TNF- Α and il-17 inhibitor compounds for the treatment of inflammatory and autoimmune diseases

Abstract

The present invention provides novel small-molecule compounds with tumor necrosis factor-alpha (TNF-α) and interleukin-17 (IL-17) inhibitory properties, intended for oral administration in the treatment of inflammatory and autoimmune diseases, such as psoriasis and rheumatoid arthritis. These compounds selectively inhibit TNF-α and IL-17, key cytokines implicated in chronic inflammation and immune dysregulation. The compounds of this invention overcome these limitations by offering an orally administrable, cost-effective alternative with high specificity and reduced toxicity. The synthetic methods for preparing the compounds, as well as pharmaceutical formulations are also disclosed.

Description

TITLE OF THE INVENTIONTNF- a AND IL- 17 INHIBITOR COMPOUNDS FOR THE TREATMENT OF INFLAMMATORY AND AUTOIMMUNE DISEASES FIELD OF THE INVENTION

[0001] The present invention relates to the field of novel antagonists, specifically to compounds with tumor necrosis factor-alpha (TNF-a) and interleukin- 17 (IL-17) inhibitory properties. These compounds are designed for oral administration in the treatment of inflammatory and autoimmune diseases such as psoriasis and rheumatoid arthritis. The compounds of the present invention specifically targets treatments for inflammatory and autoimmune diseases with a novel, cost-effective, and convenient oral drug formulation.CROSS REFERENCE TO RELATED APPLICATIONS

[0002] Applicant claims priority and the benefit of Indian Patent application 202441088493, filed 15 November 2024 (15-11-2024), said application being hereby incorporated herein in its entirety by referenceBACKGROUND OF THE INVENTION:

[0003] The immune system employs cytokines, a class of small signaling proteins (5-20 kDa), to mediate cellular communication in immune responses, inflammation, and tissue repair. Cytokines are involved in autocrine signaling, paracrine signaling and endocrine signaling as immunomodulating agents. Cytokines include chemokines, interferons, interleukins, lymphokines, and tumor necrosis factors (TNF). Key cytokines include tumor necrosis factor-alpha (TNF-a) and interleukin- 17 (IL- 17), which are involved in immune regulation and are known to play central roles in the pathogenesis of several inflammatory and autoimmune diseases. Excessive productionor dysregulation of TNF-a and IL- 17 can trigger chronic inflammation and contribute to a range of debilitating conditions, including rheumatoid arthritis, psoriasis, ankylosing spondylitis, and Crohn's disease.

[0004] Currently, treatment options for cytokine-related inflammatory diseases primarily involve biologic drugs, specifically monoclonal antibodies, and receptor inhibitors targeting TNF-a and IL-17. These biologies have significantly advanced the management of autoimmune diseases by reducing inflammation and slowing disease progression.

[0005] But anti-cytokine treatment also affects the fundamental protective functions of the body. The anti-cytokine therapy may result in latent microbial infection, demyelinating, secondary autoimmune manifestation, symptoms of lupus erythematosus and inhibition of the cytokine also results in an abundant secretion of interferon a which can lead to thromboembolism. Also, the anti-cytokine therapy is expensive and not every patient can afford this treatment. Another disadvantage with the anti-cytokine therapy is that it uses non-oral (parenteral) route for drug administration which is unsuitable for patients which are uncooperative or cannot use their mouth. The oral drugs do not get absorbed well or absorbs very slowly in patient’s body.

[0006] Most biologies for TNF-a and IL- 17 inhibition are administered intravenously or through subcutaneous injections. This route of administration can be painful, inconvenient, and is typically performed in a clinical setting, leading to increased healthcare costs and reduced patient compliance. Oral administration is generally preferred for long-term treatments, especially for chronic conditions, due to its ease of use. Biologies can interfere with fundamental protective functions of the immune system, increasing the risk of infections and adverse immune reactions. Side effects associated with anti-cytokine therapies include flu-like symptoms, fatigue, thrombocytopenia, and respiratory complications. These side effects impact patient quality of life and may limit the suitability of these therapies for long-term use.Immunogenicity is also a concern, as patients may develop antibodies against biologic drugs, reducing their efficacy over time.

[0007] Given these limitations, there is a clear unmet need for orally administrable, small-molecule inhibitors targeting TNF-a and IL- 17, which could offer safer, more accessible, and convenient therapeutic options. Such drugs could provide similar or enhanced efficacy in reducing inflammation without the drawbacks associated with biologies. Oral administration would increase patient compliance and convenience, while also lowering costs associated with healthcare provider-administered treatments.

[0008] The present invention addresses these gaps by introducing novel smallmolecule compounds with TNF-a and IL- 17 antagonists, suitable for oral administration. These compounds are designed to selectively inhibit cytokine pathways implicated in inflammation, providing a targeted therapeutic effect with reduced side effects. The invention encompasses specific molecules that demonstrate high binding affinity to TNF-a and IL- 17 receptors in preclinical studies, suggesting their potential effectiveness in modulating immune responses associated with chronic inflammatory diseases.

[0009] The compounds show high specificity for TNF-a and IL- 17, which minimizes off-target effects and potential toxicity. Early cell-line studies have indicated nontoxicity at concentrations 50- 100 times the therapeutic dose, supporting a favorable safety profile. In summary, the invention introduces novel oral TNF-a and IL- 17 inhibitors that offer a promising solution to the limitations of existing biologic therapies. This advancement could make cytokine-inhibition therapy more accessible, safer, and convenient, particularly for patients requiring long-term treatment of autoimmune and inflammatory conditions. The unique approach to synthesis, combined with the demonstrated efficacy in cytokine inhibition, establishes it as a significant improvement over existing compound.OBJECT OF INVENTION

[0010] The primary objective of this invention is to develop new small-molecule compounds that selectively inhibit TNF-a and / or IL- 17, offering a targeted treatment for various inflammatory and autoimmune diseases such as rheumatoid arthritis, psoriasis, and ankylosing spondylitis.

[0011] Yet another object of the invention seeks to allow for structural modifications of the compounds to optimize therapeutic efficacy, solubility, and bioavailability, providing flexibility in treating a broader spectrum of inflammatory conditions.SUMMARY OF THE INVENTION

[0012] The invention provides novel small molecules capable of inhibiting TNF-a and / or IL- 17 showing higher affinity and selectivity in binding assays, promising safety profiles, and potential efficacy in treating cytokine-related inflammatory conditions. Unlike existing therapies, these compounds are designed for oral administration, improving patient compliance and reducing healthcare costs.BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWINGS

[0013] The other objects, features and advantages will occur to those skilled in the art from the following description of the preferred embodiment and the accompanying drawings in which:

[0014] Figure 1-5 shows LCMS data of molecule 401

[0015] Figure 6-10 shows LCMS data of molecule 402

[0016] Figure 11-15 shows LCMS data of molecule 403

[0017] Figure 16-20 shows LCMS data of molecule 404

[0018] Figure 21-25 shows LCMS data of molecule 405

[0019] Figure 26-30 shows LCMS data of molecule 406

[0020] Figure 31 shows the binding affinity graph.DETAILED DESCRIPTION OF THE INVENTION

[0021] The present invention may be embodied in several forms, and the details of embodiments of the present invention will be described in the following content with figures. The embodiments described below with reference to the drawings are merely illustrative of the technical solutions of the present disclosure but are not to be construed as limited to the technical solutions of the present disclosure.

[0022] The terms and words used in the following description and claims are not limited to the bibliographical meanings but are merely used by the inventor to enable a clear and consistent understanding of the invention. Accordingly, it should be apparent to those skilled in the art that the following description of the present invention is provided for illustration purposes only and not for the purpose of limiting the invention as defined by the appended claims. As used in the description of the invention and the appended claims, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0023] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the relevant art and will not be interpreted in an idealized or overly formal sense unless expressly so defined herein.

[0024] An aspect of the invention relates to novel small-molecule compounds designed to selectively inhibit TNF-a and / or IL-17, key cytokines involved in inflammatory and autoimmune diseases.

[0025] Another aspect of the invention relates to orally administrable pharmaceutical composition.

[0026] Compounds as TNF-a and / or IL-17 Inhibitors: The present invention includes a set of novel compounds with specific antagonistic effects on TNF-a and / or IL- 17 cytokines. These compounds are represented by Markush structures. Each compound exhibits unique binding characteristics that enable effective modulation of TNF-a and / or IL- 17, which are central to the pathogenesis of various inflammatory and autoimmune diseases. The molecular structures of these compounds allow selective and stable binding to cytokine receptors, minimizing off-target effects which also modulates immune responses with minimal toxicity and improved patient outcomes.

[0027] The Markush structure of the compounds with specific antagonistic effects on TNF-a and / or IL- 17 cytokines are as follows:

[0028] A compound of formula Iwherein R1 and R2 are selected from the group consisting of:

[0029] A compound of formula IIwherein R1 and R2 are selected from the group consisting of:

[0030] A compound of formula III

[0031] A compound of formula IV

[0032] In a specific aspect of the invention the following compounds have antagonistic effects on TNF-a and / or IL-17 cytokines. These compounds are represented by specific examples are labeled as Compounds 401, 402, 403, 404, 405, and 406.Sr, Code Structure IUPAC Name NO1 401 3-benzyl 5-(3- hydroxyphenyl) 2- (chloromethyl)-7-oxo- o1, 5,6,7- tetrahydropyrano[3,2- Cr A no b]pyrrole-3,5- 1 HCl dicarboxylate 2 402 benzyl 2- (chloromethyl)-5- (hydroxy((4methoxyphenyl)amino)methyl)-0)“N\Cr-Wfb 7-oxo-l, 5,6,7- tetrahydropyrano[3,2- b]pyrrole-3- R- OCH3carboxylate403 HO benzyl 2- o(chloromethyl)-5-(((4- a 'Ab 0 ethoxyphenyl)amino)(O Rhydroxy)methyl)-7-ClR-OC₂H₅oxo-1, 5,6,7- tetrahydropyrano[3,2- b]pyrrole-3- carboxylate404 H3CXJJ 9-(2-(2-(lH-indol-5- yl)-4-oxochroman-7- yl)-2-hydroxyethyl)- 1, 3-dimethyl-3, 4,5,9- tetrahy dro- 1 H-purine- 2, 6-dione405 2-(2-(lH-indol-5-yl)- NH24-oxochroman-7-yl)- ° N-(6-amino-2- hydroxy-2,3 -dihydro- ■i IH-inden-l- yl)acetamide

[0033] These compounds, when administered orally, effectively inhibit TNF-a and / or IL- 17, making them suitable for chronic inflammatory and autoimmune conditions such as psoriasis, rheumatoid arthritis, ankylosing spondylitis, Crohn's disease, and ulcerative colitis.SYNTHESIS OF THE COMPOUNDSa. Preparation of Intermediate-A (IM-A) (Figure 1)■ways i- -.5,3, T-Ote&y& BOs? sssfS 4- A fesssta -S-M-L

[0034] Glycine methyl ester hydrochloride and ethyl-4-chloro-3-oxobutanoate are combined as starting materials and undergo cyclization to form a substituted pyrrole compound. The nitrogen of the pyrrole moiety is de-methylated and protected with para-toluenesulfonyl chloride (PTSC). The compound then undergoes cyclization withglyoxal to form ethyl 2-(chloromethyl)-5-formyl-7-oxo-l-tosyl-l, 5,6,7- tetrahydropyrano[3,2-b]pyrrole-3-carboxylate, which is chlorinated to yield Intermediate- A (IM- A).Figure 1b. Synthesis of Compound 401 (Figure 2)

[0035] IM-A is condensed with resorcinol to form the initial structure. This intermediate undergoes benzylation and nitrogen deprotection to yield 3-benzyl 5-(4- hydroxyphenyl) 2-(chloromethyl)-7-oxo-l,5,6,7-tetrahydropyrano[3,2-b]pyrrole- 3,5-dicarboxylate (Compound 401).Figure 2c. Synthesis of Compounds 402 and 403 (Figure 3)

[0036] IM-A is reacted with substituted anilines (e.g., p-Anisidine for Compound 402, and p-Phenetidine for Compound 403) under controlled conditions. After aniline reaction, benzylation and reduction steps are performed. The protecting group is removed to yield the final compounds.d. Synthesis of 404 (Figure 4)

[0037] IM-A on treated with l,3-dimethyl-7-(((trimethylsilyl)oxy)methyl)-3,7- dihydro-l / f-purine-2, 6-dione (synthesis procedure explained in synthetic scheme 404) followed by de-silylation to form title compound 7-(2-(2-(17 / -indol-5-yl)-4- oxochroman-7-yl)-2-hydroxyethyl)-l,3-dimethyl-3,4,5,9-tetra hydro- 1 / f-purine-2,6- dione labeled as 404.Figure 4e. Synthesis of 405 (Figure 5)

[0038] IM-A on treated with A-(6-amino-2-hydroxy-2,3-dihydro-17 / -inden-l-yl)-2- chloroacetamide (synthesis procedure explained in synthetic scheme 405) to form title compound 2-(2-(l / 7-indol-5-yl)-4-oxochroman-7-yl)- / V-(6-amino-2,3-dihydro-l / 7- inden- 1 -yl)acetamide labeled as 405.1-amino-2,3-dihydr 1-amino-6-nitro-2,3-dih A / -(6-amino-2-hydroxy-2,3-dihyd o-1H-inden-2-ol ydro-1 H-inden-2-ol dro-1 H-inden-2-ol ro-1 H-inden-1 -yl)-2-chloroaceta mideA / -(6-amino-2-hydroxy-2,3-dihyd 7-chloro-2-(1H-indol-5-yl)-2,3-di ro-1 H-inden-1 -yl)-2-chloroaceta hydro-4H-chromen-4-one mide(405) (IM-A)Figure 5f. Synthesis of 406 (Figure 6)

[0039] IM-A on treated with formylamino-3,3-dimethyl-7-oxo-4-thia-l- azabicyclo[3.2.0]heptanes-2-carboxylic acid (synthesis procedure explained in synthetic scheme 406) to form title compound 6-(2-(lH-indol-5-yl)-4-oxochromane-7- carboxamido)3,3-dimethyl-7-oxo-4-thia-l-azabicyclo[3.2.0]heptane-2-carboxylic acid labeled as 406.6-amino-3,3-di formaid 6-(formylamino)-3,3-di methyl-7-oxo-4- methyl-7-oxo-4-thia-1- thia-1-azabicycl azabicyclo[3.2.0]hepta o[3.2.0]heptane ne-2-carboxylic acid -2-carboxylic acid Figure 6

[0040] The compounds exhibit high affinity and selectivity toward TNF-a and / or IL- 17, making them suitable for managing autoimmune and inflammatory conditions.

[0041] Stability and Molecular Docking: Molecular docking simulations showed stable interactions in the TNF-a and IL- 17 binding pockets, further confirmed by molecular dynamics, which showed RMSD values indicating stable ligand-protein interaction.

[0042] Analysis of the protein ligand interaction was carried out using the Flare 9.0.0software package by Cresset BioMolecular Discovery Ltd. Molecular dynamics (MD) simulations were performed using Open FF (Version: 2.2.0) calculation method with Explicit TIP3P water solvent model. 10.0A Solvent box with the buffer of 95%water, 5% protein were used in the Truncated Octahedron shape Solvent box. Graph deviation or RMSD between 2 A suggests good stability of ligand interaction in the binding cleft of the protein. Below are the docking and dynamics results of 6 molecules of interest. (Table 1).PHARMACOLOGICAL TESTING AND RESULTS

[0044] Activity Evaluation and Binding Efficiency: Compounds demonstrated high binding affinity in Biolayer Interferometry (BLI) studies with TNF-a and IL- 17, exhibiting dissociation constants (KD). Biolayer Interferometry (BLI) is an optical technique used to measure molecular interactions in real-time binding analysis. It can be applied to study how small molecules (drugs, metabolites, etc.) interact with larger biomolecules, including proteins and nucleic acids. (Table 2 &3).Table 2Tlis below table describes toca&m of samples, sensors, cossceatratioK KD{M) of samples ns 9S well plateTable 3RESULTS SUMMARY OF SAMPLES OF INTEREST WITH KD IN MICROMOLAREDMlSIfcnW AG® Tffi® GKAFrrE GF ASSEK'iKKG® > \ \ sxKt A:Sample Cone.Sensor Type ID Target (µM) ResponseSSA (Super Streptavidin) 401 TNFα 100 0.1823 3.2 SSA (Super Streptavidin) 402 TNFα 100 0.2276 12.3 SSA (Super Streptavidin) 403 TNFα 100 0.1037 26.4 SSA (Super Streptavidin) 404 TNFα 100 1.0816 0.37 SSA (Super Streptavidin) 405 TNFα 100 0.1637 7.9 SSA (Super Streptavidin) 406 TNFα 100 0.2947 65.1 SSA (Super Streptavidin) 401 IL 17 100 0.2023 3.5 SSA (Super Streptavidin) 402 IL 17 100 0.2638 6 SSA (Super Streptavidin) 403 IL 17 100 0.1276 18.8 SSA (Super Streptavidin) 405 IL 17 100 0.1724 9.2SSA (Super Streptavidin) 406 IL 17 100 0.2969 42

[0045] Toxicity Assessment: Cell viability studies using the MTT assay showed acceptable toxicity levels at concentrations from 10 pM (micromolar) to 1000 pM (ImM ), indicating a favorable safety profile for therapeutic use. According to ISO 10993-5, percentages of cell viability above 80% are considered as non-toxic; within 80%-60% mildly toxic; 60%-40% moderately toxic and below 40% strongly toxic respectively. Results are tabulated below. (Table 4)Table 4Cells + Samples + MTT Concentration % proliferation (viability)401 10 pM 94 401 50 pM 86 401 100 pM 83 401 0.5 mM 63 401 1 mM 60 402 10 pM 89 402 50 pM 85402 100 pM 84Cells + Samples + MTT Concentration % proliferation (viability) 402 0.5 mM 60 402 1 mM 51 403 10 μM 93 403 50 μM 83 403 100 μM 75 403 0.5 mM 58 403 1 mM 51 404 10 μM 91 404 50 μM 86 404 100 μM 82 404 0.5 mM 80 404 1 mM 68 405 10 μM 100 405 50 μM 95 405 100 μM 91 405 0.5 mM 79 405 1 mM 73 406 10 μM 85 406 50 μM 81 406 100 μM 78 406 0.5 mM 71406 1 mM 63

[0046] Example 1-Pharmaceutical Formulation (10 mg)The compounds can be formulated as oral tablets or capsules, making them accessible and convenient for long-term patient use.Excipients: Lactose, microcrystalline cellulose, magnesium stearate, and hydroxypropyl methylcellulose.

[0047] Therapeutic Applications: The TNF-a and IL- 17 inhibitors are intended for treating various autoimmune and inflammatory diseases, including but not limited to:a. Psoriasis: Reducing inflammatory response by inhibiting cytokine signaling pathways.b. Rheumatoid Arthritis: Alleviating joint inflammation and associated pain. c. Crohn’s Disease and Ulcerative Colitis: Modulating immune response in inflammatory bowel diseases.

[0048] Each compound is intended for oral administration, with the recommended dose ranging from 5 mg to 100 mg, depending on disease severity, patient age, and specific medical conditions. The formulations are designed for daily administration to maintain therapeutic levels of TNF-a and IL- 17 inhibition and may be taken once or twice daily as needed.

[0049] The compounds of this invention can be modified by incorporating different substituted phenyl or alkyl groups on the pyrrole moiety to further optimize receptor binding and improve pharmacokinetic properties. Such modifications can enhance solubility, bioavailability, and efficacy, allowing for expanded therapeutic use across a broader range of inflammatory conditions.

Claims

CLAIMS1. A compound of formula I or a salt thereof2. A compound of formula II or a salt thereofwherein R1 and R2 are selected from the group consisting of:

3. A compound of formula III or a salt thereof4. A compound of formula IV or a salt thereofwherein R1 and R2 are selected from the group consisting of:

5. A compound as claimed in Claim 1 wherein the compound is3-benzyl 5-(4-hydroxyphenyl) 2-(chloromethyl)-7-oxo-l, 5,6,7- tetrahydropyrano[3,2-b]pyrrole-3,5-dicarboxylate.

6. A Compound as claimed in Claim 2 wherein the compound isR- OCH3benzyl 2-(chloromethyl)-5-(hydroxyl((4-methoxyphenyl)amino)methyl)-7- oxo-l,5,6,7-tetrahydropyrano[3,2-b]pyrrole-3-carboxylate.

7. A Compound as claimed in Claim 2 wherein the compound isbenzyl 2-(chloromethyl)-5-(((4-ethoxyphenyl)amino)(hydroxy)methyl)-7-oxo- l,5,6,7-tetrahydropyrano[3,2-b]pyrrole-3-carboxylate.

8. A Compound as claimed in Claim 3 wherein the compound isoH3CXIH3C7-(2- (2-(lH^ pman-7-yl)-2-hydroxy ethyl)- 1,3 -dimethyl- 3,4,5,9-tetraiydro- 1 H-puriit^2 fl-dione,9. A Compound as claimed in Claim 4 wherein the compound is2-(2-(lH-indol-5-yl)-4-oxochroman-7-yl)-N-(6-amino-2,3-dihydro-lH-inden- l-yl)acetamide.

10. A Compound as claimed in Claim 4 wherein the compound is6-(2-(lH-indol-5-yl)-4-oxochromane-7-carboxamido)-3,3-dimethyl-7-oxo-4- thia-l-azabicyclo[3.2.0]heptane-2-carboxylic acid.

11. The compounds as claimed in Claim 1 -4 wherein the molecules are inhibitors of TNF-a and / or IL-17 cytokines.

12. A pharmaceutical composition comprising a therapeutically effective amount of one or more compounds of Claim 1 -4 or a salt thereof and a pharmaceutically acceptable carrier, wherein the composition is formulated for oral administration as TNF-a and IL-17 inhibitor.

13. A pharmaceutical formulation comprising:a compound as claimed in Claim 1-4 as the active ingredient;one or more excipients selected from lactose, microcrystalline cellulose, magnesium stearate, and hydroxypropyl methylcellulose;wherein the formulation is provided in the form of an oral tablet or capsule.

14. A method for synthesizing a compound of formula as defined in Claims 1-4, comprising:(a) reacting glycine methyl ester hydrochloride with ethyl-4-chloro-3-oxobutanoate to form a substituted pyrrole intermediate;(b) de-methylating and protecting the nitrogen in the pyrrole moiety;(c) cyclizing the intermediate to form a compound selected from Intermediate- A or one of its derivatives; and(d) performing condensation reactions and modifications to yield the final compounds of formula as defined in Claim 1 -4.

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