Compounds for use in autoimmune conditions

Abstract

The present invention relates to the use of compounds in the treatment of autoimmune conditions, in particular in the treatment of rheumatoid arthritis.

Description

[Technical Field]

[0001] This invention relates to the treatment of autoimmune conditions. [Background technology]

[0002] Autoimmune states are characterized by chronic inflammation involving activation of the Toll-like receptor (TLR) pathway. In this pathway, any adverse stimulus triggered by injury, infection, stress, hypoxia, or cell death causes tissue damage, leading to the release of endogenous TLR ligands or "autoantigens." There are several hypotheses as to how such endogenous TLR ligands are generated and cause inflammation during autoimmunity, and many or all of these processes may occur during autoimmune diseases.

[0003] One hypothesis is that antigens normally unrecognizable by the immune system because they are inside cells may accumulate on the cell membrane due to high levels of apoptosis, becoming recognizable by the immune system and potentially acting as TLR ligands. Another hypothesis is that neoepitopes may be generated that can induce immune responses, including acting as TLR ligands. Neoepitopes may be generated by modifications of existing molecules through enzymatic cleavage, post-translational modification, or other structural modifications. These changes may be induced by environmental factors or in vivo changes, such as dysregulation of enzyme activity, including increased granzyme B activity due to apoptosis.

[0004] These ligands, also known as DAMPs (damage-associated molecular patterns), bind to Toll-like receptors (humans have 10 subtypes, called TLR1-10), leading to the activation of signaling cascades that reach peaks in inflammatory responses. Endogenous TLR ligands have been identified for at least TLR2, 3, 4, 5, 7, 8, 9, and 11 and are associated with many autoimmune diseases. In addition, microbial products that are known TLR ligands are also found in patients with autoimmune diseases and can drive the TLR signaling cascade in addition to endogenous TLR ligands.

[0005] Such a single signaling cascade activates the canonical NF-κB pathway, a critical regulator of inflammation, a central mediator of pro-inflammatory gene induction, and therefore a key driver of autoimmune pathology.

[0006] In the NF-κB pathway, binding of endogenous ligands to TLRs induces receptor dimerization. Downstream, TLRs can interact with a range of adapter proteins that mediate various signaling pathways. Myeloid differentiation primary response protein 88 (MyD88) is the most widely used TLR adapter protein and mediates signaling through all TLRs. MyD88 interacts with the threonine-serine kinase interleukin (IL)-1 receptor-associated kinase 4 (IRAK4), and when activated, phosphorylates IRAK1. Subsequently, IRAK recruits the ubiquitin ligase tumor necrosis factor receptor-associated factor 6 (TRAF-6), which polyubiquitizes and activates TAK1 kinase. TAK1 kinase activates the IKK complex, which induces the proteolysis of inhibitory κB (I-κB), an inhibitor of nuclear factor κB (NF-κB). This reveals the nuclear localization signal of NF-κB, enabling the translocation of this transcriptional complex from the cytoplasm to the nucleus, as well as the activation of a wide variety of NF-κB-responsive genes, including genes encoding pro-inflammatory cytokines and costimulatory molecules necessary for the activation of adaptive immune responses.

[0007] Thus, activation of NF-κB signaling is involved in the transcriptional induction of pro-inflammatory cytokines, chemokines, and additional inflammatory mediators in various types of immune cells. These inflammatory mediators can either directly contribute to the induction of inflammation or act indirectly by promoting the differentiation of inflammatory T cells. In this way, activation or dysregulation of TLR signaling leads to chronic inflammation, which is central to the pathogenesis of autoimmune states. [Prior art documents] [Patent Documents]

[0008] [Patent Document 1] WO2011 / 020913 [Patent Document 2] WO02 / 02596 [Patent Document 3] WO01 / 76616 [Patent Document 4] WO2004 / 084812 [Patent Document 5] WO9942125 [Non-patent literature]

[0009] [Non-Patent Document 1] Vera et al., Med. Res. Rev. 2002, 22(2), pp. 102-145. [Non-Patent Document 2] "Remington's Pharmaceutical Sciences" by EW Martin, 1995. [Overview of the Initiative] [Problems that the invention aims to solve]

[0010] Therefore, there is a need to develop new therapies to treat autoimmune conditions that are currently incurable in the majority of affected patients. In particular, there is a need to develop therapies that can stop TLR activation or prevent pathological conditions such as autoimmune conditions resulting from TLR activation. This invention addresses these needs. [Means for solving the problem]

[0011] In one aspect, the present invention is for use in treating autoimmune conditions. General formula I

[0012] [ka]

[0013] The subject is the compound, or its pharmaceutically acceptable salts or stereoisomers. (In the formula, X is selected from O and NH; Y is selected from CO and -COCH(CH3)CO-; Each n and p is independently selected from 0 and 1, and q is selected from 0, 1 and 2; Each of R1, R3, R5, R9, R11, and R15 is independently selected from hydrogen, substituted or unsubstituted C1-C6 alkyl groups, substituted or unsubstituted C2-C6 alkenyl groups, and substituted or unsubstituted C2-C6 alkynyl groups; R2 is selected from hydrogen, CORa, COORa, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, and substituted or unsubstituted C2-C6 alkynyl; Each of R4, R8, R10, R12, and R16 is independently selected from hydrogen and substituted or unsubstituted C1-C6 alkyl groups; Each R7 and R13 is independently selected from hydrogen, substituted or unsubstituted C1-C6 alkyl groups, substituted or unsubstituted C2-C6 alkenyl groups, and substituted or unsubstituted C2-C6 alkynyl groups; each R6 and R14 is independently selected from hydrogen and substituted or unsubstituted C1-C6 alkyl groups; or R6 and R7 and / or R13 and R14 together with the corresponding N and C atoms to which they are bonded can form substituted or unsubstituted heterocyclic groups; R17 is selected from hydrogen, CORa, COORa, CONHRb, COSRc, (C=NRb)ORa, (C=NRb)NHRb, (C=NRb)SRc, (C=S)ORa, (C=S)NHRb, (C=S)SRc, SO2Rc, SO3Rc, substituted or unsubstituted C1-C12 alkyl, substituted or unsubstituted C2-C12 alkenyl, substituted or unsubstituted C2-C12 alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heterocyclic groups, provided that if n, p, and q are 0, R17 is not hydrogen; Each Ra, Rb, and Rc is independently selected from hydrogen, substituted or unsubstituted C1-C12 alkyl groups, substituted or unsubstituted C2-C12 alkenyl groups, substituted or unsubstituted C2-C12 alkynyl groups, substituted or unsubstituted aryl groups, and substituted or unsubstituted heterocyclic groups.

[0014] In certain embodiments, the compound of general formula I is a PLD, or a pharmaceutically acceptable salt or stereoisomer thereof.

[0015] In another embodiment, the present invention relates to didemnin B, or a pharmaceutically acceptable salt or stereoisomer thereof.

[0016] In another aspect, the present invention also relates to a pharmaceutical composition for use according to the present invention, comprising a compound as defined herein and a pharmaceutically acceptable carrier.

[0017] In another aspect, the present invention relates to the use of compounds as defined herein in the manufacture of pharmaceuticals for the treatment of autoimmune conditions.

[0018] In another aspect, the present invention relates to a method for treating any mammal, preferably a human, for an autoimmune condition, comprising administering a therapeutically effective dose of a compound as defined herein to an individual in need.

[0019] In embodiments, the autoimmune condition is selected from systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), multiple sclerosis (MS), scleroderma, Sjögren's syndrome, autoimmune myocarditis, type 1 diabetes, and atherosclerosis. In preferred embodiments, the autoimmune condition is RA.

[0020] In a further aspect of the present invention, a kit is provided comprising a compound as defined herein, or a pharmaceutically acceptable salt or stereoisomer thereof, along with instructions for use in treating an autoimmune condition.

[0021] The following embodiments apply to all aspects of the present invention.

[0022] Autoimmune conditions can be caused by the activation of one or more Toll-like receptors (TLRs).

[0023] Autoimmune states may be characterized by increased signaling mediated by at least one or more Toll-like receptors (TLRs).

[0024] An autoimmune state may be characterized by an increase in the level of at least one pro-inflammatory cytokine.

[0025] The autoimmune state can be selected from systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), multiple sclerosis (MS), scleroderma, Sjögren's syndrome, autoimmune myocarditis, type 1 diabetes, and atherosclerosis. In a preferred embodiment, the autoimmune state is RA.

[0026] R3 and R4 may be independently selected from hydrogen and substituted or unsubstituted C1-C6 alkyl. R3 may be isopropyl and R4 may be hydrogen. R3 and R4 may be methyl (this compound is also referred to as a compound of general formula II).

[0027] R 11 may be selected from hydrogen and substituted or unsubstituted C1-C6 alkyl. R 11 may be methyl or isobutyl. R 11 may be methyl and n = 1 (this compound is also referred to as a compound of general formula III).

[0028] R1, R5, R9, and R 15 may be independently selected from hydrogen and substituted or unsubstituted C1-C6 alkyl. R1 may be selected from sec-butyl and isopropyl, R5 may be isobutyl, R9 may be p-methoxybenzyl, and R 15 may be selected from methyl and benzyl.

[0029] R8, R 10 、R 12 、及びR 16 may be independently selected from hydrogen and substituted or unsubstituted C1-C6 alkyl. R8, R 10 、及びR 12 may be methyl and R 16 may be hydrogen.

[0030] R 14R6 may be independently selected from hydrogen and substituted or unsubstituted C1-C6 alkyl groups. R6 may be selected from hydrogen and methyl groups. 14 It may be hydrogen.

[0031] R7 and R 13 R7 may be independently selected from hydrogen and substituted or unsubstituted C1-C6 alkyl groups. 13 This may be selected from hydrogen, methyl, isopropyl, isobutyl, and 3-amino-3-oxopropyl.

[0032] R6 and R7 and / or R 13 and R 14 These can, together with the corresponding N and C atoms to which they are bonded, form substituted or unsubstituted pyrrolidine groups.

[0033] R2 is hydrogen, substituted or unsubstituted C1-C6 alkyl, and COR a You may choose from R a R2 may be a substituted or unsubstituted C1-C6 alkyl group.

[0034] R 17 Hydrogen, COR a COOR a CONHR b (C=S)NHR b , and SO2R c You may choose from each R a , R b , and R c R may be independently selected from substituted or unsubstituted C1-C6 alkyl groups, substituted or unsubstituted C2-C6 alkenyl groups, substituted or unsubstituted C2-C6 alkynyl groups, substituted or unsubstituted aryl groups, and substituted or unsubstituted heterocyclic groups. 17 This can be selected from hydrogen, COO benzyl, CO benzo[b]thiophene-2-yl, SO2(p-methylphenyl), COCOCH3, and COOC(CH3)3.

[0035] X may be NH. X may be O. Y may be CO. Y may be -COCH(CH3)CO-.

[0036] The compound may be PLD, or a pharmaceutically acceptable salt or stereoisomer thereof. The compound may be PLD.

[0037] The compound may be didemnin B, or a pharmaceutically acceptable salt or stereoisomer thereof. The compound may be didemnin B.

[0038] The present invention is also further illustrated by the following non-limiting figures: [Brief explanation of the drawing]

[0039] [Figure 1] This study demonstrates that NF-κB transactivation in response to Toll-like receptor activation is inhibited by PLD. Human monocyte cells (THP-1) were stably transfected with the NF-κB-Luc plasmid. (A) Levels of NF-κB transactivation were measured in the presence and absence of PLD. (B) Compound-induced cytotoxicity was tested by the MTT cell proliferation assay. Cultures were exposed to 100 nM PLD for 6 hours. RQ-10 μg / mL reciquimod. LPS-B5-10 μg / mL lipopolysaccharide (LPS-B5) from Escherichia coli 055:B5. Poly-C-500 μg / mL polyinosine / polycytidic acid. TNF-α was used as a positive control. ***p<0.001;**p<0.01 [Figure 2] This study demonstrates that NF-κB transactivation in response to Toll-like receptor activation leads to increased secretion of inflammatory cytokines: IL-1, IL-6, IL-8, and TNF-α. Cultures were exposed to 100 nM DMSO for 6 hours. Secreted cytokines were analyzed by ELISA 6 hours post-treatment. TNF-α was used as a positive control. ***p<0.001;**p<0.01 [Figure 3]This demonstrates the ex vivo downregulation of cytokines IL-6, IL-10, and TNF-α by PLD. [Figure 4] This shows a decrease in classically activated macrophages in LPS challenge mice. [Figure 5] This image shows the effect of PLD administration on patients with bilateral pneumonia. [Figure 6] This image shows the effect of PLD administration on patients with unilateral pneumonia. [Figure 7] This shows the C-reactive protein test results for patients treated with PLD. [Figure 8] This shows the inflammatory profiles in the BALF of mice infected with influenza virus, with or without PLD treatment (PR8). [Figure 9] The effects of 1 nM, 10 nM, and 50 nM pristicepsin (APL) pretreatment on the secretion of pro-inflammatory cytokines IL6 (a), IL8 (b), IL1β (c), and TNF-α (d) are shown. (e) shows the effects of 1 nM, 10 nM, and 50 nM PLD treatment on cell viability (as a percentage of control). THP-1 cells were treated with 1 nM, 10 nM, or 50 nM APL or DMSO (0.2%) at 0:00 and stimulated with 2.5 or 5 μg / mL reciquimod 8 hours later. Cytokine and cell viability were measured at 24 hours. [Figure 10] This shows the effect of plitidepsin treatment on the production of LPS-B5-mediated pro-inflammatory cytokines, IL-6(c), IL-10(d), and TNF-α(e), in CD45+ cells isolated from bronchoalveolar lavage fluid (BALF). (a) shows the percentage of CD45+ viable cells in control, LPS-B5, and LPS-B5 and PLD-treated cells. (b) shows the cell viability in LPS-B5 and LPS-B5 and PLD-treated cells as a percentage of the control. [Figure 11] This study demonstrates the effect of pristicepsin treatment on the production of the pro-inflammatory cytokine TNF-α, which is mediated by reximod. [Figure 12]This shows the effect of pritolidepsin on alveolar macrophage recruitment in LPS-treated mice. The concentration-time curves (mean ± SD) of pritolidepsin in the plasma and lungs of mice (a), rats (b), and hamsters (c) after single intravenous administration of 1.0, 0.2, and 0.2 mg / kg, respectively. [Modes for carrying out the invention]

[0040] The following embodiments apply to all aspects of the present invention.

[0041] The present invention will now be described further. The following sections define different aspects of the present invention in more detail. Each of the aspects defined in this way may be combined with any other one or more aspects or embodiments unless explicitly indicated otherwise. In particular, any feature shown to be preferred or advantageous may be combined with any other one or more features shown to be preferred or advantageous.

[0042] This application uses many common terms and phrases, which should be interpreted as follows:

[0043] As used herein, the term “treat” means, unless otherwise indicated, to reverse, alleviate, or inhibit the progression of a disease or condition, or one or more symptoms of such disorder or condition. As used herein, the term “treat” may also include prophylactic treatments, which are treatments designed to prevent the development of an autoimmune condition or to minimize the likelihood of such a condition developing.

[0044] "Patient" includes humans, non-human mammals (e.g., dogs, cats, rabbits, cattle, horses, sheep, goats, pigs, deer, etc.), and non-mammals (e.g., birds, etc.).

[0045] Plitchidepsin (PLD) is a cyclic depsipeptide originally isolated from the marine tunicate Aplidium albicans. PLD is also known as Aplidin. PLD analogs are analogs as defined herein. In preferred embodiments, the present invention relates to the use of PLD.

[0046] PLD is (i) Inhibiting the transactivation of NF-κB induced by the activation of Toll-like receptors; (ii) Inhibiting the secretion of pro-inflammatory cytokines such as IL-1, IL-6, IL-8, and TNF-α both in vivo and ex vivo; and (iii) It was found that it inhibits macrophage activation.

[0047] These properties suggest that PLD is particularly effective in treating autoimmune conditions. References to PLD herein can be considered applicable to the compounds of the present invention (and other PLD analogues). As shown in the examples, PLD has been found to inhibit the secretion of pro-inflammatory cytokines, thereby reducing the level of inflammation, which is a major factor in the pathogenesis of autoimmune conditions. In particular, PLD has been found to inhibit the transactivation of NF-κB via Toll-like receptors (TLRs) and the subsequent secretion of pro-inflammatory cytokines. For example, PLD has been shown to inhibit the transactivation of NF-κB via the activation of TLR3, TLR4, TL7, and TLR8, all of which have been shown to be activated by endogenous ligands.

[0048] As described herein, in autoimmune states, Toll-like receptors are activated in response to numerous endogenous ligands released from damaged tissue. Binding (i.e., stimulation) of TLR ligands to Toll-like receptors (TLRs) triggers a downstream signaling cascade, ultimately leading to the activation of the transcription factor nuclear factor-kappa B (NF-κB), which regulates the induction of pro-inflammatory cytokines and chemokines. PLDs have been found to significantly block this cascade, resulting in reduced release of pro-inflammatory cytokines. Consequently, in one example, PLDs can be used to prevent autoimmune states following Toll-like receptor activation.

[0049] Furthermore, PLD was found to significantly reduce the levels of macrophage activation and / or macrophage recruitment. Activated macrophages are important mediators of inflammation, and inhibiting macrophage activation is central to the treatment of inflammation and, consequently, to the pathology of autoimmune states.

[0050] Therefore, the compounds defined herein (including PLD and didemnin B), particularly PLD, can be used to treat autoimmune conditions following activation of Toll-like receptors.

[0051] This invention may be useful in relation to the following autoimmune conditions:

[0052] Rheumatoid arthritis (RA) Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune condition characterized by progressive and irreversible destruction of the joints. RA is the most common autoimmune condition, affecting approximately 1% of the population. Currently, there is no cure, and up to 40% of the population does not respond to existing treatments. RA is characterized by persistent inflammation facilitated by the proliferation of fibroblasts in the synovial tissue, as well as the transport of T cells, B cells, neutrophils, and monocytes to the joints. Various endogenous TLR ligands, including fibrinogen, HSP60, HSP70, EDA fibronectin, HMGB1, hyaluronic acid, and HSP22, have been demonstrated to be present in the inflamed joints of RA patients and have been shown to lead to the activation of TLRs: TLR2, TLR4, TLR5, and TLR7. All of these TLR activations are responsible for the persistent expression of pro-inflammatory cytokines and activated macrophages observed in RA joints, suggesting that various TLR family members are involved in different stages of the disease. Consistent with TLR activation in the pathogenesis of RA, activated NF-κB has been detected in human synovial tissue in both the early and late stages of the disease and is thought to be involved in both the initiation and persistence of chronic inflammation. In particular, many of these ligands are thought to be induced by cell damage, degradation of the extracellular matrix, and activation of macrophage activity, all of which are prominent features of RA. Thus, the RA microenvironment may promote disease persistence and exacerbation by further release of these ligands. Interestingly, fragments of double-stranded viral RNA released by necrotic cells are effective TLR ligands and have been found in the synovial fluid of RA patients, which supports the hypothesis that microbial infections induce or maintain the TLR response in RA, leading to disease development and exacerbation.

[0053] Systemic lupus erythematosus (SLE) Systemic lupus erythematosus (SLE), or lupus, is a severe relapsing-remitting autoimmune condition that causes many symptoms in affected patients, including joint pain, skin rash, and fatigue. In some cases, the disease affects the kidneys and other organs. Patient serum has been found to contain ligands for TLRs, particularly TLR7, TLR8, and TLR9. Peripheral dendritic cells are recognized as a major driving force of RA pathology, and these cells express both TLR7 and TLR9, meaning that these cells can be activated by such ligands and trigger disease-related signaling. In particular, in SLE patients, autoreactive cells produce large amounts of autoantibodies against autonuclear antigens and form immune complexes with autonucleotides in the serum. These complexes act as TLR ligands, particularly for TLR7 and TLR9, activating the TLR pathway and causing chronic inflammation.

[0054] Similar to rheumatoid arthritis (RA), single-stranded viral RNA has been detected in lupus patients, as well as in patients with other autoimmune diseases such as scleroderma and Sjögren's syndrome, and is an effective ligand for TLR7 and TLR8. Bacterial or HSV DNA has also been found in lupus patients and is an effective ligand for TLR9. Numerous experimental systems have demonstrated that microbial TLR ligands can induce disease in experimental models of arthritis, multiple sclerosis, experimental allergic encephalomyelitis (EAE), autoimmune myocarditis, type 1 diabetes, and atherosclerosis. In these cases, too, this broadly supports the involvement of microorganisms in TLR activation that promotes autoimmune diseases.

[0055] Multiple sclerosis (MS) Multiple sclerosis (MS) is an autoimmune disease in which central nervous system (CNS) lesions result from perivascular immune cell infiltration associated with damage to myelin, oligodendrocytes, and neurons. Clinically, symptoms include numbness, weakness, loss of muscle coordination, vision, speech, and bladder control problems. The pathology of MS consists of two main stages: firstly, initial immune activation, in which an autoimmune response is triggered; and secondly, the recruitment of immune cells to the CNS, where tissue destruction and demyelination occur. Studies have shown that TLRs play a crucial role in the regulation of MS, as well as in experimental autoimmune encephalomyelitis (EAE), an animal model of MS. Interestingly, as with recruited immune cells, CNS resident microglia also express various TLRs, and the expression of these TLRs has been found to increase in response to inflammatory mediators. These cells have been shown to be essential for the establishment and exacerbation of inflammatory plaques in the CNS during MS, and may transmit the disease through the gradual activation of these receptors.

[0056] Scleroderma Scleroderma, or systemic sclerosis, is a chronic connective tissue disease commonly classified as an autoimmune rheumatic disease. Scleroderma is caused by the immune system attacking the connective tissue beneath the skin and around internal organs and blood vessels. This leads to scarring and thickening of the tissue in these areas. Some types of scleroderma are relatively mild and may eventually improve spontaneously, while others can result in severe, life-threatening problems for which there is no treatment. TLRs have been identified as crucial in the pathogenesis of scleroderma, with endogenous TLR ligands—products from damaged cells—inducing TLR signaling that promotes inflammatory and fibrotic activity. In particular, TLR signaling is thought to promote the release of TIMP, leading to fibrosis in scleroderma.

[0057] Sjögren's syndrome Sjögren's syndrome is an autoimmune disease that often coexists with rheumatoid arthritis (RA) and / or lupus, primarily affecting the salivary and lacrimal glands. These glands help the body produce moisture for the eyes and mouth in the form of saliva and tears. Therefore, in people with Sjögren's syndrome, the body is unable to produce sufficient moisture. TLRs are thought to be underlying this disorder, and numerous putative endogenous TLR ligands, such as biglycan, decorin, versican, and fibronectin, have been found in patients with Sjögren's syndrome or in mouse models of the disease. Furthermore, TLR expression has been upregulated, and peripheral blood cells have been found to be hypersensitive to ligation in patients with Sjögren's syndrome.

[0058] Autoimmune myocarditis Autoimmune myocarditis is an autoimmune disease that affects the heart. This condition is characterized by inflammation of the myocardium and does not affect other organs. Here again, TLR signaling appears to be a crucial underlying mechanism of myocarditis pathology. For example, knockout mice lacking MyD88, a canonical adapter molecule that promotes downstream TLR signaling, are protected from disease in an induced model of myocarditis. It is hypothesized that human cardiac myosin may act as an endogenous TLR ligand to trigger downstream pro-inflammatory responses via TLR2 and TLR8.

[0059] type 1 diabetes Type 1 diabetes, or insulin-dependent diabetes mellitus, is an autoimmune disease that causes the destruction of insulin-producing beta cells in the pancreas. As a result, patients are unable to produce very little or any insulin, the hormone necessary to effectively control blood glucose levels. Viral infections may trigger this cell destruction via TLR9-induced immune activation, and TLR upregulation has been shown to increase disease penetration. This highlights the crucial role of TLR signaling in the pathogenesis of type 1 diabetes.

[0060] Atherosclerosis Atherosclerosis is a condition in which fats, cholesterol, and other substances accumulate in and on the arterial walls (plaques), potentially restricting blood flow. These plaques can rupture, inducing thrombosis and leading to associated conditions such as stroke and myocardial infarction, and may particularly promote cardiovascular disease (CVD). Currently, atherosclerosis is considered an inflammatory autoimmune state, and TLRs, in particular, are thought to be important orchestrators of the disease process. There is abundant evidence supporting this from disease models, such as the fact that knockouts of MyD88, TLR2, and TLR4 can alleviate or prevent atherosclerosis in mouse models. TLR2 and TLR4 are thought to be activated during disease and may be activated by various lipopeptides (Falck-Hansen et al., 2013).

[0061] Considering the above, it is clear that TLR signaling is a fundamental driving force behind autoimmunity, whether in response to endogenous TLR ligands, microbial TLR ligands, or a combination of both, and that the disease microenvironment can often facilitate a positive feedback loop to maintain TLR signaling.

[0062] Therefore, the compounds of the present invention (including PLD) can be used to treat autoimmune conditions.

[0063] In these compounds, the groups can be selected according to the following guidelines:

[0064] Alkyl groups may be branched or unbranched and preferably have 1 to about 12 carbon atoms. One more preferred class of alkyl groups has 1 to about 6 carbon atoms. Even more preferred are alkyl groups having 1, 2, 3, or 4 carbon atoms. Methyl, ethyl, n-propyl, isopropyl, and butyl, including n-butyl, tert-butyl, sec-butyl, and isobutyl, are particularly preferred alkyl groups in the compounds of the present invention. As used herein, the term alkyl refers to both cyclic and acyclic groups unless otherwise specified, but a cyclic group includes at least three carbon ring members.

[0065] Preferred alkenyl and alkynyl groups in the compounds of the present invention may be branched or unbranched, and have one or more unsaturated bonds and 2 to about 12 carbon atoms. One more preferred class of alkenyl and alkynyl groups has 2 to about 6 carbon atoms. Even more preferred are alkenyl and alkynyl groups having 1, 2, 3, or 4 carbon atoms. As used herein, the terms alkenyl and alkynyl groups refer to both cyclic and acyclic groups unless otherwise specified, but a cyclic group contains at least three carbon ring members.

[0066] Suitable aryl groups in the compounds of the present invention include monocyclic and polycyclic compounds, and polycyclic compounds containing separate and / or fused aryl groups. Typical aryl groups contain 1 to 3 separate or fused rings and 6 to about 18 carbocyclic atoms. Preferably, the aryl group contains 6 to about 10 carbocyclic atoms. Particularly preferred aryl groups include substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, substituted or unsubstituted biphenyl, substituted or unsubstituted phenanthryl, and substituted or unsubstituted anthryl.

[0067] Suitable heterocyclic groups include heteroaromatic and heteroalicyclic groups containing 1 to 3 separate or fused rings and 5 to about 18 ring atoms. Preferably, the heteroaromatic and heteroalicyclic groups contain 5 to about 10 ring atoms, most preferably 5, 6, or 7 ring atoms. Suitable heteroaromatic groups in the compounds of the present invention include those containing one, two, or three heteroatoms selected from N, O, or S atoms, such as coumarinyl containing 8-coumarinyl, quinolyl containing 8-quinolyl, isoquinolyl, pyridyl, pyrazinyl, pyrazol-3-yl, pyrazol-4-yl and pyrazol-5-yl, pyrimidinyl, furanyl containing furan-2-yl, furan-3-yl, furan-4-yl and furan-5-yl, pyrrolyl, thienyl, thiazolyl containing thiazolyl-2-yl, thiazolyl-4-yl and thiazolyl-5-yl, isothiazolyl, thiadiazolyl containing thiadiazolyl-4-yl and thiadiazolyl-5-yl, triazolyl, tetrazolyl, isoxazole-3-yl, isoxazole This includes isoxazolyl, oxazolyl, imidazolyl, indolyl, isoindolyl, indazolyl, indolidinyl, phthalazinyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, flazanil, pyridadinyl, triazinyl, cinnolinyl, benzimidazolyl, benzofuranil, benzoflazanil, benzothiophenyl, benzothiazolyl, benzoxazolyl, imidazo[1,2-a]pyridinyl, quinazolinil, quinoxalinil, naphthilidinyl, and phlopyridyl, which include zole-4-yl and isoxazol-5-yl. Suitable heteroalicyclic groups in the compounds of the present invention contain one, two, or three heteroatoms selected from N, O, or S atoms, and include, for example, piperidinyl, morpholinyl, thiomorpholinyl, thioxanil, piperazinyl, azetidinyl, oxetanil, thietanil, homopiperidyl, and oxepani. Examples include thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridyl, 2-pyrrolinyl, 3-pyrrolinyl, dihydropyrrolyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, 3H-indolyl, and quinolidinyl.

[0068] In the above groups, one or more hydrogen atoms are one or more suitable groups, e.g., OR', =O, SR', SOR', SO2R', NO2, NHR', NR'R', =N-R', NHCOR', N(COR')2, NHSO2R', NR'C(=NR')NR'R', CN, halogen, COR', COOR', OCOR', OCONHR', OCONR'R', CONHR', CONR'R', substituted or unsubstituted C1-C 12 Alkyl, substituted, or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 The R' group may be substituted with alkynyl, substituted or unsubstituted aryl, and substituted or unsubstituted heterocyclic groups, each of which is hydrogen, OH, NO2, NH2, SH, CN, halogen, COH, COalkyl, CO2H, substituted or unsubstituted C1-C 12 Alkyl, substituted, or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 The substituents are independently selected from alkynyls, substituted or unsubstituted aryls, and substituted or unsubstituted heterocyclic groups. If the group itself is substituted, the substituents can be selected from the aforementioned list. If the substituent ends in a double bond (=O and =N-R', etc.), it replaces two hydrogen atoms on the same carbon atom.

[0069] Suitable halogen substituents in the compounds of the present invention include F, Cl, Br, and I.

[0070] The term "pharmaceutically acceptable salt" refers to any salt that can provide (directly or indirectly) the compounds described herein when administered to a patient. It will be understood that pharmaceutically unacceptable salts are also within the scope of the invention, as they may be useful in the preparation of pharmaceutically acceptable salts. Preparation of salts can be carried out by methods known in the art. For example, pharmaceutically acceptable salts of the compounds provided herein can be synthesized from parent compounds containing basic or acidic moieties by conventional chemical methods. Generally, such salts are prepared, for example, by reacting the free acidic or free base form of these compounds with a stoichiometric amount of a suitable base or acid in water, an organic solvent, or a mixture thereof. Generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Examples of acid addition salts include mineral acid addition salts, such as hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, and phosphate, and organic acid addition salts, such as acetate, trifluoroacetate, maleate, fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate, methanesulfonate, and p-toluenesulfonate. Examples of alkali addition salts include inorganic salts, such as sodium salt, potassium salt, and calcium salt, and organic alkali salts, such as ammonium salt, ethylenediamine, ethanolamine, N,N-dialkyleneethanolamine, triethanolamine, and basic amino acid salts.

[0071] The compounds of the present invention may exist in either a crystalline form as a free compound or a solvate (e.g., a hydrate, an alcoholate, and especially a methanelate), and both forms are intended to be within the scope of the present invention. Methods of solvation are generally known in the art. The compounds of the present invention may exhibit different polymorphic forms, and the present invention is intended to encompass all such forms.

[0072] Any compound referred to herein is intended to represent such a particular compound as well as a particular variant or form. In particular, the compounds referred to herein may have a chiral center and therefore may exist in different enantiomer or diastereomer forms. Thus, any compound referred to herein is intended to represent either a racemate, one or more enantiomer forms, one or more diastereomer forms, or mixtures thereof. Similarly, stereoisomerism or geometric isomerism with respect to double bonds is also possible, and therefore, in some cases, the molecule may exist as an (E)-isomer or (Z)-isomer (trans and cis isomer). If a molecule contains several double bonds, each double bond has its own stereoisomerism, which may be the same as or different from the stereoisomerism of the other double bonds in the molecule. Furthermore, the compounds referred to herein may exist as atropisomers. All stereoisomers, including enantiomers, diastereoisomers, geometricisomers and atropisomers of the compounds referred to herein, as well as mixtures thereof, are considered to be within the scope of the present invention.

[0073] In compounds of general formulas I and II, particularly preferred R1, R5, R9, R 11 and R 15 R1, R5, R9, R 11 and R 15 The group is independently selected from hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, and substituted or unsubstituted butyl, for example, substituted or unsubstituted n-butyl, substituted or unsubstituted tert-butyl, substituted or unsubstituted isobutyl, and substituted or unsubstituted sec-butyl. Preferred substituents of the group are OR', =O, SR', SOR', SO2R', NO2, NHR', NR'R', =N-R', NHCOR', N(COR')2, NHSO2R', NR'C(=NR')NR'R', CN, halogen, COR', COOR', OCOR', OCONHR', OCONR'R', CONHR', CONR'R', substituted or unsubstituted C1-C 12Alkyl, substituted, or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkynnyl, substituted or unsubstituted aryl, and substituted or unsubstituted heterocyclic groups, where each of the R' groups is hydrogen, OH, NO2, NH2, SH, CN, halogen, COH, COalkyl, CO2H, substituted or unsubstituted C1-C 12 Alkyl, substituted, or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 The substituents are independently selected from alkynyls, substituted or unsubstituted aryls, and substituted or unsubstituted heterocyclic groups. If such groups themselves are substituted, the substituents can be selected from the list above. Hydrogen, methyl, n-propyl, isopropyl, isobutyl, sec-butyl, 4-aminobutyl, 3-amino-3-oxopropyl, benzyl, p-methoxybenzyl, p-hydroxybenzyl, and cyclohexylmethyl are most preferred R1, R5, R9, R 11 and R 15 The group is. Specifically, the particularly preferred R1 is selected from sec-butyl and isopropyl, with sec-butyl being the most preferred. The particularly preferred R5 is selected from isobutyl and 4-aminobutyl, with isobutyl being the most preferred. 11 R9 is methyl and isobutyl. Particularly preferred R9 is selected from p-methoxybenzyl, p-hydroxybenzyl, and cyclohexylmethyl, with p-methoxybenzyl being the most preferred. Particularly preferred R 15 The compound is selected from methyl, n-propyl, and benzyl, with methyl and benzyl being the most preferred.

[0074] In compounds of general formula III, particularly preferred R1, R5, R9, and R 15 R1, R5, R9, and R are independently selected from hydrogen and substituted or unsubstituted C1-C6 alkyl groups. 15The group is independently selected from hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, and substituted or unsubstituted butyl, for example, substituted or unsubstituted n-butyl, substituted or unsubstituted tert-butyl, substituted or unsubstituted isobutyl, and substituted or unsubstituted sec-butyl. Preferred substituents of the group are OR', =O, SR', SOR', SO2R', NO2, NHR', NR'R', =N-R', NHCOR', N(COR')2, NHSO2R', NR'C(=NR')NR'R', CN, halogen, COR', COOR', OCOR', OCONHR', OCONR'R', CONHR', CONR'R', substituted or unsubstituted C1-C 12 Alkyl, substituted, or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkynnyl, substituted or unsubstituted aryl, and substituted or unsubstituted heterocyclic groups, where each of the R' groups is hydrogen, OH, NO2, NH2, SH, CN, halogen, COH, COalkyl, CO2H, substituted or unsubstituted C1-C 12 Alkyl, substituted, or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 The substituents are independently selected from alkynyls, substituted or unsubstituted aryls, and substituted or unsubstituted heterocyclic groups. If such a group itself is substituted, the substituents can be selected from the list above. Hydrogen, methyl, n-propyl, isopropyl, isobutyl, sec-butyl, 4-aminobutyl, 3-amino-3-oxopropyl, benzyl, p-methoxybenzyl, p-hydroxybenzyl, and cyclohexylmethyl are the most preferred R1, R5, R9, and R 15 The group is R1. Specifically, the particularly preferred R1 is selected from sec-butyl and isopropyl, with sec-butyl being the most preferred. The particularly preferred R5 is selected from isobutyl and 4-aminobutyl, with isobutyl being the most preferred. The particularly preferred R9 is selected from p-methoxybenzyl, p-hydroxybenzyl, and cyclohexylmethyl, with p-methoxybenzyl being the most preferred. 15The compound is selected from methyl, n-propyl, and benzyl, with methyl and benzyl being the most preferred.

[0075] Among the compounds of general formulas I, II, and III, R8, R8 is particularly preferred. 10 , R 12 , and R 16 R8, R8 is more preferred. 10 , R 12 , and R 16 These are independently selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, and butyl, including n-butyl, tert-butyl, isobutyl, and sec-butyl, and more preferably they are independently selected from hydrogen and methyl. Specifically, R8, R8 are particularly preferred. 10 , and R 12 R is methyl, and R is particularly preferred. 16 It is hydrogen.

[0076] In compounds of general formulas I and III, particularly preferred R3 and R4 are independently selected from hydrogen and substituted or unsubstituted C1-C6 alkyl groups. More preferred R3 and R4 are independently selected from hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, and substituted or unsubstituted butyl, for example, substituted or unsubstituted n-butyl, substituted or unsubstituted tert-butyl, substituted or unsubstituted isobutyl, and substituted or unsubstituted sec-butyl. Preferred substituents of the group are OR', =O, SR', SOR', SO2R', NO2, NHR', NR'R', =N-R', NHCOR', N(COR')2, NHSO2R', NR'C(=NR')NR'R', CN, halogen, COR', COOR', OCOR', OCONHR', OCONR'R', CONHR', CONR'R', substituted or unsubstituted C1-C 12 Alkyl, substituted, or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12Alkynnyl, substituted or unsubstituted aryl, and substituted or unsubstituted heterocyclic groups, where each of the R' groups is hydrogen, OH, NO2, NH2, SH, CN, halogen, COH, COalkyl, CO2H, substituted or unsubstituted C1-C 12 Alkyl, substituted, or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 The substituents are independently selected from alkynyls, substituted or unsubstituted aryls, and substituted or unsubstituted heterocyclic groups. If the group itself is substituted, the substituents can be selected from the list above. Hydrogen, methyl, isopropyl, and sec-butyl are the most preferred R3 and R4 groups. Specifically, the particularly preferred R3 is selected from methyl and isopropyl, and the particularly preferred R4 is methyl or hydrogen.

[0077] In one embodiment of the compounds of general formulas I, II, and III, particularly preferred R6 and R7 are independently selected from hydrogen and substituted or unsubstituted C1-C6 alkyl groups. More preferred R7 is selected from hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, and substituted or unsubstituted butyl, for example, substituted or unsubstituted n-butyl, substituted or unsubstituted tert-butyl, substituted or unsubstituted isobutyl, and substituted or unsubstituted sec-butyl. Preferred substituents of the group are OR', =O, SR', SOR', SO2R', NO2, NHR', NR'R', =N-R', NHCOR', N(COR')2, NHSO2R', NR'C(=NR')NR'R', CN, halogen, COR', COOR', OCOR', OCONHR', OCONR'R', CONHR', CONR'R', substituted or unsubstituted C1-C 12 Alkyl, substituted, or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkynnyl, substituted or unsubstituted aryl, and substituted or unsubstituted heterocyclic groups, where each of the R' groups is hydrogen, OH, NO2, NH2, SH, CN, halogen, COH, COalkyl, CO2H, substituted or unsubstituted C1-C 12 Alkyl, substituted, or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C12 R6 is independently selected from alkynyls, substituted or unsubstituted aryls, and substituted or unsubstituted heterocyclic groups. If such a group itself is substituted, the substituent can be selected from the list above. More preferred R6 is selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, and butyl, including n-butyl, tert-butyl, isobutyl, and sec-butyl. Most preferred R6 is selected from hydrogen and methyl, and most preferred R7 is methyl.

[0078] In other embodiments of the compounds of general formulas I, II, and III, it is particularly preferable that R6 and R7, together with the corresponding N and C atoms to which they are bonded, form a substituted or unsubstituted heterocyclic group. In this regard, preferred heterocyclic groups are heteroalicyclic groups containing one, two, or three heteroatoms selected from N, O, or S atoms, most preferably one N atom, and having 5 to about 10 ring atoms, most preferably 5, 6, or 7 ring atoms. Pyrrolidine groups are most preferred.

[0079] In one embodiment of compounds of general formulas I, II, and III, a particularly preferred R 13 and R 14 R is independently selected from hydrogen and substituted or unsubstituted C1-C6 alkyl groups. A more preferred R 13 The group is selected from hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted isopropyl, and substituted or unsubstituted butyl, for example, substituted or unsubstituted n-butyl, substituted or unsubstituted tert-butyl, substituted or unsubstituted isobutyl, and substituted or unsubstituted sec-butyl. Preferred substituents of the group are OR', =O, SR', SOR', SO2R', NO2, NHR', NR'R', =N-R', NHCOR', N(COR')2, NHSO2R', NR'C(=NR')NR'R', CN, halogen, COR', COOR', OCOR', OCONHR', OCONR'R', CONHR', CONR'R', substituted or unsubstituted C1-C 12 Alkyl, substituted, or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12An alkynyl, a substituted or unsubstituted aryl, and a substituted or unsubstituted heterocyclic group, each R' group being hydrogen, OH, NO2, NH2, SH, CN, halogen, COH, CO alkyl, CO2H, a substituted or unsubstituted C1-C 12 alkyl, a substituted or unsubstituted C2-C 12 alkenyl, a substituted or unsubstituted C2-C 12 They are independently selected from an alkynyl, a substituted or unsubstituted aryl, and a substituted or unsubstituted heterocyclic group. When such a group itself is substituted, the substituents can be selected from the aforementioned list. More preferred R 14 is selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, and butyl including n-butyl, tert-butyl, isobutyl, and sec-butyl. Most preferred R 13 is selected from hydrogen, methyl, isopropyl, isobutyl, and 3-amino-3-oxopropyl, and most preferred R 14 is hydrogen.

[0080] In another embodiment of the compounds of general formulas I, II, and III, R 13 and R 14 It is particularly preferred that they together with the corresponding N atom and C atom to which they are attached form a substituted or unsubstituted heterocyclic group. In this regard, preferred heterocyclic groups contain 1, 2 or 3 heteroatoms selected from N, O or S atoms, most preferably 1 N atom, and have 5 to about 10 ring atoms, most preferably 5, 6 or 7 ring atoms, and are heterocycloaliphatic groups. The pyrrolidine group is most preferred.

[0081] In the compounds of general formulas I, II, and III, particularly preferred R2 is selected from hydrogen, a substituted or unsubstituted C1-C6 alkyl, and COR a where R a is a substituted or unsubstituted C1-C six alkyl, and even more preferred R a is methyl, ethyl, n-propyl, isopropyl, and n-butyl, tert-butyl, sec-butyl and butyl including isobutyl. More preferably, R2 is hydrogen.

[0082] Among the compounds of general formulas I, II, and III, R is particularly preferred. 17 Hydrogen, COR a COOR a CONHR b (C=S)NHR b , and SO2R c Selected from, each R a , R b , and R c The group is preferably independently selected from substituted or unsubstituted C1-C6 alkyl groups, substituted or unsubstituted C2-C6 alkenyl groups, substituted or unsubstituted C2-C6 alkynyl groups, substituted or unsubstituted aryl groups, and substituted or unsubstituted heterocyclic groups. Preferred substituents of the group are OR', =O, SR', SOR', SO2R', NO2, NHR', NR'R', =N-R', NHCOR', N(COR')2, NHSO2R', NR'C(=NR')NR'R', CN, halogen, COR', COOR', OCOR', OCONHR', OCONR'R', CONHR', CONR'R', substituted or unsubstituted C1-C 12 Alkyl, substituted, or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkynnyl, substituted or unsubstituted aryl, and substituted or unsubstituted heterocyclic groups, where each of the R' groups is hydrogen, OH, NO2, NH2, SH, CN, halogen, COH, COalkyl, CO2H, substituted or unsubstituted C1-C 12 Alkyl, substituted, or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 The substituents are independently selected from alkynyls, substituted or unsubstituted aryls, and substituted or unsubstituted heterocyclic groups. If the group itself is substituted, the substituents can be selected from the list above: hydrogen, COR a COOR a , and SO2R c Most preferred R 17 The group is hydrogen, COO benzyl, CO benzo[b]thiophene-2-yl, SO2(p-methylphenyl), COCOCH3, and COOC(CH3)3 are more preferably used.

[0083] In other embodiments of the compounds of general formulas I, II, and III, Y is particularly preferably CO. In another embodiment, Y is particularly preferably -COCH(CH3)CO-.

[0084] In other embodiments of the compounds of general formulas I, II, and III, X is particularly preferably O. In other embodiments, X is particularly preferably NH.

[0085] In other embodiments of the compounds of general formulas I and II, it is particularly preferable that n, p, and q are 0. In another embodiment, it is particularly preferable that n is 1 and p and q are 0. In another embodiment, it is particularly preferable that n and p are 1 and q is 0. In another embodiment, it is particularly preferable that n, p, and q are 1. In another embodiment, it is particularly preferable that n and p are 1 and q is 2.

[0086] In another embodiment of the compound of general formula III, it is particularly preferable that p and q are 0. In another embodiment, it is particularly preferable that p is 1 and q is 0. In another embodiment, it is particularly preferable that p and q are 1. In another embodiment, it is particularly preferable that p is 1 and q is 2.

[0087] In additional preferred embodiments, the preferences described above are combined for different substituents. The present invention also relates to such combinations of preferred substitutions of formulas I, II, and III described above.

[0088] In this specification and definition, the compounds of the present invention include several groups R a , R b , and R c If they exist, it should be understood that they may be independently distinct within a given definition unless explicitly stated otherwise, i.e., R a These do not necessarily represent the same group in the given compound of the present invention.

[0089] In compounds of general formulas I, II, and III, when q takes a value of 2, the compound contains two groups R 15 and two groups R 16 Therefore, each R in a given compound exists. 15 and each R 16 It will be revealed that the base can be independently selected from the different possibilities described above for such a base.

[0090] The particularly preferred stereochemistry of compounds of general formula I is:

[0091] [ka]

[0092] And, In the formula, X, Y, n, p, q, and R1~R 17 As defined above, if Y is -COCH(CH3)CO-, the stereochemistry is as follows:

[0093] [ka]

[0094] It holds.

[0095] The particularly preferred stereochemistry of compounds of general formula II is:

[0096] [ka]

[0097] And, In the formula, X, Y, n, p, q, R1, R2, and R5~R 17 As defined above, if Y is -COCH(CH3)CO-, the stereochemistry is as follows:

[0098] [ka]

[0099] It holds.

[0100] The particularly preferred stereochemistry of compounds of general formula III is:

[0101] [ka]

[0102] And, In the formula, X, Y, p, q, R1~R 10 , and R 12 ~R 17 As defined above, if Y is -COCH(CH3)CO-, the stereochemistry is as follows:

[0103] [ka]

[0104] It holds.

[0105] Particularly preferred compounds of the present invention are the following:

[0106] [ka]

[0107] [ka]

[0108] [ka]

[0109] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0110] Compounds of general formulas I, II, and III can be prepared by any of the synthetic processes disclosed in Vera et al., Med. Res. Rev. 2002, 22(2), pp. 102-145, WO2011 / 020913 (see particularly Examples 1-5), WO02 / 02596, WO01 / 76616, and WO2004 / 084812, which are incorporated herein by reference.

[0111] Preferred compounds are PLD or its pharmaceutically acceptable salts or stereoisomers. The most preferred is PLD.

[0112] The chemical name of pritchidepsin is (-)-(3S,6R,7S,10R,11S,15S,17S,20S,25aS)-11-hydroxy-3-(4-methoxybenzyl)-2,6,17-trimethyl-15-(1-methylethyl)-7-[[(2R)-4-methyl-2-[methyl[[(2S)-1-(2-oxopropanoyl)pyrrolidine-2-yl]carbonyl]amino] Pentanoyl]amino]-10-[(1S)-1-methylpropyl]-20-(2-methylpropyl)tetradecahydro-15H-pyrrolo[2,1-f]-[1,15,4,7,10,20]dioxatetrazacyclotricosine)-1,4,8,13,16,18,21(17H)-heptone, with molecular formula C 57 H 87 N7O 15 It corresponds to [this]. Its relative molecular weight is 1110.34 g / mol, and its structure is as follows:

[0113] [ka]

[0114] References to general formulas I, II, and III include references to PLD and didemnin B. In preferred embodiments, the compound is PLD or didemnin B. PLD is the most preferred.

[0115] The present invention provides for the use of compounds as defined herein and their pharmaceutically acceptable salts or stereoisomers in the treatment of autoimmune conditions.

[0116] In one aspect of the present invention, a compound of the present invention is provided for use in the treatment of an autoimmune condition. In another aspect of the present invention, the use of the compound of the present invention in the manufacture of a pharmaceutical for the treatment of an autoimmune condition is provided. In yet another aspect of the present invention, a method for the treatment of an autoimmune condition is provided, comprising administering a therapeutically effective amount of the compound of the present invention to an individual in need thereof.

[0117] In one embodiment, the autoimmune state is caused by the activation of one or more Toll-like receptors (TLRs) and / or characterized by increased signaling via at least one TLR. The increased signaling via TLRs may be caused by increased expression in at least one TLR. In further embodiments, the autoimmune state is caused by or is caused by TLR-induced cytokine expression. In one embodiment, the TLR is TLR-3, TLR4, TLR7, or TLR8. Methods for measuring the activation of TLR signaling in response to known or possible TLR agonists are well known to those skilled in the art, but in one example, the level of NF-κB transactivation can be used as an indicator of TLR activation. As described herein, NF-κB transactivation can be measured using luciferase-tagged NF-κB transactivation as described in the examples. In another example, TLR activation can be determined by measuring one of IRAK1 (IL receptor-associated kinase), IRAK4 phosphorylation, and TAK1 activation (transforming growth factor β-activated kinase-1). Other indicators of TLR activation are known in the art (see, for example, Kawai and Akira, 2007, describing the TLR pathway).

[0118] In another embodiment, the autoimmune state is characterized by an increased level of at least one pro-inflammatory cytokine, preferably at least one of IL-1, IL-6, IL-8, IL-10, IL-12, and CCL-2, more preferably at least one of IL-1, IL-6, and IL-8.

[0119] In further embodiments, the autoimmune condition is selected from rheumatoid arthritis (RA), multiple sclerosis (MS), systemic lupus erythematosus (SLE), scleroderma, Sjögren's syndrome, autoimmune myocarditis, type 1 diabetes, or atherosclerosis. In preferred embodiments, the autoimmune condition is RA.

[0120] The compounds of the present invention can be used in pharmaceutical compositions having biological / pharmacological activity for treating the above conditions. These pharmaceutical compositions contain the compounds of the present invention together with a pharmaceutically acceptable carrier. The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle to which the active ingredient is administered together. Suitable pharmaceutically acceptable carriers are described in "Remington's Pharmaceutical Sciences" by E.W. Martin, 1995. Examples of pharmaceutical compositions include any solid (tablets, pills, capsules, granules, etc.) or liquid (solutions, suspensions, emulsions, etc.) composition for oral, topical, or parenteral administration. Pharmaceutical compositions containing the compounds of the present invention may be delivered by liposome or nanosphere encapsulation in a sustained-release formulation or by other standard delivery means.

[0121] Exemplary compositions are in the form of powders for injection solutions. For example, the compositions described in WO9942125. For example, lyophilized preparations of the compounds of the present invention containing water-soluble substances, and secondly, reconstituted solutions of mixed solvents. Specific examples include lyophilized preparations of PLD and mannitol, and reconstituted solutions of mixed solvents, such as PEG-35 castor oil, ethanol, and water for injection. For example, each vial may contain 2 mg of PLD. After reconstitution, each mL of the reconstituted solution may contain 0.5 mg of PLD, 158 mg of PEG-35 castor oil, and 0.15 mL / mL of ethanol.

[0122] Specific dosages and treatment regimens for any particular patient may vary and depend on a variety of factors, including the activity of the specific compound used, the specific formulation used, the mode of application, age, weight, general health status, sex, diet, administration time, excretion rate, drug combinations, response sensitivity, and the severity of the specific disease or condition being treated.

[0123] In further embodiments, patients can be selected for treatment with the compounds of the present invention based on clinical parameters and / or patient characteristics. Preferred parameters may be the measurements disclosed in this application.

[0124] To provide a more concise explanation, some of the quantitative expressions given herein are not qualified with the term “about.” Whether the term “about” is explicitly used or not, all quantities given herein are meant to refer to an actual given value, which also means an approximation to a given value that would be reasonably inferred based on ordinary knowledge of the art to which the invention belongs, including experimental and / or measurement equivalents and approximations of such given value.

[0125] The foregoing disclosure provides a general description of the subject matter covered within the scope of the invention, including methods and best modes thereof for constructing and using the invention; however, the following embodiments are provided to further enable those skilled in the art to practice the invention and provide a complete written description thereof. Those skilled in the art will understand, however, that the details of these embodiments should not be read as limiting the invention, and that its scope should be ascertained from the claims and equivalents attached to this disclosure. Various further aspects and embodiments of the invention will be apparent to those skilled in the art in consideration of this disclosure. [Examples]

[0126] The compounds of the present invention can be obtained by following processes described in the literature, for example, Vera et al. Med. Res. Rev. 2002, 22(2), 102-145, WO2011 / 020913 (see particularly Examples 1-5), WO02 / 02596, WO01 / 76616, and WO2004 / 084812, the contents of which are incorporated herein by reference.

[0127] The specific compounds of the present invention

[0128] [Table 1A]

[0129] [Table 1B]

[0130] That is the case.

[0131] Following the procedures described in WO02 / 02596 and the specification, and further disclosed in the previous examples:

[0132] [ka]

[0133] The following compounds are obtained.

[0134] [Table 2A]

[0135] [Table 2B]

[0136] [Table 2C]

[0137] [Table 2D]

[0138] [Table 2E]

[0139] Following the procedures described in WO02 / 02596 and the specification, and further disclosed in the previous examples:

[0140] [ka]

[0141] The following compounds are obtained.

[0142] [Table 3A]

[0143] [Table 3B]

[0144] [Table 3C]

[0145] [Table 3D]

[0146] [Table 3E]

[0147] The further compound is compound 240, known as didemnin B, and has the following structure:

[0148] [ka]

[0149] This is shown.

[0150] (Example 1) As shown in Figure 1, PLD inhibits NF-κB transactivation in vitro.

[0151] We investigated whether NFκB transcriptional activity is regulated by pritchidepsin. For this purpose, we used THP-1 cells stably transfected with an NFκB luciferase reporter plasmid. Cells were treated with 100 ng / mL TNFα (an activator of NF-κB), 500 μg / mL poly(I:C) (TLR3 ligand), 10 μg / mL LPS-B5 (TLR4 ligand), or 10 μg / mL reximod (TLR-7 / 8 ligand). Luciferase activity was quantified under each condition, using the compound alone (1A gray bar) or in combination with 100 nM pritchidepsin (1A black bar) for 6 hours. In the presence of each TLR ligand, pritchidepsin clearly inhibited luciferase production, indicating that transactivation from NF-κB was inhibited in the presence of the agent. Survival rates were analyzed using the MTT assay (gray bars (activator) and red bars (activator combined with 100 nM pristicepsin)). No cytotoxic effects were detected.

[0152] As shown in Figure 2, PLD also inhibits the secretion of pro-inflammatory cytokines IL-1, IL-6, IL-8, and TNF-α in human monocytes in vitro.

[0153] To investigate whether pritisepsin inhibits TLR-induced cytokine secretion, THP-1 cells were treated with 100 ng / mL TNFα (activator of NF-κB), 500 μg / mL poly(I:C) (TLR3 ligand), 10 μg / mL LPS-B5 (TLR4 ligand), or 10 μg / mL rexiquimod (TLR-7 / 8 ligand). Each compound was used alone (gray bars) or in combination with 100 nM pritisepsin (red bars) for 6 hours. ELISA assays were used to compare the changes in cytokine secretion in cell culture supernatant between different treatments. As shown in Figure 2, poly(I:C), LPS, and rexiquimod induced the secretion of IL-1, IL-6, IL-8, and TNFα. Furthermore, pritisepsin clearly inhibited the production of IL-1, IL-6, IL-8, and TNFα. TNFα failed to increase the secretion of IL-1 and IL-6. THP-1 cells may require other TNFα exposure times to secrete these cytokines.

[0154] In the presence of each TLR ligand, pritchidepsin clearly inhibited the secretion of pro-inflammatory cytokines IL-1, IL-6, and IL-8.

[0155] Further in vitro experiments investigated the effects of pristicepsin (APL) pretreatment on THP-1 cells. Using the THP-1 NFκB luc strain, 1, 10, or 50 nM APL or DMSO (0.2%) was added 8 hours prior to stimulation with 2.5 or 5 μg / mL rexiquimod (RQ). RQ is a TLR7 / 8 agonist that mimics ssRNA. Cytokine levels or cell viability were measured at 24 hours. As shown in Figure 9, PLD pretreatment inhibited the secretion of RQ-induced pro-inflammatory cytokines: IL6, IL8, IL1β, and TNF-α.

[0156] (Example 2) As shown in Figure 3, PLD inhibits the ex vivo secretion of pro-inflammatory cytokines, IL-6, IL-8, and TNF-α, in mice isolated from BAL.

[0157] We investigated whether pritolidepsin inhibits LPS-induced cytokine secretion in alveolar macrophages. To this end, mice were intravenously injected with pritolidepsin (1 mg / kg) or a vehicle, and bronchoalveolar lavage (BAL) fluid was collected 12 hours after administration. Cells were plated and treated ex vivo for 3 or 6 hours with or without 15 μg / mL LPS-B5, and secreted cytokines were measured. It can be seen that LPS induces the secretion of IL-6, IL-10, and TNFα (gray bars). Furthermore, in animals treated with pritolidepsin, the production of LPS-induced IL-6 and TNFα was clearly inhibited (red bars), resulting in an overall anti-inflammatory effect.

[0158] This is further shown in Figure 10. In animals treated with pritchidepsin, pritchidepsin is isolated from CD45 from bronchoalveolar lavage fluid. + In cells, LPS-B5 significantly reduced the secretion of IL-6, IL-10, and TNFα at 3 and 6 hours. As shown in Figure 10(a, b), this effect was unrelated to cell viability.

[0159] Furthermore, we investigated whether pritisepsin inhibits reximod (RQ)-induced cytokine secretion in BALF. Mice were intravenously injected with pritisepsin (1 mg / kg) or a vehicle one hour before intranasal administration of 50 μg / mouse with reximod. Bronchoalveolar lavage fluid (BALF) was collected one or three hours after intranasal administration of RQ. Cells were plated and secreted cytokines were measured. As can be seen in Figure 11, RQ induces TNFα secretion at both one and three hours post-administration. In vivo administration of PLD prevented the increase in TNFα production.

[0160] We also confirmed the effect of pritchidepsin on alveolar macrophage recruitment. Activated monocyte-derived macrophages contribute to the COVID-19 cytokine storm by releasing large amounts of pro-inflammatory cytokines. Bronchoalveolar lavage cells were stained and analyzed by flow cytometry. Pritidepsin reduced the percentage of macrophages present in the bronchoalveolar lavage fluid without cytotoxic effects.

[0161] (Example 3) As shown in Figure 4, after a single intravenous administration to mice, the number of macrophages in the BAL decreases with PLD.

[0162] To investigate whether pritisepsin reduces the percentage of alveolar macrophages in animals with acute inflammation, mice were treated with pritisepsin (1 mg / kg) IV, LPS (20 μg / kg) ip in sterile saline, or pritisepsin (1 mg / kg, IV) combined with LPS (20 μg / kg, ip). After 3 hours, bronchoalveolar lavage fluid was collected. Bronchoalveolar lavage cells were obtained by centrifugation and analyzed by flow cytometry (Figure 4b). The top panel shows the analytical strategy for the macrophage population present in the sample. The bottom right panel shows the same results expressed as a percentage of cells. The bottom left panel shows the percentage of CD45+ (leukocyte marker) viable cells. As can be seen, LPS induces the recruitment of alveolar macrophages. Treatment with pristicepsin reduces the percentage of macrophages present in bronchoalveolar lavage fluid without causing cytotoxic effects.

[0163] (Example 4) As shown in Figure 12, PLD is distributed in the lungs of nonclinical species. In addition, similar plasma exposure is achieved in mice (a nonclinical species used in pharmacological models) and patients.

[0164] Lung-to-plasma ratio in mice, rats, and hamsters ( lung AUC 0-∞ / plasma AUC 0-∞ The values ​​(calculated as ) were 133, 460, and 909, respectively, and the concentration of pristicepsin in the lungs was consistently higher than the concentration in plasma at all sampling times, thus confirming the distribution of pristicepsin in the lungs.

[0165] [Table 4]

[0166] Materials and methods Trans-activated luciferase assay. NF-κB transactivation was assayed using the Bright-Glo® Luciferase Assay System according to the manufacturer's instructions. NF-κB reporter (Luc)-THP-1 human monocytes stably transfected with the NF-κB-Luc plasmid (containing four NF-κB binding sites, a minimal promoter, and a luciferase gene) were exposed to 100 ng / mL TNFα (positive control), 500 μg / mL poly(I:C) (polyinosine-polycytidyl), 10 μg / mL LPS-B5 (lipopolysaccharide derived from E. coli O55:B5), or 10 μg / mL reciquimod. The compounds were used alone or in combination with 100 nM pristicepsin for 6 hours. Luminescence was measured using a Perkin-Elmer EnVision reader. A MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide) cell proliferation assay was performed concurrently to control for the compound's cytotoxicity. Cell viability was expressed as a percentage of control cell proliferation. The data presented are the average of three independent experiments performed three times.

[0167] ELISA assay of secreted cytokines THP1-NFκB-LUC cell cultures were treated as described above, and the culture medium was sampled 6 hours after treatment. Secreted cytokines were assayed by ELISA. The culture medium samples were stored at 4°C. The secretion of IL-8, IL-1β, IL-6, and TNFα proteins into the culture medium was quantified using ELISA kits with high specificity and sensitivity. Human IL-1β, human IL-6, human IL-8, and human TNF OptEIA® ELISA kits were obtained from BD Biosciences and performed according to the manufacturer's instructions. The data presented are the average of three independent experiments performed three times.

[0168] MTT assay Cells were seeded in 96-well microtiter plates and allowed to stand at 37°C and 5% CO2 for 24 hours before the above treatment. After 6 hours of continuous treatment, MTT was converted to its coloring reaction product, MTT formazan, and the absorbance at 540 nm was measured to estimate cell viability. The data presented here are representative from a series of three independent experiments conducted three times.

[0169] In vivo and ex vivo treatments Mice were randomly selected into groups of five and treated. Plitchidepsin (1 mg / kg) was administered intravenously (iv) to the mice, and they were euthanized 12 hours after administration. The control group received a plitchidepsin vehicle (Cremophor / ethanol / water) diluted with physiological saline. Bronchoalveolar lavage (BAL) fluid was collected from each group and centrifuged to obtain bronchoalveolar lavage cells. The cells were erythrolyzed (Roche), plated, and treated ex vivo for 3 or 6 hours with or without 15 μg / mL LPS-B5. Secreted cytokines were quantified using highly specific and sensitive ELISA kits. Mouse IL-6, mouse IL-10, and mouse TNF DuoSet ELISA kits were obtained from R&D Systems and performed according to the manufacturer's instructions. The data presented here are representative from a series of three independent experiments.

[0170] Animal inflammation models Mice were randomly selected into groups of two and treated. Mice were challenged with pritisepsin (1 mg / kg) intravenously (iv), with LPS (20 μg / kg) in sterile saline intravenously (iv), or with pritisepsin (1 mg / kg, iv) combined with LPS (20 μg / kg, ip). The control group received a pritisepsin vehicle (Cremophor / ethanol / water) diluted in saline. After 3 hours, the animals were euthanized and bronchoalveolar lavage fluid was collected (total 1.2 ml, PBS). Bronchoalveolar lavage cells were obtained by centrifugation and analyzed by flow cytometry. The data presented here are representative from a series of three independent experiments.

[0171] In another inflammation model, mice were randomly selected into groups of two and treated. Mice were challenged with pritolidepsin (1 mg / kg) intravenously (iv), followed one hour later with reciquimod (50 μg / mouse, intranasal). The control group received a pritolidepsin vehicle (Cremophor / ethanol / water) diluted in physiological saline. At one and three hours, the animals were euthanized, and bronchoalveolar lavage fluid was collected (total 1.2 ml, PBS), and TNFα was then quantified using an ELISA kit. The data presented here are representative from a series of three independent experiments.

[0172] Analysis of macrophages using flow cytometry. Bronchoalveolar lavage cells were stained with anti-F4 / 80-BV510, CD45-APC700, CD11b-BV650, CD11c-APC-Fire, CD24-PC7, and Ly6C-BV605 monoclonal antibodies (Biolegend) and the LIVE / DEAD® Fixable Green Dead Cell staining kit, 488nm excitation (Thermofisher). Macrophages (F4 / 80+) were gated with viable immune cells (CD45+ LIVE / DEAD dye), and alveolar macrophages (F4 / 80+ CD24-) were specifically gated with the CD11c+ CD11b- population from viable immune cells. Compensation and gating strategies were established using isotype controls and compensation beads.

[0173] (Example 5) A multicenter, randomized, parallel, and proof-of-concept study was conducted to evaluate the safety profiles of three doses of pritzidepsin in hospitalized COVID-19 patients. Details of the study are available at ClinicalTrials.gov identifier:NCT04382066.

[0174] Patients included in the study were randomly selected in a 1:1:1 ratio and received the following: - Group A) Administer 1.5 mg of pritisepsin once daily as a 1.5-hour infusion for 3 consecutive days (total dose is 4.5 mg). - Group B) Administer 2.0 mg of pritisepsin once daily as a 1.5-hour infusion for 3 consecutive days (total dose is 6.0 mg). - Group C) Administer 2.5 mg of pritisepsin once daily as a 1.5-hour infusion for 3 consecutive days (total dose 7.5 mg).

[0175] All patients can receive the following prophylactic medication 20-30 minutes before their pritisepsin infusion: - Diphenhydramine hydrochloride 25 mg iv (or equivalent). - Ranitidine 50 mg iv or equivalent. - Dexamethasone 6.6 mg administered intravenously. - Ondansetron 8 mg IV or equivalent administered by slow infusion over 15 minutes.

[0176] Patients included in the study will receive treatment for three days.

[0177] Pritidepsin is supplied as a powder for injection concentrate at a concentration of 2 mg / vial. Before use, reconstitute the vial with 4 ml of reconstituted solution to obtain a colorless to slightly yellowish solution containing 0.5 mg / ml pritidepsin, 25 mg / ml mannitol, 0.15 ml / ml macrogol glycerol ricinoleate oil, 0.15 ml / ml injection ethanol, and 0.70 ml / ml water. Further dilution with any suitable intravenous solution is required before injection.

[0178] Pritidepsin 2 mg is supplied in a Type I clear glass vial with a bromobutyl rubber stopper covered with an aluminum seal. Each vial contains 2 mg of pritidepsin.

[0179] Macrogol glycerol ricinoleate (polyoxyl 35 castor oil) / anhydrous ethanol / water for injection, 15% / 15% / 70% (v / v / v) reconstitution solvents are supplied in Type I colorless glass vials. The ampoule volume is 4 ml.

[0180] Pritidepsin will be labeled with the research protocol code, batch number, contents, expiration date, storage conditions, and the names of the researchers and sponsors. The investigational drug will be labeled in accordance with Annex 13 of the European Good Manufacturing Practices. Pritidepsin must be stored between 2°C and 8°C, and the vials must be stored in their outer box to protect them from light. Under these conditions, the drug is stable for 60 months.

[0181] After reconstituting a 2 mg priticinium vial with 4 ml of a reconstitution solution of macrogoal glycerol ricinoleate / ethanol / water for injection, the reconstituted solution should be diluted and used immediately after preparation. If not used immediately, the storage time and conditions until use are the responsibility of the user. The concentrated solution of the reconstituted drug has been shown to be physically, chemically, and microbiologically stable for 24 hours under refrigerated conditions (5°C ± 3°C) and, when stored in the original vial at room temperature under room light, for 6 hours. If storage is required before administration, the solution should be refrigerated and protected from light and used within 24 hours after reconstitution.

[0182] Intermediate result Patient 1 - A 50-year-old male with bilateral pneumonia. Received PLD 1.5 mg × 3. PCR COVID19 test: Positive at baseline, turned negative (no virus load) by day 4. Dramatic clinical improvement. Discharged by day 7. Dramatic clinical improvement such as all virus load removed by PLD and bilateral pneumonia treated and discharged by day 7 was achieved.

[0183] Patient 2: A 40-year-old male with bilateral pneumonia. Received PLD 1.5 mg × 3. No improvement was seen by day 6, switched to remdesivir + TOL + corticosteroid + opioid. PCR turned negative by day 15 and discharged by day 19.

[0184] Patient 3: A 53-year-old male with bilateral pneumonia. Received PLD 1.5 mg × 3. PLD prevented clinical deterioration. Discharged by day 10, PCR turned negative by day 31.

[0185] Patient 4: A 42-year-old male with bilateral pneumonia. Received PLD 2.0 mg × 3. Corticosteroid therapy was required. PCR COVID19 test: Positive at baseline and still positive on day 7, patient was PCR negative by day 15. The patient recovered to the extent of being discharged by day 10.

[0186] Patient 5: A 33-year-old woman with bilateral pneumonia at the time of enrollment. She received 1.5 mg of PLD three times. PCR COVID-19 test: Positive at baseline, turned negative (no viral load) by day 4. Bilateral pneumonia resolved by day 6 (normal Rx lungs). Major clinical improvement. Discharged by day 8. Figures 5a-5c show that the X-rays indicate the disappearance of pneumonia. Bilateral pneumonia is evident in Figure 5a. Improvement was seen on day 6 after PLD treatment. Laminar atelectasis is demonstrated in Figure 5b. In Figure 5c, the follow-up X-ray at day 15 showed a return to normal. The viral load was eliminated by day 4 with 1.5 mg of PLD three times. Major clinical improvements were achieved with PLD, including the elimination of all viral load, treatment of bilateral pneumonia, and discharge by day 8.

[0187] Patient 6: A 69-year-old woman with severe COPD. She presented with unilateral pneumonia at the time of enrollment. She received PLD 1.5 mg x 3. PCR COVID19 test: Positive at baseline, turned negative (no viral load) by day 7. Significant clinical improvement was observed. The patient was discharged by day 8. Figures 6a-6c show that X-rays indicate the progression of pneumonia. Figure 6a clearly shows unilateral pneumonia, and Figure 6b shows progression to bilateral pneumonia. Figure 6c shows improvement. As shown in Figure 6d, PLD achieved significant clinical improvement, including the elimination of all viral load, treatment of pneumonia, and discharge by day 8.

[0188] Patient 7: A 39-year-old woman with pulmonary infiltration. She received 2.0 mg of PLD three times. PCR COVID-19 test: Positive at baseline, turned negative (no viral load) by day 7. Major clinical improvement after PLD treatment. Discharged by day 8.

[0189] Patient 8: A 32-year-old male. He received 1.5 mg of PLD three times. Efficacy could not be evaluated, and he was discharged by day 4.

[0190] Patient 9: A 34-year-old male. He received PLD 2.0 mg x 3. PCR COVID-19 test: Positive at baseline and still positive on day 7. However, he showed significant clinical improvement by day 8 and was discharged.

[0191] C-reactive protein test The effect of PLD on inflammatory cytokines was also measured in patients 5, 7, and 9, and the results of the C-reactive protein test are shown in Figure 7. In patient 5 (Figure 7a), a rapid decrease was observed by day 2 after PLD administration. In patients 7 (Figure 7b) and 9 (Figure 7c), a rapid decrease was observed by day 3 after PLD administration. These data demonstrate the anti-inflammatory properties of PLD.

[0192] Upon completion of the study, 45 patients hospitalized with COVID-19 were randomly selected and administered pritisepsin at doses of 1.5, 2.0, and 2.5 mg daily for 3 days. The treatment was well-tolerated in all three dose cohorts. Treatment outcomes, assessed by discharge rate, were determined by baseline disease severity and viral load. Across all treatment cohorts, 100% (9 / 9) patients were mild, 82% (23 / 28) moderate, and 57% (4 / 7) severe, and were discharged by day 15.

[0193] (Example 6) The objective here was to evaluate the efficacy of pritisdepsin in vivo in the treatment of severe pneumonia caused by mouse adaptive A / H1N1 influenza virus infection (A / Puerto Rico / 8 / 34).

[0194] Experimental setup: To achieve this objective, high doses of PR8 influenza virus (2 x 10⁻¹⁰) were administered. 5 An in vivo model of viral pathogenesis in which severe lung infection occurred based on the administration of pfu was adopted. Next, the therapeutic effect of pritolidepsin on severe influenza virus infection in mice was evaluated. Nine-week-old female mice were anesthetized by intraperitoneal injection of ketamine-xylazine solution, and infection was induced by intranasal administration of 20 μl of PBS virus solution per nostril.

[0195] Treated mice were subcutaneously injected with 0.3 mg / kg or 0.15 mg / kg of pritzidepsin. Survival and weight loss were then monitored until day 3 post-infection. No deaths or weight loss exceeding 30% of the starting weight were recorded during the treatment period.

[0196] Control of influenza infection in the respiratory tract is mediated by enhanced inflammation in bronchoalveolar lavage fluid (BALF). Figure 8 shows the inflammatory profiles in BALF of infected mice with and without pritchidepsin treatment. Among the main pro-inflammatory cytokines, pritchidepsin significantly reduced the levels of IL-6 (Figure 8a), CCL2 (Figure 8b), IL-1α (Figure 8c), IFN-γ (Figure 8d), and TNF-α (Figure 8e). Mice that received only half the dose of the drug showed reduced defense and an intermediate phenotype.

[0197] BALF cell composition is defined as a marker of pulmonary immune response viral infection. Quantitative measurement of infiltrating cells correlated with inflammatory cytokine levels was evaluated in influenza-infected mice. Treatment with pristicepsin did not reduce the total cell composition of BALF (CD45). + x10 6 ).

[0198] Overall, these results confirm that three subsequent administrations of pritchidepsin (total dose 0.9 mg / kg) in influenza-infected mice can actively reduce inflammation, as indicated by the reduction in initial pro-inflammatory cytokines induced by the treatment.

[0199] (References) TIFF0007866504000028.tif175170

Claims

1. A pharmaceutical composition for treating an autoimmune condition, comprising pritisepsin or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the autoimmune condition is caused by the activation of one or more Toll-like receptors (TLRs).

2. The pharmaceutical composition according to claim 1, wherein the autoimmune state is characterized by increased signal transduction via at least one or more Toll-like receptors (TLRs).

3. The pharmaceutical composition according to claim 1, wherein the autoimmune state is characterized by an increased level of at least one pro-inflammatory cytokine.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the autoimmune condition is rheumatoid arthritis (RA).

5. The pharmaceutical composition according to any one of claims 1 to 3, wherein the autoimmune condition is systemic lupus erythematosus (SLE).

6. The pharmaceutical composition according to any one of claims 1 to 3, wherein the autoimmune condition is multiple sclerosis (MS).

7. The pharmaceutical composition according to any one of claims 1 to 3, wherein the autoimmune condition is scleroderma.

8. The pharmaceutical composition according to any one of claims 1 to 3, wherein the autoimmune condition is Sjögren's syndrome.

9. The pharmaceutical composition according to any one of claims 1 to 3, wherein the autoimmune state is autoimmune myocarditis.

10. The pharmaceutical composition according to any one of claims 1 to 3, wherein the autoimmune state is type 1 diabetes.

11. The pharmaceutical composition according to any one of claims 1 to 3, wherein the autoimmune condition is atherosclerosis.

12. The pharmaceutical composition according to any one of claims 1 to 11, wherein the pritchidepsin or a pharmaceutically acceptable salt or stereoisomer thereof is pritchidepsin.

13. A kit comprising plitidepsin or a pharmaceutically acceptable salt or stereoisomer thereof, along with instructions for use for treating an autoimmune condition as defined in any one of claims 1 to 12.

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