Cyclic benzimidazole derivatives as cGAS inhibitors

Abstract

The invention relates to compounds of formula I wherein R1, R2, R3, A, D, E, G, J, K and L are as defined in claim 1; as well as prodrugs, deuterated analogs and pharmaceutically acceptable salts thereof; the compounds are useful in the treatment of diseases such as systemic lupus erythematosus, systemic sclerosis (SSc), interferon lesions, non-alcoholic steatohepatitis (NASH), interstitial lung disease (ILD) and idiopathic pulmonary fibrosis (IPF). # imgabs0 #

Description

1. Background Art

[0001] 1.1 cGAS Inhibitors Innate immunity is regarded as a first-line cellular stress response that protects host cells from pathogen invasion and initiates signaling to the adaptive immune system. These processes are triggered by the sensing of conserved pathogen-associated molecular patterns (PAMPs) by different pattern recognition receptors (PRRs) and subsequent activation of cytokine and type I interferon gene expression. Major antigen-presenting cells, such as monocytes, macrophages, and dendritic cells, produce type I interferons and are crucial for initiating adaptive T cell and B cell immune system responses. Major PRRs detect abnormal nucleic acids on the cell surface, inside lysosomal membranes, or in other cellular compartments, i.e., mislocalized, immature, or unmodified nucleic acids (Barbalat et al., Annu. Rev. Immunol. 29, 185-214 (2011)).

[0002] "Cyclic GMP-AMP synthase" ( C yclic G MP- A MP S ynthase, cGAS ; UniProtKB-Q8N884)) is the major sensor for mislocalization or mishandling of abnormal double-stranded DNA (dsDNA) derived from pathogens or mislocalized or mishandled nuclear or mitochondrial cellular dsDNA (Sun et al., Science 339, 786-791 (2013); Wu et al., Science 339, 826-830 (2013); Ablasser et al., Nature 498, 380-384 (2013)). Binding of dsDNA to cGAS activates the reaction of GTP with ATP to form the cyclic dinucleotide GMP-AMP (referred to as cGAMP). Subsequently, cGAMP travels to and activates the endoplasmic reticulum membrane-anchored adaptor protein, "stimulator of interferon genes" ( St imulator of In terferon G ene, STING ). Activated STING recruits and activates TANK-binding kinase 1 ( T ANK- b inding k inase 1 , TBK1 ), which in turn phosphorylates interferon regulatory factor ( i nterferon r egulatory f actor, IRF) phosphorylates transcription factor families, inducing the expression of cytokine and type I interferon mRNAs.

[0003] The key role of cGAS in dsDNA sensing has been established in different pathogenic bacteria (Hansen et al., EMBO J. 33, 1654 (2014)), viruses (Ma et al., PNAS 112, E4306 (2015)), and retroviruses (Gao et al., Science 341, 903 - 906 (2013)). Additionally, cGAS is crucial in various other biological processes, such as cellular senescence (Yang et al., PNAS 114, E4612 (2017), Glück et al., Nat. Cell Biol. 19, 1061 - 1070 (2017)) and the recognition of broken micronuclei during the surveillance of potential cancer cells (Mackenzie et al., Nature 548, 461 - 465 (2017); Harding et al., Nature 548, 466 - 470 (2017)).

[0004] Although the cGAS pathway is crucial for host defense against pathogen invasion, cellular stress and genetic factors can also lead to the production of abnormal cellular dsDNA, for example, through nuclear or mitochondrial leakage, and thereby trigger an autoinflammatory response. Aicardi-Goutieres syndrome (AGS; Crow et al., Nat. Genet. 38, 917-920 (2006)), a lupus-like severe autoinflammatory immune-mediated disorder, is caused by loss-of-function mutations in TREX1, a major DNA exonuclease responsible for degrading abnormal DNA in the cytosol. Knockout of cGAS in TREX1-deficient mice prevents the otherwise lethal autoimmune response, thus supporting cGAS as a driver of interferonopathies (Gray et al., J. Immunol. 195, 1939-1943 (2015); Gao et al., PNAS 112, E5699-E5705 (2015)). Similarly, the embryonic lethality caused by the deficiency of DNAse2, an endonuclease responsible for degrading excess DNA in lysosomes during endocytosis, is completely rescued by additional gene knockout of cGAS (Gao et al., PNAS 112, E5699-E5705 (2015)) or STING (Ahn et al., PNAS 109, 19386-19391 (2012)). These observations support cGAS as a drug target, and inhibition of cGAS may provide a therapeutic strategy for preventing autoinflammation and treating diseases involving anti-dsDNA antibodies such as systemic lupus erythematosus (SLE) (Pisetsky et al., Nat. Rev. Rheumatol. 12, 102-110 (2016)).

[0005] 1.2 Prior Art Since it has been observed that inhibition of the cGAS pathway can provide a therapeutic strategy for preventing autoinflammation and treating autoimmune diseases, many efforts have been made to develop cGAS inhibitors.

[0006] For example, in WO 2019 / 241787, methyl 4-amino-6-(phenylamino)-1,3,5-triazine-2-carboxylate, such as CU-32 and CU-76, are disclosed as cGAS inhibitors, where the "in vitro hcGAS IC 50 value" is slightly below 1 μM (IC 50 (CU-32) = 0.66 μM and IC 50 (CU-76 = 0.27 μM).

[0007] In Hall et al., PLoS ONE 12(9); e0184843 (2017), the compound PF-06928215 has been disclosed as a cGAS inhibitor, where the "in vitro hcGAS IC 50 value" measured by fluorescence polarization is 0.049 μM. However, the compound PF-06928215 does not exhibit acceptable cellular activity as a cGAS inhibitor.

[0008] In WO 2020 / 142729 and WO2022 / 174012, (benzofuro[3,2-d]pyrimidin-4-yl)pyrrolidine-2-carboxylic acid derivatives have been disclosed as cGAS inhibitors for the treatment of autoimmune diseases such as Aicardi-Goutières syndrome (AGS), lupus erythematosus, scleroderma, inflammatory bowel disease, and non-alcoholic steatohepatitis (NASH). However, the compounds of the present invention are different from the (benzofuro[3,2-d]pyrimidin-4-yl)pyrrolidine-2-carboxylic acid derivatives of WO 2020 / 142729, and their substitution patterns at the 4-position of the pyrrolidine ring are completely different.

[0009] Recently provided cGAS inhibitors, such as the cGAS inhibitors in WO 2020 / 142729 or WO 2022 / 174012, generally exhibit insufficient cellular cGAS inhibitory potency (where the IC 50 value measured in cell assays for inhibiting the cGAS / STING pathway is generally greater than 1 μM, often greater than 5 μM). However, it is crucial to provide a therapeutic cGAS inhibitor that not only exhibits satisfactory biochemical (in vitro) inhibitory potency ("hcGAS IC 50 "), but also exhibits satisfactory cellular inhibitory potency (e.g., by showing inhibition of IFN induction in virus-stimulated THP-1 cells (THP1 (vir) IC 50 )) to ensure that the compound can exhibit a therapeutic effect in patients. Other important properties predictable for the successful development of a cGAS inhibitor as a therapeutic agent are satisfactory cGAS selectivity (versus off-target activity) and acceptable inhibitory potency in human whole blood.

[0010] Surprisingly, it has now been found that the compounds of formula I, II, or III simultaneously exhibit the following three properties: · Satisfactory "biochemical (in vitro) IC 50 value for cGAS inhibition" (where hcGAS IC 50 ≤ 100 nM, preferably ≤ 50 nM, especially ≤ 10 nM), · Satisfactory "IFN induction in THP-1 cells stimulated by virus is inhibited" (wherein THP1 IC 50(vir) ≤1 μM, preferably ≤500 nM, more preferably ≤100 nM, especially ≤50 nM) and · Satisfactory selectivity for cGAS inhibition (wherein THP1 IC 50(cGAMP) / THP1 IC 50(vir) ratio ≥10, more preferably ≥50, more preferably ≥500, especially ≥1000).

[0011] In addition, the compounds of formula I, II or III also exhibit acceptable IC 50 values for inhibiting IFN induction in a dsNAb-stimulated human whole blood assay, preferably wherein the human whole blood IC 50 value for cGAS inhibition (hWB IC 50 ) ≤5000 nM, more preferably ≤1000 nM, especially ≤100 nM.

[0012] The cGAS inhibitors of the present invention having this specific pharmacological profile, combined with excellent in vitro inhibitory potency and excellent cell inhibitory potency as well as high selectivity for cGAS inhibition, are very likely to also exhibit good therapeutic effects in patients. Due to their high cell inhibitory potency, the compounds having this specific pharmacological profile should be able to cross the cell membrane barrier and thereby reach their intracellular target positions, and due to their selectivity for specifically inhibiting cGAS activity, these compounds should not exhibit unwanted off-target effects, such as producing side effects or cytotoxic effects somewhere downstream of the signal transduction pathway of cGAS. 2. Summary of the Invention

[0013] The present invention relates to compounds of formula I wherein R 1 is selected from the group consisting of hydrogen, halogen, methyl, ethyl, -CF3, -CHF2, CFH2 and methoxy, R 2 is selected from the group consisting of hydrogen and methyl, R 3 is selected from the group consisting of hydrogen, methyl and halogen, and wherein A is selected from the group consisting of -CH2-, -O-, -CF2-, -CHF-, -N(CH3)-, -NH- and -CHCH3-; D is selected from the group consisting of -CH2-, -O-, -CF2-, -CHF- and -CHCH3-; E is selected from the group consisting of: -CH2-, -O-, -C(CH3)2-, -CHF-, CF2- and -CHCH3-; G is selected from the group consisting of: -NH-, -NCH3-, -CH2-, -O-, -CF2-, -CHF-, -CHCH3- and -C(CH3)2; J is selected from the group consisting of: -CO-, -CH2-, -O-, -CHF-, -CF2- and -CHCH3-; K is selected from the group consisting of -CH2-, -CF2-, -O- or is absent; L is selected from the group consisting of -CH2-, -O-, -CHCH3-, -CHF-, -CF2- or is absent; and their prodrugs, deuterated analogs and pharmaceutically acceptable salts.

[0014] Herein, the variables A, D, E, G, J, K and L are preferably selected in such a way that two or more heteroatoms do not directly follow each other.

[0015] In a preferred embodiment, the present invention relates to the compounds of formula I mentioned above, wherein R 1 is selected from the group consisting of hydrogen, Cl and F, R 2 is selected from the group consisting of hydrogen and methyl, R 3 is selected from the group consisting of: hydrogen, methyl, Cl and F, and wherein A is selected from the group consisting of: -CH2-, -O-, -CF2-, -N(CH3)-, D is selected from the group consisting of -CH2-, -O- and -CHCH3-, E is selected from the group consisting of -CH2-, -O- and -C(CH3)2-, G is selected from the group consisting of: -NH-, -CH2-, -O-, -CHCH3-, J is selected from the group consisting of: -CO-, -CH2-, -O- and -CHCH3-; K is selected from the group consisting of -CH2-, -CF2-, -O- or is absent; L is selected from the group consisting of -CH2-, -CF2- or is absent; and their prodrugs, deuterated analogs and pharmaceutically acceptable salts.

[0016] Herein, the variables A, D, E, G, J, K, and L are preferably selected in such a way that two or more heteroatoms do not directly follow each other.

[0017] In another preferred embodiment, the present invention relates to a compound of formula I as mentioned above, wherein L is absent; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0018] In another preferred embodiment, the present invention relates to a compound of formula I as mentioned above, wherein L is absent, and wherein A is selected from the group consisting of -CH2- and -CF2-; and its prodrugs, pharmaceutically acceptable salts, or deuterated analogs thereof.

[0019] In another preferred embodiment, the present invention relates to a compound of formula I as mentioned above, wherein L is absent, and wherein K is CF2; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0020] In another preferred embodiment, the present invention relates to a compound of formula I as mentioned above, wherein R 3 is a halogen; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0021] Herein, particularly preferably, R 3 is a halogen atom selected from the group consisting of Cl and F.

[0022] In another preferred embodiment, the present invention relates to a compound of formula I as mentioned above, wherein R 3 is Cl or F and is located at the 5-position of the benzimidazole moiety; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0023] In another preferred embodiment, the present invention relates to a compound of formula I as mentioned above, wherein R 1 is a halogen; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0024] In another preferred embodiment, the present invention relates to a compound of formula I as mentioned above, wherein R 1 is Cl or F; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0025] In another preferred embodiment, the present invention relates to a compound of formula I as mentioned above, wherein R 1 is hydrogen; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0026] In another particularly preferred embodiment, the present invention relates to a compound of formula I as mentioned above, which is selected from the group consisting of: and their prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0027] In another preferred embodiment, the present invention relates to a compound of formula I as mentioned above, wherein A is selected from the group consisting of -CH2- and -CF2-, D and E are each -CH2-, G is selected from the group consisting of -CH2- and -O-, J is selected from the group consisting of -CH2- and -O-, K is selected from the group consisting of -CF2- and -CH2-, and L is absent, and their prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0028] In another particularly preferred embodiment, the present invention relates to a compound of formula I as mentioned above, which is selected from the group consisting of: and their prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0029] In another preferred embodiment, the present invention relates to a compound of formula I as mentioned above, wherein R 2 is methyl; and their prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0030] In another particularly preferred embodiment, the present invention relates to a compound of formula II or a compound of formula III wherein R 1 is selected from the group consisting of hydrogen and halogen, R 3 is selected from the group consisting of hydrogen, methyl, and halogen, and wherein A is selected from the group consisting of: -CH2-, -O-, -CF2-, -CHF-, -N(CH3)-, -NH-, and -CHCH3-; D is selected from the group consisting of: -CH2-, -O-, -CF2-, -CHF-, and -CHCH3-; E is selected from the group consisting of: -CH2-, -O-, -C(CH3)2-, -CHF-, CF2-, and -CHCH3-; G is selected from the group consisting of: -NH-, -NCH3-, -CH2-, -O-, -CF2-, -CHF-, -CHCH3- and -C(CH3)2; J is selected from the group consisting of: -CO-, -CH2-, -O-, -CHF-, -CF2- and -CHCH3-; K is selected from the group consisting of -CH2-, -CF2-, -O- or is absent; L is selected from the group consisting of -CH2-, -O-, -CHCH3-, -CHF-, -CF2- or is absent; and its prodrugs, deuterated analogs and pharmaceutically acceptable salts.

[0031] Herein, the variables A, D, E, G, J, K and L are preferably selected in such a way that two or more heteroatoms do not directly follow each other.

[0032] More preferably, it is a compound of formula II or formula III mentioned above, wherein R 1 is selected from the group consisting of hydrogen, Cl and F, R 3 is selected from the group consisting of hydrogen, methyl, Cl and F, and wherein A is selected from the group consisting of -CH2-, -O-, -CF2- and -N(CH3)-, D is selected from the group consisting of -CH2-, -O- and -CHCH3-, E is selected from the group consisting of -CH2-, -O- and -C(CH3)2-, G is selected from the group consisting of -NH-, -CH2-, -O-, -CHCH3-, J is selected from the group consisting of -CO-, -CH2-, -O- and -CHCH3-; K is selected from the group consisting of -CH2-, -CF2-, -O- or is absent; L is selected from the group consisting of -CH2-, -CF2- or is absent; and its prodrugs, deuterated analogs and pharmaceutically acceptable salts.

[0033] Herein, it is preferred that the variables A, D, E, G, J, K and L are selected in such a way that two or more heteroatoms do not directly follow each other.

[0034] Particularly preferably, it is the compound of formula II mentioned above, wherein R 1 is selected from the group consisting of hydrogen, Cl and F, R 3 is selected from the group consisting of hydrogen, methyl, Cl and F, and wherein A is selected from the group consisting of -CH2-, -O-, -CF2- and -N(CH3)-, D is selected from the group consisting of -CH2-, -O- and -CHCH3-, E is selected from the group consisting of -CH2-, -O- and -C(CH3)2-, G is selected from the group consisting of -NH-, -CH2-, -O-, -CHCH3-, J is selected from the group consisting of -CO-, -CH2-, -O- and -CHCH3-; K is selected from the group consisting of -CH2-, -CF2-, -O- or is absent; L is selected from the group consisting of -CH2-, -CF2- or is absent; and prodrugs, deuterated analogs and pharmaceutically acceptable salts thereof.

[0035] Herein, the variables A, D, E, G, J, K and L are preferably selected such that two or more heteroatoms do not directly follow each other.

[0036] In another preferred embodiment, the present invention relates to a compound of formula II or a compound of formula III as mentioned above, wherein L is absent; and prodrugs, deuterated analogs and pharmaceutically acceptable salts thereof.

[0037] In another preferred embodiment, the present invention relates to a compound of formula II or a compound of formula III as mentioned above, wherein L is absent and wherein K is CF2; and prodrugs, deuterated analogs and pharmaceutically acceptable salts thereof.

[0038] In another preferred embodiment, the present invention relates to a compound of formula II or a compound of formula III as mentioned above, wherein R 3 is a halogen; and prodrugs, deuterated analogs and pharmaceutically acceptable salts thereof.

[0039] In another preferred embodiment, the present invention relates to a compound of formula II or a compound of formula III as mentioned above, wherein R 3 is Cl or F; and prodrugs, deuterated analogs and pharmaceutically acceptable salts thereof.

[0040] In a particularly preferred embodiment, the present invention relates to a compound of formula II or a compound of formula III as mentioned above, wherein R 3 is Cl or F and is located at the 5-position of the benzimidazole moiety; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0041] In another preferred embodiment, the present invention relates to a compound of formula II or a compound of formula III as mentioned above, wherein R 3 is hydrogen; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0042] In another preferred embodiment, the present invention relates to a compound of formula II or a compound of formula III as mentioned above, wherein R 1 is a halogen; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0043] In a particularly preferred embodiment, the present invention relates to a compound of formula II or a compound of formula III as mentioned above, wherein R 1 is selected from the group consisting of Cl or F; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0044] In another preferred embodiment, the present invention relates to a compound of formula II or a compound of formula III as mentioned above, wherein R 1 is hydrogen; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0045] In another preferred embodiment, the present invention relates to a compound of formula II or a compound of formula III as mentioned above, wherein A is selected from the group consisting of -CH2- and -CF2-, D and E are each -CH2-, G is selected from the group consisting of -CH2- and -O-, J is selected from the group consisting of -CH2- and -O-, K is selected from the group consisting of -CF2- and -CH2-, and L is absent, and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0046] In another particularly preferred embodiment, the present invention relates to a compound of formula II as mentioned above, which is selected from the group consisting of: and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

[0047] In another particularly preferred embodiment, the present invention relates to a compound of formula II or a compound of formula III as mentioned above, wherein A is selected from the group consisting of -CH2- and -CF2- D, E and G are each -CH2- J is selected from the group consisting of -CH2- and -O- K is -CF2- and L is absent and their prodrugs, deuterated analogs and pharmaceutically acceptable salts.

[0048] In another particularly preferred embodiment, the present invention relates to a compound of formula II as mentioned above, which is selected from the group consisting of: and their prodrugs, deuterated analogs and pharmaceutically acceptable salts.

[0049] The prodrug of the compound of formula I is preferably a compound of formula Ia wherein the variable R 1 、R 2 、R 3 、A, D, E, G, J, K and L are as defined above, and wherein R 4 is C 1-4 -alkyl, aryl, -CH2-aryl, NH-SO2-C 1-3 -alkyl.

[0050] Particularly preferred is the prodrug of formula Ia, wherein the variables R 1 、R 2 、R 3 、A, D, E, G, J, K and L are as defined above, and wherein R 4 is methyl.

[0051] The prodrug of the compound of formula II is preferably a compound of formula IIa wherein the variables R 1 、R 2 、R 3 、A, D, E, G, J, K and L are as defined above, and wherein R 4 is C 1-4 -alkyl, aryl, -CH2-aryl, NH-SO2-C 1-3 -alkyl.

[0052] Particularly preferred are prodrugs of formula IIa, wherein the variables R 1 、R 2 、R 3 、A, D, E, G, J, K and L are as defined above, and wherein R 4 is methyl.

[0053] The prodrug of the compound of formula III is preferably a compound of formula IIIa wherein the variables R 1 、R 2 、R 3 、A, D, E, G, J, K and L are as defined above, and wherein R 4 is C 1-4 -alkyl, aryl, -CH2-aryl, NH-SO2-C 1-3 -alkyl.

[0054] Particularly preferred are prodrugs of formula IIIa, wherein the variables R 1 、R 2 、R 3 、A, D, E, G, J, K and L are as defined above, and wherein R 4 is methyl.

[0055] In another preferred embodiment, the present invention relates to a) an intermediate compound of formula (A-I) wherein R 1 、R 2 、R 3 、A, D, E, G, J, K and L are as defined above, and wherein R13 is selected from the group consisting of hydrogen, methyl, ethyl and tert-butyl, b) an intermediate compound of formula (A-II), wherein R 1 、R 2 、R 3 、A, D, E, G, J, K and L are as defined above, and wherein R13 is selected from the group consisting of hydrogen, methyl, ethyl and tert-butyl, and wherein R is hydrogen or a protecting group selected from the group consisting of tert-butyl, methyl, ethyl and benzyl, c) an intermediate compound of formula (B-I) wherein R 1 、R 2 、R 3, A, D, E, G, J, K and L are as defined above, and wherein R13 is selected from the group consisting of hydrogen, methyl, ethyl and tert-butyl, or d) Intermediate compound of formula (B-II) Where R 1 , R 2 , R 3 , A, D, E, G, J, K and L are as defined above, and wherein R13 is selected from the group consisting of hydrogen, methyl, ethyl and tert-butyl.

[0056] In another preferred embodiment, the present invention relates to a compound of formula I, II or III or a prodrug of formula Ia, IIa or IIIa, a deuterated analogue and a pharmaceutically acceptable salt thereof as mentioned above for use in treating a disease that can be treated by inhibiting cGAS.

[0057] In another preferred embodiment, the present invention relates to a compound of formula I, II or III or a prodrug of formula Ia, IIa or IIIa, a deuterated analogue and a pharmaceutically acceptable salt thereof, for use in treating a disease selected from the group consisting of: systemic lupus erythematosus (SLE); interferonopathy; Acardi-Guterres syndrome (AGS); COPA syndrome; familial pernio lupus; age-related macular degeneration (AMD); retinopathy; glaucoma; amyotrophic lateral sclerosis (ALS); diabetes; obesity; inflammatory bowel disease (IBD); chronic obstructive pulmonary disease (COPD); Bloom's syndrome; Sjogren's syndrome (Sjogren's syndrome); Parkinson's disease; heart failure; cancer; systemic sclerosis (SSc); dermatomyositis; nonalcoholic steatohepatitis (NASH); interstitial lung disease (ILD), preferably progressive fibrosing interstitial lung disease (PF-ILD), in particular idiopathic pulmonary fibrosis (IPF); aging; muscle disorders; sepsis; rheumatoid arthritis; osteoarthritis; and COVID-19.

[0058] In another preferred embodiment, the present invention relates to a compound of formula I, II or III or a prodrug of formula Ia, IIa or IIIa, a deuterated analogue thereof and a pharmaceutically acceptable salt thereof, for use in treating a disease selected from the group consisting of systemic lupus erythematosus (SLE), interferonopathy, Acardi-Guterres syndrome (AGS), COPA syndrome, familial pernio lupus, dermatomyositis, age-related macular degeneration (AMD), amyotrophic lateral sclerosis (ALS), inflammatory bowel disease (IBD), chronic obstructive pulmonary disease (COPD), Bloom's syndrome, Sjögren's syndrome, rheumatoid arthritis and Parkinson's disease.

[0059] In another preferred embodiment, the present invention relates to a compound of formula I, II or III as mentioned above or a prodrug of formula Ia, IIa or IIIa, a deuterated analogue thereof and a pharmaceutically acceptable salt thereof for use in treating a disease selected from the group consisting of: systemic sclerosis (SSc); non-alcoholic fatty hepatitis (NASH); interferon pathology; interstitial lung disease (ILD), preferably progressive fibrosing interstitial lung disease (PF-ILD), especially idiopathic pulmonary fibrosis (IPF).

[0060] In another preferred embodiment, the present invention relates to a compound of formula I, II or III as mentioned above, or a prodrug of formula Ia, IIa or IIIa, a deuterated analogue thereof, and a pharmaceutically acceptable salt thereof for use in treating a disease selected from the group consisting of age-related macular degeneration (AMD), retinopathy, glaucoma, diabetes, obesity, aging, muscle disorders, sepsis, osteoarthritis, heart failure, COVID19 / SARS-CoV-2 infection, renal inflammation, renal fibrosis, metabolic disorders, vascular diseases, cardiovascular diseases, and cancer.

[0061] In another preferred embodiment, the present invention relates to a pharmaceutical composition comprising the above-mentioned compound of formula I, II or III or prodrug of formula Ia, IIa or IIIa, deuterated analogs and pharmaceutically acceptable salts thereof and one or more optional pharmaceutically acceptable carriers and / or excipients.

[0062] In another preferred embodiment, the present invention relates to a pharmaceutical composition comprising a compound of formula I, II or III or a prodrug of formula Ia, IIa or IIIa, a deuterated analogue thereof and a pharmaceutically acceptable salt, as mentioned above, and one or more active agents selected from the group consisting of: anti-inflammatory agents; anti-fibrotic agents; anti-allergic agents / anti-histamines; bronchodilators; β2 agonists / β mimetics; adrenergic agonists; anti-cholinergics; methotrexate; mycophenolate mofetil; leukotriene regulators; JAK inhibitors; anti-interleukin antibodies; non-specific immunotherapeutic agents such as interferons or other cytokines / chemokines; cytokine / chemokine receptor regulators; toll-like receptor agonists; immune checkpoint regulators; anti-TNF antibodies such as Humira TM ; anti-BAFF antibodies such as belimumab and etanercept; and one or more optional pharmaceutically acceptable carriers and / or excipients.

[0063] In another preferred embodiment, the present invention relates to a pharmaceutical composition comprising a compound of formula I, II or III or a prodrug of formula Ia, IIa or IIIa, a deuterated analogue thereof and a pharmaceutically acceptable salt, as mentioned above, and one or more anti-fibrotic agents selected from the group consisting of pirfenidone and nintedanib and one or more optional pharmaceutically acceptable carriers and / or excipients.

[0064] In another preferred embodiment, the present invention relates to a pharmaceutical composition comprising a compound of formula I, II or III or a prodrug of formula Ia, IIa or IIIa, a deuterated analogue thereof and a pharmaceutically acceptable salt, as mentioned above, and one or more anti-inflammatory agents selected from the group consisting of NSAIDs and corticosteroids and one or more optional pharmaceutically acceptable carriers and / or excipients.

[0065] In another preferred embodiment, the present invention relates to a pharmaceutical composition comprising a compound of formula I, II or III or a prodrug of formula Ia, IIa or IIIa, a deuterated analogue thereof and a pharmaceutically acceptable salt, as mentioned above, and one or more active agents selected from the group consisting of bronchodilators, β2 agonists / β mimetics, adrenergic agonists and anti-cholinergics, and one or more optional pharmaceutically acceptable carriers and / or excipients.

[0066] In another preferred embodiment, the present invention relates to a pharmaceutical combination comprising a compound of formula I, II or III as mentioned above, or a prodrug of formula Ia, IIa or IIIa, its deuterated analogue and a pharmaceutically acceptable salt, and one or more anti - interleukin antibodies selected from the group consisting of: anti - IL - 23 antibody, such as Risankizumab; anti - IL - 17 antibody; anti - IL - 1 antibody; anti - IL - 4 antibody; anti - IL - 13 antibody; anti - IL - 5 antibody; anti - IL - 6 antibody, such as Actemra TM ; anti - IL - 12 antibody; and anti - IL - 15 antibody.

[0067] 3. Terms and Definitions Used Unless otherwise specified, all substituents are independent of each other. For example, if multiple C 1-6 alkyl groups are possible substituents on a group, then in the case of, for example, three substituents, the C 1-6 alkyl groups can independently represent methyl, n - propyl and tert - butyl.

[0068] A cross - bond, such as the middle bond in the following butyl molecule, represents a double bond of unknown configuration (cis, trans or a mixture thereof).

[0069] The term "C 1-6 alkyl" (including those alkyl groups as part of other groups) means a branched and unbranched alkyl group having 1 to 6 carbon atoms, and the term "C 1-3 alkyl" means a branched and unbranched alkyl group having 1 to 3 carbon atoms. "C 1-4 alkyl" thus represents a branched and unbranched alkyl group having 1 to 4 carbon atoms. Preferably it is an alkyl group having 1 to 4 carbon atoms. Examples of these alkyl groups include methyl, ethyl, n - propyl, isopropyl, n - butyl, isobutyl, sec - butyl, tert - butyl, n - pentyl, isopentyl, neopentyl and hexyl. The groups mentioned above may optionally also use abbreviations such as Me, Et, n - Pr, i - Pr, n - Bu, i - Bu, t - Bu, etc. Unless otherwise specified, the definitions of propyl, butyl, pentyl and hexyl include all possible isomeric forms of said groups. Thus, for example, propyl includes n - propyl and isopropyl, and butyl includes isobutyl, sec - butyl and tert - butyl, etc.

[0070] The term "C 1-6 alkylene" (including those alkylene groups as part of other groups) means a branched and unbranched alkylene group having 1 to 6 carbon atoms, and the term "C 1-4"Alkylene" means a branched and unbranched alkylene having 1 to 4 carbon atoms. Preferably, it is an alkylene having 1 to 4 carbon atoms. Examples of these alkylenes include methylene, ethylene, propylene, 1-methylethylene, butylene, 1-methylpropylene, 1,1-dimethylethylene, 1,2-dimethylethylene, pentylene, 1,1-dimethylpropylene, 2,2-dimethylpropylene, 1,2-dimethylpropylene, 1,3-dimethylpropylene, and hexylene. Unless otherwise specified, the definitions of propylene, butylene, pentylene, and hexylene include all possible isomeric forms of the groups having the same number of carbon atoms. Thus, for example, propylene also includes 1-methylethylene and butylene includes 1-methylpropylene, 1,1-dimethylethylene, 1,2-dimethylethylene, etc.

[0071] If the carbon chain is substituted by a group that forms a carbocyclic ring having 3, 5, or 6 carbon atoms together with one or two carbon atoms of the alkylene chain, it particularly includes the following ring examples:

[0072] The term "C 2-6 alkenyl" (including those alkenyls that are part of other groups) means a branched and unbranched alkenyl having 2 to 6 carbon atoms, and the term "C 2-4 alkenyl" means a branched and unbranched alkenyl having 2 to 4 carbon atoms, provided that it has at least one double bond. Preferably, it is an alkenyl having 2 to 4 carbon atoms. Examples include: ethenyl or vinyl, propenyl, butenyl, pentenyl, or hexenyl. Unless otherwise specified, the definitions of propenyl, butenyl, pentenyl, and hexenyl include all possible isomeric forms of the groups. Thus, for example, propenyl includes 1-propenyl and 2-propenyl, and butenyl includes 1-butenyl, 2-butenyl, and 3-butenyl, 1-methyl-1-propenyl, 1-methyl-2-propenyl, etc.

[0073] The term "C 2-5 alkynyl" (including those alkynyls that are part of other groups) means a branched and unbranched alkynyl having 2 to 5 carbon atoms, and the term "C 2-4 alkynyl" means a branched and unbranched alkynyl having 2 to 4 carbon atoms, provided that it has at least one triple bond. Preferably, it is an alkynyl having 2 to 4 carbon atoms.

[0074] The term "C 2-6 alkenylene" (including those alkenylenes that are part of other groups) means a branched and unbranched alkenylene having 2 to 6 carbon atoms, and the term "C 2-4"Alkenylene" means a branched and unbranched alkylene group having 2 to 4 carbon atoms. An alkenylene group having 2 to 4 carbon atoms is preferred. Examples of these alkenylene groups include: vinylidene, propenylene, 1-methylvinylidene, butenylene, 1-methylpropenylene, 1,1-dimethylvinylidene, 1,2-dimethylvinylidene, pentenylene, 1,1-dimethylpropenylene, 2,2-dimethylpropenylene, 1,2-dimethylpropenylene, 1,3-dimethylpropenylene, and hexenylene. Unless otherwise stated, the definitions of propenylene, butenylene, pentenylene, and hexenylene include all possible isomeric forms of the groups having the same number of carbon atoms. Thus, for example, propenyl also includes 1-methylvinylidene and butenyl includes 1-methylpropenylene, 1,1-dimethylvinylidene, 1,2-dimethylvinylidene.

[0075] The term "aryl" (including aryl as part of other groups) means an aromatic ring system having 6 or 10 carbon atoms. Examples include phenyl or naphthyl, and preferably the aryl is phenyl. Unless otherwise stated, the aromatic group may be substituted with one or more groups selected from: methyl, ethyl, isopropyl, tert-butyl, hydroxy, fluorine, chlorine, bromine, and iodine.

[0076] The term "aryl-C 1-6 alkylene" (including those groups as part of other groups) means a branched and unbranched alkylene group having 1 to 6 carbon atoms, which is substituted with an aromatic ring system having 6 or 10 carbon atoms. Examples include benzyl, 1-phenylethyl or 2-phenylethyl, and 1-naphthylethyl or 2-naphthylethyl. Unless otherwise stated, the aromatic group may be substituted with one or more groups selected from: methyl, ethyl, isopropyl, tert-butyl, hydroxy, fluorine, chlorine, bromine, and iodine.

[0077] The term "heteroaryl-C 1-6 alkylene" (including those groups as part of other groups), even if it is already included under "aryl-C 1-6 alkylene", means a branched and unbranched alkylene group having 1 to 6 carbon atoms substituted with a heteroaryl group.

[0078] Unless otherwise specifically defined, such heteroaryl groups include five- or six-membered heteroaromatic groups or 5- to 10-membered bicyclic heteroaryl rings, which may contain one, two, three, or four heteroatoms selected from oxygen, sulfur, and nitrogen, and contain many conjugated double bonds forming an aromatic system. The following are examples of five- or six-membered heteroaromatic groups and bicyclic heteroaryl rings:

[0079] Unless otherwise indicated, these heteroaryls may be substituted by one or more groups selected from: methyl, ethyl, isopropyl, tert-butyl, hydroxy, amino, nitro, alkoxy, fluorine, chlorine, bromine, and iodine.

[0080] The following are heteroaryl-C 1-6 Examples of alkylene:

[0081] The term "C 1-6 haloalkyl" (including those haloalkyls that are part of other groups) means a branched or unbranched alkyl having 1 to 6 carbon atoms substituted by one or more halogen atoms. The term "C 1-4 haloalkyl" means a branched or unbranched alkyl having 1 to 4 carbon atoms substituted by one or more halogen atoms. It is preferably an alkyl having 1 to 4 carbon atoms. Examples include: CF3, CHF2, CH2F, CH2CF3.

[0082] Unless otherwise specifically defined, the term "C 3-7 cycloalkyl" (including those cycloalkyls that are part of other groups) means a cycloalkyl having 3 to 7 carbon atoms. Examples include: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. Unless otherwise indicated, the cycloalkyl may be substituted by one or more groups selected from: methyl, ethyl, isopropyl, tert-butyl, hydroxy, fluorine, chlorine, bromine, and iodine.

[0083] Unless otherwise specifically defined, the term "C 3-10 cycloalkyl" also means a monocycloalkyl having 3 to 7 carbon atoms and also means a bicycloalkyl having 7 to 10 carbon atoms, or a monocycloalkyl bridged by at least one C 1-3 carbon bridge.

[0084] Unless otherwise indicated, the term "heterocyclic ring" or "heterocycle" means a five-, six-, or seven-membered saturated, partially saturated, or unsaturated heterocycle that may contain one, two, or three heteroatoms selected from oxygen, sulfur, and nitrogen, and if there is one, the ring may be attached to the molecule through a carbon atom or through a nitrogen atom. Although included by the term "heterocyclic ring" or "heterocycle", the term "saturated heterocycle" refers to a five-, six-, or seven-membered saturated ring. Examples include:

[0085] Although included within the terms "heterocyclic" or "heterocyclic group", the term "partially saturated heterocyclic group" refers to a five-, six- or seven-membered partially saturated ring which contains one or two double bonds but not so many as to form an aromatic system, unless otherwise specifically defined. Examples include:

[0086] Although included within the terms "heterocyclic ring" or "heterocycle", unless otherwise specifically defined, the terms "heteroaromatic ring", "unsaturated heterocyclic group" or "heteroaryl" refer to a five- or six-membered heteroaromatic group or a 5- to 10-membered bicyclic heteroaryl ring which may contain one, two, three or four heteroatoms selected from oxygen, sulfur and nitrogen and which contains a number of conjugated double bonds sufficient to form an aromatic system. Examples of five- or six-membered heteroaromatic groups include:

[0087] Unless otherwise mentioned, a heterocyclic ring / heterocycle may have a ketone group. Examples include:

[0088] Although encompassed by the term "cycloalkyl", the term "bicyclic cycloalkyl" generally denotes an eight-, nine- or ten-membered bicyclic carbocyclic ring. Examples include:

[0089] Although included within the term "heterocyclic", unless otherwise specifically defined, the term "bicyclic heterocycle" generally denotes an eight-, nine- or ten-membered bicyclic ring which may contain one or more heteroatoms, preferably 1 to 4, more preferably 1 to 3, even more preferably 1 to 2, especially one heteroatom selected from oxygen, sulfur and nitrogen. The ring may be attached to the molecule through a carbon atom in the ring or through a nitrogen atom in the ring (if there is one). Examples include:

[0090] Although included within the term "aryl", the term "bicyclic aryl" denotes a 5- to 10-membered bicyclic aryl ring which contains conjugated double bonds sufficient to form an aromatic system. One example of a bicyclic aryl is naphthyl.

[0091] Although included under "heteroaryl", unless otherwise specifically defined, the term "bicyclic heteroaryl" denotes a 5-10 membered bicyclic heteroaryl ring which may contain one, two, three or four heteroatoms selected from oxygen, sulfur and nitrogen and which contains conjugated double bonds sufficient to form an aromatic system.

[0092] Although included within the terms "bicyclic cycloalkyl" or "bicyclic aryl", the terms "fused cycloalkyl" or "fused aryl" denote bicyclics in which the bridge separating the rings represents a direct single bond. Examples of fused bicyclic cycloalkyls are as follows:

[0093] Although included within the terms "bicyclic heterocycle" or "bicyclic heteroaryl", the terms "fused bicyclic heterocycle" or "fused bicyclic heteroaryl" denote 5- to 10-membered bicyclic heterocycles containing one, two, three, or four heteroatoms selected from oxygen, sulfur, and nitrogen, and in which the bridge separating the rings represents a direct single bond. "Fused bicyclic heteroaryl" additionally contains conjugated double bonds sufficient to form an aromatic system. Examples include pyrrolizine, indole, indolizine, isoindole, indazole, purine, quinoline, isoquinoline, benzimidazole, benzofuran, benzopyran, benzothiazole, benzothiadiazole, benzisothiazole, pyridopyrimidine, pteridine, pyrimidinopyrimidine,

[0094] "Halogen" within the scope of the present invention denotes fluorine, chlorine, bromine, or iodine. Unless stated to the contrary, fluorine, chlorine, and bromine are regarded as preferred halogens.

[0095] As previously mentioned, the compounds of formula I, II, or III can be converted into their salts, especially into their physiologically and pharmacologically acceptable salts for pharmaceutical use. The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, substances, compositions, and / or dosage forms that, within the scope of reasonable medical judgment, are suitable for contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications, and that are commensurate with a reasonable benefit / risk ratio. On the one hand, these salts can exist in the form of physiologically and pharmacologically acceptable acid addition salts of the compounds of formula I, II, or III with inorganic or organic acids. On the other hand, the compounds of formula I, II, or III can be converted into physiologically and pharmacologically acceptable salts by reaction with inorganic bases, where alkali metal or alkaline earth metal cations serve as counterions. Acid addition salts can be prepared, for example, using hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, p-toluenesulfonic acid, acetic acid, fumaric acid, succinic acid, lactic acid, citric acid, tartaric acid, or maleic acid. Mixtures of the acids mentioned above can also be used. For the preparation of alkali metal and alkaline earth metal salts of the compounds of formula I, II, or III, alkali metal and alkaline earth metal hydroxides and hydrides are preferably used, where the hydroxides and hydrides of alkali metals, especially sodium, potassium, magnesium, calcium, zinc, and diethanolamine, are preferred, and sodium hydroxide and potassium hydroxide are particularly preferred.

[0096] The present invention relates to said compounds, which may optionally be in the form of individual optical isomers, diastereoisomers, mixtures of diastereoisomers, individual enantiomers or mixtures of racemates; in tautomeric form; and in the form of the free base or the corresponding acid addition salts formed with pharmacologically acceptable acids, such as acid addition salts formed with hydrohalic acids (such as hydrochloric acid or hydrobromic acid) or organic acids (such as oxalic acid, fumaric acid, glycolic acid or methanesulfonic acid).

[0097] The compounds of formula I, II or III according to the invention may optionally be present in the form of a mixture of diastereoisomers, but may also be obtained in pure diastereoisomeric form. Preferred are compounds having a specific stereochemistry of formula II and III, in particular compounds having a specific stereochemistry of formula II.

[0098] 4 Synthetic Methods General Procedures The following methods are applicable for the preparation of the compounds of general formula I, II or III. Synthetic methods known to the person skilled in the art and described in the organic synthesis literature can be used to obtain the compounds according to the invention. General methods for functional group protection and deprotection steps are described, for example, in Greene, T.W. and Wuts, P.G.M. (eds.): Protective Groups in Organic Synthesis, 3rd Edition 1999; John Wiley and Sons, Inc. Preferably, these compounds are obtained analogously to the methods more fully explained below, in particular the preparation methods described in the experimental section. The compounds of general formula (I) can be prepared using several alternative synthetic routes, of which the following routes are to be given as examples.

[0099] Route A: Compounds of general formulas I, II, and III, in particular compounds of general formulas I, II, and III where A represents -CH2- or substituted -CH2-, can be obtained from compounds of formula (A-I) by standard amidation procedures. R13 can thereby represent hydrogen or a protecting group, such as tert-butyl, which can be removed by standard deprotection methods. Compounds of formula (A-I) can be prepared from compounds of formula (A-II) using standard deprotection methods. Compounds of formula (A-I) can be prepared from compounds of formula (A-II) using standard deprotection methods. Compounds of formula (II) can be prepared by reacting compounds of formula (A-III) with compounds of formula (A-IV) in the presence of a strong base such as sodium hydride. By referring to Routes B and C described below and the examples described in the experimental section, the methods suitable for preparing compound (A-III) become obvious to those skilled in the art. Compounds of formula (A-IV) can be prepared by the methods described below for synthetic intermediate B. R in accordance with formulas (A-II) and (A-III) can represent hydrogen or a protecting group selected from the group consisting of tert-butyl, methyl, ethyl, and benzyl. Route A

[0100] Route B: Compounds of general formulas I, II, and III can be prepared by reacting compounds of general formula (B-I) in the presence of a strong base such as sodium hydride. R13 can thereby represent hydrogen or a protecting group, such as tert-butyl, which can be removed by standard deprotection methods. Compounds (B-I) can be prepared by oxidizing compounds of general formula (B-II) using, for example, 3-chloro-perbenzoic acid. Compounds (B-II) can be prepared by reacting compounds of general formula (B-III) with compounds of general formula (B-IV) under standard amidation conditions. By referring to the synthesis of compound (B-VI) and Routes C and the examples described in the experimental section below, the methods suitable for preparing compound (B-III) become obvious to those skilled in the art. Compounds of formula (B-IV) can be prepared by the methods described below for synthetic intermediate B. Alternatively, compounds (B-II) can be prepared by reacting compounds of general formula (B-V) with individual enantiopure hydroxyproline, optionally in the presence of a base. Compounds (B-V) can be obtained by chlorinating compounds of general formula (B-VI) using phosphoryl chloride. Compounds (B-VI) can be prepared from compounds of general formula (B-VII) under strongly basic conditions, such as at high temperature. Compounds (B-VII) can be obtained by reacting compounds of general formula (B-VIII) with compounds of general formula (B-IX) under standard amidation procedures. Compounds of general formula (B-VIII) can be prepared by the methods exemplified below or other standard synthetic methods known to those skilled in the art. Route B

[0101] Route C: The general formula compound can be prepared from the compound of general formula (C-I) through various types of ring-closure reactions, such as (but not limited to): Heck type coupling reaction (wherein R11 represents a bromine or iodine atom and R12 contains a terminal olefin), ring-closing metathesis reaction (wherein both R11 and R12 contain terminal olefins); subsequently hydrogenating the obtained olefin; amidation (wherein R11 carries a carboxylic acid and R12 carries a primary or secondary amino group, or vice versa). R13 can thus represent hydrogen or a protecting group, such as tert-butyl, which can be removed by standard deprotection methods. By referring to Routes A and B above and the examples described in the experimental section, the method for synthesizing the compound (C-I) becomes obvious to those skilled in the art. Route C

[0102] Synthesis of intermediates Intermediate A Intermediate A-01 (racemic trans-4-amino-3-methylpiperidine-1-carboxylic acid tert-butyl ester): Step 1: To a solution of tert-butyl 3-methyl-4-oxopiperidine-1-carboxylate (175 g; 0.82 mol) in THF (1.22 L) cooled to 0 °C was added lithium tris-sec-butyl(hydrido)borate (lithium tri-sec-butylborohydride; 1 M in THF; 984 mL; 0.98 mol). The mixture was stirred for 4 h while maintaining the temperature between 0 °C and 10 °C. An aqueous sodium hypochlorite solution (10%; 525 mL) was added and the mixture was extracted with EtOAc (700 mL). The organic layer was separated, washed with brine, dried over sodium sulfate and evaporated. The crude product was purified by FC (silica gel; petroleum ether / EtOAc, 2% -> 50%). 1 H NMR (400 MHz, CDCl3) δ ppm 3.84 - 3.82 (m, 1H), 3.52 (d, J = 6.40 Hz, 2H), 3.29 (d, J = 5.40 Hz, 2H), 3.06 - 3.02 (m, 1H), 1.76 - 1.74 (m, 3H), 1.65 (s, 9H), 1.24 - 0.89 (m, 3H)

[0103] Step 2: To a solution of the product of Step 1 (330 g; 1.53 mol) in THF (1.65 L) was added triethylamine (341 g; 3.37 mol). Methanesulfonic anhydride (507 g; 2.91 mol) was added at room temperature and the mixture was stirred at room temperature for 4 h. Water (550 mL) was added and the mixture was extracted with EtOAc (1.65 L). The organic layer was separated, washed with brine, dried over sodium sulfate and concentrated under reduced pressure. The product was used in the next step without further purification. 1 H NMR (400 MHz, CDCl3) δ ppm 4.74 - 4.71 (m, 1H), 4.23 - 3.95 (m, 1H), 3.17 - 3.04 (m, 2H), 3.04 (s, 3H), 1.95 - 1.92 (m, 1H), 1.83 (s, 1H), 1.83 - 1.82 (m, 1H), 1.32 (s, 9H), 0.91 - 0.79 (m, 3H)

[0104] Step 3: The reaction was carried out under a nitrogen atmosphere. Sodium azide (186 g; 2.86 mol) was added to a solution of the product of Step 2 (420 g; 1.43 mol) in DMF (2.1 L) at room temperature. The mixture was stirred at 100 °C for 4 h and then cooled to 0 °C. Saturated sodium carbonate solution (4.0 L) was added while maintaining the temperature below 10 °C. The mixture was extracted with EtOAc, the organic layer was separated, washed with brine, dried over sodium sulfate and evaporated under reduced pressure. The product was used in the next step without further purification.

[0105] Step 4: At room temperature, a mixture of the product of Step 3 (200 g; 0.83 mol), palladium / carbon (60 g) and EtOAc (1.0 L) was stirred under a hydrogen pressure of 50 psi for 2 h. The catalyst was filtered off by suction and washed with methanol (2 L). The combined filtrates were evaporated under reduced pressure. The crude product was purified by FC (silica gel; petroleum ether / EtOAc 0% -> 100%) to give the title compound. 1 H NMR (400 MHz, CDCl3) δ ppm 7.85 (s, 1H) 3.99 - 3.74 (m, 2H), 2.81 - 2.79 (m, 2H), 2.71 (s, 1H), 2.72 - 2.70 (m, 1H), 2.64 - 2.52 (m, 1H), 2.25 - 2.16 (m, 2H), 1.66 - 1.57 (m, 1H), 1.35 - 1.30 (m, 2H), 1.30 - 1.27 (m, 8H), 1.17 - 1.06 (m, 2H), 0.83 - 0.78 (m, 3H), 0.75 - 0.71 (m, 1H)

[0106] Intermediate B Intermediate B-01: Step 1: Stir a mixture of Intermediate A-01 (2. g; 9.97 mmol), 5-chloro-2-fluoronitrobenzene (1.21 mL; 9.97 mmol), DIPEA (3.43 mL; 19.9 mmol) and DMF (17 mL) at 60 °C until the reaction control by RP HPLC indicates that the starting materials have been significantly converted (10 h). Pour the mixture onto ice water. Then add water and extract the mixture three times with ethyl acetate. Wash the combined organic layers with water and then with brine, separate, dry over magnesium sulfate, filter and concentrate. Purify the crude product by FC (silica gel; cyclohexane / EtOAc 20% -> 100%) to obtain (racemic-trans)-tert-butyl 4-[(4-chloro-2-nitrophenyl)amino]-3-methylpiperidine-1-carboxylate. ESI-MS: 392 [M+Na] + R t (HPLC): 0.83 min (Method A)

[0107] Step 2: In a Parr apparatus, at room temperature, under a hydrogen pressure of 50 psi, shake the product of Step 1 (3.25 g; 7.90 mmol), Raney nickel (410 mg) and THF (60 mL) for 5 h. Then add Raney nickel (160 mg) several more times after every 10 - 15 h until the reaction control by RP HPLC indicates that the starting materials have been highly converted (here: the catalyst is added 3 times). Filter out the catalyst and evaporate the filtrate. The crude product is used for the next step. ESI-MS: 340 [M+H] + R t (HPLC): 0.63 min (Method A)

[0108] Step 3: Stir a mixture of the product of Step 2 (2.16 g; 6.37 mmol), 1-(1H-imidazole-1-thiocarbonyl)-1H-imidazole (1.47 g; 8.25 mmol) and DMF (34 mL) at room temperature for 2 h. Slowly add ice water (200 mL) to the reaction mixture while stirring and continue stirring for 10 min. Filter out the precipitate, wash with water, dry at 50 °C overnight and use it for the next step without further purification. ESI-MS: 382 [M+H] + R t (HPLC): 0.71 min (Method A)

[0109] Step 4: React under an argon atmosphere. Potassium tert-butoxide (1.42 g; 12.7 mmol) was added to a solution of the product from Step 3 (2.57 g; 6.26 mmol) in anhydrous DMF. The mixture was stirred at room temperature for 15 min, then methyl iodide (597 μL; 9.49 mmol) was added. The mixture was stirred at room temperature until the reaction control by RP HPLC indicated that the starting material had been nearly completely converted (2 h). Ice water (100 mL) was added and the mixture was kept at 6 °C overnight. The formed precipitate was filtered out, dissolved in EtOAc, concentrated under reduced pressure and dried by co-evaporation with toluene to give tert-butyl (racemic trans)-4-[5-chloro-2-(methylthio)-1H-1,3-benzodiazol-1-yl]-3-methylpiperidine-1-carboxylate. ESI-MS: 396 [M+H] + R t (HPLC): 0.73 min (Method A)

[0110] Step 5: The mixture of enantiomers from Step 4 was separated by preparative SFC (instrument: Sepiatec 2Prep SFC 100; column: Lux Cellulose-2 (21.2 mm * 250 mm, 5 μm); mobile phase: A for CO2 and B for IPA; gradient: B% = 20% isocratic elution mode; flow rate: 60 mL / min; wavelength: 220 nm; column temperature: 40 °C; system back pressure: 150 bar).

[0111] Based on the method described by D. J. Patel et al., PNAS 2019, 11946 - 11955 (doi.org / 10.1073 / pnas.1905013116), the absolute configurations of the two separated enantiomers were assigned based on the co-crystal structure of Example 1.01 with the human cGAS protein. 11H NMR (of the first eluted isomer) (400 MHz, DMSO-d6) δ ppm 7.46 - 7.69 (m, 1H), 7.10 - 7.26 (m, 1H), 7.17 (br d, J = 6.84 Hz, 1H), 3.87 - 4.22 (m, 3H), 2.87 - 3.06 (m, 1H), 2.57 - 2.79 (m, 4H), 2.31 - 2.45 (m, 1H), 2.13 - 2.30 (m, 1H), 1.80 (br d, J = 11.79 Hz, 1H), 1.45 (s, 9H), 0.58 (d, J = 6.46 Hz, 3H)

[0112] This first eluted isomer is used in the next step. Intermediate B-02 is prepared from the second eluted isomer.

[0113] Step 6: To a solution of the first eluted isomer from Step 5 (811 mg; 2.05 mmol) in DCM (11.8 mL) was added 3-chloroperoxybenzoic acid (77%; 0.987 g; 4.40 mmol). The mixture was stirred for 2 h, then diluted with additional DCM and washed with aqueous potassium carbonate solution (15%). The aqueous layer was re-extracted with DCM and the combined organic layers were washed with water, separated, dried over sodium sulfate, filtered and concentrated under reduced pressure to give the title compound. ESI-MS: 428 [M+H] + R t (HPLC): 0.74 min (Method A)

[0114] In a manner similar to Intermediate 1.01 described above, the following intermediates were prepared from the designated starting materials. Thus, the reaction temperature in Step 1 was adjusted according to the reactivity of the respective nitrobenzene starting materials. The absolute configuration of the selected intermediates was assigned according to the method described by D. J. Patel et al., PNAS 2019, 11946 - 11955 (doi.org / 10.1073 / pnas.1905013116) based on the co-crystal structures of the exemplary compounds prepared from each intermediate with the human cGAS protein. The absolute configurations of the other intermediates and examples were assigned based on the assumption that the eutomers share the same absolute stereochemistry in all cases.

[0115] Intermediate B-12 (racemic trans) According to General Procedure D, the BOC protecting group of the product of Step 4 of synthetic Intermediate B-08 was removed. ESI-MS: 262 [M+H] + R t (HPLC): 0.47 min (Method E)

[0116] Intermediate B-14 (racemic trans) A mixture of Intermediate B-08 (700 mg; 1.78 mmol) and dioxane containing hydrochloric acid (4 M; 1.33 mL; 5.45 mmol) was stirred for 2 h. Tert-butyl methyl ether (20 mL) was added and the mixture was allowed to stand for 1 h without stirring. The precipitate was filtered off and dissolved in methanol. The resulting solution was evaporated under reduced pressure to give the title compound. ESI-MS: 294 [M+H] + R t (HPLC): 0.60 min (Method E)

[0117] Intermediate I Intermediate I.01: Intermediate N-R was reacted with Intermediate B-5 according to General Procedure A to give the title compound (3S,4S)-4-(2-{[(3S,5S)-1-{4-bromo-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2,4,6,9,11-hexaen-6-yl}-5-[(tert-butoxy)carbonyl]pyrrolidin-3-yl]oxy}-1H-1,3-benzodiazol-1-yl)-3-methylpiperidine-1-carboxylic acid tert-butyl ester. ESI-MS: 747 [M+H] + R t (HPLC): 1.27 min (Method B)

[0118] Intermediate I.02: Step 1: The reaction was carried out under an argon atmosphere. The solvent was degassed and dried by adding molecular sieves. DMSO (10 mL), methyl bromodifluoroacetate (266 μL; 2.42 mmol), and copper powder (291 mg; 4.58 mmol) were added to intermediate I.01 (721 mg; 0.916 mmol) in a round-bottom flask. The mixture was stirred overnight, and then 1 equivalent of methyl bromodifluoroacetate and another 2 equivalents of copper were added. After further stirring for 72 h, the mixture was diluted with EtOAc, a potassium dihydrogen phosphate solution (1.27 M; 20 mL) was added, and the mixture was stirred for an additional 30 min, after which it was filtered. The organic layer of the filtrate was washed with water, concentrated, and the crude product was purified by FC (silica gel; CH / EtOAc 10% -> 100%) to give (3S,4S)-4-(2-{[(3S,5S)-5-[(tert-butoxy)carbonyl]-1-[4-(1,1-difluoro-2-methoxy-2-oxoethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2,4,6,9,11-hexaen-6-yl]pyrrolidin-3-yl]oxy}-1H-1,3-benzodiazol-1-yl)-3-methylpiperidine-1-carboxylic acid tert-butyl ester.

[0119] Step 2: Sodium borohydride (244 mg; 6.40 mmol) was added portionwise to anhydrous ethanol (dried over molecular sieves) containing the product of Step 1 (585 mg; 0.647 mmol) over 1 h. The reaction was quenched by adding water, the mixture was extracted three times with EtOAc, and the combined organic layers were washed successively with water and brine, separated, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by FC (silica gel; CH / EtOAc 20% -> 100%) to give (3S,4S)-4-(2-{[(3S,5S)-5-[(tert-butoxy)carbonyl]-1-[4-(1,1-difluoro-2-hydroxyethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2,4,6,9,11-hexaen-6-yl]pyrrolidin-3-yl]oxy}-1H-1,3-benzodiazol-1-yl)-3-methylpiperidine-1-carboxylic acid tert-butyl ester.

[0120] Step 3: To a solution of the product of Step 2 (269 mg; 0.359 mmol) and allyl bromide (220 μL; 2.52 mmol) in DMA (1.67 mL) was added sodium hydride (55% in mineral oil; 47 mg; 1.08 mmol). The mixture was stirred for 10 min and then quenched by the addition of water. The mixture was extracted three times with EtOAc, and the combined organic layers were washed sequentially with water and brine, separated, dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by FC (silica gel; CH / EtOAc 20% -> 100%) to give the title compound (3S,4S)-4-(2-{[(3S,5S)-5-[(tert-butoxy)carbonyl]-1-{4-[1,1-difluoro-2-(prop-2-en-1-yloxy)ethyl]-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2,4,6,9,11-hexaen-6-yl}pyrrolidin-3-yl]oxy}-1H-1,3-benzodiazol-1-yl)-3-methylpiperidine-1-carboxylic acid tert-butyl ester. ESI-MS: 789 [M+H] + R t (HPLC): 1.29 min (Method B)

[0121] Intermediate I.03: (racemic mixture of trans isomers) To a solution of 3-allyloxypropanoic acid (38 μL; 0.303 mmol) in DMF (500 μL) was added EDC hydrochloride (70 mg; 0.364 mmol) and N,N-diisopropylethylamine (105 μL; 0.606 mmol). The mixture was stirred for 10 min, then Intermediate B-14 (100 mg; 0.303 mmol) was added and the mixture was stirred for 90 min, then diluted with methanol (2 mL) and purified by RP HPLC (Sunfire C18; ACN / water, modifier: TFA). ESI-MS: 406 [M+H] + R t (HPLC): 0.88 min (Method E)

[0122] Intermediate N The synthetic procedures described below were carried out in part according to the general procedures indicated below. General Procedure Int-A: Hydrogenation using Pd / C catalyst (see Intermediate N-A, Step 4) General procedure Int-B: Ring closure under basic conditions (see: Intermediate N-A, step 5) General procedure Int-C: Chlorination using phosphorus oxychloride (see: Intermediate N-A, step 7) General procedure Int-D: Use of S of hydroxyproline ester N Ar (see: Intermediate N-A, step 8) General procedure Int-E: Ester cleavage using lithium hydroxide (see: Intermediate N-F, step 2) General procedure Int-F: Amidation using PFTU (see: Intermediate N-F, step 3) General procedure Int-G: Heck-type coupling (see: Intermediate N-J, step 1)

[0123] Intermediate N-A: Step 1: A mixture of tert-butyl 3-(2-oxoethoxy)propionate (7.25 g; 38.5 mmol) and methyl (triphenylphosphoranylidene)acetate (13.1 g; 38.5 mmol) prepared as described in EP2409977 in DCM (200 mL) was stirred overnight at room temperature. The mixture was evaporated under reduced pressure and dissolved in CH / EtOAc (3:1). The insoluble material was removed by filtration and the filtrate was evaporated. The crude product was purified by FC (silica gel; CH / EtOAc 10% -> 45%) to give the product in the form of a mixture of cis and trans isomers.

[0124] Step 2: The product of step 1 (1.50 g; 6.14 mmol) was stirred into a mixture of DCM (15 mL) and TFA (10 mL) overnight. The mixture was evaporated and dissolved in methanol (10 mL). Polymer-bound tetraalkylammonium carbonate (2 weight equivalents) was added and the mixture was stirred for 90 min. The insoluble material was filtered off and the filtrate was evaporated.

[0125] Step 3: To a solution of the product of Step 2 (5.00 g; 21.3 mmol) in ACN (140 mL) was added 1-chloro-N,N,2-trimethylallylamine (4.45 mL; 33.6 mmol). The mixture was stirred at room temperature for 10 min, then pyridine (5.10 mL; 63.0 mmol) and 3-aminobenzofuran-2-carboxamide (3.70 g; 21.0 mmol) were added. The mixture was stirred at room temperature overnight, then water was added. The mixture was extracted with DCM and the organic layer was separated and evaporated. The crude product was purified first by FC (silica gel; petroleum ether / EtOAc 40% -> 80%) and then by RP HPLC (Sunfire C18, ACN / water, modifier: TFA). ESI-MS: 347 [M+H] + R t (HPLC): 0.80 min (Method E)

[0126] Step 4: General Procedure Int-A: A mixture of the product of Step 3 (3.40 g; 9.73 mmol), palladium / carbon (10%; 350 mg) and ethanol (500 mL) was shaken under a hydrogen pressure of 50 psi until the reaction control by RP HPLC indicated that the starting material had been converted (here: 90 min). The catalyst was filtered off and the filtrate was evaporated to dryness. ESI-MS: 349 [M+H] + R t (HPLC): 0.81 min (Method E)

[0127] Step 5: General Procedure Int-B: A mixture of the product of Step 4 (4.55 g; 13.1 mmol) and aqueous sodium hydroxide solution (4 M; 100 mL; 400 mmol) was stirred at 60 °C until the reaction control by RP HPLC indicated that the starting material had been converted (here: 60 min). The mixture was cooled to room temperature and then acidified by adding aqueous hydrochloric acid solution (4 M). The precipitate was collected and dried at 60 °C. ESI-MS: 317 [M+H] + R t (HPLC): 0.73 min (Method E)

[0128] Step 6: At room temperature, oxalyl chloride (1.24 mL; 14.5 mmol) was added to a mixture of the product of step 5 (3.53 g; 11.2 mmol), DCM (60 mL) and a few drops of DMF. The mixture was stirred at room temperature for 3 h, then methanol was added and stirring was continued for 60 min. The mixture was extracted with water, the organic layer was separated and evaporated to dryness. ESI-MS: 331 [M+H] + R t (HPLC): 0.82 min (method E)

[0129] Step 7: General procedure Int-C: A mixture of the product of step 6 (3.70 g; 11.2 mmol) and phosphorus oxychloride (70 mL) was stirred at 90 °C for 4 h. The excess phosphorus oxychloride was removed by distillation, and water was carefully added. The resulting mixture was extracted with EtOAc, the organic layer was separated and evaporated. The crude product was used for the next step. ESI-MS: 349 [M+H] + R t (HPLC): 1.02 min (method E)

[0130] Step 8: General procedure Int-D: A mixture of the product of step 7 (300 mg; 0.896 mmol), (2S,4S)-tert-butyl 4-hydroxypyrrolidine-2-carboxylate hydrochloride (253 mg; 1.08 mmol), potassium carbonate (300 mg; 2.06 mmol) and DMF (7.0 mL) was stirred overnight at room temperature. Water was added and the mixture was acidified by adding aqueous hydrochloric acid (1 M). The mixture was extracted with EtOAc, the organic layer was evaporated, and the crude product was purified by FC (silica gel; petroleum ether / EtOAc 40% -> 75%). ESI-MS: 500 [M+H] + R t (HPLC): 0.75 min (method E)

[0131] Step 9: The product of step 8 was reacted according to general procedure Int-E to give the title compound with ester cleavage. ESI-MS: 486 [M+H] + R t (HPLC): 0.70 min (method E)

[0132] Intermediate N-B: Prepared in step 3 by applying 3-amino-6-chloro-1-benzofuran-2-carboxamide in a sequence similar to that described for the synthesis of intermediate N-A. ESI-MS: 520 [M+H] + R t (HPLC): 0.50 min (method A)

[0133] Intermediate N-C: Prepared from intermediate N-V and 2-(but-3-en-1-yloxy)acetic acid using a two-step sequence: Step 1: According to the general procedure Int-G Step 2: According to the general procedure Int-A, the title compound was obtained. ESI-MS: 504 [M+H] + R t (HPLC): 0.45 min (method A)

[0134] Intermediate N-D: Prepared in a sequence similar to that described for the synthesis of intermediate N-A, starting from tert-butyl 2-(3-oxopropoxy)acetate prepared as described in EP1939201. ESI-MS: 486 [M+H] + R t (HPLC): 0.71 min (method E)

[0135] Intermediate N-E: Step 1: A mixture of 3-aminobenzofuran-2-carboxamide (2.00 g; 11.4 mmol), ethyl 2-[2-(2-ethoxy-2-oxoethoxy)ethoxy]acetate (5.32 g; 22.7 mmol) and 1H,2H,3H,4H,6H,7H,8H-[1,3]diazino[1,2-a]pyrimidine (6.45 g; 45.4 mmol) was heated to 120 °C for 150 min. After cooling to room temperature, aqueous hydrochloric acid (1 M; 70 mL) was added. The precipitate was filtered off with suction, washed with water and dried in vacuo at 60 °C to give 2-[2-({6-oxo-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7A mixture of ethyl 2-({6-oxo-8-oxa-3,5-diazatricyclo[7.4.0.0

[0136] Step 2: For the re-esterification of the acid, the mixture from Step 1 (2.8 g) was dissolved in DCM (200 mL). 2 drops of DMF were added, followed by oxalyl chloride (392 μL, 4.57 mmol). The mixture was stirred overnight, then ethanol (10 mL) was added and the mixture was stirred for an additional 2 h. The mixture was concentrated under reduced pressure. Methyl tert-butyl ether was added and the resulting precipitate was washed with methyl tert-butyl ether and dried at 50 °C to give ethyl 2-[2-({6-oxo-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(9),2(7),3,10,12-pentaen-4-yl}methoxy)ethoxy]acetate.

[0137] The product of Step 2 was further reacted according to the First General Procedure Int-C and then Int-D in a 2-step sequence to give the title compound. ESI-MS: 516 [M+H] + R t (HPLC): 0.80 min (Method E)

[0138] Intermediate N-F: Step 1: The zinc powder used was washed with 2% aqueous hydrochloric acid, water and acetone before use. The purified powder was dried under high vacuum and stored under argon. Zinc powder (2.44 g; 36.9 mmol), nickel(II) chloride hexahydrate (0.754 g; 3.14 mmol), THF (35.0 mL) and 3 drops of water were charged to a flask and the mixture was stirred at room temperature for 10 min. Subsequently, tert-butyl hex-5-enoate (4.63 g; 18.5 mmol; prepared as described in WO2010 / 15447) was added in one portion and ethyl iodo fluoroacetate (2.80 mL; 18.5 mmol) was added dropwise (an exothermic reaction occurred during the addition) while maintaining the temperature below 30 °C. After the addition was complete, the reaction mixture was stirred at 60 °C for 4 h. The reaction mixture was poured into a mixture of saturated ammonium chloride solution (100 mL) and diethyl ether (100 mL) and stirred for 10 min. Subsequently, the mixture was filtered through a pad of diatomaceous earth and after phase separation, the aqueous phase was extracted with diethyl ether. The combined organic layers were washed with water, dried over sodium sulfate, filtered and evaporated. The crude product was purified by FC (silica gel; CH / DCM 10% -> 100%). ESI-MS: 312 [M+NH4] + R t (HPLC): 0.78 min (Method A)

[0139] Step 2: General Procedure Int-E: Lithium hydroxide (276 mg; 11.0 mmol) was added to a solution of the product of Step 1 in 2:1 THF / H2O and the reaction mixture was stirred at room temperature until the reaction control by RP HPLC indicated exhaustion of the starting material (here: 2.5 h). Volatiles were removed in vacuo, the residue was acidified to pH = 1 by addition of 5.50 mL of 1 N aqueous hydrochloric acid, and the mixture was extracted three times with EtOAc. The combined organic phases were dried over sodium sulfate, filtered, and evaporated. ESI-MS: 211 [M-isobutene + H] + R t (HPLC): 0.59 min (Method A)

[0140] Step 3: General Procedure Int-F: At room temperature, PFTU (2.40 g; 5.60 mmol) was added to a stirred solution of the product of Step 2 (1.40 g; 5.09 mmol) and DIPEA (970 μL; 5.60 mmol) in DMF (21.0 mL), and the mixture was stirred for 30 min. Subsequently, 3-aminobenzofuran-2-carboxamide (1.01 g; 5.60 mmol) and additional DIPEA (970 μL; 5.60 mmol) were added and the mixture was stirred at room temperature for 10 min. Subsequently, the reaction mixture was heated to 50 °C and stirred at this temperature for 16 h. Since the reaction control by RP HPLC indicated incomplete exhaustion of the starting material, additional DIPEA (441 μL; 2.80 mmol) and PFTU (0.86 g; 2.04 mmol) were added and stirring was continued at 50 °C for 2.5 h. The reaction mixture was diluted with water, acidified with TFA, filtered, and purified by RP HPLC (XBridge C18; ACN / water; modifier: TFA). ESI-MS: 425 [M+H] + R t (HPLC): 0.72 min (Method A)

[0141] Step 4: At room temperature, chlorotrimethylsilane (4.05 mL; 30.3 mmol) was slowly added to a solution of the product of step 3 (950 mg; 2.13 mmol) and TEA (13.0 mL; 92.4 mmol) in 1,2-dichloroethane (28.5 mL). After the addition was complete, the reaction mixture was heated to 85 °C and stirred at this temperature for 24 h. The reaction mixture was poured into 30 mL of 4 M hydrochloric acid (pH = 1) and extracted twice with DCM. The combined organic phases were washed with water, dried over sodium sulfate, filtered and evaporated to give tert-butyl 7,7-difluoro-7-{6-oxo-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2(7),3,9,11-pentaen-4-yl}heptanoate. ESI-MS: 405 [M-H] - R t (HPLC): 0.44 min (method F)

[0142] Step 5: Using the general procedure Int-C, the product of step 4 was converted into the chlorinated product, which was then purified by RP-HPLC (XBridge C18; ACN / water; modifier: TFA). ESI-MS: 367 [M-H] - R t (HPLC): 0.64 min (method A)

[0143] Step 6: The product of step 6 was reacted according to the general procedure Int-D to give the title compound. ESI-MS: 520 [M+H] + R t (HPLC): 0.61 min (method A)

[0144] Intermediate N-G: Prepared in step 3 using 3-amino-6-fluorobenzofuran-2-carboxamide, analogous to the reaction sequence described for the synthesis of intermediate N-F.

[0145] Intermediate N-H: Prepared in step 3 using 3-amino-6-chlorobenzofuran-2-carboxamide, analogous to the reaction sequence described for the synthesis of intermediate N-F.

[0146] Intermediate N-I: Step 1: To a solution of magnesium powder (45.0 g; 1.85 mol) in THF (285 mL) was added iodine (1.00 g; 3.94 mmol), and then a solution of 1-bromo-3-butene (111 g; 821 mmol) in THF (850 mL) was added dropwise. The temperature was thus maintained below 50 °C. The resulting mixture was cooled to -75 °C, and a solution of diethyl oxalate (100 g; 684 mmol; 93.5 mL) in THF (1.89 L) was added dropwise at -75 °C. The mixture was further stirred at -75 °C for 4 h. The reaction mixture was quenched by adding saturated aqueous ammonium chloride solution (900 mL) at 0 °C, and then the pH was adjusted to pH 3 by adding aqueous hydrochloric acid solution (1 M). The mixture was extracted three times with EtOAc (500 mL). The combined organic layers were washed with brine (900 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure to give a residue, which was purified by FC (silica gel; petroleum ether / ethyl acetate 0% -> 100%). 1 H NMR: (400 MHz, CDCl3) δ = 5.85 - 5.72 (m, 1H), 5.11 - 4.92 (m, 2H), 4.37 - 4.24 (m, 2H), 2.92 (t, J = 7.3 Hz, 2H), 2.36 (q, J = 7.1 Hz, 2H), 1.24 - 1.21 (m, 3H)

[0147] Step 2: At 0 °C, bis-(2-methoxyethyl)aminosulfur trifluoride (Deoxofluor) (120 g, 544 mmol, 119 mL) and ethanol (2.95 g, 64.0 mmol) were added to a solution of the product of Step 1 (50.0 g, 320 mmol) in DCM (1000 mL). The mixture was stirred at 25 °C for 12 h. The reaction mixture was quenched by adding 500 mL of saturated aqueous sodium bicarbonate solution, and then extracted three times with DCM (500 mL). The combined organic layers were washed with aqueous hydrochloric acid solution (1 M; 200 mL) and brine (200 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The crude product was vacuum distilled (30 °C, 0.09 MPa / oil pump). 1 H NMR: (400 MHz, CDCl3): δ = 5.80 (br dd, J = 10.3, 16.9 Hz, 1H), 5.13 - 5.00 (m, 2H), 4.39 - 4.28 (m, 2H), 2.30 - 2.12 (m, 4H), 1.43 - 1.34 (m, 3H)

[0148] Step 3: To a mixture of zinc powder (8.79 g, 134 mmol) and THF (30.0 mL) was added nickel(II) chloride hexahydrate (804 mg, 3.38 mmol). The mixture was stirred at -65 °C for 5 min. Subsequently, ethyl difluoroiodoacetate (12.0 g, 48.0 mmol) and the product of Step 2 (6.00 g, 33.7 mmol) were added dropwise at -65 °C. The mixture was stirred at 25 °C for 12 h. The reaction mixture was quenched by addition of saturated aqueous ammonium chloride solution (60.0 mL) at 0 °C and then extracted three times with DCM (60.0 mL). The combined organic layers were washed with brine (60.0 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. 1 1H NMR: (400 MHz, chloroform-d): δ = 4.33 (q, J = 7.2 Hz, 4H), 2.18 - 1.98 (m, 4H), 1.56 (td, J = 3.8, 8.0 Hz, 4H), 1.36 (t, J = 7.2 Hz, 6H)

[0149] Step 4: To a solution of the product of Step 3 (11.0 g, 36.4 mmol) in dioxane (30.0 mL) was added 1,5,7-triazabicyclo[4.4.0]dec-5-ene (14.2 g, 102 mmol) and 3-amino-1-benzofuran-2-carboxamide (4.50 g, 25.5 mmol). The mixture was stirred at 110 °C for 2 h, then diluted with water (30.0 mL) and extracted three times with EtOAc (30.0 mL). The combined organic layers were washed with brine (30.0 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by RPHPLC (ACN / water, modifier: TFA). 1 1H NMR: (400 MHz, DMSO-d): δ = 8.09 (d, J = 7.7 Hz, 1H), 7.88 (d, J = 8.4 Hz, 1H), 7.72 (t, J = 7.8 Hz, 1H), 7.53 (t, J = 7.5 Hz, 1H), 2.40 - 2.37 (m, 2H), 2.12 – 2.07 (m, 2H), 1.54 - 1.48 (m, 4H).

[0150] Step 5: Chlorinate the product of Step 4 using the general procedure Int-C.

[0151] Step 6: React the product of Step 5 using the general procedure Int-D to give the title compound. ESI-MS: 556 [M+H] + R t (HPLC): 0.64 min (Method A)

[0152] Intermediate N-J: Prepared from Intermediate N-R using the following sequential 3-step procedure: Step 1: General Procedure Int-G: Dissolve Intermediate N-R (300 mg; 0.69 mmol) and methyl 5-hexenoate (298 μL; 2.07 mmol) in DMF (10 mL; 123 mmol). Add triethylamine (0.39 mL; 2.76 mmol) and degas the mixture with argon. Add palladium(II) acetate (31 mg; 0.14 mmol) and tri-o-tolylphosphine (84 mg; 0.28 mmol) under argon. Stir the sealed vial overnight at 95 °C. Dilute the mixture with ACN / water, acidify with TFA, and filter through a syringe filter. Purify by preparative RP HPLC (Sunfire C18, ACN / water; modifier: TFA) to give tert-butyl (2S,4S)-4-hydroxy-1-{4-[6-methoxy-6-oxohex-1-en-1-yl]-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2,4,6,9,11-hexaen-6-yl}pyrrolidine-2-carboxylate.

[0153] Step 2: Hydrogenate the product of Step 1 according to General Procedure Int-A to give tert-butyl (2S,4S)-4-hydroxy-1-[4-(6-methoxy-6-oxohexyl)-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2,4,6,9,11-hexaen-6-yl]pyrrolidine-2-carboxylate. ESI-MS: 484 [M+H] + R t (HPLC): 0.65 min (Method B)

[0154] Step 3: React the product of Step 2 according to General Procedure Int-E to give the title compound with ester cleavage.

[0155] Intermediate N-K: The preparation is carried out from intermediate N-R and methyl 3-(but-3-en-2-yloxy)propionate using the following three-step sequence (the starting material methyl 3-(but-3-en-2-yloxy)propionate is prepared from the respective tert-butyl esters by acidic ester cleavage followed by methyl ester formation (methanol, thionyl chloride)): Step 1: Heck-type coupling according to general procedure Int-G Step 2: Hydrogenation according to general procedure Int-A Step 3: Ester cleavage according to general procedure Int-E to give the title compound. ESI-MS: 500 [M+H] + R t (HPLC): 1.82 min (method G)

[0156] Intermediate N-L: 2-(4-{6-[(2S,4S)-2-[(tert-butoxy)carbonyl]-4-hydroxypyrrolidin-1-yl]-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2,4,6,9,11-hexaen-4-yl}-4,4-difluorobutoxy)acetic acid Step 1: Equip a dry reaction vessel with a magnetic stir bar and charge with tert-butyl 2-allyloxyacetate (1.00 g; 5.81 mmol), anhydrous nickel(II) chloride (0.038 g; 0.290 mmol), and sodium carbonate (0.61 g; 5.81 mmol). Subsequently, briefly evacuate the reaction vessel and backfill with argon (this sequence is repeated a total of three times). Add anhydrous DMF (40 mL), ethyl bromodifluoroacetate (1.5 mL, 11.6 mmol), and phenylsilane (2.9 mL; 23.2 mmol) to the reaction vessel sequentially via syringe. Heat the vessel in an oil bath at 70 °C and stir until TLC monitoring indicates depletion of the starting material (here: overnight). Dilute the reaction mixture with 30 mL of EtOAc, and wash the organic layer with 80 mL of saturated aqueous sodium chloride. Thereafter, dry the organic layer over sulfate and concentrate under reduced pressure, and further purify by FC (silica gel; hexane / EtOAc) to give ethyl 5-(2-tert-butoxy-2-oxo-ethoxy)-2,2-difluoro-pentanoate.

[0157] Step 2: At room temperature, 1,5,7-triazabicyclo[4.4.0]dec-5-ene (0.79 g; 5.68 mmol) was added to a solution of 3-aminobenzofuran-2-carboxamide (0.25 g; 1.42 mmol) in 1,4-dioxane (1 mL). Subsequently, the product of step 1 (0.84 g; 2.84 mmol) was added, the temperature was raised to 120 °C and stirring was continued until TLC indicated almost complete conversion (here: 18 h). The reaction mixture was diluted with water (15 mL) and extracted with DCM (3 × 7 mL). The pH of the resulting aqueous layer was adjusted to between 4 and 5 and the precipitated solid was filtered off. ESI-MS: 353 [M+H] + R t (HPLC): 1.61 min (method H)

[0158] Step 3: The product of step 2 was reacted according to the general procedure Int-C. ESI-MS: 371 [M+H] + R t (HPLC): 1.82 min (method H)

[0159] Step 4: The product of step 2 was reacted according to the general procedure Int-D to give the title compound. ESI-MS: 522.6 [M+H] + R t (HPLC): 2.96 min (method G)

[0160] Intermediate N-M: Prepared analogously to the procedure described for the synthesis of intermediate N-L, using 3-amino-6-chloro-1-benzofuran-2-carboxamide (prepared as described in EP1710233) as starting material in step 2.

[0161] Intermediate N-N: Prepared from intermediate N-R and methyl 4,4-dimethylhept-6-enoate analogously to the reaction sequence described for the synthesis of intermediate N-J.

[0162] Intermediate N-O: Prepared from intermediate N-R and ethyl 6-heptenoate analogously to the reaction sequence described for the synthesis of intermediate N-J.

[0163] Intermediate N-P: Intermediate N-U is reacted according to the general procedure Int-E to obtain the title compound. ESI-MS: 414 [M+H] + R t (HPLC): 0.55 min (Method B)

[0164] Intermediate N-Q: (2S,4S)-1-[4-(7-Ethoxy-7-oxohept-1-en-2-yl)-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2,4,6,9,11-hexaen-6-yl]-4-hydroxypyrrolidine-2-carboxylic acid tert-butyl ester, which is formed as a by-product in Step 1 of the synthesis of Intermediate N-O, is separated by RP HPLC and further reacted according to the reaction sequence described for the preparation of Intermediate N-O to obtain the title compound.

[0165] Intermediate N-R: (2S,4S)-1-{4-Bromo-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2(7),3,5,9,11-hexaen-6-yl}-4-hydroxypyrrolidine-2-carboxylic acid tert-butyl ester Step 1: 4,6-Dibromo-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2(7),3,5,9,11-hexaene A mixture of 1H-benzo[4,5]furo[3,2-d]pyrimidine-2,4-dione (11.6 g; 0.0573 mol) and phosphorus tribromide (40.8 mL, 0.402 mol) is heated at 150 °C for 3 h. The mixture is cooled to room temperature, the pH is adjusted to pH = 7 using saturated aqueous sodium bicarbonate while cooling at 0 °C, and the mixture is extracted with EtOAc (3 × 50 mL). The organic layers are combined, dried over sodium sulfate, and evaporated under reduced pressure. The remaining residue is purified by dissolving it in a mixture of DCM (3V relative to the weight of the crude material) and EtOAc (3V relative to the weight of the crude material) with stirring. After stirring at room temperature for 30 min, the mixture is filtered and the supernatant is evaporated under reduced pressure. ESI-MS: 327 / 329 / 331 [M+H]+ R t (HPLC): 0.67 min (Method A)

[0166] Step 2: (2S,4S)-1-{4-bromo-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2(7),3,5,9,11-hexaen-6-yl}-4-hydroxypyrrolidine-2-carboxylic acid tert-butyl ester To a mixture of 4,6-dibromo-8-oxa-3,5-diazatricyclo[7.4.0.02,7]trideca-1(13),2(7),3,5,9,11-hexaene (5.56 g, 16.9 mmol) in 93 mL of DMF was added (2S,4S)-4-hydroxypyrrolidine-2-carboxylic acid tert-butyl ester hydrochloride (4.17 g, 16.6 mmol) and potassium carbonate (7.03 g, 50.8 mmol). After stirring overnight at room temperature, the reaction mixture was poured into water and neutralized with 4 M aqueous HCl. The precipitate was collected by filtration and dried in vacuo. ESI-MS: 434 / 436 [M+H] + R t (HPLC): 0.64 min (Method A)

[0167] Intermediate N-S: Step 1: Dissolve 2-allyloxyacetic acid (9.02 g; 73.8 mmol) in DCM with a drop of DMF and cool to 0 °C. Add oxalyl chloride (23.4 g; 184 mmol) dropwise. Stir the reaction mixture at 0 °C for 2 h. Then evaporate the solvent under reduced pressure. Dissolve the residue in DMF and add it to a stirred solution of 3-aminobenzofuran-2-carboxamide (13.0 g; 73.8 mmol) in DMF. After 2 h, pour the mixture into 100 mL of water and stir for 5 min. Collect the solid formed, dissolve it in DCM and dry over magnesium sulfate. Evaporate the volatiles and triturate the solid with tert-butyl methyl ether to give 3-[2-(prop-2-en-1-yloxy)acetamido]-1-benzofuran-2-carboxamide.

[0168] Step 2: Stir a suspension of the product from Step 1 in 4 M aqueous sodium hydroxide at 70 °C for 2 h. Acidify the mixture by adding aqueous hydrochloric acid and collect the precipitate formed and dry to give 4-[(prop-2-en-1-yloxy)methyl]-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7]13-1(9),2(7),3,10,12-penten-6-one.

[0169] The product of step 2 was further reacted in 2 steps according to the general procedures Int-C and then Int-D to give the title compound. ESI-MS:426[M+H] + R t (HPLC):0.49min (Method A)

[0170] Intermediate NT: Step 1: Sodium hydride (60% in mineral oil; 1.73 g, 44.2 mmol) was added portionwise to sodium 2-chloro-2,2-difluoroacetate (4.5 g, 29.5 mmol) and prop-2-en-1-ol (2.14 g, 36.8 mmol) in THF (30 mL) at 0 ° C under nitrogen over a period of 2 minutes. The resulting suspension was stirred at 65 ° C for 16 hours. The reaction mixture was cooled and diluted with aqueous hydrochloric acid (2M) until pH = 5 to 6 was reached, and the aqueous layer was then extracted twice with DCM (30 mL). The combined organic layers were washed with brine, dried over sodium sulfate, filtered and evaporated to give the crude product. The crude product was purified by FC (silica gel; petroleum ether / EtOAc 0%->30%) to give 2-(allyloxy)-2,2-difluoroacetic acid. 1 H NMR (400MHz, CDCl3) δ9.34 (s, 1H), 6.01-5.91 (m, 1H), 5.45-5.36 (m, 1H), 5.33-5.26 (m, 1H), 4.50 (dt, J = 5.8, 1.2Hz, 2H)

[0171] Step 2: At 0 ° C under a nitrogen atmosphere, 3-amino-1-benzofuran-2-carboxamide (4.63 g, 26 mmol) was added to a solution of the product of step 1 (5 g, 33 mmol) in pyridine (100 mL), followed by dropwise addition of phosphorus oxychloride (15.3 g, 0.1 mol). The resulting mixture was stirred at room temperature overnight. The mixture was diluted with water (200 mL) and extracted three times with EtOAc (200 mL). The combined organic layers were washed with brine, dried over sodium sulfate and concentrated. The residue was purified by FC (silica gel; petroleum ether / EtOAc 15%) to give 3-(2-(allyloxy)-2,2-difluoroacetamido)benzofuran-2-carboxamide. ESI-MS: 311[M+H] +

[0172] Step 3: The product from Step 2 (3 g, 9.7 mmol) was added to an aqueous sodium hydroxide solution (14.5 mL, 4 M, 58.2 mmol), and then THF (1.5 mL) was added. The reaction mixture was stirred at 70 °C for 4 h. The cooled (RT) reaction mixture was acidified to pH = 5 to 6 with aqueous hydrochloric acid (2 M), and the formed precipitate was collected and dried to give crude 2-((allyloxy)difluoromethyl)benzofuro[3,2-d]pyrimidin-4(3H)-one, which was used in the next step without further purification.

[0173] Step 4: DMF (1.36 g, 18.6 mmol) and DCM (250 mL) were charged into a 0.5 L three-necked flask equipped with a thermometer and a nitrogen balloon. The flask was cooled to 0 °C, and then a solution of oxalyl chloride (3.54 g, 27.9 mmol) in DCM (5 mL) was added dropwise over 5 min while maintaining the temperature at 0 °C to 5 °C. The mixture was stirred at ambient temperature for 0.5 h. The reaction mixture was cooled to 0 °C in an ice bath and the product from Step 3 (1.8 g, 6.2 mmol) was added portionwise. The reaction mixture was stirred at room temperature for 15 min and then at 40 °C for 2 h. After cooling to room temperature, the reaction mixture was poured into ice, neutralized with aqueous sodium bicarbonate, and extracted twice with DCM (100 mL). The combined organic phases were washed with water, dried over sodium sulfate and concentrated. The crude product was purified by FC (silica gel; petroleum ether / EtOAc 2% -> 10%) to give the title compound 2-((allyloxy)difluoromethyl)-4-chlorobenzofuro[3,2-d]pyrimidine. 1 H NMR (400 MHz, DMSO-d6) δ 8.41–8.31 (m, 1H), 7.79 (m, 2H), 7.60–7.54 (m, 1H), 7.26 (s, 1H), 6.06 (dq, J = 10.8, 6.0 Hz, 1H), 5.45 (dd, J = 17.2, 1.2 Hz, 1H), 5.30 (dd, J = 10.4, 1.0 Hz, 1H), 4.70 (d, J = 6.0 Hz, 2H)

[0174] Step 5: The product from Step 4 was reacted according to the general procedure Int-D to give the title compound. ESI-MS: 462 [M+H] + R t (HPLC): 0.68 min (Method A)

[0175] Intermediate N-U: (2S,4S)-1-[4-(2-Ethoxy-2-oxoethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]-4-hydroxypyrrolidine-2-carboxylic acid tert-butyl ester Step 1: Ethyl 2-{6-oxo-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(9),2(7),3,10,12-pentaen-4-yl}acetate To a mixture of ethyl 3-aminobenzofuran-2-carboxylate (5.00 g; 24.4 mmol) in 4 M aqueous hydrochloric acid (50.00 mL; 200 mmol) at room temperature was added ethyl cyanoacetate (5.19 mL; 48.7 mmol). The mixture was heated at 100 °C for 4 h. After cooling to room temperature, 2.6 mL (24.4 mmol) of ethyl cyanoacetate was added and heating was continued at 100 °C for 48 h. The solvent was evaporated and the crude residue was diluted with 100 mL of MeOH, filtered and dried. The crude product was used directly in the next step. ESI-MS: 273 [M+H] + R t (HPLC): 0.63 min (Method B)

[0176] Step 2: Ethyl 2-{6-chloro-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(9),2(7),3,5,10,12-hexaen-4-yl}acetate A mixture of ethyl 2-{6-oxo-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(9),2(7),3,10,12-pentaen-4-yl}acetate (3.35 g; 12.3 mmol) in phosphorus oxychloride (50.0 mL; 547 mmol) was heated at 110 °C for 1.5 h. The reaction mixture was cooled to room temperature and added dropwise with stirring over 30 min to an ice bath (500 mL). Ethyl acetate was added and the layers were separated. Saturated bicarbonate solution was added slowly to the organic layer and the phases were separated. The organic layer was washed with water and brine, dried over sodium sulfate, filtered and concentrated in vacuo. ESI-MS: 291 [M+H] + R t (HPLC): 0.43 min (Method A)

[0177] Step 3: (2S,4S)-1-[4-(2-Ethoxy-2-oxoethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]-4-hydroxypyrrolidine-2-carboxylic acid tert-butyl ester To 30 mL of NMP containing ethyl 2-{6-chloro-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(9),2(7),3,5,10,12-hexaen-4-yl}acetate (8.17 mmol; 1.00 eq, 2.50 g) was added (2S,4S)-tert-butyl 4-hydroxypyrrolidine-2-carboxylate hydrochloride (8.99 mmol; 2.01 g) and DIPEA (27.0 mmol; 4.64 mL), and the resulting mixture was stirred at 70 °C for 1.5 h. The reaction mixture was cooled to room temperature and slowly added to 300 mL of ice water. The precipitate was filtered, washed several times with water and dried. ESI-MS: 442 [M+H] + R t (HPLC): 0.67 min (Method B)

[0178] Intermediate N-V: Similar to the reaction sequence described for the synthesis of Intermediate N-R, it was prepared from 11-fluoro-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2(7),9,11-tetraene-4,6-dione. Similar to the synthesis of 11-chloro-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2(7),9,11-tetraene-4,6-dione described in WO2019059577, the starting material 11-fluoro-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2(7),9,11-tetraene-4,6-dione was prepared.

[0179] Preparation of the Final Compounds The absolute configuration of the piperidine moiety of the compound assigned as R2 being methyl has been assigned in part (e.g., for Example 1.01) according to the method described by D.J. Patel et al., PNAS 2019, 11946 - 11955 (doi.org / 10.1073 / pnas.1905013116) based on the co-crystal structure with human cGAS protein. In other cases, the absolute configuration has been assigned based on the assumption that the more potent diastereomer always has the (S,S,S,S)-configuration.

[0180] Example 1.01 (General Procedures A, B, C, D) Step 1 (General Procedure A): In a dried glassware under argon atmosphere, sodium hydride (55% in mineral oil; 73.0 mg; 1.67 mmol) was added to Intermediate N-A (216 mg; 0.418 mmol) and Intermediate B-01 (188 mg; 0.439 mmol) dissolved in DMA (4.05 mL). After the bubbling had subsided, the reaction mixture was warmed to 35 °C and stirred for 25 min. The reaction was then quenched by adding ice water, diluted with EtOAc and acidified by adding aqueous hydrochloric acid (1 M; 1.7 mL). The mixture was extracted three times with EtOAc, and the combined organic layers were washed with water and brine and evaporated to dryness. The crude reaction mixture was purified by FC (silica gel; cyclohexane / EtOAc 25% -> 100%). ESI-MS: 833 [M+H] + R t (HPLC): 0.92 min (Method B)

[0181] Step 2 (General Procedure B): TosOH (124 mg; 0.653 mmol) was added to a mixture of the Intermediate from Step 1 (272 mg; 0.326 mmol) and ACN (10 mL), and the mixture was stirred in a sealed vial at room temperature for 3 days. Reaction control by HPLC indicated a low conversion rate. TosOH (45.0 mg; 0.237 mmol) was added again and the mixture was further heated at 40 °C until reaction control by HPLC indicated a high conversion rate of the desired product. DMF and a drop of water were added, and then the mixture was filtered and purified by HPLC (Sunfire C18; ACN / water, with TFA as a modifier). ESI-MS: 733 [M+H] + R t (HPLC): 0.62 min (Method B)

[0182] Step 3 (General Procedure C): In a dried glassware, under argon, with vigorous stirring, a solution of the intermediate from Step 2 (145 mg; 0.198 mmol) in DMF (12 mL) was slowly added via a syringe pump to a solution of HATU (81.2 mg; 0.214 mmol) in DMF (4 mL). The mixture was further stirred until the reaction control by HPLC indicated a high conversion of the desired product. Subsequently, the reaction was quenched by adding water, extracted three times with EtOAc, and the combined organic layers were then washed with water and brine and evaporated to dryness. The crude reaction mixture was purified by FC (cyclohexane / EtOAc 50% -> 100%, then EtOAc / MeOH 0% -> 10%). ESI-MS: 715 [M+H] + R t (HPLC): 0.80 min (Method B)

[0183] Step 4 (General Procedure D): A mixture of the intermediate from Step 3 (79.0 mg; 0.110 mmol), DCM (1.06 ml), and TFA (852 μL) was stirred at room temperature for 90 min. Since the reaction control by HPLC indicated a low conversion, the temperature was raised to 37 °C and stirring was continued for 3 h. Since the reaction control by HPLC still indicated a low conversion, the temperature was raised to 45 °C and stirring was continued until the reaction control by HPLC indicated a high conversion of the desired product. The mixture was evaporated and the crude product was purified by preparative HPLC (Sunfire C18; ACN / water, with TFA as the modifier). ESI-MS: 659 [M+H] + R t (HPLC): 0.83 min (Method E)

[0184] Similar to Example 1.01 above, the following compounds were prepared following General Procedures A, B, C, and D:

[0185] Example 2.01: React intermediate B-09 according to General Procedure D to obtain the TFA salt of racemic-trans-7-chloro-2-(methylsulfonyl)-1-(3-methylpiperidin-4-yl)-1H-1,3-benzodiazole

[0186] Step 2: React the product of Step 1 with 1.2 equivalents of 4-{[(tert-butoxy)carbonyl]amino}butyric acid according to General Procedure C

[0187] Step 3: React the product of Step 2 with intermediate N-P according to General Procedure A to obtain 2-{6-[(2S,4S)-2-[(tert-butoxy)carbonyl]-4-({1-[1-(4-{[(tert-butoxy)carbonyl]amino}butanoyl)-3-methylpiperidin-4-yl]-7-chloro-1H-1,3-benzodiazol-2-yl}oxy)pyrrolidin-1-yl]-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(9),2(7),3,5,10,12-hexaen-4-yl}acetic acid (with racemic trans configuration at the piperidine moiety).

[0188] Step 4: At room temperature, deprotect the BOC group of the product of Step 3 by reacting it with dioxane containing 1.5 equivalents of hydrochloric acid (4 M in dioxane) overnight. The crude product is purified by RP HPLC (Sunfire C18; ACN / water / modifier: TFA) to obtain the TFA salt of 2-{6-[(2S,4S)-4-({1-[1-(4-aminobutanoyl)-3-methylpiperidin-4-yl]-7-chloro-1H-1,3-benzodiazol-2-yl}oxy)-2-[(tert-butoxy)carbonyl]pyrrolidin-1-yl]-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(9),2(7),3,5,10,12-hexaen-4-yl}acetic acid (with racemic trans configuration at the piperidine moiety).

[0189] Step 5: React the product of Step 4 according to General Procedure C to obtain (12S,14S)-4-chloro-37-methyl-29,34-dioxo-11,18-dioxa-2,9,15,26,30,35,40-heptaazaoctacyclo[33.2.2.1 12,15 .1 16,27 .0 2,10 .0 3,8 .0 17 ,25 .0 19,24tert-Butyl 41-3,5,7,9,16(40),17(25),19(24),20,22,26-decene-14-carboxylate (with racemic trans configuration at the piperidine moiety).

[0190] Step 6: React the product of Step 5 according to General Procedure D to obtain the title compound (12S,14S)-4-chloro-37-methyl-29,34-dioxo-11,18-dioxa-2,9,15,26,30,35,40-heptaazaoctacyclo[33.2.2.1 12,15 .1 16,27 .0 2 ,10 .0 3,8 .0 17,25 .0 19,24 41-3,5,7,9,16(40),17(25),19(24),20,22,26-decene-14-carboxylic acid (with racemic trans configuration at the piperidine moiety). ESI-MS: 672 [M+H] + R t (HPLC): 0.88 min (Method E)

[0191] Example 3.01: Remove the BOC protecting group from Intermediate I.02 according to General Procedure B to obtain the TFA salt of (2S,4S)-1-{4-[1,1-difluoro-2-(prop-2-en-1-yloxy)ethyl]-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2,4,6,9,11-hexaen-6-yl}-4-({1-[(3S,4S)-3-methylpiperidin-4-yl]-1H-1,3-benzodiazol-2-yl}oxy)pyrrolidine-2-carboxylic acid tert-butyl ester. ESI-MS: 689 [M+H] + R t (HPLC): 0.62 min (Method A)

[0192] Step 2: General Procedure G: At 0 °C, DCC (1 M; 165 μL; 0.165 mmol) was added dropwise to a solution of the product of step 1 (105 mg; 0.127 mmol), vinylacetic acid (14.5 μL; 0.165 mmol), and DMAP (2.0 mg; 0.016 mmol) in DCM (1.05 mL). With stirring, the mixture was warmed to room temperature and then stirred for an additional hour. The mixture was evaporated to dryness and the residue was subjected to FC (CH / EtOAc 40% -> 100%). The resulting impure product was further purified by RP HPLC (Sunfire C18; ACN / water, modifier: TFA) to give tert-butyl (2S,4S)-4-({1-[(3S,4S)-1-(but-3-enoyl)-3-methylpiperidin-4-yl]-1H-1,3-benzodiazol-2-yl}oxy)-1-{4-[1,1-difluoro-2-(prop-2-en-1-yloxy)ethyl]-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2,4,6,9,11-hexaen-6-yl}pyrrolidine-2-carboxylate. ESI-MS: 757 [M+H] + R t (HPLC): 0.77 min (method A)

[0193] Step 3: General procedure H: In a dried glass vessel, Grubbs II catalyst (5.0 mg) was added to a solution of the product of step 2 (by azeotropic co-distillation with toluene; 77.0 mg; 0.102 mmol) in 1,2-dichloroethane (degassed; 9.0 mL). The mixture was stirred at 80 °C for 24 h, then the same amount of catalyst was added again and the mixture was stirred for another 24 h. The reaction was quenched by the addition of imidazole (10 mg), and then it was cooled to room temperature. The mixture was concentrated and subjected to FC (silica gel; CH / EtOAc 20% -> 100%) to give tert-butyl (1S,12S,14S,32E,38S)-28,28-difluoro-38-methyl-35-oxo-11,18,30-trioxa-2,9,15,26,36,41-hexaazaoctacyclo[34.2.2.1 12,15 .1 16,27 .0 2,10 .0 3,8 .0 17,25 .0 19,24 tetracont-3(8),4,6,9,16(41),17(25),19,21,23,26,32-undecaene-14-carboxylate. ESI-MS: 729 [M+H] + R t (HPLC): 0.73 min (Method A)

[0194] Step 4: In a Parr apparatus, a mixture of the product from Step 3 (22.0 mg; 0.0302 mmol), ethanol (1.5 mL), and 10% Pd / C was kept under hydrogen (5 psi) overnight. The mixture was filtered, evaporated, and used for the next step. ESI-MS: 731 [M+H] + R t (HPLC): 1.09 min (Method E)

[0195] Step 5: The product from Step 4 (22.1 mg) was reacted according to General Procedure D. The product was further purified by RP HPLC (Sunfire C18, ACN / water, modifier: TFA). ESI-MS: 675 [M+H] + R t (HPLC): 0.60 min (Method A)

[0196] Example 4.01: Step 1: To a degassed solution of Intermediate I.02 (150 mg; 0.190 mmol) in ethyl acrylate (1.56 mL; 14.3 mmol) was added Grubbs II catalyst (8.1 mg; 0.0095 mmol). The mixture was stirred for 24 h, then evaporated and subjected to FC (CH / EtOAc 30% -> 100%) to give (3S,4S)-4-(2-{[(3S,5S)-5-[(tert-butoxy)carbonyl]-1-[4-(2-{[4-ethoxy-4-oxobut-2-en-1-yl]oxy}-1,1-difluoroethyl)-8-oxa-3,5-diazatricyclo 2,7 trideca-1(13),2,4,6,9,11-hexaen-6-yl]pyrrolidin-3-yl]oxy}-1H-1,3-benzodiazol-1-yl)-3-methylpiperidine-1-carboxylic acid tert-butyl ester. ESI-MS: 861 [M+H] + R t (HPLC): 1.28 min (Method B)

[0197] Step 2: General procedure E In a Parr apparatus, at room temperature, under hydrogen (50 psi), a mixture of the product of Step 1 (140 mg; 0.163 mmol), 10% Pd / C (28 mg), and ethanol (2.0 mL) was reacted until HPLC control indicated that the starting material had been converted (4 h). The mixture was filtered, concentrated under reduced pressure and purified by RP HPLC (XBridge C18, ACN / water, modifier: TFA) to give tert-butyl (3S,4S)-4-(2-{[(3S,5S)-5-[(tert-butoxy)carbonyl]-1-{4-[2-(4-ethoxy-4-oxobutoxy)-1,1-difluoroethyl]-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2,4,6,9,11-hexaen-6-yl}pyrrolidin-3-yl]oxy}-1H-1,3-benzodiazol-1-yl)-3-methylpiperidine-1-carboxylate. ESI-MS: 863 [M+H] + R t (HPLC): 1.28 min (Method B)

[0198] Step 3: General procedure F The product of Step 2 (118 mg; 0.137 mmol) was dissolved in a mixture of aqueous lithium hydroxide (2 M; 153 μL; 0.306 mmol), MeOH (0.80 mL), and THF (4.0 mL). The mixture was stirred at 40 °C until HPLC analysis indicated conversion (7 h), concentrated under reduced pressure, diluted with water and acidified with an equimolar amount of aqueous hydrochloric acid. One drop of methanol was added and the mixture was extracted with DCM. The organic layer was separated, concentrated under reduced pressure and used in the next step.

[0199] Step 4: The product of Step 3 was reacted according to General procedure B to give 4-(2-{6-[(2S,4S)-2-[(tert-butoxy)carbonyl]-4-({1-[(3S,4S)-3-methylpiperidin-4-yl]-1H-1,3-benzodiazol-2-yl}oxy)pyrrolidin-1-yl]-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2,4,6,9,11-hexaen-4-yl}-2,2-difluoroethoxy)butanoic acid.

[0200] Step 5: React the product of Step 4 according to General Procedure C to obtain (1S,12S,14S,37S)-28,28-difluoro-37-methyl-34-oxo-11,18,30-trioxa-2,9,15,26,35,40-hexaazaoctacyclo[33.2.2.1 12,15 .1 16,27 .0 2, 10 .0 3,8 .0 17,25 .0 19,24 tetratriaconta-3,5,7,9,16,19,21,23,25,27(40)-decene-14-yl tert-butyl carboxylate.

[0201] Step 6: React the product of Step 5 according to General Procedure D to obtain the title compound ESI-MS: 661 [M+H] + R t (HPLC): 0.90 min (Method B)

[0202] Example 5.01: Using intermediates N-S and B-11 as starting materials, perform the synthesis using the following reaction sequence: Step 1: Nucleophilic aromatic substitution using General Procedure A (reagents: N-S and B-11) Step 2: Removal of the BOC protecting group using General Procedure B Step 3: Reaction with 4-pentenoic acid using General Procedure G Step 4: Metathesis using General Procedure H Step 5: Hydrogenation using General Procedure E Step 6: Removal of the tert-butyl ester protecting group using General Procedure D ESI-MS: 643 [M+H] + R t (HPLC): 0.58 min (Method A)

[0203] Example 6.01: Using intermediates N-T and B-02 as starting materials, perform the synthesis using the following reaction sequence: Step 1: Nucleophilic aromatic substitution using General Procedure A (reagents: N-T and B-02) Step 2: Metathesis using a modified version of General Procedure H with methyl 3-butenoate as the second alkene (reaction in DCM at room temperature, 15 equivalents of methyl 3-butenoate were applied, and an additional 15 equivalents of methyl 3-butenoate and an additional 0.3 equivalents of catalyst were added portionwise after 24 h). Step 3: Hydrogenation using General Procedure E (solvent: EtOAc) Step 4: Ester saponification using General Procedure F (aqueous lithium hydroxide / THF, 50 °C, 5 h) Step 5: Removal of the BOC protecting group using General Procedure B Step 6: Ring-closing amidation using General Procedure C (solvent: DMA) Step 7: Removal of the tert-butyl ester protecting group using General Procedure D ESI-MS: 695 [M+H] + R t (HPLC): 0.72 min (Method A)

[0204] Example 7.01: Starting from intermediate N-R and intermediate B-10, the following reaction sequence was applied for the synthesis: Step 1: Nucleophilic aromatic substitution using General Procedure A (reagents: N-R and B-10) Step 2: Removal of the BOC protecting group using General Procedure B Step 3: Amidation using General Procedure C with (3R)-3-(prop-3-en-1-yloxy)butanoic acid Step 4: To a degassed solution of the product from Step 3 (162 mg; 0.213 mmol), cesium carbonate (174 mg; 0.533 mmol), and silver iodide (55.1 mg; 0.235 mmol) in dioxane (under argon) was added the catalyst [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (21.3 mg; 0.0320 mmol). The mixture was stirred at 120 °C for 24 h and then cooled to room temperature. The mixture was diluted with ACN / methanol, filtered, and purified by preparative RP HPLC (XBridge C18, ACN / water, modifier: TFA) to give (12S,14S,32R)-32-methyl-34-oxo-11,18,31-trioxa-2,9,15,26,35,40-hexaazaoctacyclo[33.2.2.1 12,15 .1 16,27 .0 2,10 .0 3,8 .017,25 .0 19,24 tert-Butyl 41-3(8),4,6,9,16(40),17(25),19(24),20,22,26,28-undeca-14-enoate ESI-MS: 679 [M+H] + R t (HPLC): 0.60 min (Method A) Step 5: Hydrogenation using General Procedure E Step 6: Removal of the tert-butyl ester protecting group using General Procedure D ESI-MS: 625 [M+H] + R t (HPLC): 0.47 min (Method A)

[0205] Example 8.01: Using intermediates N-S and B-10 as starting materials, the following reaction sequence was applied for synthesis: Step 1: Nucleophilic aromatic substitution using General Procedure A (reagents: N-S and B-10) Step 2: Reaction under argon. At room temperature, ethyl bromodifluoroacetate (209 mg; 1.00 mmol) and phenylsilane (223 mg; 2.00 mmol) were added to a degassed mixture of the product from Step 1 (381 mg; 0.500 mmol), nickel(II) chloride (3.3 mg; 0.025 mmol), sodium carbonate (53 mg; 0.50 mmol) and anhydrous DMF (4.77 ml). The mixture was stirred at 70 °C for 14 h in a sealed container. The mixture was poured into saturated brine and the resulting mixture was extracted twice with EtOAc. The combined organic layers were dried over sodium sulfate, filtered and evaporated. The crude product was purified by preparative RP HPLC (XBridge C18; ACN / water, modifier: TFA) to give tert-butyl 4-(2-{[(3S,5S)-5-[(tert-butoxy)carbonyl]-1-(4-{[(5-ethoxy-4,4-difluoro-5-oxopentyl)oxy]methyl}-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2,4,6,9,11-hexaen-6-yl)pyrrolidin-3-yl]oxy}-1H-1,3-benzodiazol-1-yl)piperidine-1-carboxylate ESI-MS: 649 [M+H] + R t(HPLC): 0.80 min (Method A) Step 3: Ester saponification using General Procedure F (aqueous lithium hydroxide / THF, room temperature, 2.5 h) Step 4: Removal of the BOC protecting group using General Procedure B Step 5: Cyclization amidation using General Procedure C, using DMF containing HATU Step 6: Removal of the tert-butyl ester protecting group using General Procedure D ESI-MS: 647 [M+H] + R t (HPLC): 0.53 min (Method A)

[0206] Example 9.01: Starting from intermediate N-R and intermediate B-10, the following reaction sequence was applied for synthesis: Step 1: Nucleophilic aromatic substitution using General Procedure A (reagents: N-R and B-10) Step 2: General Procedure I: Under argon, palladium(II) acetate (9.0 mg; 0.040 mmol) and tri-o-tolylphosphine (24.4 mg; 0.080 mmol) were added to a degassed solution of the product of Step 1 (326 mg; 0.400 mmol), allyloxyacetic acid (133 μL; 1.20 mmol), and triethylamine (112 μL; 0.800 mmol) in DMF (6.52 mL). The vial was sealed and the reaction mixture was stirred at 115 °C for 2.5 h. The reaction mixture was diluted with ACN / H2O, filtered, and purified directly by preparative RP HPLC (XBridge C18; 30 - 100% ACN / H2O, modifier: ammonia) to give a mixture of E / Z isomers of 2-{[(3-{6-[(2S,4S)-2-[(tert-butoxy)carbonyl]-4-[(1-{1-[(tert-butoxy)carbonyl]piperidin-4-yl}-1H-1,3-benzodiazol-2-yl)oxy]pyrrolidin-1-yl]-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2,4,6,9,11-hexaen-4-yl}prop-2-en-1-yl]oxy}acetic acid. ESI-MS: 769 [M+H] + R t (HPLC): 0.57 / 0.58 min (Method F) Step 3: Hydrogenation using General Procedure E (solvent: EtOAc) Step 4: Removal of the BOC protecting group using General Procedure B Step 5: Intramolecular amide cyclization using General Procedure C with DMF containing HATU Step 6: Removal of the tert-butyl ester protecting group using General Procedure D ESI-MS: 597 [M+H] + R t (HPLC): 0.42 min (Method A)

[0207] Examples 10.01 and 10.02: Starting from intermediate N-R and intermediate I.03, the following reaction sequence was applied for synthesis: Step 1: Nucleophilic aromatic substitution using General Procedure A (reagents: N-R and I.03) Step 2: Intramolecular Heck coupling using General Procedure I (reaction time 17 h; 100 °C) Step 3: Hydrogenation using General Procedure E (catalyst: Raney-Ni; solvent: methanol) Step 4: Removal of the tert-butyl ester protecting group using General Procedure D Step 5: The resulting diastereomeric mixture was separated by preparative SFC (instrument: Sepiatec PrepSFC50; column: CHIRALART Cellulose-SC (10 mm × 250 mm, 5 μm); mobile phase: A for CO2 and B for IPA [+20 mM NH3]; gradient: B% = 25% isocratic elution mode; flow rate: 15 mL / min; wavelength: 220 nm; column temperature: 40 °C; system back pressure: 150 bar). Example 10.01 (first eluate) ESI-MS: 625 [M+H] + R t (HPLC): 0.804 min (Method E) and Example 10.02 (second eluate). ESI-MS: 625 [M+H] + R t (HPLC): 0.814 min (Method E)

[0208] Example 11.01: Starting from intermediate N-R and intermediate B-10 as starting materials, the synthesis is carried out using the following reaction sequence: Step 1: Nucleophilic aromatic substitution using General Procedure A (reagents: N-R and B-10) Step 2: Stir a mixture of the product of Step 1 (500 mg; 0.682 mmol), 1,4-butanediol (1.22 mL; 13.6 mmol), potassium tert-butoxide (229 mg; 2.04 mmol), and ACN (10 mL; dried over molecular sieves) in a sealed vial at 80 °C overnight. Evaporate the mixture and purify the residue by RP HPLC (Sunfire C18, ACN / water, modifier: TFA) to obtain tert-butyl 4-(2-{[(3S,5S)-5-[(tert-butoxy)carbonyl]-1-[4-(4-hydroxybutoxy)-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(13),2,4,6,9,11-hexaen-6-yl]pyrrolidin-3-yl]oxy}-1H-1,3-benzodiazol-1-yl)piperidine-1-carboxylate. ESI-MS: 743 [M+H] + R t (HPLC): 0.70 min (Method A) Step 3: Stir a mixture of the product of Step 2 (143 mg; 0.193 mmol), 4-nitrophenyl chloroformate (87.1 mg; 0.424 mmol), pyridine (103 μL; 1.27 mmol), and DCM (6.0 mL) at room temperature for 4 h. Evaporate the volatiles and use the crude product without further purification in the next step. Step 4: Carry out acid removal of the BOC protecting group using the conditions of General Procedure D. Step 5: For the formation of a closed loop for the carbamate, dissolve the crude product of Step 4 in an excess of DIPEA and stir at 70 °C for 90 min. Evaporate the mixture and use it without purification in the next step. Step 6: Step 4: Carry out removal of the tert-butyl ester protecting group using General Procedure D ESI-MS: 613 [M+H] + R t (HPLC): 0.50 min (Method A)

[0209] Example 12.01 (racemic mixture of the trans isomers): Step 1: To a mixture of tert-butyl 3-(2-hydroxyethoxy)propionate (1.45 g; 7.62 mmol), pyridine (663 mg; 8.38 mmol) and DCM (15 mL) cooled to 0 °C was added dropwise a solution of 4-nitrophenyl chloroformate (1.54 g; 7.62 mmol) in DCM (15 mL). The mixture was stirred overnight at room temperature, then diluted with DCM and extracted with water. The organic layer was evaporated and the crude product was used in the next step.

[0210] Step 2: A mixture of the crude product from Step 1 (686 mg), intermediate B-12 (500 mg), diisopropylethylamine (742 μL) and THF (8.0 mL) was refluxed for 2 h. The mixture was evaporated, dissolved in EtOAc and extracted twice with aqueous sodium hydroxide solution (1 M), then washed with water and brine. The organic layer was evaporated to dryness and the crude reaction product was purified by FC (silica gel; petroleum ether / EtOAc 5% -> 35%) to give racemic trans tert-butyl 2-{3-[3-methyl-4-[2-(methylthio)-1H-1,3-benzodiazol-1-yl]piperidine-1-carbonyloxy]propoxy}acetate. ESI-MS: 478 [M+H] + R t (HPLC): 0.85 min (Method E)

[0211] Step 3: Apply general procedure D and perform acidic ester cleavage without chromatographic purification.

[0212] Step 4: To a solution of the product from Step 3 (579 mg) in ACN (10 mL) at room temperature was added 1-chloro-N,N,2-trimethylpropenylamine (259 μL; 3.92 mmol). The mixture was stirred overnight, evaporated, dissolved in DCM and extracted with water. The organic layer was evaporated and passed through FC (silica gel; petroleum ether / EtOAc 45% -> 95%). ESI-MS: 580 [M+H] + R t (HPLC): 0.81 min (Method E)

[0213] Step 5: Suspend the product of Step 4 (650 mg; 1.12 mmol) in aqueous sodium hydroxide solution (4 M; 15 mL). Stir the mixture at 60 °C for 1 h, then cool it to room temperature and acidify by adding aqueous hydrochloric acid (4 M). Decant the supernatant and purify the solid by RP HPLC (Sunfire C18, ACN / water, modifier: TFA). ESI-MS: 562 [M+H] + R t (HPLC): 0.76 min (Method E)

[0214] Step 6: Suspend the product of Step 5 (390 mg; 0.694 mmol) in phosphorus oxychloride (6 mL). Stir the mixture at 90 °C for 1 h, then evaporate. Carefully add water to the residue and extract the resulting mixture with EtOAc. Separate the organic layer and evaporate to dryness to obtain racemic trans-3-({6-chloro-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(9),2(7),3,5,10,12-hexaen-4-yl}methoxy)propyl-3-methyl-4-[2-(methylthio)-1H-1,3-benzodiazol-1-yl]piperidine-1-carboxylate. ESI-MS: 580, 582 [M+H] + R t (HPLC): 0.89 min (Method E)

[0215] Step 7: Stir a mixture of the product of Step 6 (300 mg; 0.517 mmol), (2S,4S)-tert-butyl 4-hydroxypyrrolidine-2-carboxylate hydrochloride (142 mg; 0.621 mmol), potassium carbonate (173 mg; 1.19 mmol) and DMF (7.0 mL) at room temperature overnight. Add water, collect the precipitate and purify by RP HPLC (Sunfire C18, ACN / water, modifier: TFA) to obtain 3-({6-[(2S,4S)-2-[(tert-butoxy)carbonyl]-4-hydroxypyrrolidin-1-yl]-8-oxa-3,5-diazatricyclo[7.4.0.0 2 ,7 trideca-1(9),2(7),3,5,10,12-hexaen-4-yl}methoxy)propyl-3-methyl-4-[2-(methylthio)-1H-1,3-benzodiazol-1-yl]piperidine-1-carboxylate (the piperidine moiety has a racemic trans configuration). ESI-MS: 731 [M+H]+ R t (HPLC): 0.77 min (Method E)

[0216] Step 8: Dissolve the product of Step 7 (239 mg; 0.327 mmol) in DCM (8.0 mL). Add meta-chloroperbenzoic acid (77%; 161 mg; 0.719 mmol) and stir the mixture for 2 h, then wash with sodium bicarbonate solution. Evaporate the organic layer to obtain 3-({6-[(2S,4S)-2-[(tert-butoxy)carbonyl]-4-hydroxypyrrolidin-1-yl]-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(9),2(7),3,5,10,12-hexaen-4-yl}methoxy)propyl 4-(2-methylsulfonyl-1H-1,3-benzodiazol-1-yl)-3-methylpiperidine-1-carboxylate (racemic trans configuration with piperidine moiety). ESI-MS: 763 [M+H] + R t (HPLC): 0.85 min (Method E)

[0217] Step 9: Apply General Procedure A, cyclization by aromatic nucleophilic substitution.

[0218] Step 10: Apply acidic tert-butyl ester cleavage of General Procedure D to obtain the title compound. ESI-MS: 627 [M+H] + R t (HPLC): 0.83 min (Method E)

[0219] Example 13.01: Step 1: Add benzyl chloroformate (774 μL; 5.27 mmol) dropwise to a cold solution of N-methyl-(pent-4-en-1-yl)amine (500 mg; 4.79 mmol) in aqueous sodium carbonate solution (2 M; 4.79 mL; 9.58 mmol). Stir the mixture at 0 °C for 1 h, then dilute with water and extract three times with EtOAc. Wash the combined organic layers with water and then with brine, dry over magnesium sulfate, filter and concentrate under reduced pressure. Purify the crude product by FC (silica gel; cyclohexane / EtOAc 25% -> 100%). ESI-MS: 234 [M+H] + R t(HPLC): 0.67 min (Method A)

[0220] Step 2: The reaction is carried out in a dried glassware under argon. Triethylamine (180 μL; 1.30 mmol) is added to a mixture of the product of Step 1 (191 mg; 0.778 mmol), Intermediate I.01 (200 mg; 0.259 mmol), 1,1'-bis(di-tert-butylphosphino)ferrocene dichloropalladium (20.2 mg; 0.0311 mmol) and previously degassed DMA (2.4 mL). The reaction mixture is degassed and then heated to 115 °C in a sealed vial for 10 h. The mixture is evaporated and the residue is purified by preparative RP HPLC (XBridge C18, ACN / water, modifier: ammonia) to obtain (3S,4S)-4-(2-{[(3S,5S)-1-{4-[5-{[(benzyloxy)carbonyl](methyl)amino}pent-1-en-1-yl]-8-oxa-3,5-diazatricyclo[7.4.0.0 2 ,7 trideca-1(9),2(7),3,5,10,12-hexaen-6-yl}-5-[(tert-butoxy)carbonyl]pyrrolidin-3-yl]oxy}-1H-1,3-benzodiazol-1-yl)-3-methylpiperidine-1-carboxylic acid tert-butyl ester. Step 3: Apply General Procedure E for hydrogenation (including removal of the Cbz protecting group) without chromatographic purification.

[0221] Step 4: Triethylamine (125 μL; 1.04 mmol) and 4-nitrophenyl chloroformate (53 mg; 0.26 mmol) are added to a solution of the crude product of Step 3 (199 mg) in DCM (3 mL). The mixture is stirred at room temperature for 1 h. The mixture is evaporated and purified by FC (cyclohexane / EtOAc 20% -> 100%) to obtain (3S,4S)-4-(2-{[(3S,5S)-5-[(tert-butoxy)carbonyl]-1-[4-(5-{methyl[(4-nitrophenoxy)carbonyl]amino}pentyl)-8-oxa-3,5-diazatricyclo[7.4.0.0 2,7 trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]pyrrolidin-3-yl]oxy}-1H-1,3-benzodiazol-1-yl)-3-methylpiperidine-1-carboxylic acid tert-butyl ester.

[0222] Step 5: Apply the BOC protecting group removal of General Procedure B

[0223] Step 6: Over 25 minutes, the crude product from Step 5 was added dropwise via an injection pump to a solution of N,N-diisopropylethylamine (49 μL) in THF (1.5 mL). The mixture was stirred at 80 °C for 10 h. The mixture was passed through a sodium bicarbonate cartridge and evaporated. N-Methylpyrrolidone (3.0 mL) and DIPEA (100 μL) were added and the mixture was heated in a microwave oven for 30 min to 180 °C. The mixture was purified by RP HPLC (XBridge C18, ACN / water, modifier: TFA). ESI-MS: 695 [M+H] + R t (HPLC): 0.61 min (Method A)

[0224] Step 7: Apply the tert-butyl ester cleavage of General Procedure D to obtain the title compound. ESI-MS: 638 [M+H] + R t (HPLC): 0.55 min (Method A)

[0225] General Technical Remarks The terms "ambient temperature" and "room temperature" are used interchangeably and refer to a temperature of about 20 °C, e.g., 15 °C to 25 °C.

[0226] Generally, the 1 1H NMR spectra and / or mass spectra of the prepared compounds have been obtained. Unless otherwise stated, all chromatographic operations are carried out at room temperature.

[0227] List of Abbreviations ACN Acetonitrile aq. Aqueous solution BOC tert-Butyloxycarbonyl °C Degree Celsius CH Cyclohexane DCM Dichloromethane DIPEA N,N-Diisopropylethylamine DMA Dimethylacetamide DMAP 4-Dimethylaminopyridine DMF N,N-Dimethylformamide DMSO Dimethyl sulfoxide EDC ({[3-(Dimethylamino)propyl]imino}methylene)(ethyl)amine ESI-MS Electrospray ionization mass spectrometry (m / z) EtOAc Ethyl acetate FC Flash chromatography, using SiO2 if no other details are given Grubbs II Dichloro[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene]-(benzylidene)-(tricyclohexylphosphine) Ruthenium (II) h hour HCl Hydrochloric acid HATU [Dimethylamino-(1,2,3-triazolo[4,5-b]pyridin-3-yloxy)-methylene]-dimethyl-ammonium hexafluorophosphate HPLC High Performance Liquid Chromatography IPA Isopropyl alcohol MeOH Methanol min Minute mL Milliliter M Mole n.d. Not determined Pd / C(x%) Palladium (x wt%) / Carbon PFTU Pentafluorophenyl - tetramethylurea hexafluorophosphate rac Racemic RP Reverse Phase RT Room temperature (ca. 20 °C) SFC Supercritical Fluid Chromatography TFA Trifluoroacetic acid THF Tetrahydrofuran TosOH p - Toluenesulfonic acid monohydrate R t Retention time in minutes Vol% Volume percentage

[0228] Analytical Methods (HPLC / SFC): HPLC Method A: Column: XBridge BEH C18_2.1×30mm_1.7μm (Waters); CT: 60 °C HPLC Method B: Column: Sunfire C18_3.0×30mm_2.5μm (Waters); CT: 60 °C HPLC Method C: Column: XSelect HSS PFP_2.1×30mm_1.8μm (Waters); CT: 60 °C HPLC Method D: Column: Zorbax StableBond C18_3.0×30mm_1.8μm (Agilent); CT: 60 °C HPLC method E: Column: Sunfire C18_3.0×30mm_2.5μm (Waters); CT: 60 °C HPLC method F: Column: XBridge BEH C18_2.1×30mm_2.5μm (Waters); CT: 60 °C HPLC method G: Column: Kinetex XB-C18 2.6μm (4.6×50mm), CT: 25 °C HPLC method H: Column: Acquity UPLC BEH C18 1.7μm (2.1×100mm); CT: 40 °C HPLC method I: Column: Kinetex XB-C18 2.6μm (4.6×50mm), CT: 25 °C 4. Examples 5.1 Example Compounds of Formulas I, II or III of the Present Invention The following Example compounds of Formula I, II or III outlined in Table 1 have been synthesized and tested for their pharmacological properties with respect to their potency in inhibiting cGAS activity.

[0230] Specifically, the “biochemical (in vitro) IC 50 value” (hcGAS IC 50 ) for cGAS inhibition; the “IC 50 value for inhibiting IFN induction in virus-stimulated THP1 cells” (THP (vir) IC 50 ); the “IC 50 value for inhibiting IFN induction in cGAMP-stimulated THP1 cells” (THP (cGAMP) IC 50) and "Regarding the IFN-induced IC in human whole blood inhibited by dsDNA stimulation 50 value" (hWB IC 50 ). The results are summarized in Table 1.

[0231] The exemplified compounds of Formula I, II or III summarized in Table 1 simultaneously exhibit the following three properties: · A satisfactory "biochemical (in vitro) IC regarding cGAS inhibition 50 value" (where hcGAS IC 50 ≤ 100 nM, preferably ≤ 50 nM, especially ≤ 10 nM), · A satisfactory "cellular IC regarding cGAS inhibition 50 value" (where THP1 (vir) IC 50 ≤ 1 μM, preferably ≤ 500 nM, more preferably ≤ 100 nM, especially ≤ 50 nM) and · A satisfactory selectivity for cGAS inhibition (where THP1 (cGAMP) IC 50 / THP1 (vir) IC 50 ratio ≥ 10, more preferably ≥ 50, more preferably ≥ 500, especially ≥ 1000).

[0232] In addition, the exemplified compounds of Formula I, II or III also exhibit an acceptable IC regarding the IFN induction in human whole blood inhibited by dsDNA stimulation 50 value (hWB IC 50 ). Table 1: Pharmacological properties of the exemplified compounds of Formula I, II or III of the present invention

[0233] 5.2 Comparison of Example Compounds of Formulas I, II or III with Compounds of the Prior Art 5.2.1 Compounds of WO 2020 / 142729 In WO 2020 / 142729, cGAS inhibitors with partially similar structures have been disclosed.

[0234] On pages 44 and 45 of WO 2020 / 142729, the "biochemical (in vitro) IC 50 value" (corresponding to "hcGAS IC 50 ") regarding cGAS inhibition is disclosed. Herein, compounds with a "biochemical (in vitro) IC 50 value" less than 100 nM have been assigned to "Group A", compounds with a "biochemical (in vitro) IC 50 value" greater than 100 nM and less than 500 nM have been assigned to "Group B", compounds with a "biochemical (in vitro) IC 50 value" greater than 500 nM and less than 1 μM have been assigned to "Group C", compounds with a "biochemical (in vitro) IC 50 value" greater than 1 μM and less than 10 μM have been assigned to "Group D", and compounds with a "biochemical (in vitro) IC 50 value" greater than 10 μM have been assigned to "Group E" (see page 44 of WO 2020 / 142729).

[0235] On page 45 of WO 2020 / 142729, it is disclosed that only Compound No. 25 can be assigned to "Group A" with a "biochemical (in vitro) IC 50 value" less than 100 nM. All other example compounds of WO 2020 / 142729 exhibit a "biochemical (in vitro) IC 50 value" exceeding 100 nM.

[0236] Selected prior art compounds of WO 2020 / 142729, including Compound No. 25, have been synthesized and subsequently tested for their pharmacological properties regarding their potency in inhibiting the cGAS / STING pathway using the exact same assays as those used to test the compounds of the present invention. Specifically, the "biochemical (in vitro) IC 50 value" (hcGASIC 50 ) regarding cGAS inhibition of the structurally closest example of WO2020 / 142729 has been determined experimentally according to the assay methods described in Chapter 6 below; the "cellular IC 50 value" regarding the inhibition of IFN induction in virus-stimulated THP1 cells (THP1 (vir) IC 50 ); the "cellular IC 50 value" regarding the inhibition of IFN induction in cGAMP-stimulated THP1 cells (THP1 (cGAMP) IC 50 ); and the "IC 50 value" regarding the inhibition of IFN induction in human whole blood (hWB) (see Table 2). Table 2: Pharmacological properties of a series of example compounds of WO 2020 / 142729

[0237] The pharmacological properties of the example compounds of the present invention outlined in Table 1 and the respective pharmacological properties of the compounds of WO 2020 / 142729 outlined in Table 2 can be compared with each other because these compounds were determined experimentally according to the same assay procedures described in Chapter 6 below.

[0238] From the data shown in Table 2, it is evident that all example compounds of WO 2020 / 142729 exhibit a "biochemical (in vitro) IC 50 value" (= hcGAS IC 50 ) significantly greater than 100 nM, with the sole exception being Example No. 25 of WO 2020 / 142729 (in WO 2020 / 142729, designated as Group A with a "biochemical (in vitro) IC 50 value" (= hcGAS IC 50 ) less than 100 nM). In contrast, the "biochemical (in vitro) IC 50 values" (hcGASIC 50 ) of the example compounds of the present invention are all less than 100 nM. However, Example No. 25 of WO 2020 / 142729 with a "biochemical (in vitro) IC 50 value" (hcGAS IC 50 ) of 55 nM does not fully meet the selection criterion of "satisfactory cell growth inhibitory potency" exhibited by THP1 (vir) IC 50 below 1 μM, because the THP1 (vir) IC 50 of Example No. 25 of WO 2020 / 142729 is 17 μM.

[0239] 5.2.2 Compounds of WO 2022 / 174012 In WO 2022 / 174012, cGAS inhibitors with partially similar structures have been disclosed.

[0240] The "biochemical (in vitro) IC 50 value" regarding cGAS inhibition has been disclosed on page 65 of WO 2022 / 174012 and the "cellular IC 50Value (THP-1 stimulation for IFNβ ELISA). Compound 5 (BBL0100455) of WO 2022 / 174012 seems to be the only compound in WO 2022 / 174012 that may meet the selection criteria of the present invention, which means it has a) "Biochemical (or enzymatic) (in vitro) IC 50 value" less than 100 nM (in the "enzyme assay of WO 2022 / 174012", Compound 5 was measured to belong to "Group B", which represents an "enzyme IC 50 value" of 50 nM to 100 nM. See Table 2, page 65 of WO 2022 / 174012) b) And has a "cellular IC 50 value" less than 1 μM (THP-1 stimulation for IFNβ ELISA) (in the "cellular assay of WO 2022 / 174012", Compound 5 was measured to belong to "Group A", which represents a "cellular IC 50 value" of < 1 μM. See pages 67 and 68, Table 3 of WO 2022 / 174012).

[0241] However, both the biochemical / enzyme assay and the cellular assay of WO 2022 / 174012 are different from the respective "biochemical / enzyme assay and cellular assay" of the present invention. Therefore, the biochemical / enzyme IC 50 value and the cellular IC 50 value measured in WO 2022 / 174012 cannot be compared with the respective IC 50 values measured for the compounds of the present invention. Therefore, Compound 5 of WO 2022 / 174012 has been synthesized and subsequently tested for its pharmacological properties regarding the potency of inhibiting the cGAS / STING pathway using exactly the same assays as those used to test the compounds of the present invention and described in Chapter 6 below. Table 3: Pharmacological properties of Compound 5 of WO 2022 / 174012

[0242] As shown in the data of Table 3, Compound No. 5 (BBL0100455) of WO 2022 / 174012 has an acceptable biochemical / enzyme IC 50 value (hcGAS IC 50 = 55 nM), but the cellular IC 50 value is greater than 10000 nM (THP1 (vir) IC 50 = 10000 nM). Therefore, the compounds of the present invention in terms of their biochemical / enzyme IC 50is equivalent to Compound No. 5 of WO 2022 / 174012 in terms of value, but is significantly superior to Compound No. 5 of WO 2022 / 174012 in terms of its cellular IC 50 values (all of which are less than 1000 nM of the compounds of Formula I, II or III of the present invention).

[0243] 5.3 Prodrugs Esters of active agents known to have a carboxylic acid group can represent viable prodrugs. For example, compared to the respective active agents, they can exhibit improved oral absorption / bioavailability. Commonly used prodrugs of active agents having a carboxylic acid group are, for example, methyl esters, ethyl esters, isopropyl esters, etc. (See Beaumont et al., Current Drug Metabolism, 2003, Vol. 4, No. 6, 461-485).

[0244] In addition, Nakamura et al., Bioorganic & Medicinal Chem., Vol. 15, No. 24, pp. 7720-7725 (2007) describe that N-acylsulfonamide derivatives and N-acylsulfonylurea derivatives of certain active agents having a free carboxylic acid group may also be viable prodrugs.

[0245] Furthermore, experiments have suggested that the methyl esters of the example compounds of Formula I, II or III also represent viable prodrugs of the cGAS inhibitors of Formula I, II or III.

[0246] Both PCT / EP2022 / 062480 and PCT / EP2022 / 062496 (both not publicly available yet) disclose structurally similar cGAS inhibitors as the cGAS inhibitors of the present invention, all of which also contain a carboxylic acid group linked to a pyrrolidine moiety. In both PCT / EP2022 / 062480 and PCT / EP2022 / 062496, it has been experimentally shown that the methyl ester derivatives of these cGAS inhibitors carrying a carboxylic acid group linked to a pyrrolidine moiety are viable prodrugs of the cGAS inhibitors having a free carboxylic acid group.

[0247] Compounds P01, P02, P03 and P04 of PCT / EP2022 / 062480 are methyl ester derivatives and putative prodrugs of the respective example compounds 4.04, 1.10, 1.12 and 3.14 of PCT / EP2022 / 062480 (all of which have a free carboxyl group and are active cGAS inhibitors having lower biochemical IC 50 values and lower cellular IC 50 values).

[0248] The compounds P01, P02 and P03 of PCT / EP2022 / 062496 are methyl ester derivatives and putative prodrugs of Examples compounds 2.12, 1.13 and 1.05 of PCT / EP2022 / 062496, all of which have a free carboxyl group and are active cGAS inhibitors with a lower biochemical IC 50 value and a lower cellular IC 50 value).

[0249] In both PCT / EP2022 / 062480 and PCT / EP2022 / 062496, the "active cGAS inhibitors / Example compounds with their free carboxylic acids" and their "respective methyl ester derivatives / putative prodrugs" have been synthesized and tested for their pharmacological properties regarding the potency of inhibiting the cGAS / STING pathway.

[0250] This comparison between the properties of the Example compounds of PCT / EP2022 / 062480 and PCT / EP2022 / 062496 with their free carboxylic acids on the one hand and the properties of their corresponding methyl ester derivatives / putative prodrugs on the other hand shows that the "biochemical IC 50 value (hcGAS IC 50 value)" of the Example compounds is always approximately or even less than 10 nM, while the "biochemical IC 50 value" (hcGAS IC 50 value) of the corresponding methyl ester derivatives / prodrugs is always extremely high, i.e., generally greater than 7000 nM. The large difference between the IC 50 value of the Example compounds on the one hand and the IC 50 value of their corresponding methyl ester derivatives / prodrugs on the other hand has never been observed in the respective cellular IC 50 values (THP1 (vir) IC 50 values), and the cellular IC 50 values between the Example compounds and their corresponding prodrugs more or less always remain in the same range (see Table 4 below).

[0251] A possible explanation for this observation is that the Example compounds all have a free carboxyl group, which seems to be crucial for inhibiting cGAS activity, while in all "methyl ester derivatives / prodrugs", the carboxyl group is masked by a carboxyl-methyl group. Therefore, the methyl ester derivatives / prodrugs lose their inhibitory potency in the "in vitro human cGAS enzyme assay" (see Chapter 6.1 below), because in this assay, the intracellular enzymes that cleave the carboxyl-methyl group are absent, and thus the key free carboxylic acid group cannot be restored in the biochemical assay. Therefore, the prodrugs show extremely high "biochemical (in vitro) IC 50 value" (= hcGAS IC50 ), while the corresponding example compounds (which initially have a free carboxylic acid group) exhibit a smaller "biochemical (in vitro) IC 50 value" (= hcGAS IC 50 ).

[0252] In the cell assay (= "human cGAS cell and counter cell assay", see Chapter 6.2 below), there are endogenous cellular enzymes that cleave the carboxy - methyl ester group. Thus, not only do the example compounds of PCT / EP2022 / 062480 and PCT / EP2022 / 062496 themselves (which already carry a free carboxylic acid group) exhibit a smaller THP1 (vir) IC 50 value, but the corresponding methyl ester derivatives / prodrugs also exhibit a relatively smaller "THP1 (vir) IC 50 value" because in this "human cGAS cell assay", the carboxy - methyl ester group of the prodrug can be cleaved by endogenous intracellular enzymes, thereby releasing the "active example compound with a free carboxylic acid group", which in turn exhibits cGAS inhibitory potency.

[0253] This explanation, together with the measurement results shown in Table 4, indicates that the carboxy - methyl ester derivatives of the structurally similar example compounds of PCT / EP2022 / 062480 and PCT / EP2022 / 062496 actually seem to represent viable prodrugs of the respective example compounds with a free carboxylic acid group (which themselves do not have inhibitory potency regarding in vitro human biochemical cGAS inhibition). However, after cleavage of the carboxy - methyl ester by the endogenous intracellular enzymes present in the cell assay, the "active example compound" is restored, which again exhibits inhibitory potency regarding the cGAS / STING pathway.

[0254] Since the example compounds of formula I, II or III of the present invention have a free carboxylic acid linked to the pyrrolidine moiety that is extremely identical to that of the example compounds of PCT / EP2022 / 062480 or PCT / EP2022 / 062496, it is expected that the carboxy - methyl ester derivatives of these compounds of formula I, II or III will also act as prodrugs. Table 4: Comparison between selected cGAS inhibitor compounds disclosed in PCT / EP2022 / 062480 and PCT / EP2022 / 062496 and their respective methyl ester prodrugs:

[0255] 5. Biological Experiments The activity of the compounds of the present invention can be demonstrated using the following in vitro cGAS enzyme and cell assays:

[0256] 6.1 Method: Human cGAS Enzyme Assay (hcGASIC 50 (in vitro)) Human cGAS enzyme was incubated in the presence of 45 base pair double-stranded DNA to activate the enzyme, with GTP and ATP as substrates. The compound activity was determined by measuring the effect of the compound on the formation of the enzyme reaction product cGAMP, which was measured by mass spectrometry.

[0257] Enzyme Preparation: Human cGAS (amino acids 1 - 522) with an N-terminal 6x-His tag and a SUMO tag was expressed in Escherichia coli (E. coli) BL21(DE3)pLysS (Novagen) cells at 18 °C for 16 h. The cells were lysed in a buffer containing 25 mM Tris (pH 8), 300 mM NaCl, 10 mM imidazole, 10% glycerol, protease inhibitor cocktail (cOmplete TM , EDTA-free, Roche) and DNase (5 μg / mL). The cGAS protein was separated by affinity chromatography on Ni-NTA agarose resin and further purified by size exclusion chromatography using a Superdex200 column (GE Healthcare) equilibrated in 20 mM Tris (pH 7.5), 500 mM KCl and 1 mM TCEP. The purified protein was concentrated to 1.7 mg / mL and stored at -80 °C.

[0258] Assay Methods The compounds were delivered into a 10 mM DMSO solution, serially diluted and transferred to a 384-well assay plate (Greiner #781201) using an Echo acoustic dispenser. Usually 8 concentrations were used, with the highest concentration in the final assay volume being 10 μM, followed by an approximate 1:5 dilution step. The DMSO concentration was set to 1% in the final assay volume. The 384-well assay plate contained 22 test compounds (rows 1 - 22) and DMSO in rows 23 and 24.

[0259] After compound transfer, 15 μL of the enzyme-DNA working solution (12 nM cGAS, 0.32 μM 45 base pair DNA in assay buffer, 10 mM Tris pH 7.5 / 10 mM KCl / 5 mM MgCl2 / 1 mM DTT) was added to each well of rows 1 - 23 by a MultiDrop Combi dispenser. In row 24, 15 μl of assay buffer without enzyme / DNA was added as a low control.

[0260] Subsequently, the culture dish was pre-incubated at room temperature for 60 min.

[0261] Thereafter, 10 μL of the assay buffer of the GTP (ThermoFisher #R0461)-ATP (Promega #V915B) mixture was added to the assay dish (rows 1 - 24, 30 μM final concentration each) using a Multidrop Combi.

[0262] The culture dish was incubated again at room temperature for 90 min.

[0263] After incubation, the reaction was stopped by adding 80 μL of the assay buffer containing 5 nM cyclic-di-GMP (Sigma #SML1228) in 0.1% formic acid, which served as an internal standard for mass spectrometry. The total volume / well was 105 μL.

[0264] RapidfireMS Detection Each dish was centrifuged at 4000 rpm at 4 °C for 5 min.

[0265] The RapidFire autosampler was connected to a binary pump (Agilent 1290) and a Triple Quad 6500 (ABSciex, Toronto, Canada). The system was equipped with a 10 μL loop, a C18 [12 μL bed volume] cartridge (Agilent, part number G9210A), which contained 10 mM NH4Ac(aq) in water (pH 7.4) as eluent A (pump 1 at 1.5 mL / min, pump 2 at 1.25 mL / min) and a v / v / v 47.5 / 47.5 / 5 ACN / MeOH / H2O of 10 mM NH4Ac (pH 7.4) as eluent B (pump 3 at 1.25 mL / min). Aspiration time: 250 ms; loading time: 3000 ms; elution time: 3000 ms; wash volume: 500 μL.

[0266] MS was operated in positive ion mode using an HESI ion source, where the source temperature was 550 °C, curtain gas = 35, gas 1 = 65, gas 2 = 80. Unit mass resolution was obtained in the SRM mode. The following transitions and MS parameters (DP: declustering potential and CE: collision energy) were determined for cGAMP and DicGMP: Analyte: cGAMP at 675.1 / 524, DP = 130, CE = 30, and Internal standard: cyclic-di-GMP at 690.1 / 540, DP = 130, CE = 30.

[0267] Monitor the formation of cGAMP and evaluate the ratio to cyclic-di-GMP.

[0268] Data Evaluation and Calculation: For data evaluation and calculation, the measurement results of the low control group were set to 0% control, while those of the high control were set to 100% control. The IC 50 value was calculated using the standard four-parameter logistic regression formula: [y = (a - d) / (1 + (x / c)^b) + d], where a = low value, d = high value; x = concentration M; c = IC50 M; b = slope.

[0269] 6.2 Methods: Human cGAS cell assay and reverse cell assay stimulated by cGAMP (THP1 (vir) IC 50 and THP1 (cGAMP) IC 50 ) Use THP1-Dual cells (InvivoGen#thpd-nfis) expressing the IRF-dependent Lucia luciferase reporter gene as the basis for both assays. For the detection of cellular cGAS activity, cells were stimulated by infection with a baculovirus (pFastbac-1, Invitrogen, no coding insert) delivering cGAS enzyme to stimulate double-stranded DNA (measuring THP1 TM IC (vir) ) 50 )

[0270] For the reverse assay, cells were stimulated with cGAMP (SigmaAldrich#SML1232) to activate the same pathway that is independent of cGAS and directly downstream of cGAS (measuring THP1 (cGAMP) IC 50 )

[0271] Pathway activity was monitored by measuring the Lucia luciferase activity induced by DNA-stimulated cGAS enzyme activity (measuring THP1 (vir) IC 50 ) or directly by cGAMP (measuring THP1 (cGAMP) IC 50 , reverse assay).

[0272] Assay Methods Compounds were delivered in a 10 mM DMSO solution, serially diluted and transferred to a 384-well assay plate (Greiner#781201) using an Echo acoustic dispenser. Usually 8 concentrations were used, with the highest concentration in the final assay volume being 10 μM, followed by an approximately 1:5 dilution step. The DMSO concentration was set to 1% in the final assay volume. The 384-well assay plate contained 21 test compounds (rows 1 - 22) and DMSO in rows 23 and 24.

[0273] Cells cultured according to the manufacturer's conditions were collected by centrifugation at 300 g / 10 min and then resuspended and diluted to 1.66E5 cells / ml in fresh cell culture medium (RPMI 1640 (Gibco#A10491-01), 10% FCS (Gibco#10500), 1×GlutaMax (Gibco#35050-061), 1×Pen / Strep solution (Gibco#15140-122), 100 μg / ml Normocin (InvivoGen#ant-nr), 100 μg / ml Zeocin (InvivoGen#ant-zn), 10 μg / ml Blasticidin S (LifeTechnologies#A11139-03)). Then the baculovirus solution was added to the cells at 1:200 (varies depending on the virus batch) (measure THP1 (vir) IC 50 ). Alternatively, for the reverse assay, cGAMP was added to the cells at a final concentration of 10 μM (measure THP1 (cGAMP) IC 50 ). 30 μL of the cell / virus mixture was added to each well (5000 cells / well) in rows 1-23 of the compound plate using a MultiDrop Combi dispenser. In row 24, 30 μl / 5000 cells / well (without virus) was added as a low control group.

[0274] The culture plates were then incubated at 37 °C for 18 h in a humidified incubator.

[0275] Thereafter, 15 μL of QuantiLuc detection reagent (InvivoGen#rep-qlcg5) was added to each well using a MultiDrop Combi. Measurements were taken immediately after addition using an EnVision reader (US - luminescence reading mode).

[0276] Data Evaluation and Calculation: For data evaluation and calculation, the measurement result of the low control group was set as 0% control, and the measurement result of the high control was set as 100% control. The IC 50 value was calculated using the standard 4-parameter logistic regression formula. Calculation: [y = (a - d) / (1 + (x / c)^b) + d], a = low value, d = high value; x = concentration M; c = IC50 M; b = slope.

[0277] 6.3 Method: Human Whole Blood Determination (Human WBIC 50 ) For the detection of cellular cGAS activity, double-stranded DNA was used to stimulate human whole blood by transfection. Pathway activity was monitored by measuring the production of IFNα2α.

[0278] Assay Methods The compounds were delivered as 10 mM DMSO solutions and serially diluted, transferred to 96-well cell culture plates (Corning #3595) using an Echo acoustic dispenser, with each well pre-filled with 20 μl of OptiMEM (Gibco, #11058-021). Usually 8 concentrations were used, with the highest concentration in the final assay volume being 10 μM, followed by an approximate 1:5 dilution step. The DMSO concentration was set at 0.1% in the final assay volume. The 96-well assay plates contained 10 test compounds and DMSO in the control wells.

[0279] Human whole blood was collected simultaneously in the form of sodium citrate blood (e.g., 3.8% in Monovettes from Sarstedt) from 3 or more healthy donors (male or female, off medications for 7 days, except birth control and thyroxine). After collection, the whole blood was kept at room temperature for up to 3 hours until used in the assay.

[0280] 160 μl of whole blood samples were transferred to each well of the 96-well assay plates filled with compounds / OptiMEM. All assay plates were prepared in duplicate with blood from different donors. The blood plates were kept at room temperature for 60 minutes and continuously shaken at 450 rpm, covered with a lid but not sealed.

[0281] A DNA-Fugene mixture (Herring DNA, Sigma Aldrich #D6898-1G; Fugene (5×1 mL), Promega #E2312) was prepared in OptiMEM and incubated at room temperature for 10 min (125 ng DNA / 20 μl, Fugene ratio 9.6:1). 20 μl of the DNA Fugene mixture was added to each well, resulting in 125 ng DNA / well / 200 μl, Fugene ratio 9.6:1. 20 μl of OptiMEM and 9.6:1 Fugene were added to all low control wells.

[0282] After covering the assay plates with an aera seal and lid, the blood plates were kept at room temperature for 30 minutes and continuously shaken at 450 rpm, then incubated overnight at 37 °C for 22 h in an incubator without shaking.

[0283] To detect IFNα-2α in human plasma, according to the manufacturer's instructions, the biotinylated capture antibody (antibody set IFNA2, Meso Scale Diagnostics #B21VH-3, including coating and capture antibodies) was diluted 1:17.5 in diluent 100 (Meso Scale Diagnostics #R50AA-4). The U-Plex MSD GOLD 96-well streptavidin SECTOR plate (Meso Scale Diagnostics #L45SA-5) was coated with 25 μl of the diluted capture antibody. The coated plate was incubated for 60 min at room temperature with continuous shaking at 700 rpm. The MSD IFNα-2α plate was washed three times with 150 μl of wash buffer (1×HBSS, 0.05% Tween).

[0284] After blocking each plate for 60 min at room temperature with 100 μl of blocking solution / well (1×HBSS, containing 0.2% Tween, 2% BSA) with continuous shaking at 700 rpm, the plates were emptied as dry as possible by pouring, and then human plasma was used continuously.

[0285] The whole blood assay plates were centrifuged at 1600 rpm for 10 minutes. 25 μl of the supernatant was transferred from each whole blood plate to the corresponding IFNα-2α plate using a pipetting robot. Each plate was sealed with a microplate seal and incubated again for two hours at room temperature with continuous shaking at 700 rpm.

[0286] Subsequently, the MSD IFNα-2α plate was washed three times with 150 μl of wash buffer (1×HBSS, 0.05% Tween), and then 25 μl of MSD SULFO-TAG IFNα-2α antibody solution (diluted 1:100 in diluent 3 (Meso Scale Diagnostics #R50AP-2)) was added to each well of the plate.

[0287] Subsequently, each plate was sealed with a microplate seal and incubated again for two hours at room temperature with continuous shaking at 700 rpm. Finally, the MSD IFNα-2α plate was washed three times with 150 μl of wash buffer (1×HBSS, 0.05% Tween). 150 μl of 2× read buffer was added to each well, and each plate was immediately measured using an MSD Sector S600 reader with the supplier barcode.

[0288] Data Evaluation and Calculation: For data evaluation and calculation, the control % of each well was calculated based on the average of the high (DNA stimulation control) and the average of the low (unstimulated control) controls using the following formula: [(Count (sample) - Count (low)) / (Count (high) - Count (low))] * 100 Calculate the IC using the standard 4-parameter logistic regression formula 50 value. Calculate: [y = (a - d) / (1 + (x / c)^b) + d], where a = low value, d = high value; x = concentration M; c = IC 50 M; b = slope.

[0289] 6. Indications As has been found, the compounds of formula I, II or III are characterized by their scope of application in the field of therapy. In particular, mention should be made of those applications of the compounds of formula I, II or III according to the invention which are preferably used on the basis of their pharmaceutical activity as cGAS inhibitors. Although the cGAS pathway is crucial for host defense against pathogen invasion, such as viral infections and the invasion of some intracellular bacteria, cellular stress and genetic factors can also lead to the production of abnormal cellular dsDNA, for example through nuclear or mitochondrial leakage, and thereby trigger an autoinflammatory response. Therefore, cGAS inhibitors have strong therapeutic potential for the treatment of various autoinflammatory and autoimmune diseases.

[0290] An et al., Arthritis Rheumatol. April 2017; 69(4):800 - 807 disclosed that the cGAS expression in peripheral blood mononuclear cells (PBMC) of patients with the autoimmune disease systemic lupus erythematosus (SLE) was significantly higher than that in the normal control group. By targeted measurement of cGAMP by tandem mass spectrometry, cGAMP was detected in 15% of the tested SLE patients, but not in the normal or rheumatoid arthritis control groups. SLE patients with cGAMP had a higher disease activity compared to SLE patients without cGAMP. Although the higher cGAS expression may be the result of exposure to type I interferon (IFN), the detection of cGAMP in SLE patients with increased disease activity indicates that the cGAS pathway may be involved in disease expression.

[0291] Park et al., Ann Rheum Dis. October 2018; 77(10):1507 - 1515 also disclosed that the cGAS pathway is involved in the development of SLE.

[0292] Thim-Uam et al., iScience September 4, 2020; 23(9), 101530 (doi:10.1016 / j.isci.2020.101530) disclosed that the STING pathway mediates lupus through the activation of conventional dendritic cell maturation and plasmacytoid dendritic cell differentiation.

[0293] Gao et al., Proc. Natl. Acad. Sci. USA. 2015 Oct 20;112(42):E5699-705 describe that cGAS causes certain autoimmune diseases, such as interferonopathy, through activation of self-DNA.

[0294] Tonduti et al., Expert Rev. Clin. Immunol. 2020 Feb; 16(2): 189-198 disclose that cGAS inhibitors have particular therapeutic potential in Icardi-Guterres syndrome and familial lupus chilblains, which are lupus-like severe autoinflammatory immune-mediated disorders.

[0295] Steiner et al., Nat Commun. 2022 Apr 28;13(1):232;doi:10.1038 show that deficiency of coatomer complex I causes aberrant activation of STING signaling and COPA syndrome, and that cGAS is required to drive type I IFN signaling in a cell model of COPA syndrome.

[0296] Li et al. showed that extracellular vesicles containing plasma-derived DNA induced STING-mediated proinflammatory responses in dermatomyositis (Theranostics. 2021; 11(15): 7144-7158). Zhou et al. (J Clin Lab Anal. 2022 Oct; 36(10): e24631) described the correlation between cGAS-STING pathway activation and muscle fiber atrophy / necrosis in dermatomyositis.

[0297] In Yu et al., Cell 2020 Oct 29; 183(3): 636-649, a link between TDP-43-triggered activation of mitochondrial DNA and the cGAS / STING pathway in amyotrophic lateral sclerosis (ALS) is described.

[0298] Ryu et al., Arthritis Rheumatol. 2020 Nov; 72(11): 1905-1915 also showed that bioactive plasma mitochondrial DNA is associated with disease progression in specific fibrotic diseases, such as systemic sclerosis (SSc) or interstitial lung disease (ILD), progressive fibrosing interstitial lung disease (PF-ILD), and idiopathic pulmonary fibrosis (IPF).

[0299] In Schuliga et al., Clin. Sci. (Lond). 2020 Apr 17;134(7):889-905, it is described that self-DNA perpetuates senescence of IPF lung fibroblasts in a cGAS-dependent manner.

[0300] Other scientific hints linking the etiology of other fibrotic diseases, such as non-alcoholic steatohepatitis (NASH), to the cGAS / STING pathway have been described in Yu et al., J. Clin. Invest. February 1, 2019; 129(2):546-555 and Cho et al., Hepatology. October 2018; 68(4):1331-1346.

[0301] Nascimento et al., Sci. Rep. October 16, 2019; 9(1):14848 disclosed that self-DNA release and STING-dependent sensing drive inflammation in mice due to cigarette smoke, suggesting a link between the cGAS-STING pathway and chronic obstructive pulmonary disease (COPD).

[0302] Ma et al., Sci. Adv. May 20, 2020; 6(21):eaaz6717 disclosed that ulcerative colitis and inflammatory bowel disease (IBD) can be restricted by controlling cGAS-mediated inflammation.

[0303] Gratia et al., J. Exp. Med. May 6, 2019; 216(5):1199-1213 showed that the Bloom syndrome protein restricts the innate immune sensing of micronuclei through cGAS. Thus, cGAS inhibitors have therapeutic potential for treating Bloom syndrome.

[0304] Kerur et al., Nat. Med. January 2018; 24(1):50-61 described that cGAS plays an important role in the atypical inflammasome activation in age-related macular degeneration (AMD).

[0305] Visitchanakun et al., Int J Mol Sci. October 23, 2021; 22(21):11450 showed that in the cecal ligation puncture (CLP) and lipopolysaccharide (LPS) injection sepsis models, the severity of GAS-deficient mice was lower than that of wild-type mice.

[0306] Wang et al., Mediators Inflamm. 2015; 2015:192329, described that cGAS is required for cell proliferation and inflammatory cytokine production in rheumatoid arthritis synoviocytes. It has also been reported that in a mouse model of inflammatory arthritis, cGAS deficiency inhibits the interferon response, inflammatory cell infiltration, and joint swelling (Willemsen et al., Cell Rep. November 9, 2021; 37(6):109977).

[0307] Guo et al., Osteoarthritis Cartilage. August 2021; 29(8):1213-1224 described damaged DNA as a key pathological factor in osteoarthritis (OA), which may be mediated by the cGAS / STING pathway, as STING knockdown alleviated the instability of medial meniscus-induced OA development in mice.

[0308] Mao et al., Arterioscler Thromb Vasc Biol (2017) 37(5):920-929 showed that in diet-induced obesity, the cGAS / STING pathway mediated endothelial inflammation in response to free fatty acid-induced mitochondrial disruption, indicating that cGAS inhibitors also have the potential to treat obesity and diabetes.

[0309] Kerur et al., Nat Med. January 2018; 24(1):50-61 described elevated cGAS levels in the retinal pigment epithelium of the eyes of people with geographic atrophy, and cGAS-driven atypical inflammasome activation in age-related macular degeneration.

[0310] cGAS promotes cellular senescence and the senescence-associated secretory phenotype (Yang et al., Proc Natl Acad Sci USA June 6, 2017; 114:E4612-E4620). Cytoplasmic chromatin triggers inflammation during senescence through cGAS / STING, and STING null mice have reduced tissue inflammation and senescence (Dou et al., Nature. 2017 550:402-406). In addition, variants within the human STING gene are associated with healthy aging, most likely due to reduced inflammation (Hamann et al., Gerontology 2019; 65:145-154). Overall, STING inhibitors will reduce senescence-associated inflammation and senescent cell accumulation, and will improve senescence-associated diseases, such as aging, muscle disorders, and fibrosis.

[0311] In addition, cGAS inhibitors of Formula I, II, or III have therapeutic potential in the treatment of cancer (see Hoong et al., Oncotarget. July 28, 2020; 11(30):2930-2955, and Chen et al., Sci. Adv. October 14, 2020; 6(42):eabb8941).

[0312] In addition, cGAS inhibitors of Formula I, II, or III also have therapeutic potential in the treatment of heart failure (Hu et al., Am. J. Physiol. Heart Circ. Physiol. June 1, 2020; 318(6): H1525 - H1537).

[0313] There are also other scientific hints of the correlation between Parkinson's disease and the cGAS / STING pathway (Sliter et al., Nature. September 2018; 561(7722): 258 - 262) and between Sjogren's syndrome and the cGAS / STING pathway (Papinska et al., J. Dent. Res. July 2018; 97(8): 893 - 900).

[0314] Furthermore, cGAS inhibitors of Formula I, II, or III also have therapeutic potential in the treatment of COVID - 19 / SARS - CoV - 2 infection, as shown in the following literature: Di Domizio et al., Nature. January 19, 2022, doi:10.1038 / s41586 - 022 - 04421 - w: “The cGAS - STING pathway drives type I IFN immunopathology in COVID - 19”, and Neufeldt et al., Commun Biol. January 12, 2022; 5(1): 45. doi:10.1038 / s42003 - 021 - 02983 - 5: “SARS - CoV - 2 infection induces a pro - inflammatory cytokine response through cGAS - STING and NF - kappaB”.

[0315] In addition, the cGAS inhibitors of Formula I, II or III have therapeutic potential in the treatment of kidney inflammation and renal fibrosis, as shown in the following literature: Chung et al., Cell Metab. 2019 30:784-799: "Mitochondrial Damage and Activation of the STING Pathway Lead to Renal Inflammation and Fibrosis", and Maekawa et al., Cell Rep. 2019 29:1261-1273: "Mitochondrial Damage Causes Inflammation via cGAS-STING Signaling in Acute Kidney Injury".

[0316] Furthermore, the cGAS inhibitors of Formula I, II or III have therapeutic potential in the treatment of cancer, as shown in the following literature: Bakhoum et al., Nature. January 25, 2018; 553(7689):467-472: "Chromosomal instability drives metastasis through a cytosolic DNA response", and Liu et al., Nature. November 2018; 563(7729):131-136: "Nuclear cGAS suppresses DNA repair and promotes tumorigenesis".

[0317] In addition, because STING gt animals exhibit reduced macrophage infiltration in adipose tissue after subchronic high-calorie intake (HFD), and the deficiency of STING gt and IRF3 causes reduced blood glucose and insulin and weight loss, the cGAS inhibitors of Formula I, II or III have therapeutic potential in the treatment of metabolic disorders (Mao et al., Arterioscler Thromb Vasc Biol, 2017; 37(5):920-929).

[0318] In addition, because mitochondrial DNA is released into the cytosol of endothelial cells, it causes cGAS / STING pathway activation and suppresses endothelial cell proliferation, so cGAS inhibitors of Formula I, II or III have therapeutic potential in treating vascular diseases and cause vascular repair / regeneration. In addition, gene knockout of the cGAS gene restores endothelial repair / regeneration in a mouse model of inflammatory lung injury (Huang et al., Immunity, March 2020, 2017; 52(3):475-486.e5.doi:10.1016 / j.immuni.2020,02.002).

[0319] In addition, cGAS inhibitors of Formula I, II or III have therapeutic potential in treating aging-related and obesity-related cardiovascular diseases (Hamann et al., Immun Ageing, 2020 Mar 14;17:7; doi:10.1186 / s12979-020-00176-y.eCollection 2020).

[0320] Therefore, compounds of Formula I, II or III that are cGAS inhibitors are useful for treating autoinflammatory and autoimmune diseases, such as systemic lupus erythematosus (SLE), interferonopathy, Acardi-Guterres syndrome (AGS), COPA syndrome, familial lupus pernio, age-related macular degeneration (AMD), amyotrophic lateral sclerosis (ALS), inflammatory bowel disease (IBD), chronic obstructive pulmonary disease (COPD), Bloom's syndrome, Sjögren's syndrome, rheumatoid arthritis and Parkinson's disease.

[0321] In addition, compounds of formula I, II or III as cGAS inhibitors can be used to treat fibrotic diseases, such as systemic sclerosis (SSc), interferon pathology, non-alcoholic steatohepatitis (NASH), interstitial lung disease (ILD), preferably progressive fibrosing interstitial lung disease (PF-ILD), especially idiopathic pulmonary fibrosis (IPF).

[0322] In addition, compounds of Formula I, II or III that are cGAS inhibitors can be used to treat age-related macular degeneration (AMD), retinopathy, glaucoma, diabetes, obesity, aging, muscle disorders, sepsis, osteoarthritis, heart failure, COVID-19 / SARS-CoV-2 infection, kidney inflammation, renal fibrosis, metabolic disorders, vascular diseases, cardiovascular diseases and cancer.

[0323] 7. Combinations The compounds of Formula I, II or III may be administered to a patient alone or in combination with one or more other pharmacologically active agents.

[0324] In a preferred embodiment of the present invention, the compound of formula I, II or III can be combined with one or more pharmacological active agents selected from the group consisting of: anti-inflammatory agents; anti-fibrotic agents; anti-allergic agents / anti-histamines; bronchodilators; β2 agonists / β mimetics; adrenergic agonists; anticholinergics; methotrexate; mycophenolate mofetil; leukotriene regulators; JAK inhibitors; anti-interleukin antibodies; non-specific immunotherapeutic agents, such as interferons or other cytokines / chemokines; cytokine / chemokine receptor regulators (i.e., cytokine receptor agonists or antagonists); toll-like receptor agonists (= TLR agonists); immune checkpoint regulators; anti-TNF antibodies (Humira TM ); and anti-BAFF antibodies (belimumab and etanercept).

[0325] The anti-fibrotic agent is preferably selected from pirfenidone or tyrosine kinase inhibitors, such as nintedanib, with nintedanib being particularly preferred.

[0326] Preferred examples of anti-inflammatory agents are NSAIDs and corticosteroids.

[0327] The NSAIDs are preferably selected from ibuprofen, naproxen, diclofenac, meloxicam, celecoxib, acetylsalicylic acid (Aspirin TM ), indomethacin, mefenamic acid and etoricoxib.

[0328] The corticosteroids are preferably selected from flunisolide, beclomethasone, triamcinolone, budesonide, fluticasone, mometasone, ciclesonide, rofleponide and dexamethasone.

[0329] The anti-allergic agents / anti-histamines are preferably selected from epinastine, cetirizine, azelastine, fexofenadine, levocabastine, loratadine, ebastine, desloratidine and mizolastine.

[0330] The β2-agonist / β-mimetic can be a long-acting β2-agonist (LABA) or a short-acting β-agonist (SABA). Particularly preferred β2-agonists / β-mimetics are selected from Bambuterol, Bitolterol, Carbuterol, Clenbuterol, Fenoterol, Formoterol, Hexoprenalin, Ibuterol, Pirbuterol, Procaterol, Reproterol, Salmeterol, Sulfonterol, Terbutalin, Tolubuterol, Olodaterol, and Salbutamol, especially Olodaterol.

[0331] The anticholinergic is preferably selected from ipratropium salts, tiotropium salts, glycopyrronium salts, and theophylline, with tiotropium bromide being particularly preferred.

[0332] The leukotriene regulator is preferably selected from Montelukast, Pranlukast, Zafirlukast, Ibudilast, and Zileuton.

[0333] The JAK inhibitor is preferably selected from Baricitinib, Cerdulatinib, Fedratinib, Filgotinib, Gandotinib, Lestaurtinib, Momelotinib, Pacritinib, Peficitinib, Ruxolitinib, Tofacitinib, and Upadacitinib.

[0334] The anti-interleukin antibody is preferably selected from anti-IL23 antibodies (such as risankizumab), anti-IL17 antibodies, anti-IL1 antibodies, anti-IL4 antibodies, anti-IL13 antibodies, anti-IL-5 antibodies, anti-IL-6 antibodies (such as Actemra TM) Anti-IL-12 antibody, anti-IL-15 antibody.

[0335] 8. Formulations The compounds of the present invention can be administered by any suitable route of administration, including both systemic and topical administration. Systemic administration includes oral administration, parenteral administration, transdermal administration, rectal administration, and administration by inhalant. Parenteral administration refers to routes of administration other than enteral, transdermal, or by inhalant, and is usually by injection or infusion. Parenteral administration includes intravenous, intramuscular, and subcutaneous injection or infusion. Inhalation refers to administration to the lungs of a patient, whether by inhalation through the mouth or through the nasal passages. Topical administration includes application to the skin. The compounds of the present invention can be administered by eye drops to treat Sjögren's syndrome.

[0336] Forms suitable for administration are, for example, tablets, capsules, solutions, syrups, emulsions, or inhalable powders or aerosols. In each case, the content of the pharmaceutically effective compound should be in the range of 0.1 to 90% by weight, preferably 0.5 to 50% by weight, of the total composition, i.e., an amount sufficient to achieve the dosage range specified below.

[0337] The preparation can be administered orally in the form of tablets, powders, powders in capsules (e.g., hard gelatin capsules), solutions, or suspensions. When administered by inhalation, the active substance combination can be administered as a powder, aqueous solution, or aqueous ethanol solution, or using a propellant gas preparation.

[0338] Therefore, preferably, the pharmaceutical preparation is characterized by the content of one or more compounds of formula I, II, or III according to the above preferred embodiments.

[0339] Oral administration of the compounds of formula I, II, or III is particularly preferred, and administration once or twice a day is also particularly preferred. Suitable tablets can be obtained, for example, by mixing the active substance with known excipients (e.g., inert diluents such as calcium carbonate, calcium phosphate, or lactose; disintegrants such as corn starch or alginic acid; binders such as starch or gelatin; lubricants such as magnesium stearate or talc; and / or agents for delayed release such as carboxymethyl cellulose, cellulose acetate phthalate, or polyvinyl acetate)). Tablets can also comprise several layers.

[0340] Therefore, coated tablets can be prepared by coating a core similar to that produced by tablets with substances commonly used for tablet coating (e.g., polyvinylpyrrolidone (kollidone) or shellac, gum arabic, talc, titanium dioxide, or sugar). To achieve delayed release or prevent incompatibility, the core can also consist of multiple layers. Similarly, the excipients mentioned above with respect to tablets can be used, and the tablet coating can consist of multiple layers to achieve delayed release.

[0341] According to the invention, a syrup containing an active substance or a combination thereof may additionally contain sweeteners and flavor enhancers such as saccharin, cyclamate, glycerol or sugars, and flavoring agents such as vanilla essence or orange extract. It may also contain suspending adjuvants or thickeners such as sodium carboxymethylcellulose; wetting agents such as condensation products of fatty alcohols and ethylene oxide; or preservatives such as parabens.

[0342] Capsules containing one or more active substances or combinations of active substances can be prepared, for example, by mixing the active substances with an inert carrier such as lactose or sorbitol and filling them into gelatin capsules. Suitable suppositories can be manufactured, for example, by mixing carriers provided for this purpose, such as neutral fats or polyethylene glycols or their derivatives.

[0343] Excipients that can be used include: for example, water; pharmaceutically acceptable organic solvents such as paraffin (such as petroleum fractions), vegetable oils (such as peanut oil or sesame oil), monofunctional or polyfunctional alcohols (such as ethanol or glycerol); carriers such as natural mineral powders (such as kaolin, clay, talc, chalk), synthetic mineral powders (such as highly disperse silicic acid and silicates), sugars (such as sucrose, lactose and glucose), emulsifiers (such as lignin, sulfite waste liquor, methylcellulose, starch and polyvinylpyrrolidone) and lubricants (such as magnesium stearate, talc, stearic acid and sodium lauryl sulfate). Of course, for oral administration, in addition to the above carriers, tablets may also contain additives such as sodium citrate, calcium carbonate and dibasic calcium phosphate, as well as various additives such as starch (preferably potato starch), gelatin and the like. In addition, lubricants such as magnesium stearate, sodium lauryl sulfate and talc can be used simultaneously in the tabletting process. In the case of aqueous suspensions, the active substances can be combined with various flavor enhancers or colorants in addition to the excipients mentioned above.

Claims

1. A compound of formula I wherein R 1 is selected from the group consisting of hydrogen, halogen, methyl, ethyl, -CF3, -CHF2, -CFH2, and methoxy, R 2 selected from the group consisting of hydrogen and methyl, R 3 selected from the group consisting of hydrogen, methyl and halogen, and wherein A is selected from the group consisting of: -CH2-, -O-, -CF2-, -CHF-, -N(CH3)-, -NH- and -CHCH3-; D is selected from the group consisting of: -CH2-, -O-, -CF2-, -CHF- and -CHCH3-; E is selected from the group consisting of: -CH2-, -O-, -C(CH3)2-, -CHF-, CF2- and -CHCH3-; G is selected from the group consisting of: -NH-, -NCH3-, -CH2-, -O-, -CF2-, -CHF-, -CHCH3- and -C(CH3)2; J is selected from the group consisting of: -CO-, -CH2-, -O-, -CHF-, -CF2- and -CHCH3-; K is selected from the group consisting of -CH2-, -CF2-, -O- or is absent; L is selected from the group consisting of -CH2-, -O-, -CHCH3-, -CHF-, -CF2- or is absent; and prodrugs, deuterated analogs and pharmaceutically acceptable salts thereof.

2. The compound of formula I according to claim 1, wherein R 1 selected from the group consisting of hydrogen, Cl and F, R 2 selected from the group consisting of hydrogen and methyl, R 3 selected from the group consisting of hydrogen, methyl, Cl and F, and wherein A is selected from the group consisting of: -CH2-, -O-, -CF2- and -N(CH3)-, D is selected from the group consisting of -CH2-, -O- and -CHCH3-, E is selected from the group consisting of -CH2-, -O- and -C(CH3)2-, G is selected from the group consisting of: -NH-, -CH2-, -O-, -CHCH3-, J is selected from the group consisting of: -CO-, -CH2-, -O- and -CHCH3-; K is selected from the group consisting of -CH2-, -CF2-, -O- or is absent; L is selected from the group consisting of -CH2-, -CF2- or is absent; and prodrugs, deuterated analogs and pharmaceutically acceptable salts thereof.

3. The compound of formula I according to at least one of claims 1 or 2, wherein L is absent; and prodrugs, deuterated analogs and pharmaceutically acceptable salts thereof.

4. The compound of formula I according to at least one of claims 1 or 2, wherein L is absent, and wherein A is selected from the group consisting of -CH2- and -CF2-, and prodrugs, pharmaceutically acceptable salts or deuterated analogs thereof.

5. The compound of formula I according to claim 1 or 2, wherein L is absent, and wherein K is CF2; and prodrugs, deuterated analogs and pharmaceutically acceptable salts thereof.

6. The compound of formula I according to at least one of claims 1 to 5, wherein R 3 is halogen; and its prodrugs, deuterated analogs and pharmaceutically acceptable salts.

7. The compound of formula I according to claim 6, wherein R 3 is Cl or F; and prodrugs, deuterated analogs and pharmaceutically acceptable salts thereof.

8. The compound of formula I according to claim 7, wherein R 3 is Cl or F and is in the 5-position of the benzimidazole moiety; and prodrugs, deuterated analogs, and pharmaceutically acceptable salts thereof.

9. The compound of formula I according to at least one of claims 1 to 8, wherein R 1 is halogen; and its prodrugs, deuterated analogs and pharmaceutically acceptable salts.

10. The compound of formula I according to claim 9, wherein R 1 is Cl or F; and prodrugs, deuterated analogs, and pharmaceutically acceptable salts thereof.

11. The compound of formula I according to at least one of claims 1 to 8, wherein R 1 is hydrogen; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

12. The compound of formula I according to at least one of claims 1 or 2, which is selected from the group consisting of: and prodrugs, deuterated analogs and pharmaceutically acceptable salts thereof.

13. The compound of formula I according to at least one of claims 1 or 2, wherein A is selected from the group consisting of -CH2- and -CF2-, D and E are each -CH2-, G is selected from the group consisting of -CH2- and -O-, J is selected from the group consisting of -CH2- and -O-, K is selected from the group consisting of -CF2- and -CH2-, and L is absent, and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

14. The compound of formula I according to claim 13, which is selected from the group consisting of: and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

15. The compound of formula I according to claim 1, wherein R 2 is methyl; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

16. The compound of formula II according to claim 15 or the compound of formula III and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

17. The compound of formula II according to claim 16 and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

18. The compound of formula II according to at least one of claims 16 or 17 or the compound of formula III according to claim 16, wherein L is absent; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

19. The compound of formula II according to at least one of claims 16 or 17 or the compound of formula III according to claim 16, wherein L is absent and wherein K is CF2; and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

20. The compound of formula II according to at least one of claims 16 or 17 or the compound of formula III according to claim 16, wherein R 3 is halogen; and its prodrugs, deuterated analogs and pharmaceutically acceptable salts.

21. The compound of formula II or formula III according to claim 20, wherein R 3 is Cl or F; and prodrugs, deuterated analogs, and pharmaceutically acceptable salts thereof.

22. The compound of formula II or formula III according to claim 21, wherein R 3 is Cl or F and is located at the 5-position of the benzimidazole moiety; and prodrugs, deuterated analogs, and pharmaceutically acceptable salts thereof.

23. The compound of formula II according to at least one of claims 16 or 17 or the compound of formula III according to claim 16, wherein R 3 is hydrogen; and its prodrugs, deuterated analogs and pharmaceutically acceptable salts.

24. The compound of formula II according to at least one of claims 16 or 17 or the compound of formula III according to claim 16, wherein R 1 is halogen; and its prodrugs, deuterated analogs and pharmaceutically acceptable salts.

25. The compound of formula II or formula III according to claim 24, wherein R 1 is selected from the group consisting of Cl or F; and prodrugs, deuterated analogs, and pharmaceutically acceptable salts thereof.

26. The compound of formula II according to at least one of claims 16 or 17 or the compound of formula III according to claim 16, wherein R 1 is hydrogen; and its prodrugs, deuterated analogs and pharmaceutically acceptable salts.

27. The compound of formula II according to at least one of claims 16 or 17 or the compound of formula III according to claim 16, wherein A is selected from the group consisting of -CH2- and -CF2-, D and E are each -CH2-, G is selected from the group consisting of -CH2- and -O-, J is selected from the group consisting of -CH2- and -O-, K is selected from the group consisting of -CF2- and -CH2-, and L is absent, and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

28. The compound of formula II according to claim 27, which is selected from the group consisting of: and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

29. The compound of formula II or formula III according to claim 27, wherein A is selected from the group consisting of -CH2- and -CF2-, D, E, and G are each -CH2-, J is selected from the group consisting of -CH2- and -O-, K is -CF2-, and L is absent, and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

30. The compound of formula II according to claim 29, which is selected from the group consisting of: and its prodrugs, deuterated analogs, and pharmaceutically acceptable salts.

31. An intermediate compound having formula (A-I) wherein R 1 , R 2 , R 3 , A, D, E, G, J, K and L are as defined above, and wherein R13 is selected from the group consisting of: hydrogen, methyl, ethyl and tert-butyl, formula (A-II), wherein R 1 , R 2 , R 3 , A, D, E, G, J, K and L are as defined above, and wherein R13 is selected from the group consisting of: hydrogen, methyl, ethyl and tert-butyl, and wherein R is hydrogen or a protecting group selected from the group consisting of tert-butyl, methyl, ethyl, and benzyl, formula (B-I) wherein R 1 and R 2 and R 3 A, D, E, G, J, K and L are as defined above, and wherein R13 is selected from the group consisting of hydrogen, methyl, ethyl and tert-butyl, or formula (B-II) wherein R 1 , R 2 , R 3 , A, D, E, G, J, K and L are as defined above, and wherein R13 is selected from the group consisting of: hydrogen, methyl, ethyl and tert-butyl.

32. The compound of formula I according to one or more of claims 1 to 15 or the compound of formula II or formula III according to one or more of claims 16 to 30, which is used for treating diseases that can be treated by inhibiting cGAS.

33. A compound of formula I according to one or more of claims 1 to 15 or a compound of formula II or formula III according to one or more of claims 16 to 30 for use in the treatment of a disease selected from the group consisting of: systemic lupus erythematosus (SLE); interferonopathy; Aicardi-Goutières syndrome syndrome, AGS); COPA syndrome; familial pernio lupus; age-related macular degeneration (AMD); amyotrophic lateral sclerosis (ALS); retinopathy; glaucoma; diabetes; obesity; inflammatory bowel disease (IBD); chronic obstructive pulmonary disease (COPD); Bloom's syndrome; dermatomyositis; Sjögren's syndrome; Parkinson's disease; heart failure; cancer; aging; muscle disorders; sepsis; rheumatoid arthritis; osteoarthritis; COVID-19; systemic sclerosis (SSc); non-alcoholic steatohepatitis (NASH); interstitial lung disease (ILD), preferably progressive fibrosing interstitial lung disease (PF-ILD), in particular idiopathic pulmonary fibrosis (IPF).

34. A compound of formula I according to one or more of claims 1 to 15 or a compound of formula II or formula III according to one or more of claims 16 to 30 for use in the treatment of a disease selected from the group consisting of systemic lupus erythematosus (SLE), interferonopathy, Acardi-Guterres syndrome (AGS), COPA syndrome, familial lupus pernio, dermatomyositis, age-related macular degeneration (AMD), amyotrophic lateral sclerosis (ALS), inflammatory bowel disease (IBD), chronic obstructive pulmonary disease (COPD), Bloom's syndrome, Sjögren's syndrome, rheumatoid arthritis and Parkinson's disease.

35. A compound of formula I according to one or more of claims 1 to 15 or a compound of formula II or formula III according to one or more of claims 16 to 30 for use in the treatment of a disease selected from the group consisting of: systemic sclerosis (SSc); non-alcoholic steatohepatitis (NASH); interferonopathy; interstitial lung disease (ILD), preferably progressive fibrosing interstitial lung disease (PF-ILD), in particular idiopathic pulmonary fibrosis (IPF).

36. A compound of formula I according to one or more of claims 1 to 15 or a compound of formula II or formula III according to one or more of claims 16 to 30 for use in the treatment of a disease selected from the group consisting of age-related macular degeneration (AMD), retinopathy, glaucoma, diabetes, obesity, aging, muscle disorders, sepsis, osteoarthritis, heart failure, COVID19 / SARS-CoV-2 infection, kidney inflammation, renal fibrosis, metabolic disorders, vascular diseases, cardiovascular diseases and cancer.

37. A pharmaceutical composition comprising a compound of formula I according to one or more of claims 1 to 15 or a compound of formula II or formula III according to one or more of claims 16 to 30, and optionally one or more pharmaceutically acceptable carriers and / or excipients.

38. A pharmaceutical composition comprising a compound of formula I according to one or more of claims 1 to 15 or a compound of formula II or formula III according to one or more of claims 16 to 30, which is used in combination with one or more active agents selected from the group consisting of: anti-inflammatory agents; anti-fibrotic agents; anti-allergic agents / antihistamines; bronchodilators; β2 agonists / β mimetics; adrenergic agonists; anticholinergics; methotrexate; mycophenolate mofetil; leukotriene regulators; JAK inhibitors; anti-interleukin antibodies; non-specific immunotherapeutic agents such as interferons or other cytokines / chemokines; cytokine / chemokine receptor regulators; Toll-like receptor agonists; immune checkpoint regulators; anti-TNF antibodies such as Humira TM ; anti-BAFF antibodies such as Belimumab and Etanercept; and optionally one or more pharmaceutically acceptable carriers and / or excipients.

39. A pharmaceutical composition comprising a compound of formula I according to one or more of claims 1 to 15 or a compound of formula II or formula III according to one or more of claims 16 to 30, which is used in combination with one or more anti-fibrotic agents selected from the group consisting of Pirfenidon and Nintedanib, and optionally one or more pharmaceutically acceptable carriers and / or excipients.

40. A pharmaceutical composition comprising a compound of formula I according to one or more of claims 1 to 15 or a compound of formula II or formula III according to one or more of claims 16 to 30, which is used in combination with one or more anti-inflammatory agents selected from the group consisting of NSAIDs and corticosteroids, and optionally one or more pharmaceutically acceptable carriers and / or excipients.

41. A pharmaceutical composition comprising a compound of formula I according to one or more of claims 1 to 15 or a compound of formula II or formula III according to one or more of claims 16 to 30, which is used in combination with one or more active agents selected from the group consisting of bronchodilators, β2 agonists / β mimetics, adrenergic agonists, and anticholinergics; and optionally one or more pharmaceutically acceptable carriers and / or excipients.

42. A pharmaceutical combination comprising a compound of formula I according to one or more of claims 1 to 15 or a compound of formula II or formula III according to one or more of claims 16 to 30 and one or more anti - interleukin antibodies selected from the group consisting of: anti - IL - 23 antibodies, such as Risankizumab; anti - IL - 17 antibodies; anti - IL - 1 antibodies; anti - IL - 4 antibodies; anti - IL - 13 antibodies; anti - IL - 5 antibodies; anti - IL - 6 antibodies, such as Actemra TM ; anti - IL - 12 antibodies; and anti - IL - 15 antibodies.

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