TLR7 inhibitors combined with prednisolone or hydroxychloroquine to treat cutaneous lupus erythematosus

Abstract

A method for treating a patient having cutaneous lupus erythematosus is disclosed, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a second agent selected from a therapeutically effective dose of prednisolone and hydroxychloroquine or a pharmaceutically acceptable salt thereof to the patient.

Description

Technology Field

[0001] Cross-reference

[0002] This application claims priority to Indian provisional application serial number 202011024586 filed on June 11, 2021, the full text of which is incorporated herein.

[0003] explanation

[0004] The present invention relates to a method for treating a patient having cutaneous lupus erythematosus, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a second agent selected from a therapeutically effective dose of prednisolone and hydroxychloroquine, or a pharmaceutically acceptable salt thereof. Background Technology

[0005] Toll-like receptors (TLRs) are a family of pattern recognition receptors that recognize highly conserved components of various pathogens and induce innate and adaptive immune responses (Akira S, Takeda K, T. Kaisho, T., Nat Immunol. 2001;2:675-680; Pandey S, Agrawal DK., Immunol Cell Biol. 2006;84:333-341; Kawai T, Akira S. J Biochem. 2007;141:137-145). TLR7 and TLR8 have been found to be expressed in endosome compartments and recognize ssRNA molecular patterns. TLR7 is expressed in B cells and plasmacytoplasmic dendritic cells (pDCs); TLR8 is expressed in monocytes and myeloid dendritic cells (mDCs) (Chuang TH, Ulevitch RJ. Eur Cytokine Net. 2000;11:372-378; Iwasaki A, Medzhitov R. Nat Immunol. 2004;5:987-995).

[0006] Cutaneous lupus erythematosus (CLE) is a skin disorder that can occur as an isolated cutaneous sign or in combination with concomitant systemic lupus erythematosus (SLE) (Okon LG, Werth VP. Best Pract Res Clin Rheumatol. 2013;27:391-404; Cohen MR, Crosby D. J Rheumatol. 1994;21:1665-1669).

[0007] CLE is often associated with the presence of autoantibodies with varying frequencies. Similar to SLE, dysregulation of type I interferon (IFN) is suggested to be a major pathogenic driver in CLE. CLE patients often exhibit IFN signatures in blood and skin lesions, and blood IFN signatures correlate with skin disease activity scores (Braunstein I, Klein R, Okawa J, et al., Br J Dermatol. 2012;166:971-975; Meller S, Winterberg F, Gilliet M, et al., Arthritis Rheum. 2005;52:1504-1516, Wenzel J, Zahn S, Mikus S, et al., Br J Dermatol. 2007;157:752-757).

[0008] Treatment with cipalimumab (anti-IFNα mAb) and anifrolumab (anti-IFNAR mAb) demonstrated improvement in skin disease activity scores in SLE patients presenting with skin disease in Phase II clinical trials, which further supports the contribution of type I IFN to the pathogenesis of CLE (Furie R, Khamashta M, Merrill JT, et al. Arthritis Rheumatol. 2017;69:376-86; Petri M, Wallace DJ, Spindler A, et al. Arthritis Rheum. 2013;65:1011-1021). Plasma cell-like dendritic cells (pDCs) are specialized cells that accumulate in the skin of CLE patients and account for 5-10% of immune infiltrates, and have been suggested to be the major source of type I IFN in CLE skin lesions. pDCs robustly secrete IFNα in response to stimulation of TLR7 and TLR9 by nucleic acid ligands, whereas TLR8 induces various bone marrow cells to produce inflammatory cytokines, such as TNFα and IL-6 (Siegal FP, Kadowaki N, Shodell M, et al. Science 1999;284:1835-1837; Tomasini D, Mentzel T, Hantschke M, et al., J Cutan Pathol. 2010;37:1132-1139; Vermi W, Lonardi S, Morassi M, et al., Immunobiology 2009;214:877-86).

[0009] Apoptotic fragments induced by ultraviolet radiation, a widely known environmental trigger for CLE, cause the accumulation of ribonucleic acid (RNA) and deoxyribonucleic acid (DNA) fragments that activate TLR7 and TLR9, respectively, leading to pDC activation and type I IFN production (Wenzel J, Proelss J, Wiechert A, et al., J Am Acad Dermatol. 2007;56: 648-650).

[0010] A new method to treat CLE is desired.

[0011] A method for treating cutaneous lupus erythematosus is disclosed herein, comprising administering to a patient a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a therapeutically effective dose of prednisolone or a pharmaceutically acceptable salt thereof.

[0012] Additionally, a method for treating cutaneous lupus erythematosus is disclosed herein, comprising administering to a patient a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a therapeutically effective dose of hydroxychloroquine or a pharmaceutically acceptable salt thereof.

[0013] Summary of the Invention

[0014] The present invention provides a method for treating a patient having cutaneous lupus erythematosus, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a second agent selected from a therapeutically effective dose of prednisolone and hydroxychloroquine or a pharmaceutically acceptable salt thereof.

[0015] The present invention provides a method for treating a patient having cutaneous lupus erythematosus, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a therapeutically effective dose of prednisolone or a pharmaceutically acceptable salt thereof to the patient.

[0016] The present invention provides a method for treating a patient having cutaneous lupus erythematosus, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a therapeutically effective dose of hydroxychloroquine or a pharmaceutically acceptable salt thereof to the patient.

[0017] These and other features of the present invention will be described in an expanded form as the disclosure continues. Brief explanation of the drawing

[0018] The present invention is illustrated with reference to the attached drawings described below. Figure 1 shows the inhibition of skin lesions after treatment with compound (I) or prednisolone in female MRL / lpr mice. A: Neck lesions; B: Head lesions; C: Facial lesions; D: Ear lesions; E: Total lesions. Figure 2 shows the reduction of erythema following treatment of skin lesions with compound (I) or prednisolone in female MRL / lpr mice. A: Cervical erythema; B: Ear erythema; C: Total erythema. Figure 3 shows the inhibition of circulating antibody titers, including anti-dsDNA, anti-smRNP, and anti-Ro antibody titers, after treatment with compound (I) or prednisolone. Figure 4 shows the reduction in proteinuria and urinary NGAL after treatment with compound (I) or prednisolone. Figure 5 shows that compound (I) inhibits TLR7-induced IL-6 in an in vitro setting after treatment with compound (I) or prednisolone. Figure 6 shows total lesions in female MRL / lpr mice in established disease treated with vehicle, compound (I), prednisolone, hydroxychloroquine, compound (I) + prednisolone, and compound (I) + hydroxychloroquine. Figure 7 shows the cervical erythema scores for female MRL / lpr mice treated with vehicle, compound (I), prednisolone, hydroxychloroquine, compound (I) + prednisolone, and compound (I) + hydroxychloroquine. Figure 8 shows the ear erythema scores for female MRL / lpr mice in established disease treated with vehicle, compound (I), prednisolone, hydroxychloroquine, compound (I) + prednisolone, and compound (I) + hydroxychloroquine. Figure 9 shows the total erythema scores for female MRL / lpr mice in established disease treated with vehicle, compound (I), prednisolone, hydroxychloroquine, compound (I) + prednisolone, and compound (I) + hydroxychloroquine. Figure 10 shows the proteinuria levels in female MRL / lpr mice in an established disease treated with vehicle, compound (I), prednisolone, hydroxychloroquine, compound (I) + prednisolone, and compound (I) + hydroxychloroquine. Figure 11 shows urinary NGAL levels in female MRL / lpr mice in established disease treated with vehicle, compound (I), prednisolone, hydroxychloroquine, compound (I) + prednisolone, and compound (I) + hydroxychloroquine. Figure 12 shows total lesions in female MRL / lpr mice in progressive disease treated with vehicle, compound (I), prednisolone, hydroxychloroquine, compound (I) + prednisolone, and compound (I) + hydroxychloroquine. Figure 13 shows the cervical erythema scores for female MRL / lpr mice treated with vehicle, compound (I), prednisolone, and compound (I) + prednisolone. Figure 14 shows the cervical erythema scores for female MRL / lpr mice treated with vehicle, hydroxychloroquine, and compound (I) + hydroxychloroquine. Figure 15 shows the ear erythema scores for female MRL / lpr mice in progressive disease treated with vehicle, compound (I), prednisolone, hydroxychloroquine, compound (I) + prednisolone, and compound (I) + hydroxychloroquine. Figure 16 shows the total erythema scores for female MRL / lpr mice in progressive disease treated with vehicle, compound (I), prednisolone, hydroxychloroquine, compound (I) + prednisolone, and compound (I) + hydroxychloroquine. Specific details for implementing the invention

[0019] definition

[0020] To facilitate a better understanding of the contents described herein, specific terms are defined first. Additional definitions are provided throughout the detailed description.

[0021] "TR7 inhibitors" inhibit the function of TLR7. TLR7 inhibitors can associate with TLR7 reversibly or irreversibly and include antibodies, small molecules, and millimolecule compounds.

[0022] The compound of chemical formula (I) is a TLR7 inhibitor and has the following structure:

[0023] .

[0024] The chemical name for the compound of formula (I) is 2-(4-(2-(7,8-dimethyl-[1,2,4]triazolo[1,5-a]pyridine-6-yl)-3-isopropyl-1H-indole-5-yl)piperidine-1-yl)acetamide. The discovery and synthesis of the compound of formula (I) are described in WO2018 / 005586 A1.

[0025] Prednisolone is a corticosteroid drug having the chemical name: 11,17-dihydroxy-17-(2-hydroxyacetyl)-10,13-dimethyl-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydrocyclopenta[a]phenanthren-3-one. Prednisolone may be administered orally as a tablet, orally disintegrating tablet, solution, or suspension. Prednisolone for ophthalmic use is available as a solution (liquid) and a suspension (eye drops). Prednisolone is available as a sodium salt. Prednisolone may also be administered as the prodrug prednisone. As used herein, "Prednisolone" includes both prednisolone and prednisone. As used herein, "Pred" refers to prednisolone.

[0026] Hydroxychloroquine has the following structure.

[0027] .

[0028] "HCQ" as used herein refers to hydroxychloroquine. Hydroxychloroquine is used to treat or prevent autoimmune diseases, including lupus and rheumatoid arthritis. It can reduce skin problems in lupus, including cutaneous lupus erythematosus, and prevent swelling / pain in arthritis. Hydroxychloroquine is available as a sulfate salt, and 200 mg of the sulfate salt is equivalent to 155 mg of the base.

[0029] Unless otherwise defined, scientific and technical terms used in connection with the present invention shall have the meanings commonly understood by a person skilled in the art. Additionally, unless otherwise required by the context, singular terms shall include the plural, and plural terms shall include the singular.

[0030] As used herein, the terms “treat,” “treating,” and “treatment” refer to any type of intervention or process performed on a patient, or the administration of an activator to a patient, for the purpose of reversing, alleviating, improving, suppressing, slowing, or preventing the progression, occurrence, severity, or relapse of symptoms, complications, conditions, or biochemical signs associated with the disease. Treatment includes curative treatments and prophylactic or preventive measures, the purpose of which is to prevent or alleviate the targeted condition or disorder.

[0031] The term “therapeutic effective dose” or “therapeutic effective dosage” of a drug or therapeutic agent refers to an amount of drug effective in treating a disease or disorder in a patient. In certain embodiments, the effective dose refers to an amount effective in achieving the desired therapeutic or preventive result for the required duration at the required dosage. The ability of a therapeutic agent to promote disease regression or to suppress the onset or recurrence of disease may be evaluated by testing the activity of said therapeutic agent using various methods known to a skilled practitioner, for example, in human subjects during clinical trials, in animal model systems that predict efficacy in humans, or in in vitro assays.

[0032] The therapeutically effective dose of TLR7 inhibitors, prednisolone, and hydroxychloroquine may vary depending on factors such as the patient's disease status, age, sex, and weight, as well as the ability of TLR7 inhibitors, prednisolone, and hydroxychloroquine to elicit a desired response in the patient. The therapeutically effective dose of TLR7 inhibitors, prednisolone, and hydroxychloroquine encompasses an amount in which the therapeutically beneficial effect outweighs any toxic or adverse effects.

[0033] The terms "administering" and "administering" refer to the physical introduction of a composition containing a therapeutic agent into a patient using any of the various methods and delivery systems known to those skilled in the art. Routes of administration for TLR7 inhibitors and second agonists include enteral, topical, and mucosal administration, e.g., oral, topical, sublingual, rectal, intranasal, and intravenous administration, and parenteral administration, e.g., intravenous, intramuscular, and subcutaneous injection.

[0034] Administration in combination with one or more additional therapeutic agents includes simultaneous (co-) and sequential (sequential) administration in any order. For example, the patient may swallow the oral formulation of the TLR7 inhibitor and the oral formulation of the second agonist in any order (sequentially); or swallow the two oral formulations together (co-).

[0035] The term "patient" includes human and other mammalian subjects receiving any one therapeutic treatment.

[0036] definition

[0037] The features and benefits of the present invention may be more readily understood by those skilled in the art by reading the following detailed description. For clarity, it should be recognized that specific features of the present invention described above and below in relation to individual embodiments may also be combined to form a single embodiment. Conversely, for the sake of brevity, various features of the invention described in the context of a single embodiment may also be combined to form a sub-combination thereof. Embodiments identified herein as exemplary or preferred are intended to be illustrative, not limiting.

[0038] One or more methods for treating patients with cutaneous lupus erythematosus are provided herein.

[0039] One embodiment provides a method for treating a patient having cutaneous lupus erythematosus, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a second agent selected from a therapeutically effective dose of prednisolone and hydroxychloroquine or a pharmaceutically acceptable salt thereof to the patient.

[0040] One embodiment provides a method for treating a patient having cutaneous lupus erythematosus, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a therapeutically effective dose of prednisolone or a pharmaceutically acceptable salt thereof to the patient. A method in which the TLR7 inhibitor is a compound of formula (I) is included in this embodiment.

[0041] One embodiment provides a method for treating a patient having cutaneous lupus erythematosus, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a therapeutically effective dose of prednisone or a pharmaceutically acceptable salt thereof to said patient. A method in which the TLR7 inhibitor is a compound of formula (I) is included in this embodiment.

[0042] One embodiment provides a method for treating a patient having cutaneous lupus erythematosus, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a therapeutically effective dose of hydroxychloroquine or a pharmaceutically acceptable salt thereof to the patient. A method in which the TLR7 inhibitor is a compound of formula (I) is included in this embodiment.

[0043] One embodiment provides a method for treating a patient having cutaneous lupus erythematosus, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a second agent selected from a therapeutically effective dose of prednisolone and hydroxychloroquine or a pharmaceutically acceptable salt thereof, wherein the second agent is administered simultaneously with the TLR7 inhibitor. A method in which the TLR7 inhibitor is a compound of formula (I) is included in this embodiment.

[0044] One embodiment provides a method for treating a patient having cutaneous lupus erythematosus, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a second agent selected from a therapeutically effective dose of prednisolone or a pharmaceutically acceptable salt thereof, wherein the second agent is administered simultaneously with the TLR7 inhibitor. A method in which the TLR7 inhibitor is a compound of formula (I) is included in this embodiment.

[0045] One embodiment provides a method for treating a patient having cutaneous lupus erythematosus, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a second agent selected from a therapeutically effective dose of hydroxychloroquine or a pharmaceutically acceptable salt thereof, wherein the second agent is administered simultaneously with the TLR7 inhibitor. A method in which the TLR7 inhibitor is a compound of formula (I) is included in this embodiment.

[0046] One embodiment provides a method for treating a patient having cutaneous lupus erythematosus, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a second agonist selected from a therapeutically effective dose of prednisolone and hydroxychloroquine or a pharmaceutically acceptable salt thereof, wherein the second agonist is administered sequentially with the TLR7 inhibitor. An embodiment in which the TLR7 inhibitor is administered before the administration of the second agonist is included in this embodiment. An embodiment in which the TLR7 agonist is administered after the second agonist is also included in this embodiment. Additionally, an embodiment in which the TLR7 inhibitor is a compound of formula (I) is included in this embodiment.

[0047] One embodiment provides a method for treating a patient with cutaneous lupus erythematosus, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a second agonist selected from a therapeutically effective dose of prednisolone or a pharmaceutically acceptable salt thereof, wherein the second agonist is administered sequentially with the TLR7 inhibitor. An embodiment in which the TLR7 inhibitor is administered before the administration of the second agonist is included in this embodiment. An embodiment in which the TLR7 agonist is administered after the second agonist is also included in this embodiment. Additionally, an embodiment in which the TLR7 inhibitor is a compound of formula (I) is included in this embodiment.

[0048] One embodiment provides a method for treating a patient having cutaneous lupus erythematosus, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a second agonist selected from a therapeutically effective dose of hydroxychloroquine or a pharmaceutically acceptable salt thereof, wherein the second agonist is administered sequentially with the TLR7 inhibitor. A method in which the TLR7 inhibitor is administered before the administration of the second agonist is included in this embodiment. A method in which the TLR7 agonist is administered after the second agonist is also included in this embodiment. Additionally, a method in which the TLR7 inhibitor is a compound of formula (I) is included in this embodiment.

[0049] In one embodiment, the therapeutic effective dose of the compound of formula (I) is in the range of 0.1 to 100 mg.

[0050] In one embodiment, the therapeutic effective dose of the prednisolone compound is in the range of 0.5 to 50 mg.

[0051] In one embodiment, the therapeutically effective dose of a compound of hydroxychloroquine is in the range of 1 to 20 mg.

[0052] The therapeutically effective dose of a TL7 inhibitor may be administered as a single dose (qd) per day, divided into two doses per day (bid), or divided into three or more doses per day.

[0053] The therapeutically effective dose of prednisolone may be administered as a single dose (qd) per day, divided into two doses per day (bid), or divided into three or more doses per day.

[0054] The therapeutically effective dose of hydroxychloroquine may be administered as a single dose (qd) per day, divided into two doses per day (bid), or divided into three or more doses per day.

[0055] In one embodiment, the therapeutically effective dose of a TLR7 inhibitor is administered as a single daily dose.

[0056] In one embodiment, a therapeutically effective dose of the compound of formula (I) is administered as a single daily dose.

[0057] In one embodiment, a therapeutically effective dose of prednisolone is administered as a single daily dose.

[0058] In one embodiment, the therapeutically effective dose of hydroxychloroquine is administered as a single daily dose.

[0059] In one embodiment, a therapeutically effective dose of a TLR7 inhibitor is administered as a single daily dose, and a therapeutically effective dose of a second agonist selected from prednisolone and hydroxychloroquine is administered as a single daily dose. A method of administering the TLR7 inhibitor and the second agonist once daily is included in this embodiment, wherein the TLR7 inhibitor and the second agonist are administered simultaneously, or the TLR7 inhibitor is administered immediately before or immediately after the administration of the second agonist. Additionally, in this embodiment, a method is included in which the TLR7 inhibitor is a compound of formula (I). Furthermore, a method is included in this embodiment in which the second agonist is prednisolone. Additionally, a method is included in this embodiment in which the second agonist is hydroxychloroquine.

[0060] In one embodiment, a therapeutically effective dose of prednisolone is administered as a single daily dose.

[0061] In one embodiment, the therapeutically effective dose of hydroxychloroquine is administered as a single daily dose.

[0062] In one embodiment, a therapeutically effective dose of the TLR7 inhibitor is administered twice daily, and a therapeutically effective dose of the second agonist is administered once or twice daily. A method in which the TLR7 inhibitor is a compound of formula (I) is included in this embodiment.

[0063] In one embodiment, a therapeutically effective dose of a TLR7 inhibitor is administered twice daily, and a therapeutically effective dose of a second agonist selected from prednisolone and hydroxychloroquine is administered twice daily. This embodiment includes a method in which the TLR7 inhibitor and the second agonist are administered twice daily, wherein the TLR7 inhibitor and the second agonist are administered simultaneously twice daily, or the TLR7 inhibitor is administered immediately before or immediately after the administration of the second agonist twice daily. Additionally, this embodiment includes a method in which the TLR7 inhibitor is a compound of formula (I). Furthermore, this embodiment includes a method in which the second agonist is prednisolone. Additionally, this embodiment includes a method in which the second agonist is hydroxychloroquine.

[0064] In another embodiment, a method for treating a patient having cutaneous lupus erythematosus is provided, comprising administering a therapeutically effective dose of a TLR7 inhibitor or a pharmaceutically acceptable salt thereof to a patient in combination with a second agent selected from a therapeutically effective dose of prednisolone and hydroxychloroquine or a pharmaceutically acceptable salt thereof, and in combination with one or more additional third agents. Examples of suitable third agents include corticosteroids, rolipram, calfostin, cytokine-inhibiting anti-inflammatory drugs (CSAIDs), interleukin-10, glucocorticoids, salicylates, nitric oxide, and other immunosuppressants; nuclear translocation inhibitors, e.g., deoxyspergualin (DSG); non-steroidal anti-inflammatory drugs (NSAIDs), e.g., ibuprofen, celecoxib, and rofecoxib; steroids, e.g., dexamethasone; Antiproliferative agents, such as methotrexate, leflunomide, FK506 (tacrolimus, PROGRAF®); cytotoxic drugs, such as azatiphrine and cyclophosphamide; TNF-α inhibitors, such as Tenidab, anti-TNF antibodies or soluble TNF receptors, and rapamycin (sirolimus or rapamune®) or derivatives thereof. When the third agent is used in combination with a combination of compound (I) and the second agent, it may be used, for example, in an amount as indicated in the Physicians' Desk Reference (PDR) or otherwise determined by a person skilled in the art. In the method of the present invention, one or more third agents may be administered before, simultaneously with, or after the administration of compound (I) or the second agent.

[0065] Cutaneous lupus erythematosus study in mice

[0066] Materials and Methods

[0067] All animal experiment procedures were reviewed and approved by the Institutional Animal Ethics Committee (IAEC) and performed in accordance with the procedures established by the Committee for the Purpose of Control and Supervision on Experiments on Animals (CPCSEA). Mice were housed in groups at the Syngene Laboratory Animal Research Facility (SLAR, Bangalore, India; AAALAC accredited) and maintained under a normal 12-hour light / 12-hour dark cycle with free access to food and water. At the end of the study, animals were euthanized by CO2 asphyxiation for plasma and tissue collection.

[0068] Lupus skin model

[0069] Female MRL / lpr mice aged 12 to 14 weeks were screened and randomized based on the titers of anti-dsDNA antibodies and urinary neutrophil gelatinase-associated lipocalin (NGAL). Mice were orally treated once daily for 8 weeks with a vehicle (10% ethanol; 45% PEG 300; 5% Pluronic F-68; 40% 20 mM citrate buffer) or different doses of the test compound (Compound (I)) or reference compound (Prednisolone). The effects of the vehicle, test, or reference compounds on skin disease severity were evaluated by individually assessing skin lesions on the neck, head, face, and ears. The severity of skin lesions was assessed weekly by counting the number of erythema as well as the lesions (on a scale of 0 to 4: 0-none; 1-mild; 2-moderate; 3-moderate to severe; and 4-very significant and severe). Total erythema was determined by summing the erythema scores of lesions on the neck, head, ears, and face. Since nephritis also occurred in female MRL / lpr mice in addition to skin lesions, the effects of the vehicle, test, or reference compounds on renal disease were evaluated by measuring proteinuria, urinary NGAL, and autoantibody titers, such as anti-Ro, anti-smRNP, and anti-dsDNA Ab titers. One week prior to the end of the experiment, blood was drawn from mice at various time points to capture the complete pharmacokinetic profiles of the test or reference compounds and the degree of inhibition of gardiquimod-induced IL-6 in an in vitro whole blood assay. Spleen samples were collected at the end of the study to measure the effects of treatment on IL-6 and IFNα-producing cell populations. Blood samples were analyzed for gene expression by qPCR and for TLR7 target coverage using an in vitro blood assay for Gardiquimod-induced IL-6 production. Statistical analysis was performed using one-way ANOVA with Dunnett's test to calculate the significance of the test or reference compound treatment groups compared to the vehicle control group.For the test or reference compound treatment group, the percentage reduction in disease severity relative to the vehicle control group for each parameter was calculated.

[0070] Suppression of skin signs in the MRL / lpr model of lupus

[0071] Female MRL / lpr mice spontaneously induce skin lesions, making them a widely accepted preclinical murine model of cutaneous lupus. In addition to skin signs, these mice also exhibited renal damage. We investigated the effects of compounds on both skin lesion and renal damage markers in this model. In this study, mice were randomized based on urinary NGAL and anti-dsDNA Ab titers. Immediately after recruitment, mice were treated with compound (I) and prednisolone for 8 weeks.

[0072] Fig. 1: Inhibition of skin lesions in female MRL / lpr mice after treatment with compound (I). Mice were administered orally for 8 weeks. Subsequently, skin lesions were evaluated on the neck (a), head (b), face (c), and ears (d). The effect of treatment on total lesions was also presented by summing the number of lesions present on the face, neck, head, and ears. (e) Data are from one experiment using 13 to 15 mice per group. *P < 0.01, **P < 0.001, ***P < 0.0001 (Vehicle vs. Vehicle, by two-way ANOVA with Bonferroni test).

[0073] Fig. 2: Treatment of female MRL / lpr mice with compound (I) inhibited erythema of skin lesions. Mice were administered orally for 8 weeks. Erythema of skin lesions was evaluated on the neck (a) and ears (b). The effect of treatment on total erythema was also presented by evaluating the erythematous scores of skin lesions present on the back, face, neck, ears, and head (c). Data are from one experiment using 13 to 15 mice per group. *P < 0.01, **P < 0.001, ***P < 0.0001 (vehicle versus, by binary ANOVA with Bonferroni test).

[0074] Fig. 3: Inhibition of autoantibodies in female MRL / lpr mice treated with compound (I). Mice were administered orally for 8 weeks with each treatment. After 8 weeks of administration, anti-dsDNA (a), anti-Ro (b), and anti-smRNP antibody titers (c) were evaluated. Data are from one experiment using 13 to 15 mice per group. Percent inhibition relative to the vehicle group is shown above each bar. *P < 0.01, **P < 0.001, ***P < 0.0001 relative to the vehicle, by one-way ANOVA with Dunnett test.

[0075] Fig. 4: Inhibition of renal injury markers in female MRL / lpr mice treated with compound (I). Mice were administered orally for 8 weeks with each treatment. After 8 weeks of administration, proteinuria (a) and urinary NGAL (b) were evaluated. Data are from one experiment using 13 to 15 mice per group. Percent inhibition relative to the vehicle group is shown above each bar. *P < 0.01, **P < 0.001, ***P < 0.0001 relative to vehicle, by one-way ANOVA with Dunnett test.

[0076] Fig. 5: Compound (I) inhibited TLR7-dependent IL-6 induction in an in vitro assay in a female MRL / lpr model of lupus. Mice were administered orally for 7 weeks with each treatment. After 7 weeks of administration, mouse whole blood was collected at different time points and challenged with gardiquimod for 16 to 18 hours. IL-6 inhibition was evaluated by ELISA. Data are from one experiment in which measurements were performed on 3 mice per group at each time point.

[0077] Treatment with compound (I) improved the survival of mice in a female MRL / lpr model of lupus compared to the vehicle.

[0078] Table 1

[0079]

[0080] Abbreviation: NA, Not applicable.

[0081] The data in Table 1 indicate that treatment with compound (I) improved the survival of mice in this study. While only 87% of vehicle mice survived for 8 weeks, mice administered 0.25 and 0.75 mg / kg doses of compound (I) achieved 100% survival.

[0082] As shown in Fig. 1, compound (I) exhibited significant and robust dose-dependent inhibition of skin signs, as assessed by measuring the number of lesions on the face, neck, head, and ears. The effect of treatment on total lesions was also presented by summing the number of lesions present on the face, neck, head, and ears. Additionally, the effect of compound (I) was evaluated by assessing erythema on the neck and ears, and presented individually. Furthermore, total erythema was evaluated after assessing the erythema of skin lesions present on the back, face, neck, ears, and head. A significant reduction in erythema was observed upon treatment with compound (I) (Fig. 2). Representative images of lesions in mice treated with compound (I) and vehicle mice at the 8-week time point are shown in Fig. 3.

[0083] Dose-dependent inhibition of skin lesions, including erythema of the lesions as well as the number of lesions, was observed in mice with treatment with compound (I). Greater efficacy was observed at doses of 0.25 mg / kg and 0.75 mg / kg of compound (I). These two doses improved the survival rate of vehicle-treated mice to 100% compared to 87%. At these doses, compound (I) demonstrated inhibition of lupus nephritis, as assessed by kidney damage markers such as proteinuria and urinary NGAL. Additionally, at these two doses, compound (I) demonstrated significant inhibition of anti-dsDNA, anti-RO, and anti-smRNP auto-antibody titers and inhibited type I IFN-responsive gene expression.

[0084] Combinational research

[0085] Table 2

[0086]

[0087] 1. Combination study of compound (I) and prednisolone and compound (I) and hydroxychloroquine (HQC) in an MRL / lpr model of lupus-established disease

[0088] Female MRL / lpr mice aged 15 weeks were treated for 5 weeks. Mice were mobilized when they exhibited a group mean of a total skin lesion score of 1.5 to 2. The total skin lesion score includes the cumulative scores of cervical / dorsal, facial, head, and ear lesions.

[0089] Table 3

[0090]

[0091] Fig. 6: Treatment with compound (I) inhibited the total number of skin lesions in mice with established disease compared to treatment with vehicle. Monotherapy with prednisolone and hydroxychloroquine showed inhibition of skin lesions, but less than treatment with compound (I). Treatment with the combination of compound (I) + prednisolone and the combination of compound (I) and hydroxychloroquine showed increased inhibition compared to monotherapy with hydroxychloroquine and 1 mpk prednisolone.

[0092] Fig. 7: Treatment with compound (I) inhibited erythema of skin lesions on the neck of mice in established disease compared to vehicle. Monotherapy with prednisolone and hydroxychloroquine showed inhibition of erythema of skin lesions on the neck of mice. Treatment with the combination of compound (I) + prednisolone and the combination of compound (I) and hydroxychloroquine showed inhibition of erythema of skin lesions on the neck of mice.

[0093] Fig. 8: Treatment with compound (I) inhibited erythema of skin lesions on the ears of mice in established disease compared to vehicle. Monotherapy with prednisolone and hydroxychloroquine showed inhibition of erythema of skin lesions on the ears of mice, but less than treatment with compound (I). Treatment with the combination of compound (I) + prednisolone and the combination of compound (I) and hydroxychloroquine showed increased inhibition of erythema of skin lesions compared to monotherapy with hydroxychloroquine and 1 mpk prednisolone.

[0094] Fig. 9: Treatment with compound (I) inhibited total erythema of skin lesions on mice in established disease compared to vehicle. Monotherapy with prednisolone and hydroxychloroquine showed inhibition of total erythema of skin lesions in mice. Treatment with the combination of compound (I) + prednisolone and the combination of compound (I) and hydroxychloroquine showed inhibition of total erythema of skin lesions in mice.

[0095] Fig. 10: Treatment with compound (I) showed inhibition of the renal damage marker proteinuria and urinary neutrophil gelatinase-associated lipocalin (NGAL) in established disease. Monotherapy with prednisolone or hydroxychloroquine showed inhibition of the renal damage marker proteinuria. The combination of compound (I) and prednisolone and the combination of compound (I) and hydroxychloroquine showed increased inhibition of the renal damage marker proteinuria.

[0096] Fig. 11: Treatment with compound (I) showed inhibition of the renal damage marker urinary NGAL in established disease. Monotherapy with prednisolone or hydroxychloroquine showed inhibition of the renal damage marker urinary NGAL. The combination of compound (I) and prednisolone and the combination of compound (I) and hydroxychloroquine showed increased inhibition of the renal damage marker urinary NGAL.

[0097] 2. Combination study of compound (I) and prednisolone and compound (I) and hydroxychloroquine (HQC) in an MRL / lpr model of lupus-progressive disease

[0098] 16-week-old female MRL / lpr mice were treated for 5 weeks. Mice were mobilized when they exhibited a group mean of a total skin lesion score of 2 to 2.5. The total skin lesion score includes cumulative scores for cervical / dorsal, facial, head, and ear lesions. Table 2 shows the treatment, dose, and number of mice in each treatment group.

[0099] Table 4

[0100]

[0101] Fig. 12: Treatment with compound (I) suppressed the total number of skin lesions in mice with progressive disease compared to treatment with vehicle. Monotherapy with prednisolone and hydroxychloroquine showed suppression of skin lesions, but less than treatment with compound (I). Treatment with the combination of compound (I) + prednisolone and the combination of compound (I) and hydroxychloroquine showed increased suppression compared to monotherapy with hydroxychloroquine and 1 mpk prednisolone.

[0102] Fig. 13: Treatment with compound (I) inhibited erythema of skin lesions on the neck of mice with progressive disease compared to vehicle. Monotherapy with prednisolone showed inhibition of erythema of skin lesions on the neck of mice. Treatment with the combination of compound (I) + prednisolone showed inhibition of erythema of skin lesions on the neck of mice.

[0103] Fig. 14: Treatment with compound (I) inhibited erythema of skin lesions on the neck of mice with progressive disease compared to vehicle. Monotherapy with hydroxychloroquine showed inhibition of erythema of skin lesions on the neck of mice. Treatment with compound (I) and treatment with the combination of compound (I) + hydroxychloroquine showed increased inhibition of erythema of skin lesions on the neck of mice compared to treatment with hydroxychloroquine.

[0104] Fig. 15: Treatment with compound (I) inhibited erythema of skin lesions on the ears of mice with progressive disease compared to vehicle. Monotherapy with prednisolone and hydroxychloroquine showed inhibition of erythema of skin lesions on the ears of mice, but less than treatment with compound (I). Treatment with the combination of compound (I) + prednisolone and the combination of compound (I) and hydroxychloroquine showed increased inhibition of erythema of skin lesions compared to monotherapy with hydroxychloroquine and 1 mpk prednisolone.

[0105] Fig. 16: Treatment with compound (I) inhibited total erythema of skin lesions on mice in progressive disease compared to vehicle. Monotherapy with prednisolone and hydroxychloroquine showed inhibition of total erythema of skin lesions in mice. Treatment with the combination of compound (I) + prednisolone and the combination of compound (I) and hydroxychloroquine showed inhibition of total erythema of skin lesions in mice compared to monotherapy with hydroxychloroquine and 1 mpk prednisolone.

Claims

Claim 1 A pharmaceutical composition comprising a TLR7 inhibitor or a pharmaceutically acceptable salt thereof for use in a method for treating a patient with cutaneous lupus erythematosus, wherein the method comprises administering to the patient a therapeutically effective dose of the TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a second agonist selected from a therapeutically effective dose of prednisolone and hydroxychloroquine or a pharmaceutically acceptable salt thereof, wherein the TLR7 inhibitor Or a pharmaceutical composition that is a salt permitted by his pharmaceutical regulations. Claim 2 A pharmaceutical composition according to claim 1, wherein the method comprises administering to the patient a therapeutically effective dose of the TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a therapeutically effective dose of prednisolone or a pharmaceutically acceptable salt thereof. Claim 3 A pharmaceutical composition according to claim 1, wherein the method comprises administering to the patient a therapeutically effective dose of the TLR7 inhibitor or a pharmaceutically acceptable salt thereof in combination with a therapeutically effective dose of hydroxychloroquine or a pharmaceutically acceptable salt thereof. Claim 4 A pharmaceutical composition according to claim 1, wherein the therapeutic effective dose of the TLR7 inhibitor is 0.1 to 100 mg per day. Claim 5 A pharmaceutical composition according to claim 1, wherein the therapeutic effective dose of the prednisolone is 0.5 to 50 mg per day. Claim 6 A pharmaceutical composition according to claim 1, wherein the therapeutic effective dose of the hydroxychloroquine is 1 to 20 mg per day. Claim 7 A pharmaceutical composition according to claim 1, wherein the TLR7 inhibitor and the second agonist are administered jointly. Claim 8 A pharmaceutical composition according to claim 1, wherein the TLR7 inhibitor and the second agonist are administered sequentially. Claim 9 A pharmaceutical composition according to claim 1, wherein the TLR7 inhibitor is administered prior to the administration of the second agonist. Claim 10 A pharmaceutical composition according to claim 1, wherein the second agent is administered prior to the administration of the TLR7 inhibitor. Claim 11 delete Claim 12 delete Claim 13 delete

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