1 / M2 marker mRNAs indicates that the subject is responsive.

[0566] In addition, the amount of proinflammatory cytokines IL1α, IL1β and TNF-α secreted by macrophages and / or fibroblasts in macrophage / fibroblast co-cultures was found to have a particular predictive effect on identifying healed skin wounds or unhealed skin wounds and monitoring wound healing. In particular, it was found that the secretion of these cytokines was higher in the presence of WE from unhealed wounds than in the presence of WE from healed wounds. The cytokines IL1α, IL-1β and TNF-α are proteins well known to the skilled person, preferably human proteins. IL1α (also known as interleukin-1α or IL-1alpha), IL-1β (also known as interleukin-1β or IL-1beta) and TNF-α (also known as tumor necrosis factor α or TNF-alpha) can be determined by methods known in the art, for example by using immunological detection, even more preferably by using ELISA detection as described in the "Examples", or by determining IL1α, IL1β and TNF-α mRNA expression. IL1α, IL1β and TNF-α are known to be proinflammatory cytokines.

[0567] Therefore, in a more preferred embodiment of the present invention, one, two, three or four of step iiia) and / or the following steps iiib) to iiie) are additionally performed:

[0568] iiia) measuring the proliferation of keratinocytes in the presence of:

[0569] (1) a wound exudate sample or a wound biofilm sample obtained from a skin wound of the subject, and

[0570] (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof;

[0571] iiib) measuring the amount of one or more M1 markers and one or more M2 markers in the supernatant of macrophages incubated with:

[0572] (1) a wound exudate sample or a wound biofilm sample obtained from the skin wound, and

[0573] (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof,

[0574] Macrophages were co-cultured with fibroblasts, and

[0575] wherein the one or more M1 markers are selected from CXCL10 and IL-23p19, and the one or more M2 markers are selected from CCL22 and CCL18; iiic) measuring the amount and / or frequency distribution of the one or more M1 cell surface markers and the one or more M2 cell surface markers on macrophages incubated with:

[0576] (1) a wound exudate sample or a wound biofilm sample obtained from the skin wound, and

[0577] (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof,

[0578] Macrophages were co-cultured with fibroblasts, and

[0579] wherein the one or more M1 cell surface markers are selected from CD38, CD64 and CD197, and wherein the one or more M2 cell surface markers are selected from CD200 receptor, CD206 and CD209;

[0580] iiid) measuring the expression level of one or more M1 marker mRNAs and one or more M2 marker mRNAs in macrophages incubated with:

[0581] (1) a wound exudate sample or a wound biofilm sample obtained from the skin wound, and

[0582] (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof,

[0583] Macrophages were co-cultured with fibroblasts, and

[0584] wherein the one or more M1 marker mRNAs are selected from CD38, CD64, CD197, CXCL10 and IL-23p19, and the one or more M2 marker mRNAs are selected from CD200 receptor (CD200R), CD206, CD209, CCL22 and CCL18;

[0585] iiie) measuring the amount of one or more cytokine markers in the supernatant of macrophages incubated with:

[0586] (1) a wound exudate sample or a wound biofilm sample obtained from the skin wound, and

[0587] (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof,

[0588] Macrophages were co-cultured with fibroblasts, and

[0589] wherein the one or more cytokine markers are selected from IL-1α, IL-1β and TNF-α, and

[0590] wherein the subject is identified as responsive to treatment with at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof if the value of fibroblast proliferation measured in step i) and / or the value of fibroblast-derived matrix formation of fibroblasts measured in step ii) and / or the value of keratinocyte proliferation in step iiia) is at least 20% higher than a control value established in the absence of the at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2), and, optionally, if the value of the amount of at least one IL-1 cytokine marker in the supernatant of the fibroblasts obtained in step i) is lower than a control value established in the absence of the at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2),

[0591] and / or if one or more of the following applies:

[0592] -iiib) the ratio of the amount of one or more M1 markers to the amount of one or more M2 markers obtained is lower than a control value established in the absence of the at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2),

[0593] -iiic) to the amount and / or frequency distribution of one or more M1 cell surface markers to the amount and / or frequency of one or more M2 cell surface markers is lower than a control value established in the absence of at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2), in particular, wherein the ratio is selected from the group consisting of CD38 / CD209 ratio, CD197 / CD209 ratio and CD197 / CD206 ratio,

[0594] -iiid) is lower than a control value established in the absence of at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2),

[0595] -iiie) is lower than a control value established in the absence of the at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2).

[0596] Preferably, a subject is identified as responding to treatment with a SEGRM if the value of fibroblast proliferation measured in step i) and / or the value of fibroblast-derived matrix formation of fibroblasts measured in step ii) and / or the value of keratinocyte proliferation in step iiia) is at least 20% higher than a control value established in the absence of the at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2), and, optionally, if the value of the amount of at least one IL-1 cytokine marker in the supernatant of the fibroblasts obtained in step i) is lower, for example at least 5%, 10%, 15%, 20%, 30%, 40%, 50% or more lower than a control value established in the absence of the at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2),

[0597] and / or if:

[0598] -iiie) is lower than a control value established in the absence of at least one selective glucocorticoid receptor modulator (SEGRM) of (2) or a pharmaceutically acceptable salt thereof, for example, at least 5%, 10%, 15%, 20%, 30%, 40%, 50% or more lower.

[0599] In a more preferred embodiment, the cytokine marker in iiie) is selected from IL-1α and IL-1β, even more preferably the cytokine marker in iiie) is IL-1β.

[0600] The present inventors have found that the following M1 cell surface marker / M2 cell surface marker ratios can also predict responsiveness: a CD38 / CD209 ratio, a CD197 / CD209 ratio, or a CD197 / CD206 ratio that is lower than the control value established in the absence of SEGRM of (2) indicates that the patient responds to SEGRM treatment.

[0601] Therefore, in another preferred embodiment, the ratio of amount and / or frequency distribution is selected from the group consisting of CD38 / CD209 ratio, CD197 / CD209 ratio and CD197 / CD206 ratio.

[0602] Frequency distribution can be determined by determining the percentage of cells within a population that are positive for a given marker, which is the most commonly used readout in FACS analysis. Alternatively, the amount can be determined by determining the amount of cell surface expression (e.g., by mean fluorescence intensity measurement) as a surrogate for the number of labeled molecules on the cell surface of each individual cell when a labeled binding agent is used as a marker.

[0603] In a preferred embodiment, measuring the amount of one or more M1 markers and one or more M2 markers in the supernatant of macrophages incubated with a wound exudate sample or a wound biofilm sample obtained from a skin wound comprises the following steps:

[0604] (i) Primary human monocytes were co-cultured with (a) human dermal fibroblasts in 2D cell culture or (b) fibroblast-derived matrix,

[0605] (ii) incubating the cells until macrophage differentiation is achieved, optionally wherein CD163 is used as a cell surface marker for macrophage differentiation,

[0606] (iii) contacting the cells with a wound exudate sample or a wound biofilm sample (optionally diluted) with the SEGRM of (2), and

[0607] (iv) determining the amount of one or more M1 markers and one or more M2 markers in the cell culture supernatant,

[0608] Preferably, wherein the one or more M1 markers are selected from CXCL10 and IL-23p19, and / or the one or more M2 markers are selected from CCL22 and CCL18, more preferably, wherein the markers are determined by using immunological detection, even more preferably by using ELISA detection.

[0609] In a preferred embodiment of the present invention, the sample is a wound exudate sample. In another preferred embodiment, the sample is a wound biofilm sample. In a more preferred embodiment, the sample is a wound exudate sample.

[0610] For example, primary human monocytes can be co-cultured with human dermal fibroblasts in 2D cell culture or with fibroblast-derived matrix. The method for producing fibroblast-derived matrix is ​​as described above and in the embodiment. Subsequently, cells are incubated until macrophage differentiation is reached. For example, CD163 can be used as a cell surface marker for macrophage differentiation. Further, cells are contacted with wound exudate samples or wound biofilm samples (which are optionally diluted) and the SEGRM of (2), for example, by pipetting the sample to the cells, and optionally gentle mixing. Compounds are added after macrophage differentiation (for example, 4 to 7 days later). Further, cells are incubated, preferably 1 hour to 100 hours. Subsequently, the amount of one or more M1 markers and one or more M2 markers in the cell culture supernatant is determined. For this purpose, supernatants are usually harvested and markers are determined using suitable detection (for example, immunological detection). For example, ELISA can be used.

[0611] In another preferred embodiment, measuring the amount and / or frequency distribution of one or more M1 cell surface markers and one or more M2 cell surface markers on macrophages incubated with a wound exudate sample or a wound biofilm sample obtained from a skin wound comprises the following steps:

[0612] (i) Primary human monocytes were co-cultured with (a) human dermal fibroblasts in 2D cell culture or (b) fibroblast-derived matrix,

[0613] (ii) incubating the cells until macrophage differentiation is achieved, optionally wherein CD163 is used as a cell surface marker for macrophage differentiation,

[0614] (iii) contacting the cells with a wound exudate sample or a wound biofilm sample (optionally diluted) with the SEGRM of (2),

[0615] (iv) determining the amount and / or frequency distribution of one or more M1 cell surface markers and one or more M2 cell surface markers on the cell surface of macrophages.

[0616] In a preferred embodiment of the present invention, the sample is a wound exudate sample. In another preferred embodiment, the sample is a wound biofilm sample. In a more preferred embodiment, the sample is a wound exudate sample.

[0617] For example, primary human monocytes can be co-cultured with human dermal fibroblasts in 2D cell culture or with fibroblast-derived matrix. The method for producing fibroblast-derived matrix is ​​as described above and in the embodiments. Subsequently, cells are incubated until macrophage differentiation is reached. For example, CD163 can be used as a cell surface marker for macrophage differentiation. Further, cells are contacted with wound exudate samples or wound biofilm samples (which are optionally diluted) and SEGRM of (2), for example, by pipetting the sample to the cells, and optionally gentle mixing. Compounds are added after macrophage differentiation (for example, 4 to 7 days later). Further, cells are incubated, preferably 1 hour to 100 hours. Subsequently, the amount and / or frequency distribution of one or more M1 cell surface markers and one or more M2 cell surface markers on the cell surface of macrophages are determined. For example, cells can be harvested and FACS analysis can be performed, and gates are set for monocyte / macrophage populations. The geometric mean of mean fluorescence intensity can be used to quantify surface marker expression.

[0618] Preferably, one or more M1 cell surface markers are selected from CD38, CD64 and CD197, and / or one or more M2 cell surface markers are selected from CD200 receptor, CD206 and CD209, more preferably wherein the amount and / or frequency distribution of the cell surface markers are determined by immunological detection and / or fluorescence detection, in particular by FACS analysis.

[0619] The present inventors have found that the following M1 cell surface marker / M2 cell surface marker ratios can also be predicted for determining responsiveness: CD38 / CD209 ratio, CD197 / CD209 ratio, and CD197 / CD206 ratio. A CD38 / CD209 ratio, CD197 / CD209 ratio, or CD197 / CD206 ratio that is lower than the control value established in the absence of SEGRM of (2) indicates that the patient has responded to treatment with SEGRM.

[0620] Therefore, in another preferred embodiment, the ratio of amount and / or frequency distribution is selected from the group consisting of CD38 / CD209 ratio, CD197 / CD209 ratio and CD197 / CD206 ratio.

[0621] Therefore, in another preferred embodiment, one or more M1 cell surface markers are selected from CD38 and one or more M2 cell surface markers are selected from CD209, or one or more M1 cell surface markers are selected from CD197 and one or more M2 cell surface markers are selected from CD209 and CD206.

[0622] In a preferred embodiment, step (iv) comprises contacting the macrophages with a binding agent (preferably an antibody that specifically recognizes one or more M1 surface markers and one or more M2 surface markers), wherein the binding agent is optionally labeled, in particular labeled with a fluorescent marker; and determining the amount of binding molecules bound to the macrophages, in particular by determining the mean fluorescence intensity; thereby determining the amount of cell surface markers. For example, antibodies that specifically recognize surface markers and comprise a fluorescent marker can be used.

[0623] In another preferred embodiment, step (iv) comprises contacting the macrophages with a binding agent (preferably an antibody that specifically recognizes one or more M1 surface markers and one or more M2 surface markers), wherein the binding agent is optionally labeled, in particular labeled with a fluorescent marker; and determining the percentage of cells in the cell population that are positive for one or more M1 surface markers and one or more M2 surface markers, respectively, in particular wherein a FACS analysis is performed; thereby determining the frequency distribution of cell surface markers. For example, antibodies that specifically bind to surface markers and contain fluorescent markers can be used.

[0624] Any of a variety of known methods for identifying and obtaining proteins that specifically interact with proteins or polypeptides (e.g., yeast two-hybrid screening system) can be used to determine whether a protein is a binder of a marker protein. A binder that specifically recognizes a marker preferably has at least 10 affinity to its corresponding target molecule. 7 l / mol affinity. A binding agent that specifically recognizes a marker preferably has 10 8 l / mol affinity, or more preferably 10 9 l / mol. As will be appreciated by the skilled artisan, the term specificity is used to denote a binding agent to which other biomolecules present in the sample do not significantly bind to the specific recognition marker. Preferably, the level of binding to biomolecules other than the target marker molecule results in a binding affinity that is only 10% or less, more preferably only 5% or less, of the affinity to the target marker molecule, respectively. Preferred specific binding agents will meet the above minimum criteria for affinity and specificity.

[0625] The binding agent of specific recognition marker is preferably an antibody that reacts with the marker. The term antibody refers to polyclonal antibodies, monoclonal antibodies, antigen-binding fragments of such antibodies, single-chain antibodies, and gene constructs comprising antibody binding domains. The term "antibody" includes polyclonal antibodies, monoclonal antibodies, fragments thereof such as F(ab')2 and Fab fragments, and any naturally occurring or recombinantly produced binding partners (molecules that specifically bind to marker proteins). Any antibody fragment retaining the above-mentioned criteria of a specific binding agent can be used.

[0626] In order to measure, under the conditions suitable for forming a binding agent label-complex, the sample obtained from the individual is incubated with the binding agent of the label discussed by specific recognition. Such conditions do not need to be described in detail, because the technician does not need any creative effort to easily determine such suitable incubation conditions. The amount of the binding agent label-complex is measured and used in the methods and applications of the present invention. The technician will understand that there are many methods to measure the amount of the specific binding agent label-complex, all of which are described in detail in relevant textbooks (see, for example, Tijssen P., see above, or Diamandis, EP and Christopoulos, TK (eds.), Immunoassay, Academic Press, Boston (1996)).

[0627] In particular, monoclonal antibodies directed against a marker are used in quantitative (determine the amount or concentration of the marker) immunoassays.

[0628] For example, the marker can be detected in a sandwich type detection format. In such a detection, a first specific binding agent is used on one side to capture the marker in question, and a second specific binding agent (e.g., a second antibody) is used on the other side to be labeled as directly or indirectly detectable. The second specific binding agent can include a detectable reporter portion or label, such as an enzyme, a dye, a radionuclide, a luminescent group, a fluorescent group, or biotin, etc. Any reporter molecule portion or label can be used in the method of the present invention, as long as its signal is directly related to or proportional to the amount of the binding agent retained on the support after washing. The amount of the second binding agent that is retained in conjunction with the solid support is then determined using a method suitable for a specific detectable reporter molecule portion or label. For radioactive groups, scintillation counting or autoradiography methods are generally suitable. Antibody-enzyme conjugates can be prepared using a variety of coupling techniques. Spectroscopic methods can be used to detect dyes (including, for example, colorimetric products of enzyme reactions), luminescent groups, and fluorescent groups. Biotin can be detected using avidin or streptavidin coupled to different reporter groups (usually radioactive or fluorescent groups or enzymes). Enzyme reporter groups can usually be detected by adding substrate, usually for a specific period of time, followed by spectroscopic, spectrophotometric or other analysis of the reaction products. Standards and standard additions can be used to determine the level of antigen in a sample using well-known techniques.

[0629] Immunoassays for measuring the marker proteins of the invention include, for example, ELISA, enzyme immunoassay (EIA), and electrochemiluminescent immunoassay (ECLIA) for quantitative determination of the marker proteins described herein.

[0630] In another preferred embodiment, measuring the expression levels of one or more M1 marker mRNAs and one or more M2 marker mRNAs in macrophages incubated with a wound exudate sample or a wound biofilm sample obtained from a skin wound comprises the following steps: (i) co-culturing primary human monocytes with (a) human dermal fibroblasts in 2D cell culture or (b) fibroblast-derived matrix, (ii) incubating the cells until macrophage differentiation is achieved, optionally wherein CD163 is used as a cell surface marker for macrophage differentiation, (iii) contacting the cells with the wound exudate sample or the wound biofilm sample (optionally diluted) with the SEGRM of (2), and (iv) determining the expression levels of one or more M1 marker mRNAs and one or more M2 marker mRNAs in the macrophages.

[0631] In a preferred embodiment of the present invention, the sample is a wound exudate sample. In another preferred embodiment, the sample is a wound biofilm sample. In a more preferred embodiment, the sample is a wound exudate sample.

[0632] Preferably, one or more M1 marker mRNAs are selected from CD38, CD64, CD197, CXCL10 and IL-23p19, and / or one or more M2 marker mRNAs are selected from CD200 receptor (CD200R), CD206, CD209, CCL22 and CCL18, more preferably wherein the method comprises contacting a probe that specifically binds to the marker mRNA (wherein the probe is optionally labeled) with macrophage RNA under conditions that are favorable for hybridization, and detecting the hybridized probe.

[0633] For example, primary human monocytes can be co-cultured with human dermal fibroblasts in 2D cell culture or with fibroblast-derived matrix. The method for producing fibroblast-derived matrix is ​​as described above and in the embodiments. Subsequently, cells are incubated until macrophage differentiation is reached. For example, CD163 can be used as a cell surface marker for macrophage differentiation. Further, cells are contacted with wound exudate samples or wound biofilm samples (which are optionally diluted) and SEGRM of (2), for example, by pipetting the sample to the cells, and optionally gentle mixing. Compounds are added after macrophage differentiation (for example, 4 to 7 days later). Further, cells are incubated, preferably 1 hour to 100 hours. Subsequently, the expression levels of one or more M1 marker mRNAs and one or more M2 marker mRNAs in macrophages are determined. For example, cells can be harvested and mRNA expression levels can be determined using suitable probes. For example, the expression level of a housekeeping gene (such as actin or GAPDH) can be determined, and the expression level of M1 or M2 marker RNA can be determined as the expression level relative to the housekeeping gene.

[0634] In another preferred embodiment, measuring the amount of one or more cytokine markers selected from IL-1α, IL-1β and TNF-α in the supernatant of macrophages incubated with a wound exudate sample or a wound biofilm sample obtained from a skin wound comprises the following steps:

[0635] (i) Primary human monocytes were co-cultured with (a) human dermal fibroblasts in 2D cell culture or (b) fibroblast-derived matrix,

[0636] (ii) incubating the cells until macrophage differentiation is achieved, optionally wherein CD163 is used as a cell surface marker for macrophage differentiation,

[0637] (iii) contacting the cells with a wound exudate sample or a wound biofilm sample (optionally diluted) with the SEGRM of (2), and

[0638] (iv) determining the amount of one or more cytokine markers selected from the group consisting of IL-1α, IL-1β and TNF-α in the cell culture supernatant,

[0639] Preferably, wherein the cytokine markers are determined by using an immunological assay, more preferably by using an ELISA assay.

[0640] In a preferred embodiment of the present invention, the sample is a wound exudate sample. In another preferred embodiment, the sample is a wound biofilm sample. In a more preferred embodiment, the sample is a wound exudate sample.

[0641] For example, primary human monocytes can be co-cultured with human dermal fibroblasts in 2D cell culture or with fibroblast-derived matrix. The method for producing fibroblast-derived matrix is ​​as described above and in the embodiment. Subsequently, cells are incubated until macrophage differentiation is reached. For example, CD163 can be used as a cell surface marker for macrophage differentiation. Further, cells are contacted with wound exudate samples or wound biofilm samples (which are optionally diluted) and the SEGRM of (2), wherein contact can be carried out, for example, by pipetting the sample to the cells, and optionally gentle mixing. Compounds are added after macrophage differentiation (for example, 4 to 7 days later). Further, cells are incubated, preferably 1 hour to 100 hours. Subsequently, the amount of one or more of IL-1α, IL-1β and TNF-α in the cell culture supernatant is determined. For this purpose, supernatant is usually harvested and markers are determined using suitable detection (for example, immunological detection). For example, ELISA can be used. In a preferred embodiment, the sample is a wound exudate sample.

[0642] The amount of IL-1α, IL-1β and TNF-α in the supernatant of macrophages indicates the responsiveness of the patient to treatment with the compound of (2). Therefore, if the values ​​obtained for the amounts of IL-1α, IL-1β and TNF-α are lower than the control values ​​established in the absence of the compound of (2), the patient is determined to be responsive to treatment with the compound of (2).

[0643] The SEGRM for therapeutic or prophylactic use or contained in the kit or set of parts of the present invention is preferably in a pharmaceutical composition. The pharmaceutical composition contains the respective active agent, and optionally one or more pharmaceutically acceptable excipients and / or pharmaceutically acceptable excipients. The active agent is a SEGRM or a pharmaceutically acceptable salt thereof.

[0644] "Pharmaceutically acceptable carrier" refers to a carrier or diluent that does not cause significant irritation to the organism and does not eliminate the biological activity and characteristics of the active agent being administered. The carrier used can be, for example, a solid, a liquid or a gas. Examples of solid carriers include lactose, terra alba, sucrose, talc, gelatin, agar, pectin, gum arabic, magnesium stearate and stearic acid. Examples of liquid carriers are syrup, peanut oil, olive oil and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

[0645] "Pharmaceutically acceptable excipients" refer to inert substances added to pharmaceutical compositions to further facilitate administration of a compound. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and starches, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.

[0646] In a preferred embodiment of any of the above aspects of the invention, the SEGRM is formulated for systemic administration, preferably oral or intravenous administration; or is formulated for local administration, in particular topical, mucosal, ocular, intradermal or subcutaneous administration. For example, topical formulations for the administration of mapracorat or a pharmaceutically acceptable salt thereof are known in the art and are described in detail in US 8,282,909. In addition, the skilled person is aware of the technology for providing other formulations for topical administration, in particular for topical, mucosal, ocular, intradermal or subcutaneous administration. For example, the SEGRM or a pharmaceutically acceptable salt thereof can be formulated for incorporation into a wound dressing or bandage, or made into a gel, semisolid gel, cream, lotion, ointment, spray, foam, dispersion, salves, liposomes or nanoparticle formulations or for microneedle application.

[0647] Therefore, in another preferred embodiment of the present invention, a selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof is

[0648] (i) formulated for systemic administration, preferably oral or intravenous administration, or

[0649] (ii) formulated for topical administration, in particular for topical, mucosal, ocular, intradermal or subcutaneous administration.

[0650] In a more preferred embodiment of the present invention, the SEGRM or a pharmaceutically acceptable salt thereof for a particular use of the present invention is formulated for topical administration, in particular for external, mucosal, ocular, intradermal or subcutaneous administration.

[0651] Topical formulations for the administration of mapracorat or a pharmaceutically acceptable salt thereof are known in the art and are described in detail in US8,282,909. Such formulations comprise at least one selective glucocorticoid receptor modulator (SEGRM) and a) oleyl alcohol, b) cetearyl octanoate and c) a vegetable oil. In a more preferred embodiment, the SEGRM is a SEGRM as disclosed in US8,282,909, in particular mapracorat or a pharmaceutically acceptable salt thereof.

[0652] Therefore, in yet another preferred embodiment of the present invention, the selective glucocorticoid receptor (SEGRM) or a pharmaceutically acceptable salt thereof is formulated for topical administration, wherein the pharmaceutical formulation comprises at least one selective glucocorticoid receptor modulator (SEGRM) and a) oleyl alcohol, b) stearyl octyl cetyl alcohol and c) vegetable oil.

[0653] In a preferred embodiment, the pharmaceutical formulation comprises at least one selective glucocorticoid receptor modulator (SEGRM) and a) 2 to 50% by weight of oleyl alcohol, b) 2 to 50% by weight of cetearyl octyl ester and c) 2 to 50% by weight of vegetable oil,

[0654] Preferably, wherein:

[0655] Vegetable oils are soybean, olive, sesame, castor, or peanut oil, or

[0656] The pharmaceutical preparation further contains d) propylene glycol and e) glycerol, or

[0657] The pharmaceutical preparation contains 3 to 15% (by weight) of oleyl alcohol, or

[0658] Pharmaceutical preparations containing 2 to 15% (by weight) of cetearyl octanoate

[0659] The pharmaceutical preparation contains 3 to 15% (by weight) of vegetable oil, or

[0660] The pharmaceutical preparation further contains medium chain triglycerides, mineral oil, cyclomethicone, stearyl alcohol, butylated hydroxytoluene, polyethylene glycol glyceryl hydroxystearate, povidone, acrylic acid copolymer, hydroxyethylcellulose, acrylic acid and / or tromethamine, or

[0661] The pharmaceutical preparation further contains d) silicon dioxide, or

[0662] The pharmaceutical preparation further contains d) aluminum stearate,

[0663] The pharmaceutical preparation further contains d) propylene glycol, e) glycerol, and f) a propellant,

[0664] The pharmaceutical preparation contains 2% to 10% (by weight) of a propellant, or

[0665] The propellant is a hydrocarbon or a fluoroalkane, or

[0666] At least one pharmaceutically active compound has a solubility of 20 mg / l in water at 20°C or below, or

[0667] The pharmaceutical preparation contains 2 to 50% (by weight) of oleyl alcohol, 3 to 20% (by weight) of cetearyl octanoate, and 5 to 20% (by weight) of vegetable oil, or

[0668] The pharmaceutical preparation contains 3 to 15% (by weight) of oleyl alcohol, 2 to 15% (by weight) of cetearyl octanoate, and 3 to 15% (by weight) of vegetable oil, or

[0669] The pharmaceutical preparation further contains medium chain triglycerides, mineral oil, cyclomethicone, stearyl alcohol, butylated hydroxytoluene, polyethylene glycol glyceryl hydroxystearate, povidone, acrylic acid copolymer, hydroxyethylcellulose, acrylic acid and / or tromethamine, or

[0670] The pharmaceutical preparation also contains silicon dioxide and aluminum stearate, or

[0671] The pharmaceutical formulation contains 6 to 8% (by weight) of a propellant, preferably wherein the propellant is propane, butane, isobutylene, heptafluoropropane, tetrafluoroethane, dimethyl ether or a mixture thereof.

[0672] In another preferred embodiment, the pharmaceutical formulation comprises at least one selective glucocorticoid receptor modulator (SEGRM) and a) 2 to 50% by weight of oleyl alcohol, b) 2 to 50% by weight of cetearyl octyl ester and c) 2 to 50% by weight of vegetable oil, more preferably wherein:

[0673] The pharmaceutical preparation further contains d) propylene glycol and e) glycerol, or

[0674] The pharmaceutical preparation contains 3 to 15% (by weight) of oleyl alcohol, 2 to 15% (by weight) of cetearyl octanoate, and 3 to 15% (by weight) of vegetable oil, or

[0675] The pharmaceutical preparation further contains medium chain triglycerides, mineral oil, cyclomethicone, stearyl alcohol, butylated hydroxytoluene, polyethylene glycol glyceryl hydroxystearate, povidone, acrylic acid copolymer, hydroxyethylcellulose, acrylic acid and / or tromethamine, or

[0676] The pharmaceutical preparation also contains silicon dioxide and aluminum stearate, or

[0677] The pharmaceutical preparation further contains d) propylene glycol, e) glycerol, and f) a propellant, or

[0678] The pharmaceutical preparation contains 2 to 10% (by weight) of a propellant, more preferably wherein the propellant is a hydrocarbon or a fluoroalkane, or wherein the propellant is propane, butane, isobutylene, heptafluoropropane, tetrafluoroethane, dimethyl ether or a mixture thereof; or the pharmaceutical preparation contains 6 to 8% (by weight) of a propellant, and wherein the propellant is propane, butane, isobutylene, heptafluoropropane, tetrafluoroethane, dimethyl ether or a mixture thereof, or

[0679] at least one pharmaceutically active compound has a solubility of 20 mg / l in water at 20°C or below,

[0680] Or the pharmaceutical preparation further contains (R)-1,1,1-trifluoro-4-(5-fluoro-2,3-benzodihydrofuran-7-yl)-4-methyl-2-{[-(2-methyl-5-quinolyl)amino]methyl}pentan-2-ol, or

[0681] The pharmaceutical preparation contains 2 to 50% (by weight) of oleyl alcohol, 3 to 20% (by weight) of cetearyl octanoate, and 5 to 20% (by weight) of vegetable oil.

[0682] The cream, foam and oleogel formulations explicitly disclosed in US 8,282,909 are particularly preferred topical formulations. These cream, foam and oleogel formulations are explicitly incorporated by reference:

[0683]

[0684]

[0685]

[0686]

[0687] Continued from the table:

[0688]

[0689] In another embodiment, the present invention relates to an in vitro method for identifying a subject with impaired skin wound healing responsive to treatment with a glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof, comprising performing steps i) and / or ii):

[0690] i) measuring the proliferation of fibroblasts and, optionally, the amount of at least one IL-1 cytokine marker in the supernatant of the fibroblasts in the presence of,

[0691] (1) a wound exudate sample or a wound biofilm sample obtained from a skin wound of the subject, and

[0692] (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof;

[0693] ii) Measuring fibroblast-derived matrix formation by fibroblasts in the presence of:

[0694] (1) a wound exudate sample or a wound biofilm sample obtained from a skin wound of the subject, and

[0695] (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof.

[0696] Conditions in which a subject is identified as responsive to treatment with a selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof are:

[0697] if the value of fibroblast proliferation measured in step i) and / or the value of fibroblast-derived matrix formation of fibroblasts measured in step ii) is at least 20% higher than a control value established in the absence of the at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2),

[0698] And, optionally, if the value of the amount of at least one IL-1 cytokine marker in the supernatant of the fibroblasts obtained in step i) is lower than a control value established in the absence of at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2).

[0699] In a preferred embodiment of the in vitro method of the present invention, step iiia) and / or one, two, three or four of the following steps iiib) to iiie) are additionally performed:

[0700] iiia) measuring the proliferation of keratinocytes in the presence of:

[0701] (1) a wound exudate sample or a wound biofilm sample obtained from a skin wound of the subject, and

[0702] (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof;

[0703] iiib) measuring the amount of one or more M1 markers and one or more M2 markers in the supernatant of macrophages incubated with:

[0704] (1) a wound exudate sample or a wound biofilm sample obtained from the skin wound, and

[0705] (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof,

[0706] Macrophages were co-cultured with fibroblasts, and

[0707] wherein the one or more M1 markers are selected from CXCL10 and IL-23p19, and the one or more M2 markers are selected from CCL22 and CCL18; iiic) measuring the amount and / or frequency distribution of the one or more M1 cell surface markers and the one or more M2 cell surface markers on macrophages incubated with:

[0708] (1) a wound exudate sample or a wound biofilm sample obtained from the skin wound, and

[0709] (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof,

[0710] Macrophages were co-cultured with fibroblasts, and

[0711] wherein the one or more M1 cell surface markers are selected from CD38, CD64 and CD197, and wherein the one or more M2 cell surface markers are selected from CD200 receptor, CD206 and CD209;

[0712] iiid) measuring the expression level of one or more M1 marker mRNAs and one or more M2 marker mRNAs in macrophages incubated with:

[0713] (1) a wound exudate sample or a wound biofilm sample obtained from the skin wound, and

[0714] (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof,

[0715] Macrophages were co-cultured with fibroblasts, and

[0716] wherein the one or more M1 marker mRNAs are selected from CD38, CD64, CD197, CXCL10 and IL-23p19, and the one or more M2 marker mRNAs are selected from CD200 receptor (CD200R), CD206, CD209, CCL22 and CCL18;

[0717] iiie) measuring the amount of one or more cytokine markers selected from the group consisting of IL-1α, IL-1β and TNF-α in the supernatant of macrophages incubated with:

[0718] (1) a wound exudate sample or a wound biofilm sample obtained from the skin wound, and

[0719] (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof,

[0720] as well as

[0721] Wherein, the subject is identified as responsive to treatment with a selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof if: the value of fibroblast proliferation measured in step i) and / or the value of fibroblast-derived matrix formation of fibroblasts measured in step ii) and / or the value of keratinocyte proliferation in step iiia) is at least 20% greater than a control value established in the absence of the at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2), and / or if one or more of the following is true:

[0722] -iiib) the ratio of the amount of one or more M1 markers to the amount of one or more M2 markers obtained is lower than a control value established in the absence of the at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2),

[0723] -iiic) to the amount and / or frequency distribution of one or more M1 cell surface markers to the amount and / or frequency of one or more M2 cell surface markers is lower than a control value established in the absence of at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2), in particular, wherein the ratio is selected from the group consisting of CD38 / CD209 ratio, CD197 / CD209 ratio and CD197 / CD206 ratio,

[0724] -iiid) is lower than a control value established in the absence of at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2),

[0725] -iiie) is lower than a control value established in the absence of the at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2).

[0726] In a preferred embodiment, the in vitro method of the invention is characterized by the features of the preferred embodiments of the selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof for a specific use of the invention.

[0727] In another embodiment, the present invention relates to a kit or a set of parts comprising:

[0728] (a) a pharmaceutical composition comprising at least one glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof, and

[0729] (b) a diagnostic kit comprising one or more of the following:

[0730] i) fibroblasts,

[0731] ii) a support having a plurality of defined areas or cavities, wherein a subset of the areas or cavities are (i) coated with an adhesion enhancing agent, and / or (ii) filled with a fibroblast-derived matrix (FDM),

[0732] iii) Matrix boosting supplements.

[0733] In a preferred embodiment, the kit or set of parts of the invention features the features of the preferred embodiments of the selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof for a specific use of the invention.

[0734] Therefore, it should be understood that preferred embodiments described in the context of other embodiments of the invention are also applicable to this embodiment of the invention.

[0735] The pharmaceutical compositions, cells and matrix-enhancing supplements may be provided in containers, vials, syringes, ampoules, and the like.

[0736] b) The diagnostic kit optionally further comprises one or more of the following substances:

[0737] iv) keratinocytes,

[0738] v) Matrix boosting supplements

[0739] vi) monocytes, and

[0740] vii) a binding agent, preferably an antibody that specifically recognizes one or more M1 markers and one or more M2 markers; and / or a binding agent, preferably an antibody that specifically recognizes one or more M1 surface markers and one or more M2 surface markers; and / or a probe that specifically recognizes one or more M1 marker mRNAs and one or more M2 marker mRNAs

[0741] viii) a binding agent, preferably, an antibody that specifically recognizes one or more cytokine markers selected from IL-1α, IL-1β and TNF-α, or a hybridization probe that specifically hybridizes with a cytokine marker mRNA selected from IL-1α, IL-1β and TNF-α.

[0742] In a more preferred embodiment, the diagnostic kit of b) further comprises viii) a binding agent, preferably an antibody that specifically recognizes one or more cytokine markers selected from IL-1α, IL-1β and TNF-α, or a hybridization probe that specifically hybridizes with the mRNA of a cytokine marker selected from IL-1α, IL-1β and TNF-α.

[0743] In a more preferred embodiment, the cytokine marker is selected from IL-1α, IL-1β, in particular IL-1β.

[0744] Preferred M1 and M2 markers, cell surface markers and / or marker mRNA are as described above.

[0745] In a preferred embodiment, the binding agent, preferably the antibody of vii) above, is a binding agent, preferably an antibody that specifically recognizes one or more M1 cell surface markers and one or more M2 cell surface markers, wherein the one or more M1 cell surface markers are selected from CD38, CD64 and CD197, and wherein the one or more M2 cell surface markers are selected from CD200 receptor, CD206 and CD209, and, optionally:

[0746] The binding agent is preferably an antibody that specifically recognizes one or more M1 markers and one or more M2 markers and / or a probe that specifically recognizes one or more M1 marker mRNAs and one or more M2 marker mRNAs, wherein the one or more M1 markers are selected from CXCL10 and IL-23p19, and the one or more M2 markers are selected from CCL22 and CCL18, and wherein the one or more M1 marker mRNAs are selected from CD38, CD64, CD197, CXCL10 and IL-23p19, and the one or more M2 marker mRNAs are selected from CD200 receptor (CD200R), CD206, CD209, CCL22 and CCL18.

[0747] Therefore, in another more preferred embodiment, one or more M1 cell surface markers are selected from CD38, and one or more M2 cell surface markers are selected from CD209; or one or more M1 cell surface markers are selected from CD197, and one or more M2 cell surface markers are selected from CD209 and CD206.

[0748] In a preferred embodiment, the keratinocytes are selected from HaCaT cells and primary keratinocytes, in particular human primary keratinocytes.

[0749] In a more preferred embodiment, the keratinocytes used in the present invention are HaCaT cells.

[0750] Fibroblast-derived matrix (FDM) can be obtained by the following steps: (i) seeding primary human dermal fibroblasts on a support pre-coated with an adhesion enhancer (e.g., gelatin), (ii) culturing the cells on the support, preferably until confluence is achieved, and (iii) contacting the cells with a matrix-promoting supplement (e.g., vitamin C or a physiologically acceptable salt thereof, or 2-phospho-L-ascorbic acid or a physiologically acceptable salt thereof, or a combination of EGF and insulin). The FDM can be formed in situ or can be transferred to the support after formation.

[0751] Furthermore, the support is preferably a chip, which allows the in vitro method of the invention or the method steps of the medical use of the invention to be performed. For example, a chip can be provided, which allows the identification of subjects that respond to treatment with SEGRM or a pharmaceutically acceptable salt thereof for impaired wound healing.

[0752] Therefore, in another preferred embodiment, the present invention relates to a kit or a kit of parts according to the present invention, wherein the support ii) of the diagnostic kit (b) is suitable for performing the method according to the present invention or the method steps for the medical use according to the present invention, wherein the support comprises a plurality of defined areas or cavities, and wherein:

[0753] a) a subset of the area or cavity is coated with an adhesion enhancer,

[0754] b) a subset of regions or cavities are coated with an adhesion enhancer and / or filled with a fibroblast-derived matrix (FDM),

[0755] c) a subset of the region or cavity is not treated,

[0756] d) optionally:

[0757] d1) a subset of the regions or cavities contain a binding agent, preferably an antibody that specifically recognizes one or more M1 markers, and

[0758] d2) a subset of the regions or cavities contain a binding agent, preferably an antibody that specifically recognizes one or more M2 markers,

[0759] e) preferably:

[0760] e1) a subset of the regions or cavities contain a binding agent, preferably an antibody that specifically recognizes one or more M1 surface markers, and

[0761] e2) a subset of the regions or cavities contain a binding agent, preferably an antibody that specifically recognizes one or more M2 surface markers,

[0762] f) optionally:

[0763] f1) a subset of the regions or cavities contain probes that specifically recognize one or more M1 marker mRNAs, and

[0764] f2) a subset of the regions or cavities contain probes that specifically recognize one or more M2 marker mRNAs, and

[0765] g) Optionally: a subset of the regions or cavities contain a binding agent, preferably an antibody that specifically recognizes one or more cytokine markers selected from IL-1α, IL-1β and TNF-α

[0766] where subsets a) to g) do not overlap,

[0767] Preferably

[0768] (x) at least some of the regions or cavities according to a) further contain fibroblasts, and / or

[0769] (xi) at least some of the regions or cavities according to (x) or b) further contain mononuclear cells, and / or

[0770] (xii) at least some of the regions or cavities according to c) further contain fibroblasts, and / or

[0771] (xiii) at least some of the regions or cavities according to c) further contain keratinocytes,

[0772] wherein the regions or cavities according to (xii) and (xiii) do not overlap.

[0773] In a preferred embodiment, the one or more M1 markers are selected from CXCL10 and IL-23p19, and the one or more M2 markers are selected from CCL22 and CCL18.

[0774] In a preferred embodiment, the one or more M1 cell surface markers are selected from CD38, CD64 and CD197, and wherein the one or more M2 cell surface markers are selected from CD200 receptor, CD206 and CD209.

[0775] In a preferred embodiment, the one or more M1 marker mRNAs are selected from CD38, CD64, CD197, CXCL10 and IL-23p19, and the one or more M2 marker mRNAs are selected from CD200 receptor (CD200R), CD206, CD209, CCL22 and CCL18.

[0776] In a more preferred embodiment, the support of the kit or set of parts is a chip, an array (eg a microarray or a nanoarray), a plate (eg a multiwell plate) or a culture dish, and / or the support is a plastic support.

[0777] The solid support of the kit or kit preferably contains a plurality of defined cavities. The cavity can fill space and thus allow 3D cell culture. For example, a multiwell plate or a microarray or nanoarray containing a plurality of defined cavities can be used. In the embodiments, a multiwell plate was successfully used. Preferably, the solid support does not substantially interfere with the viability of the cells and / or is suitable for culturing cells, for example, the support can be a plastic support. For 3D cell culture, the solid support can contain a plurality of defined holes. For example, a multiwell plate can be used. In a preferred embodiment, the support comprises 2, 3, 4, 5, 6, 7, 8, 9, 10 or more defined areas or cavities, for example 2, 3, 4, 5, 6, 7, 8, 9 or 10 to 10 5 ; 2, 3, 4, 5, 6, 7, 8, 9 or 10 to 10 4 ; 2, 3, 4, 5, 6, 7, 8, 9 or 10 to 10 3 ; or 2, 3, 4, 5, 6, 7, 8, 9 or 10 to 10 2 a defined area or cavity.

[0778] In yet another embodiment, the present invention is directed to a method of preventing or treating impaired skin wound healing in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of a SEGRM or a pharmaceutically acceptable salt thereof.

[0779] "Effective amount" refers to an amount sufficient to induce a desired biological, pharmacological or therapeutic result in a subject. A therapeutically effective amount of a compound can be used as a zwitterion or a pharmaceutically acceptable salt. A therapeutically effective amount refers to a sufficient amount of a compound to treat or prevent impaired skin wound healing at a reasonable benefit / risk ratio applicable to any medical treatment. However, it should be understood that the total daily dosage of the compounds and compositions of the present invention will be determined by the attending physician within the scope of reasonable medical judgment. The specific therapeutically effective dosage level for any particular patient will depend on a variety of factors, including the disease being treated and the severity of the disease; the activity of the specific compound used; the specific composition used; the patient's age, weight, general health, sex and diet; the administration time, route of administration and excretion rate of the specific compound used; the duration of treatment; drugs used in combination or concomitantly with the specific compound used; and factors well known in the medical field. For example, it is well within the technical scope of the art to start the dose of the compound at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dose until the desired effect is achieved. BRIEF DESCRIPTION OF THE DRAWINGS

[0780] Figure 1 : Analysis of the active SEGRM compound BI-653048 in a human dermal fibroblast proliferation assay (2D) with or without wound exudate from patients 1-3 is shown compared to its inactive analog BI-3047. Fibroblast proliferation in culture medium in the absence of WE is shown in 1D. Filled circles: gradient concentrations of BI-653048; open triangles: gradient concentrations of BI-3047. The active compound reversed the inhibition of fibroblast proliferation induced by wound exudate (WE), while the inactive analog had no effect.

[0781] Figure 2 : Shown is the effect of the active SEGRM compound BI-653048 on fibroblast proliferation (2A) and IL-1β secretion in the supernatant (2B) compared to its inactive analog BI-3047 in the presence of WE-1. Filled circles: gradient concentrations of BI-653048; open triangles: gradient concentrations of BI-3047. The active compound dose-dependently reversed the inhibition of wound exudate (WE)-induced fibroblast proliferation and inhibited IL-1β secretion at the same concentration. The inactive analog had no effect on proliferation and had a much smaller effect on IL-1β secretion.

[0782] Figure 3: Shown is the effect of the active SEGRM compound BI-653048 on collagen 1 and 3 mRNA expression in a fibroblast proliferation assay in the presence of WE-2 compared to its inactive analog BI-3047. Solid bars (3A and 3B): 72 hours incubation in the presence of WE-2; open bars (3C and 3D): 72 hours incubation in culture medium in the absence of WE. Both compounds were used at a concentration of 1 μM. The active compound enhanced the expression of collagen 1 and collagen 3 mRNA in the presence of WE, but not in the culture medium.

[0783] Figure 4 : Shown is the effect of gradient concentrations of SEGRM BI-653048 on the formation of fibroblast-derived matrices of WE-1 in 3D fibroblast culture in the presence and absence of a glucocorticoid receptor antagonist (mifepristone) (concentration 1 μM). Addition of mifepristone abolished the matrix-promoting effects of BI-653048, indicating that these effects are mediated by glucocorticoid receptors.

[0784] Figure 5 : Shows the effect of SEGRM compounds (mapracorat and BI-53048) on fibroblast proliferation in the presence of WE-4. Closed circles: gradient concentrations of BI-53048; Closed squares: gradient concentrations of mapracorat. Both compounds dose-dependently reversed the inhibition of wound exudate (WE)-induced fibroblast proliferation, albeit at different concentrations.

[0785] Figure 6 : Shows the effect of gradient concentrations of mapracorat on the expression of collagen 1 mRNA in fibroblast proliferation assay in the presence of WE-1. Solid bars (6A): 72 hours incubation in the presence of WE-1; open bars (6B): 72 hours incubation in culture medium in the absence of WE. Mapracorat dose-dependently enhances collagen 1 mRNA expression in the presence of WE, but not in culture medium.

[0786] Figure 7 : Shows the effect of gradient concentrations of mapracorat on the expression of collagen 3 mRNA in a fibroblast proliferation assay in the presence of WE-1. Solid bars (7A): 72 hours incubation in the presence of WE-1; open bars (7B): 72 hours incubation in culture medium in the absence of WE. Mapracorat enhanced collagen 3 mRNA expression at all three concentrations in the presence of WE, but not in culture medium.

[0787] Figure 8 : Shows the effect of gradient concentrations of mapracorat on the expression of IL-1β mRNA in fibroblast proliferation assay in the presence of WE-1. Solid bars (8A): 72 hours incubation in the presence of WE-1; open bars (8B): 72 hours incubation in the medium in the absence of WE. Mapracorat dose-dependently inhibits IL-1β mRNA expression in the presence of WE. No IL-1β induction in the medium.

[0788] Fig. 9 : Shown are the effects of SEGRM compounds (ZK216348 and HY14234) on fibroblast proliferation (9A) and IL-1β secretion in the supernatant (9B) in the presence of WE-1. Filled circles: gradient concentrations of HY14234; filled triangles: gradient concentrations of ZK216348. All tested compounds resulted in a dose-dependent partial reversal of the inhibition of wound exudate (WE)-induced fibroblast proliferation, and they inhibited IL-1β secretion at the same concentrations. ZK216348 was less effective than HY14234.

[0789] Fig.10 : Shows the effect of SEGRM compounds (ZK216348 and HY14234) on fibroblast proliferation (10A) and IL-1β secretion in the supernatant (10B) in the presence of WE-2. Solid circles: gradient concentrations of HY14234; solid triangles: gradient concentrations of ZK216348. All tested compounds dose-dependently reversed the inhibition of wound exudate (WE)-induced fibroblast proliferation, while the inhibition of IL-1β secretion was negligible in the context of WE-2.

[0790] Fig.11 : Shows the effect of SEGRM compounds (ZK216348 and HY14234) on fibroblast proliferation (11A) and IL-1β secretion in supernatant (11B) in the presence of WE-3. Solid circles: gradient concentrations of HY14234; solid triangles: gradient concentrations of ZK216348. All tested compounds dose-dependently reversed the inhibition of wound exudate (WE)-induced fibroblast proliferation and inhibited IL-1β secretion at the same concentration. ZK216348 was less effective than HY14234.

[0791] Fig.12: Shown are the effects of multiple SEGRMs in 3D fibroblast cultures of WE from patient #92. AZD7594 was the most active of the SEGRMs tested in 3D culture, followed by mapracorat and BI653048. This is consistent with their potency on glucocorticoid receptor activation (EC 50 The inactive compound BI3047 did not induce matrix formation. Solid diamonds: AZD7594; solid circles: mapracorat; solid squares: HY14234; solid triangles: BI653048; open triangles: BI3047; x: ZK216348.

[0792] Fig.13 : The effect of mapracorat in a porcine model of delayed wound healing is shown. In the presence of the TLR7 / 8 agonist R848 ( Fig.13 AC) or human serum alone as a control ( Fig.13 Wounds were induced on the back of pigs for 5 days using human chronic wound exudates WE-01 and WE-02 or normal human serum in the presence of D). Compound treatment started on day 6 and continued until day 10. Total wound scores (wound appearance, size, contents, pus, scab, erythema, erythema width, swelling, necrosis) were determined daily until day 12.

[0793] 10mM and 1mM Mapracorat (MAPRA) were combined with WE-01 ( Fig.13 A) WE-02( Fig.13 B) and R848( Fig.13 C) Reduced wound scores from day 6 to day 12. In the presence of human serum, Mapracorat did not have any negative effect on the healing of control wounds ( Fig.13 D).

[0794] Although the overall effects of mapracorat were similar in both pigs, the extent of wound score reduction varied depending on the individual pig and the stimulus used (WE and / or R848). Pig #2 responded better than Pig #1, and mapracorat improved WE-02 better than WE-01. In a preferred embodiment, a personalized medicine approach can be performed for individual patients by performing the methods described herein, including pre-testing of the patient's exudate for a specific drug as described above.

[0795] Fig.14Translocation of the glucocorticoid receptor from the cytoplasm of fibroblasts (C) to the nucleus (N) is shown as a measure of activity. Cells were incubated with graded concentrations of compounds and the localization of the glucocorticoid receptor within the cells was determined by indirect immunofluorescence using antibodies against the receptor.

[0796] Both mapracorat and active BI653048 showed nuclear translocation at low concentrations (1-10 nM), whereas BI3047, the inactive stereoisomer of BI653048, was inactive up to 100 nM. Clobetasol, a positive control, also showed nuclear staining.

[0797] Fig.15 : The table shows the effect of 1 μM mapracorat on fibroblast proliferation in the presence of a number of different wound exudates from chronic non-healing skin wounds of human patients. The level of proliferation in the presence of the respective WE was set to 100% and formed the basis for calculating the effect of mapracorat. All values ​​> 120% (mean + 2SD of WE control) were considered to promote growth. In a preferred embodiment of the invention, the method of the invention is used to preselect patients who are most likely to respond effectively to treatment in a personalized manner, as described above.

[0798] Fig.16 Inhibition of LPS-induced IL-8 secretion in U937 cells (A) and spontaneous IL-8 secretion in human monocytes (B) in response to the active SEGRMs mapracorat (filled circles), HY14234 (filled triangles) and BI653048 (filled squares), the inactive compound BI3047 (open squares) and the glucocorticoid dexamethasone as a comparator (filled diamonds) are shown. The active SEGRMs inhibit LPS-induced and spontaneous IL-8 secretion in U937 and primary human monocytes, respectively, while the inactive stereoisomer shows induction of IL-8 secretion even at the highest concentration. The comparator corticosteroid (dexamethasone) inhibits IL-8 in a similar manner to the SEGRMs, confirming the anti-inflammatory properties of these compounds. Example

[0799] Example 1: Detection used in the present invention

[0800] abbreviation

[0801] Abbreviation

[0802] DMSO Dimethyl sulfoxide

[0803] FACS fluorescence activated cell sorting

[0804] FCS Fetal calf serum

[0805] FDM Fibroblast-Derived Matrix

[0806] HaCaT human keratinocyte cell line

[0807] HBSS Hank's Balanced Salt Solution

[0808] HDF Human dermal fibroblasts

[0809] HGF Hepatocyte Growth Factor

[0810] M-CSF Macrophage colony stimulating factor

[0811] PBS Phosphate buffered saline

[0812] RPMI Rossville Parker Memorial Institute Medium

[0813] SRB Sulforhodamine B

[0814] TGFbeta Transforming Growth Factor Beta (TGF-β)

[0815] WE wound exudate

[0816] The assays described in Examples 1.1 and 1.2 represent a predictive model for skin wound healing. Most non-healing wound exudates (WE) obtained from various patients inhibited the proliferation of primary human fibroblasts (HDFs) in the assay described in Example 1.1 and also inhibited the formation of fibroblast-derived matrices (FDMs) in 3D as described in Example 1.2.

[0817] Example 1.1: Fibroblast proliferation assay: In fibroblasts obtained from skin wounds (especially chronic human skin wounds) Measuring fibroblast proliferation and IL-1β secretion in the presence of wound exudate samples

[0818] Primary human dermal fibroblasts (HDF) were purchased from CELLnTEC, Bern. They were routinely grown in Dulbecco's modified Eagle's medium (DMEM) containing 10% FCS, 2 mM glutamine and 100 U / ml penicillin / 100 μg / ml streptomycin. Culture medium, antibiotics and glutamine were purchased from Lonza. Cells were used at passages 5-15. Cells were trypsinized and seeded at 2500 cells / well in 30 μl into the inner wells of a 384-well plate in the absence or presence of gradient compound concentrations in the culture medium, with or without different dilutions of sterile filtered WE. For control samples, 30 μl of culture medium was added instead of the specific stimulus. The outer wells were filled with sterile water. Cells were incubated at 37°C for 72 hours.

[0819] At the end of the culture period, the supernatant was removed for IL-1β determination, the cells were fixed with 4% paraformaldehyde (Morphisto) for 15 min at room temperature, and washed three times with PBS. After overnight cell attachment (day 1), control plates were fixed to determine the starting cell number.

[0820] Total cellular protein was determined as a measure of cell number by staining fixed cells with sulforhodamine B (SRB, Sigma). A 0.4% SRB in 1% acetic acid solution was added to the wells for 30 minutes. The wells were then washed with 1% acetic acid until the wash solution remained colorless. After drying, the dye was eluted with 10 mM Tris·HCl, pH 8.5, and the absorbance was measured at 550 or 492 nm for lower and higher cell densities, respectively. The average absorbance of the sample representing the starting cell number on day 1 was subtracted from the absorbance values ​​of the WE-treated cells.

[0821] IL-1β levels were determined using a commercial ELISA kit. The amount of IL-1β contained in the wound exudate added to the cells was subtracted from the total IL-β in the supernatant to determine the cytokine secreted by the cells.

[0822] For each sample and concentration, all experiments were performed in triplicate and the mean ± standard deviation (SD) was used to evaluate the experiments. Results are expressed as a percentage of the control value of unstimulated cells.

[0823] Fig.15 The table in shows the effect of 1 μM mapracorat on fibroblast proliferation in the presence of a number of different wound exudates from chronic, non-healing skin wounds of human patients. The proliferation level in the presence of the respective WE was set to 100% and formed the basis for calculating the effect of mapracorat. All values ​​> 120% (mean + 2SD of WE control) were considered growth promoting.

[0824] Further results are shown in Table 1 below:

[0825] WE AZD1594 WE-04 100 1139 WE-05 100 203 WE-37 100 171 WE-03 100 161 WE-02 100 196 WE-77 100 128

[0826] Table 1

[0827] The table shows the effect of 1 μM AZD7594 on fibroblast proliferation in the presence of a variety of different wound exudates. The level of proliferation in the presence of the respective WE was set to 100% and formed the basis for calculating the effect of AZD7594 as a percentage of the proliferation of the respective WE control. All values ​​> 120% (mean + 2SD of WE control) were considered growth promoting. The efficacy of AZD7594 was comparable to that of mapracorat (see Fig.15 ).

[0828] Example 1.2: Measurement of fibroblast-derived matrix formation (FDM) of fibroblasts: In the The fibroblastic proliferation rate was measured in the presence of wound exudate samples obtained from chronic human skin wounds. Cell-derived matrix formation

[0829] Human dermal fibroblasts (HDF) were seeded at 1250 cells / well on day -3 into 384-well tissue culture plates that had been pre-coated with 0.2% gelatin solution (Sigma) for 1 hour at 37°C. When the cells reached confluence (= day 0), matrix-promoting supplements (vitamin C: 2-phospho-L-ascorbic acid trisodium salt, 100 μg / ml; Sigma) were added as described for the HDF proliferation assay together with test samples containing TGF-β1 or graded concentrations of compound- / +WE. After 4 days, the culture medium was replaced with fresh vitamin C and stimulants and medium containing compounds, maintaining the conditions starting on day 0. TGF-β1 was included as a positive control to promote the formation of FDM. After a total incubation time of 7 to 8 days, FDM production was measured and evaluated in fixed cultures by SRB staining as described above. In some cases, the experiment was stopped and evaluated already on day 4.

[0830] For example, the experimental results are as follows Fig.12 shown. Fig.12 The effects of multiple SEGRMs in 3D fibroblast cultures of WE from patient #92 are shown. AZD7594 was the most active of the SEGRMs tested in 3D culture, followed by mapracorat and BI653048. This is consistent with their potency on glucocorticoid receptor activation (EC 50 The inactive BI3047 (which is not a SEGRM) did not induce matrix formation.

[0831] Example 1.3: Keratinocyte proliferation assay: In the case of cells obtained from skin wounds (especially chronic human skin wounds) Measuring keratinocyte proliferation in the presence of wound exudate samples

[0832] The HaCaT keratinocyte cell line was routinely cultured in DMEM containing 10% FCS, 2 mM glutamine and 100 U / ml penicillin / 100 μg / ml streptomycin. Proliferation assays were performed as described for HDF cells. Primary human keratinocytes were grown in KBM medium (Lonza) containing 0.06 mM calcium and supplemented with growth factors (Lonza) on plastic coated with rat tail collagen (40 μg / ml; Gibco) or gelatin (0.2%; Sigma). No antibiotics were used. Proliferation assays were performed as described for HDF cells.

[0833] Example 1.4: Primary human macrophage stimulation assay: measuring cytokine production

[0834] Primary human macrophages are differentiated from monocytes, which are isolated from peripheral blood mononuclear cells (PBMC). PBMCs were isolated from buffy coats obtained from the Vienna Red Cross using LymphoPrep (Technoclone). 30 ml of buffy coat concentrate was diluted 1:2 with PBS, gently primed with 15 ml Lymphoprep in a 50 ml Falcon tube, and centrifuged at 1800 rpm for 25 minutes at 21°C. The interphase was carefully transferred to a new Falcon tube and filled to 50 ml with ice-cold PBS. After centrifugation again (10 minutes, 1200 rpm, 4°C), the cell pellet was washed 3 times with PBS, resuspended in RPMI medium containing 20% ​​FCS and 10% DMSO, and frozen in liquid nitrogen. Monocytes were generated from frozen aliquots by positive selection using CD14 Beads-Kit (Miltenyi) on an autoMACS-Sorter (Miltenyi) according to the manufacturer's instructions.

[0835] For the culture and differentiation into macrophages, monocytes were cultured at 3-5x10 6 100 mononuclear cells / well were seeded in 6-well plates (Nunc) and incubated with 20 ng / ml M-CSF (R&D Systems) in RPMI supplemented with 10% FCS, 2 mM glutamine and 100 U / ml penicillin / 100 μg / ml streptomycin in a total volume of 5 ml per well. After 2 days, 2 ml of supernatant was removed and replaced with 2.5 ml / well of fresh medium containing 20 ng / ml M-CSF. On the third day, microscopic examination showed differentiation into adherent, frequently elongated cells.

[0836] Macrophages were harvested and replated in 200 μl or 50 μl serum-free medium on 96-well or 384-well plates, respectively, and cells were combined with graded concentrations of test compounds in the absence or presence of various dilutions of sterile-filtered WE.

[0837] A combination of 100 ng / ml LPS (Sigma) and 50 ng / ml IFN-γ (PeproTech) was used as a positive control for induction of cytokine secretion. For negative control samples, culture medium was added instead of the specific stimulus.

[0838] After 24 hours, supernatants were transferred to fresh plates and frozen at -20°C for future cytokine analysis (IL-1α, IL-1β, TNF-α). Cytokine concentrations of WE input were subtracted from supernatant levels to calculate WE-induced cytokine stimulation.

[0839] Example 1.5: Human monocyte-dermal fibroblast co-culture as a reflection of macrophage behavior in human skin In vitro model:

[0840] (a) measuring the amount of one or more M1 markers and one or more M2 markers in the supernatant of macrophages incubated with a wound exudate sample obtained from a skin wound, wherein the macrophages are co-cultured with fibroblasts; and (b) measuring the amount and / or frequency distribution of one or more M1 cell surface markers and one or more M2 cell surface markers on macrophages incubated with a wound exudate sample obtained from a skin wound, wherein the macrophages are co-cultured with fibroblasts; (c) measuring the expression level of one or more M1 marker mRNA and one or more M2 marker mRNA in macrophages incubated with a wound exudate sample obtained from a skin wound, wherein the macrophages are co-cultured with fibroblasts; and (d) the amount of one or more cytokine markers selected from IL-1α, IL-1β and TNF-α in the supernatant of macrophages incubated with a wound exudate sample obtained from a skin wound, wherein the macrophages are co-cultured with fibroblasts.

[0841] CD14 isolated from PBMCs of healthy donors by magnetic bead separation + Monocytes were incubated alone or in the presence of primary human dermal fibroblasts (CellNTec) or fibroblast-derived matrix (FDM). Primary human dermal fibroblasts were incubated for 3 weeks with growth supplements vitamin C or insulin and EGF (vitamin C: 2-phospho-L-ascorbic acid trisodium salt, 100 μg / ml; human EGF, 5 ng / ml; human insulin, 5 μg / ml) to generate FDM. Alternatively, fibroblast monolayer cultures can also be used. After allowing macrophages to differentiate for 4 days to one week in the presence or absence of M-CSF (25 ng / ml), the cultures were stimulated overnight with gradient concentrations of test compounds in the absence or presence of various dilutions of sterile filtered WE. IFN-γ (50 ng / ml), LPS (100 ng / ml) and IL-4 (25 ng / ml) or their combinations were used as controls for M1 and M2 macrophage induction. For negative control samples, culture medium was added instead of specific stimuli. WE with and without compounds were added to the culture medium at dilutions ranging from 1:25 to 1:100 for overnight stimulation.

[0842] Supernatants were collected and frozen for cytokine detection by ELISA, and cells were collected and subjected to FACS analysis, gating on the monocyte population. The geometric mean of mean fluorescence intensity (MFI) was used to quantify surface marker expression.

[0843] By measuring the mean fluorescence intensity, there are two possibilities for the assessment: a) the percentage of cells in the population that are positive for a given marker, which is the most commonly used readout in FACS analysis, or b) the amount of cell surface expression (as a surrogate for the number of labeled molecules on the cell surface per cell).

[0844] Specific mRNA levels are determined as ratios compared to housekeeping genes; the resulting values ​​are relative to the expression of the housekeeping genes.

[0845] The following readouts were used:

[0846] FACS: CD38, CD64 and CD197 are used for M1 macrophages, CD200 receptor (CD200R), CD206 and CD209M2 are used for M2 macrophages, and CD163 is used as a marker for macrophage differentiation. The ratio of M1 / M2 cell surface marker expression was calculated.

[0847] ELISA: CXCL10 and IL-23p19 for M1 macrophages, CCL22 and CCL18 as M2 macrophage markers, IL-1α, IL-1β, and TNF-α as pro-inflammatory markers indicative of the M1 phenotype.

[0848] mRNA: CD38, CD64 CD38, CD64 and CD197 are used for M1 macrophages, CD200 receptor (CD200R), CD206 and CD209 are used for M2 macrophages, and CD163 is used as a marker for macrophage differentiation.

[0849] Example 1.6: Determination of CCL18

[0850] CCL18 in WE and macrophage supernatants was determined using the hCCL18 / PARC DuoSet ELISA kit from R&D Systems (#DY394) in F96 Maxisorp Nunc immunoplates (Nunc, #439454) according to the manufacturer's instructions. Enzyme reactions and measurements were performed as described for IL-1α.

[0851] Example 1.7: Analysis of macrophage surface markers by flow cytometry

[0852] Cells were harvested and resuspended in FACS buffer (PBS containing 2% FCS). Nonspecific antibody binding was prevented by incubation on ice for 10 minutes with human Trustain FCR blocking solution (Biolegend, #422302). The following fluorescent dye-coupled antibodies from eBioscience (now ThermoFisher Scientific) were used to detect specific surface markers by staining on ice for 30 minutes: CD38-PerCPeFluor710 (#46-0388-42), CD197-APC (#17-1979-42), CD206-AF488 (#53-2069-42), CD209-PerCP Cy5.5 (#45-2099-42). When analyzed from co-culture, CD45 eFluor (#506 69-0459-42) co-staining was used to distinguish macrophages from primary human fibroblasts. After washing with FACS buffer, cells were fixed with 1% paraformaldehyde in PBS and stored at 4°C in the dark until data acquisition on Beckman Coulter Gallios flow cytometer and analysis using Kaluza analysis software 1.3.

[0853] Example 1.8: Analysis of mRNA expression in fibroblast cultures

[0854] Cells were seeded into 24-well plates and incubated with compounds for 72 hours in the presence or absence of wound exudate. Total RNA was isolated using the RNeasy Mini Kit (QIAGEN #74106) according to the manufacturer's protocol. RNA integrity and concentration of each sample were confirmed and measured using a Qubit fluorometer. Each RNA sample was then diluted to 2 ng / μl in nuclease-free double distilled water.

[0855] 20 ng of total RNA was reverse transcribed into cDNA and immediately subjected to PCR amplification using SuperScript III Platinum One-Step Quantitative RT-PCR System (Invitrogen #11745) containing ROX. qRT-PCR amplification was performed in a 20 μl reaction containing 9 μl 2X Reaction Mix containing ROX, 0.4 μl SuperScript III RT / Platinum Taq Mix and 1 μl primer (final: 900 nM, Taqman Gene Expression Assay, Applied Biosystems).

[0856] The program included reverse transcription at 48° C. for 30 minutes, initial denaturation at 95° C. for 5 minutes, followed by 40 cycles of denaturation at 95° C. for 15 seconds and annealing at 60° C. for 60 seconds. Reactions were set up in 96-well format PCR plates (Peqlab #732-2879) and performed in a Mx3005P Real-Time PCR Detection System (Stratagene).

[0857] TaqMan Gene Expression Assays (Applied Biosystems) were used, which included pre-optimized probes and primer sets specific for the gene to be validated. The sequences and ID numbers are listed below:

[0858]

[0859] The housekeeping gene elongation factor 1a (EF1a) was used as a reference gene. Normalized expression was calculated using the comparative Ct (or ΔCt) method, with fold changes derived from the 2-ΔΔCt value of each gene. Graphs were prepared using relative Ct values ​​calculated by subtracting the EF1a Ct value from the corresponding Ct value of the gene being measured.

[0860] Example 1.9: Delayed wound healing in a porcine model

[0861] Animal experiments were approved by government authorities. Two young farm pigs (10-12 kg) were anesthetized and full-thickness excisional wounds were created using a 6-mm biopsy punch. For the first 5 days after wounding, the wounds were stimulated with: a) 0.05% R848 (resiquimod) and 50 μl of human wound exudate, b) 0.05% R848 and human serum or c) human serum alone. R848 was applied in the morning and 6 hours later the exudate and serum were added to a 50 μl solution and gelled with 2% HPMC. On days 6-10, the wounds were treated daily with mapracorat or vehicle (50% PG / 47.5% H 2 Wounds were treated once with 1% 2% tween (0.2% Tween 80 / 0.5% Tween 80 / 2% HPMC). Wounds were clinically scored daily. The total wound score (maximum score = 21) included wound appearance (dry vs. wet), size, wound contents, pus, scab, erythema intensity, erythema width, swelling, and necrosis. The reported values ​​are individual values ​​for each of the 2 pigs.

[0862] Example 1.10: Translocation of glucocorticoid receptor from cytoplasm to nucleus in primary human fibroblasts

[0863] As described in Example 1.1, primary human fibroblasts were seeded into 384-well plates using 2500 cells / well in a total volume of 50 μl. After 24 hours of attachment, the cells were serum starved overnight and then incubated at 37°C for 45 minutes with gradient concentrations of SEGRM or corticosteroids as positive controls. The cells were then fixed with 4% paraformaldehyde for 10 minutes at room temperature and then permeabilized with 0.5% Triton X100 in 1% BSA in PBS for 10 minutes at room temperature. They were stained with mouse anti-glucocorticoid receptor monoclonal antibodies (CellSignaling #47411) or small IgG1-κ isotype controls (e-Bioscience #14-4714) and developed with Alexa Fluor donkey anti-mouse IgG (Molecular Probes #A21202). Cells were examined under an Olympus CKX53 fluorescence microscope and assessed based on the localization of glucocorticoid receptor: predominantly nuclear (N), nuclear + cytoplasmic (N / C), or predominantly cytoplasmic (C).

[0864] The results are shown in Table 2 below:

[0865] Table 2

[0866] 100nM 10nm 1nM 0.1nM AZD7594 N N N N / C CORT108297 N N N / C C HY14234 N N N N / C Mapracorat N N N N / C ZK216348 N N N N / C BI353048 N N N C BI3047 C / N C C C

[0867] The table shows the translocation of the glucocorticoid receptor from the cytoplasm (C) to the nucleus (N) of fibroblasts as a measure of activity. Cells were incubated with graded concentrations of compounds and the localization of the glucocorticoid receptor within the cells was determined by indirect immunofluorescence using antibodies against the receptor. The localization of the glucocorticoid receptor is represented by N (nucleus), N / C (nucleus > cytoplasm), C / N (cytoplasm > nucleus) and C (cytoplasm) for compound concentrations ranging from 0.1 to 100 nM.

[0868] Example 1.11: Detection of cytokine secretion in primary human monocytes

[0869] Primary human monocytes were prepared from peripheral blood mononuclear cells (PBMC). PBMCs were isolated from buffy coats obtained from the Vienna Red Cross using LymphoPrep (Technoclone). 30 ml of buffy coat concentrate was diluted with PBS 1:2, gently primed with 15 ml of Lymphoprep in a 50 ml Falcon tube, and centrifuged at 1800 rpm for 25 minutes at 21°C. The interphase was carefully transferred to a new Falcon tube and filled to 50 ml with ice-cold PBS. After centrifugation again (10 minutes, 1200 rpm, 4°C), the cell pellet was washed 3 times with PBS, resuspended in RPMI medium containing 20% ​​FCS and 10% DMSO, and frozen in liquid nitrogen. Positive selection was performed on autoMACS-Sorter (Miltenyi) using CD14 Beads-Kit (Miltenyi) according to the manufacturer's instructions to generate monocytes from frozen aliquots.

[0870] The cells were plated at 8x10 5 Cells were seeded in 384-well plates at 50 μl / well and mixed with graded concentrations of test compounds. After 16 hours, supernatants were transferred to fresh plates and frozen at -20°C for IL-8 analysis using a commercial ELISA kit.

[0871] Example 1.12: U937 cell stimulation assay

[0872] The human monocytic cell line U937 (ATCC CRL 1593) was routinely grown in RPMI 1640 supplemented with 10% FCS and 2 mM glutamine and 100 U / ml penicillin / 100 μg / ml streptomycin. Cells were plated at 4×10 5 / ml, 50μl / well were seeded in 384-well plates and cells were mixed with gradient concentrations of test compounds and LPS at 100ng / ml. After 16 hours, supernatants were transferred to fresh plates and frozen at -20°C for IL-8 analysis using a commercial ELISA kit.

[0873] Wound exudate

[0874] Wound exudate (WE) from venous, arterial, pressure and diabetic ulcers and surgical wounds was collected after negative pressure therapy in a gel-free container or by swabbing with a nylon flocked swab (Copan #502CS01). Informed consent was obtained from the patients in accordance with the Declaration of Helsinki.

[0875] Test compound

[0876] Low molecular weight compounds (see Table 3 for a list) were dissolved in DMSO (Cell Culture Bioreagents, Sigma) at 10 mM or 100 mM and diluted at least 1:1000 in culture medium for cell-based assays (final DMSO concentration ≤ 0.1%). Compounds were typically tested in a logarithmic (1:10) or semi-logarithmic (1:3.33) dilution series, starting with 10 μM or 100 μM as the highest compound concentration.

[0877] Cells were incubated with compounds for 72 hours in the proliferation assay and up to 8 days in the FDM assay (refreshed after 4 days). When compounds were tested for their effect on WE stimulation, incubation of cells with compounds was started and ended simultaneously with WE-incubation.

[0878] Table 3 List of low molecular weight compounds used in cell assays

[0879] Compound source Dexamethasone 21-acetate Sigma-Aldrich BI-653048 Boehringer Ingelheim BI-3047 Boehringer Ingelheim Mifepristone MedChem Express Prednisolone Sigma-Aldrich Mapracorat MedChem Express ZK216348 Axon Medchem HY14234 MedChem Express AZD7594 MedChem Express Sequence Listing <110> Acris Biopharmaceuticals Ltd. <120> Selective glucocorticoid receptor modifiers for the treatment of impaired cutaneous wound healing <130> 21F-1859-CNP <150> EP19153026.0 <151> 2019-01-22 <150> EP19170117.6 <151> 2019-04-18 <160> 4 <170> PatentIn version 3.5 <210> 1 <211> 25 <212> DNA <213> Artificial sequence <220> <223> EF1a probe <400> 1 aagtgctaac atgccttggt tcaag 25 <210> 2 <211> 25 <212> DNA <213> Artificial sequence <220> <223> Collagen 1a probe <400> 2 aagacgaaga catcccacca atcac 25 <210> 3 <211> 25 <212> DNA <213> Artificial sequence <220> <223> Collagen 3a probe <400> 3 gaactcaaga gtggagaata ctggg 25 <210> 4 <211> 25 <212> DNA <213> Artificial sequence <220> <223> IL1β probe <400> 4 cagatgaagt gctccttcca ggacc 25

Claims

1. Use of a selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof in the preparation of a pharmaceutical composition for treating impaired skin wound healing in a subject, wherein the SEGRM is selected from (i) or a pharmaceutically acceptable salt thereof; (ii) or a pharmaceutically acceptable salt thereof; (iii) compound (R)-2-(4-((5-(ethylsulfonyl)-1H-pyrrolo[2,3-c]pyridin-2-yl)methyl)-5,5,5-trifluoro-4-hydroxy-2-methylpentan-2-yl)-5-fluorobenzamide, or a pharmaceutically acceptable salt thereof; (iv) or a pharmaceutically acceptable salt thereof; and (v) or a pharmaceutically acceptable salt thereof.

2. The use according to claim 1, wherein the skin wound is selected from: a wound of a diabetic patient; a skin wound infected by at least one microorganism, an ischemic wound, a wound of a patient suffering from insufficient blood supply or venous stasis, an ulcer, a neuropathic wound, a surgical wound, a burn, a fissure, and a bullous skin disease.

3. The use according to claim 2, wherein the ulcer is selected from diabetic ulcers, venous ulcers, arterial ulcers, mixed ulcers, pressure ulcers, leg ulcers, tumor ulcers and rare ulcers.

4. The use according to claim 3, wherein the lower extremity ulcer is a lower extremity arterial ulcer.

5. The use according to claim 2, wherein the bullous skin disease is epidermolysis bullosa.

6. The use according to claim 1, wherein the subject suffers from at least one comorbidity associated with impaired skin wound healing, and / or wherein the subject is treated with at least one immunosuppressive drug.

7. The use according to claim 6, wherein the comorbidity is diabetes.

8. The use according to claim 1, wherein the subject suffers from at least one diabetic ulcer, and / or wherein the subject: (i) have undergone a transplant, and / or (ii) Access to immunosuppressive therapy.

9. The use according to claim 1, wherein the subject is identified as responsive to treatment for impaired skin wound healing by performing steps i) and / or ii): i) Measuring the proliferation of fibroblasts in the presence of: (1) a wound exudate sample or a wound biofilm sample obtained from a skin wound of the subject, and (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof; ii) Measuring fibroblast-derived matrix formation by fibroblasts in the presence of: (1) a wound exudate sample or a wound biofilm sample obtained from a skin wound of the subject, and (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof.

10. The use according to claim 9, wherein step i) further comprises measuring the amount of at least one IL-1 cytokine marker in the supernatant of the fibroblasts.

11. The use of claim 9, wherein the subject is identified as responsive to treatment with a selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof for impaired skin wound healing: If the value of fibroblast proliferation measured in step i) and / or the value of fibroblast-derived matrix formation of fibroblasts measured in step ii) is at least 20% higher than a control value established in the absence of the at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2).

12. The use of claim 10, wherein the subject is identified as responsive to treatment with a selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof for impaired skin wound healing: If the value of the fibroblast proliferation measured in step i) and / or the value of the fibroblast-derived matrix formation of the fibroblasts measured in step ii) is at least 20% higher than a control value established in the absence of at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2), and if the value of the amount of at least one IL-1 cytokine marker in the supernatant of the fibroblasts obtained in step i) is lower than a control value established in the absence of at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2).

13. The use according to claim 9, wherein one, two, three or four of step iiia) and / or the following steps iiib) to iiie) are additionally performed: iiia) Measuring the proliferation of keratinocytes in the presence of: (1) a wound exudate sample or a wound biofilm sample obtained from a skin wound of the subject, and (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof; iiib) measuring the amount of one or more M1 markers and one or more M2 markers in the supernatant of macrophages incubated with: (1) a wound exudate sample or a wound biofilm sample obtained from the skin wound, and (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof, wherein the macrophages are co-cultured with fibroblasts, and wherein the one or more M1 markers are selected from CXCL10 and IL-23p19, and the one or more M2 markers are selected from CCL22 and CCL18; iiic) measuring the amount and / or frequency distribution of one or more M1 cell surface markers and one or more M2 cell surface markers on macrophages incubated with: (1) a wound exudate sample or a wound biofilm sample obtained from the skin wound, and (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof, wherein the macrophages are co-cultured with fibroblasts, and wherein the one or more M1 cell surface markers are selected from CD38, CD64 and CD197, and wherein the one or more M2 cell surface markers are selected from CD200 receptor, CD206 and CD209; iiid) measuring the expression level of one or more M1 marker mRNAs and one or more M2 marker mRNAs in macrophages incubated with: (1) a wound exudate sample or a wound biofilm sample obtained from the skin wound, and (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof, wherein the macrophages are co-cultured with fibroblasts, and wherein the one or more M1 marker mRNAs are selected from CD38, CD64, CD197, CXCL10 and IL-23p19, and the one or more M2 marker mRNAs are selected from CD200 receptor (CD200R), CD206, CD209, CCL22 and CCL18; iiie) measuring the amount of one or more cytokine markers in the supernatant of macrophages incubated with: (1) a wound exudate sample or a wound biofilm sample obtained from the skin wound, and (2) at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof, wherein the macrophages are co-cultured with fibroblasts, and wherein the one or more cytokine markers are selected from IL-1α, IL-1β and TNF-α, and in, The subject is identified as responsive to treatment with at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof if the value of fibroblast proliferation measured in step i) and / or the value of fibroblast-derived matrix formation of fibroblasts measured in step ii) and / or the value of keratinocyte proliferation in step iiia) is at least 20% higher than a control value established in the absence of the at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2), and, when step i) further comprises measuring the amount of at least one IL-1 cytokine marker in the supernatant of fibroblasts, if the value of the amount of at least one IL-1 cytokine marker in the supernatant of fibroblasts obtained in step i) is lower than a control value established in the absence of the at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2), and / or if one or more of the following applies: -iiib) the ratio of the amount of the one or more M1 markers to the amount of the one or more M2 markers obtained is lower than a control value established in the absence of the at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2), -iic) to the amount and / or frequency distribution of one or more M1 cell surface markers obtained in (2) and (3) to the amount and / or frequency of one or more M2 cell surface markers is lower than a control value established in the absence of at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2), in particular, wherein the ratio is selected from the group consisting of CD38 / CD209 ratio, CD197 / CD209 ratio and CD197 / CD206 ratio, -iiid) is lower than a control value established in the absence of at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2), -iiie) is lower than a control value established in the absence of the at least one selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof of (2).

14. The method of any one of claims 1 to 13, wherein the selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof is (i) formulated for systemic administration; or (ii) formulated for topical administration.

15. The use according to claim 14, wherein the systemic administration is oral or intravenous administration.

16. The use according to claim 14, wherein the local administration is topical, mucosal, ocular, intradermal or subcutaneous administration.

17. The use according to claim 14, wherein the selective glucocorticoid receptor modulator (SEGRM) or a pharmaceutically acceptable salt thereof is formulated for topical administration, wherein the pharmaceutical formulation comprises at least one selective glucocorticoid receptor modulator (SEGRM) and a) oleyl alcohol, b) cetearyl octanoate and c) vegetable oil.

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