PYRIDINE DERIVATIVES WITH N-LINKED CYCLIC SUBSTITUENTS AS cGAS INHIBITORS

Abstract

The invention relates to new proline derivatives of formula (I) as cGAS inhibitors,

##STR00001##

wherein

wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11 and G are defined as in claim 1,

and prodrugs or pharmaceutically acceptable salts of these compounds

for the treatment of diseases such as systemic lupus erythematosus, systemic sclerosis (SSc), non-alcoholic steatotic hepatitis (NASH), interstitial lung disease (ILD) and idiopathic pulmonary fibrosis (IPF).

Claims

1. A compound of formula (I), ##STR00422## wherein R.sup.1 is selected from methyl, ethyl, halomethyl, haloethyl and halogen, wherein G is selected from O, NR.sup.8, CH.sub.2, C and CR.sup.8R.sup.9, wherein R.sup.2 is selected from H, halogen, cyclopropyl, C.sub.1-3-alkyl, —C.sub.2-5-alkynyl, —S-methyl and CN, or wherein R.sup.2 is a cyclic group, wherein this cyclic group is selected from the group consisting of a phenyl or a five- to six-membered heteroaryl comprising 1, 2, 3 or 4 heteroatoms each independently selected from N, S and O, wherein this cyclic group is substituted by one or two, identical or different substituents R.sup.10, wherein R.sup.3 is H or methyl, R.sup.4 is H or methyl, R.sup.5 is selected from H, methyl, —CN, -methylene-OH and —CF.sub.3, or R.sup.5 may be absent, R.sup.6 is selected from H, methyl, —CN, -methylene-OH and —CF.sub.3, or R.sup.5 and R.sup.6 together with the C-atoms in between form a ring selected from oxetane, tetrahydrofurane and cyclopropane, R.sup.7 is selected from H, halogen, (C.sub.1-3)-alkyl and halo-(C.sub.1-3)-alkyl, R.sup.8 is selected from CN, H and methyl, R.sup.9 is selected from H, methyl and halogen or R.sup.9 may be absent, wherein each R.sup.10 is independently selected from the group consisting of hydrogen, halogen, haloalkyl, -methyl, -ethyl, —NH—CO-methyl, —N(CH.sub.3).sub.2, —CH.sub.2—OH, —NH(CH.sub.3), —O—(C.sub.1-3-alkyl), —CN, —S—CH.sub.3, —CO—NH.sub.2, —CH.sub.2—NH(CH.sub.3), —CH.sub.2—NH.sub.2, —SO—(CH.sub.3), cyclopropyl and —O—R.sup.11, wherein R.sup.11 is a five- or six-membered heterocycle with one or two heteroatoms each independently selected from N, O and S, or G is CR.sup.8R.sup.9, R.sup.5 and R.sup.9 are absent, and R.sup.8 and R.sup.6 and the two C-atoms in between R.sup.8 and R.sup.6 form an annulated five-membered aromatic or non-aromatic heterocycle comprising one, two or three heteroatoms each independently selected from N, S and O, or G is CR.sup.8R.sup.9 and R.sup.8 and R.sup.9 form together with the C-atom in between R.sup.8 and R.sup.9 a diazirine ring; and prodrugs or pharmaceutically acceptable salts thereof.

2. The compound according to claim 1 of formula (I′), ##STR00423## wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11 and G are defined as in claim 1; and prodrugs or pharmaceutically acceptable salts thereof.

3. The compound of formula (I) according to claim 1, wherein R.sup.7 is selected from H, F, Cl, methyl, ethyl, halomethyl and haloethyl; and prodrugs or pharmaceutically acceptable salts thereof.

4. The compound of formula (I) according to claim 1, wherein R.sup.1 is selected from halomethyl, haloethyl and methyl; and prodrugs or pharmaceutically acceptable salts thereof.

5. The compound according to claim 3, wherein R.sup.1 is a fluoromethyl selected from the group consisting of —CF.sub.3, —CHF.sub.2 and —CH.sub.2F; and prodrugs or pharmaceutically acceptable salts thereof.

6. The compound of formula (I) according to claim 1, wherein at least one of R.sup.3 and R.sup.4 is methyl; and prodrugs or pharmaceutically acceptable salts thereof.

7. The compound of formula (I) according to claim 1, wherein one of R.sup.3 and R.sup.4 is methyl and the other one is H; and prodrugs or pharmaceutically acceptable salts thereof.

8. The compound of formula (I) according to claim 1, wherein G is O; and prodrugs or pharmaceutically acceptable salts thereof.

9. The compound of formula (I) according to claim 1, wherein G is O, and wherein one of R.sup.3 or R.sup.4 is methyl and the other one is H; and prodrugs or pharmaceutically acceptable salts thereof.

10. The compound according to claim 8, wherein R.sup.4 is methyl and R.sup.3 is H, wherein R.sup.5 and R.sup.6 together with the C-atoms in between form an oxetane ring; and prodrugs or pharmaceutically acceptable salts thereof.

11. The compound of formula (I) according to claim 1, wherein R.sup.2 is selected from the group consisting of H, ethynyl, 1-propynyl, —S— methyl, halogen; and prodrugs or pharmaceutically acceptable salts thereof.

12. The compound of formula (I) according to claim 1, wherein R.sup.2 is ethynyl; and prodrugs or pharmaceutically acceptable salts thereof.

13. The compound of formula (I) according to claim 1, wherein R.sup.4 is methyl and R.sup.3 is H, wherein G is O, wherein R.sup.5 and R.sup.6 together form an oxetane ring wherein R.sup.2 is selected from the group consisting of H, ethynyl, 1-propynyl, —S— methyl, halogen; and prodrugs or pharmaceutically acceptable salts thereof.

14. The compound of formula (I) according to claim 1, wherein R.sup.2 is a cyclic group wherein the cyclic group is selected from the group consisting of a phenyl or a five- to six-membered heteroaryl comprising 1, 2 or 3 heteroatoms selected from N, S and O, wherein this cyclic group is substituted by one or two, identical or different substituents R.sup.10, wherein each R.sup.10 is independently selected from the group consisting of hydrogen, halogen, haloalkyl, -methyl, -ethyl, —NH—CO-methyl, —N(CH.sub.3).sub.2, —CH2-OH, —NH(CH.sub.3), —O—CH.sub.3, —CN, —S—CH.sub.3, —CO—NH.sub.2, —CH.sub.2—NH(CH.sub.3), —CH.sub.2—NH.sub.2, —SO—(CH.sub.3), cyclopropyl and —O—R.sup.11, wherein each R.sup.11 is selected from a five- or six-membered heterocycle with one or two heteroatoms each independently selected from N and O; and prodrugs or pharmaceutically acceptable salts thereof.

15. The compound according to claim 14, wherein R.sup.2 is a cyclic group selected from the group consisting of pyrazolyl, pyridinyl, imidazolyl, phenyl and isoxazolyl, wherein this cyclic group is substituted by one or two, identical or different substituents R.sup.10, wherein each R.sup.10 is independently selected from the group consisting of hydrogen, halogen, haloalkyl, -methyl, -ethyl, —NH—CO-methyl, —N(CH.sub.3).sub.2, —CH2-OH, —NH(CH.sub.3), —O—CH.sub.3, —CN, —S—CH.sub.3, —CO—NH.sub.2, —CH.sub.2—NH(CH.sub.3), —CH.sub.2—NH.sub.2, —SO—(CH.sub.3), cyclopropyl and —O—R.sup.11, wherein each R.sup.11 is tetrahydropyrane; and prodrugs or pharmaceutically acceptable salts thereof.

16. The compound according to claim 15, wherein G is O, wherein one of R.sup.3 or R.sup.4 is methyl and the other one is H; and prodrugs or pharmaceutically acceptable salts thereof.

17. The compound according to claim 16, wherein R.sup.4 is methyl and R.sup.3 is H, wherein R.sup.5 and R.sup.6 together with the C-atom in between form an oxetane ring; and prodrugs or pharmaceutically acceptable salts thereof.

18. The compound of formula (I) according to claim 1, wherein G is CR.sup.8R.sup.9 and wherein R.sup.8 and R.sup.6 and the two C-atoms in between R.sup.8 and R.sup.6 form an annulated five-membered aromatic heterocycle comprising one or two heteroatoms each independently selected from N and O which is selected from an annulated isoxazolyl ring, an annulated pyrazolyl ring, an annulated pyrrolyl ring and an annulated furanyl ring and wherein R.sup.9 and R.sup.5 are absent; and prodrugs or pharmaceutically acceptable salts thereof.

19. The compound of formula (I) according to claim 1, which is selected from the group consisting of ##STR00424## ##STR00425## ##STR00426## ##STR00427## ##STR00428## ##STR00429## ##STR00430## ##STR00431## ##STR00432## ##STR00433## ##STR00434## ##STR00435## ##STR00436## ##STR00437## ##STR00438## ##STR00439## ##STR00440## ##STR00441## ##STR00442## ##STR00443## and prodrugs or pharmaceutically acceptable salts thereof.

20. An intermediate of formula (IV) ##STR00444## or formula (V) ##STR00445## or formula (X) ##STR00446## or formula (XI) ##STR00447## wherein R.sup.1 is selected from methyl, ethyl, halomethyl, haloethyl and halogen, wherein G is selected from O, NR.sup.8, CH.sub.2, C and CR.sup.8R.sup.9, wherein R.sup.2 is selected from H, halogen, cyclopropyl, C.sub.1-3-alkyl, —C.sub.2-5-alkynyl, —S-methyl and CN, or wherein R.sup.2 is a cyclic group, wherein this cyclic group is selected from the group consisting of a phenyl or a five- to six-membered heteroaryl comprising 1, 2, 3 or 4 heteroatoms each independently selected from N, S and O, wherein this cyclic group is substituted by one or two, identical or different substituents R.sup.10, wherein R.sup.3 is H or methyl, R.sup.4 is H or methyl, R.sup.5 is selected from H, methyl, —CN, -methylene-OH and —CF.sub.3, or R.sup.5 may be absent, R.sup.6 is selected from H, methyl, —CN, -methylene-OH and —CF.sub.3, or R.sup.5 and R.sup.6 together with the C-atoms in between form a ring selected from oxetane, tetrahydrofurane and cyclopropane, R.sup.7 is selected from H, halogen, (C.sub.1-3)-alkyl and halo-(C.sub.1-3)-alkyl, R.sup.8 is selected from CN, H and methyl, R.sup.9 is selected from H, methyl and halogen or R.sup.9 may be absent, wherein each R.sup.10 is independently selected from the group consisting of hydrogen, halogen, haloalkyl, -methyl, -ethyl, —NH—CO-methyl, —N(CH.sub.3).sub.2, —CH.sub.2—OH, —NH(CH.sub.3), —O—(C.sub.1-3-alkyl), —CN, —S—CH.sub.3, —CO—NH.sub.2, —CH.sub.2—NH CH.sub.3), —CH.sub.2—NH.sub.2, —SO—CH.sub.3), cyclopropyl and —O—R.sup.11, wherein R.sup.11 is a five- or six-membered heterocycle with one or two heteroatoms each independently selected from N, O and S, or G is CR.sup.8R.sup.9, R.sup.5 and R.sup.9 are absent, and R.sup.8 and R.sup.6 and the two C-atoms in between R.sup.8 and R.sup.6 form an annulated five-membered aromatic or non-aromatic heterocycle comprising one, two or three heteroatoms each independently selected from N, S and O, or G is CR.sup.8R.sup.9 and R.sup.8 and R.sup.9 form together with the C-atom in between R.sup.8 and R.sup.9 a diazirine ring, and wherein X is F or NO.sub.2, and wherein PG is a protecting group selected from the group consisting of tert-butoxycarbonyl (BOC), benzyloxycarbonyl (Cbz), fluorenylmethylenoxycarbonyl (Fmoc) and allyloxycarbonyl (Alloc).

21. A prodrug of any of the compounds of formula (I) as defined in claim 1 which falls into the scope of formula (A) ##STR00448## or of formula (A′) ##STR00449## wherein R.sup.12 is C.sub.1-4-alkyl, aryl, —CH.sub.2-aryl, NH—SO.sub.2—C.sub.1-3-alkyl.

22. The prodrug of formula (A) or of formula (A′) according to claim 21, wherein R.sup.12 is methyl.

23. A method of treating in a subject a disease that can be treated by the inhibition of cGAS, said method comprising administering to the subject a compound of formula (I) according to claim 1.

24. A method of treating in a subject a disease selected from the group consisting of systemic lupus erythematosus (SLE), interferonopathies, Aicardi-Goutières syndrome, age-related macular degeneration (AMD), amyotrophic lateral sclerosis (ALS), inflammatory bowel disease (IBD), chronic obstructive pulmonary disease (COPD), Bloom's syndrome, Sjogren's syndrome, Parkinsons disease, heart failure and cancer, systemic sclerosis (SSc), non-alcoholic steatotic hepatitis (NASH), interstitial lung disease (ILD), progressive fibrosing interstitial lung disease (PF-ILD), and idiopathic pulmonary fibrosis (IPF), said method comprising administering to the subject a compound of formula (I) according to claim 1.

25. A method of treating in a subject a disease selected from the group consisting of systemic lupus erythematosus (SLE), interferonopathies, Aicardi-Goutières syndrome, age-related macular degeneration (AMD), amyotrophic lateral sclerosis (ALS), inflammatory bowel disease (IBD), chronic obstructive pulmonary disease (COPD), Bloom's syndrome, Sjogren's syndrome and Parkinsons disease, said method comprising administering to the subject a compound of formula (I) according to claim 1.

26. A method of treating in a subject a fibrosing disease selected from the group consisting of systemic sclerosis (SSc), non-alcoholic steatotic hepatitis (NASH), interferonopathies, interstitial lung disease (ILD), progressive fibrosing interstitial lung disease (PF-ILD), and idiopathic pulmonary fibrosis (IPF), said method comprising administering to the subject a compound of formula (I) according to claim 1.

27. A method of treating in a subject a disease selected from the group consisting of, age-related macular degeneration (AMD), heart failure, COVID-19/SARS-CoV-2 infection, renal inflammation, renal fibrosis, dysmetabolism, vascular diseases, cardiovascular diseases and cancer, said method comprising administering to the subject a compound of formula (I) according to claim 1.

28. A pharmaceutical composition comprising a compound of formula (I) according to claim 1 and optionally one or more pharmaceutically acceptable carriers and/or excipients.

29. A pharmaceutical composition comprising a compound of formula (I) according to claim 1 in combination with one or more active agents selected from the group consisting of anti-inflammatory agents, anti-fibrotic agents, anti-allergic agents/anti-histamines, bronchodilators, beta 2 agonists/betamimetics, adrenergic agonists, anticholinergic agents, methotrexate, mycophenolate mofetil, leukotriene modulators, JAK inhibitors, anti-interleukin antibodies, non-specific immunotherapeutics such as interferons or other cytokines/chemokines, cytokine/chemokine receptor modulators, toll-like receptor agonists, immune checkpoint regulators, an anti-TNF antibody, and an anti-BAFF antibody.

30. The pharmaceutical composition according to claim 29, wherein the compound of formula (I) is combined with one or more anti-fibrotic agents selected from the group consisting of Pirfenidon and Nintedanib.

31. The pharmaceutical composition according to claim 29, wherein the compound of formula (I) is combined with one or more anti-inflammatory agents selected from the group consisting of NSAIDs and corticosteroids.

32. The pharmaceutical composition according to claim 29, wherein the compound of formula (I) is combined with one or more active agents selected from the group of bronchodilators, beta 2 agonists/betamimetics, adrenergic agonists and anticholinergic agents.

33. The pharmaceutical composition according to claim 29, wherein the compound of formula (I) is combined with one or more anti-interleukin antibodies selected from the group consisting of anti-IL-23 antibodies, anti-IL-17 antibodies, anti-IL-1 antibodies, anti-IL-4 antibodies, anti-IL-13 antibodies, anti-IL-5 antibodies, anti-IL-6 antibodies, anti-IL-12 antibodies and anti-IL-15 antibodies.

Description

PREPARATION OF FINAL COMPOUNDS

Example 1.01 (General Route)

(2S,4S)-4-{[5-Chloro-3-(2-cyanomorpholin-4-yl)pyridin-2-yl]oxy}-1-[4-(trifluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.SUP.2,7.]trideca-1(13),2,4,6,9,11-hexaen-6-yl]pyrrolidine-2-carboxylic acid

[0393] ##STR00183##

[0394] To (2S,4S)-4-hydroxy-1-[4-(trifluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.sup.2,7]trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]pyrrolidine-2-carboxylic acid (INTERMEDIATE 1.3.I, 58.0 mg, 0.15 mmol) in 2.00 mL DMA were added 4-(5-chloro-2-nitropyridin-3-yl)morpholine-2-carbonitrile (INTERMEDIATE 2.6.111, 80.6 mg, 0.30 mmol, 2.0 eq) and NaH (18.0 mg, 0.45 mmol, 3.0 eq). The reaction mixture was stirred for 20 min at 80° C. The reaction mixture was diluted with ACN/water, acidified with TFA, filtered and purified by HPLC (ACN/H2O/TFA). The product was obtained as a mixture of two diastereoisomers.

[0395] ESI-MS: 589 [M+H].sup.+

[0396] R.sub.t (H PLC): 1.05 min (method H)

[0397] The compounds listed in the table below were prepared according to the general procedure (EXAMPLE 1.01) described above. Where indicated in the table, the EXAMPLE compounds were either isolated as diastereomeric mixtures (ds-mix) or pure diasteroisomers (R.sub.t are given for both isolated diastereoisomers (EXAMPLE and 2.sup.nd diasteroisomer (2.sup.nd ds)).

TABLE-US-00014 R.sub.t (HPLC) [min] Starting Reaction (method) Ex. materials Structure conditions ESI-MS Example 2.sup.nd ds 1.02 Int.1.3.II + Int.2.6.IV (2.0 eq) [00184] solvent: DMA, NaH: 2.0 eq, RT, 1 h 585 [M + H].sup.+ 0.52 (D) ds-mix — 1.03 Int.1.3.I + Int.2.6.IV (2.0 eq) [00185] solvent: DMA, NaH: 2.0 eq, RT, 1 h 603 [M + H].sup.+ 0.57 (D) ds-mix — 1.04 Int.1.3.II + Int.2.6.V (2.0 eq) [00186] solvent: DMA, NaH: 2.0 eq, RT, 1 h 583 [M + H].sup.+ 0.63 (D) ds-mix — 1.05 Int.1.3.I + Int.2.6.V (2.0 eq) [00187] solvent: DMA, NaH: 2.0 eq, RT 1 h 601 [M + H].sup.+ 0.67 (D) ds-mix — 1.06 Int.1.3.I + Int.2.6.VI (1.0 eq) [00188] solvent: DMA, NaH: 3.0 eq, 85° C., 45 min 592/ 594 [M + H].sup.+ 2.72 (L) 2.81 (L) 1.07 Int.1.3.I + Int.7.1.I (1.3 eq) [00189] solvent: NMP, NaH: 5.0 eq, RT, 1 h 601 [M + H].sup.+ 0.74 (A) — 1.08 Int.1.3.II + Int.7.1.I (1.3 eq) [00190] solvent: NMP, NaH: 5.0 eq, RT 1 h 583 [M + H].sup.+ 0.69 (A) — 1.09 Int.1.3.II + Int.2.6.II [00191] solvent: NMP, NaH: 5.0 eq, 80° C. 1 h 602/ 604 [M + H].sup.+ 0.70 (A) 0.74 (A) 1.10 Int.1.3.I + Int.2.6.I (1.3 eq) [00192] solvent: DMA, NaH: 3.0 eq, 80° C. 10 min, RT overnight 664/ 666 [M + H].sup.+ 1.07 (B) 1.13 (B) 1.11 Int.1.3.II + Int.2.7.I (1.0 eq) [00193] solvent: NMP, NaH: 5.0 eq, RT, 10 min 592 [M + H].sup.+ 0.67 (A) 0.71 (A) 1.12 Int.1.3.I + Int.2.7.I (1.0 eq) [00194] solvent: NMP, NaH: 5.0 eq, RT 15 min 610 [M + H].sup.+ 0.74 (A) 0.79 (A) 1.13 Int.1.3.I + Int.2.6.II (2.0 eq) [00195] solvent: NMP, NaH: 5.0 eq 80° C., 1 h 620/ 622 [M + H].sup.+ 0.75 (A) 0.79 (A) 1.14 Int.1.3.III + Int.2.7.I (1.0 eq) [00196] solvent: NMP, NaH: 5.0 eq, RT, 25 min, 606 [M + H].sup.+ 0.69 (A) 0.73 (A)- 1.15 Int.1.3.III + Int.2.7.I (1.0 eq) [00197] solvent: NMP, NaH: 5.0 eq, RT 25 min, 606 [M + H].sup.+ 0.73 (A) 0.69 (A) 1.16 Int.1.3.IV + 2.7.I (1.0 eq) [00198] solvent: NMP, NaH: 5.0 eq, RT, 20 min 626 [M + H].sup.+ 1.14 (B) 1.65 (B) 1.17 Int.1.3.V + Int.2.7.I (1.0 eq) [00199] solvent: NMP, NaH: 5.0 eq, RT, 30 min 660 [M + H].sup.+ 1.04 (C) 1.09 (C) 1.18 Int.1.3.II + Int.11.2 (1.3 eq) [00200] solvent: DMF, NaH: 3.0 eq, RT, 1.25 h 628 [M + H].sup.+ 1.12 (C) — 1.19 1.3.II + 2.7.IV (1.5 eq) [00201] solvent: DMA, NaH: 3.0 eq, RT, 30 min 548 [M + H].sup.+ 0.71 (H) — 1.20 Int.1.3.II + Int.2.6.XVI (1.0 eq) [00202] solvent: DMF, NaH: 3.0 eq, RT, 15 min 574 [M + H].sup.+ 1.13 (C) — 1.21 Int.1.3.II + Int. 2.6.XVII (2.0 eq) [00203] solvent: NMP, NaH: 5.0 eq, 80° C., 1 h 600/ 602 [M + H].sup.+ 0.74 (A) 0.80 (A) 1.22 Int.1.3.I + Int.2.7.VI (1.0 eq) [00204] solvent: NMP, NaH: 5.0 eq, 80° C., 1 h 608 [M + H].sup.+ 0.78 (A) 0.85 (A) 1.23 Int.1.3.II + Int.2.7.VI (1.0 eq) [00205] solvent: NMP, NaH: 5.0 eq, 80° C.1 h 590 [M + H].sup.+ 0.73 (A) 0.80 (A) 1.24 Int.1.3.II + Int.2.6.IX (2.0 eq) [00206] solvent: DMA NaH: 3.0 eq 80° C., 10 min 632 [M + H].sup.+ 1.03 min (H) — 1.25 Int.1.3.II + Int.2.6.X [00207] solvent: DMA, 80° C. 10 min 578 [M + H].sup.+ 0.96 (H) — 1.26 Int.1.3.II + Int.2.6.XI [00208] solvent: DMA, RT overnight 618 [M + H].sup.+ 1.028 (C) — 1.27 Int.1.3.II + Int.2.6.XII [00209] solvent: DMA, 10 min 80° C. 560 [M + H].sup.+ 0.53 (D) — 1.28 Int.1.3.II + Int.2.6.XIII [00210] solvent: DMA, 10 min 80° C. 604 [M + H].sup.+ 1.05 (H) — 1.29 Int.1.3.VI + Int.2.6.XIV [00211] solvent: DMF, 30 min RT 514 [M + H].sup.+ 0.86 (C) — 1.30 Int.1.3.I + Int.2.6.XIV [00212] solvent: DMF, 30 min RT 568 [M + H].sup.+ 1.16 (C) — 1.31 Int.1.3.II + Int.2.6.XIV [00213] solvent: DMA, 45 min RT 550 [M + H].sup.+ 1.18 (C) — 1.32 Int.1.3.VII + Int.2.6.XIV [00214] solvent: DMA, 45 min RT 534 [M + H].sup.+ 1.11 (C) — 1.33 Int.1.3.I + Int.2.6.XIX (1.1 eq) [00215] solvent: NMP 1.5 h, RT 648/ 650 [M + H].sup.+ 0.79 (A) 1.34 Int.1.3.I + Int.6.1 (1.2 eq) [00216] solvent: NMP 30 min, RT 604 [M + H].sup.+ 0.70 (A) 0.75 (A)

[0398] The absolute stereochemistry of Example 1.10 was confirmed via small molecule X-ray as illustrated below:

##STR00217##

[0399] The absolute stereochemistry of Example 1.28 was confirmed via small molecule X-ray as illustrated below:

##STR00218##

Example 2.01 (General Route)

(2S,4S)-1-[4-(Difluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.SUP.2,7.]trideca-1(13),2,4,6,9,11-hexaen-6-yl]-4-{[5-ethynyl-3-(morpholin-4-yl)pyridin-2yl]oxy}-pyrrolidine-2-carboxylic acid

[0400] ##STR00219##

[0401] To 6-chloro-4-(difluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.sup.2,7]trideca-1(9),2(7),3,5,10,12-hexaene (INTERMEDIATE 1.2.11, 30.0 mg, 0.12 mmol) in 1.50 mL DMSO were added (2S,4S)-4-{[5-ethynyl-3-(morpholin-4-yl)pyridin-2-yl]oxy}pyrrolidine-2-carboxylic acid (INTERMEDIATE 8.1, 55.9 mg, 0.13 mmol) and DI PEA (60.8 μL, 0.35 mmol). The reaction mixture was stirred for 1 h at 110° C. The reaction mixture was diluted with ACN, acidified with TEA, filtered and purified by HPLC (ACN/H2O/TFA).

[0402] ESI-MS: 536 [M+H].sup.+

[0403] R.sub.t (H PLC): 1.08 min (method C)

[0404] The following EXAMPLES were prepared according to the general procedure (EXAMPLE 2.1) described above:

TABLE-US-00015 R.sub.t (HPLC) Reaction [min] Ex. Starting materials Structure conditions ESI-MS (method) 2.02 Int. 1.2.I + Int. 8.1 (1.1 eq) [00220] solvent: DMSO, DIPEA: 3.0 eq, 110° C., 1 h 554 [M + H].sup.+ 1.14 (C) 2.03 [00221]+ Int. 9.2 (1.0 eq) [00222] 4 eq. K.sub.2CO.sub.3 solvent: DMF, RT, 1 h, 50° C. 30 min 556 [M + H].sup.+ 0.50 (A) 2.04 1.2.VI + Int. 9.2 [00223] 4 eq. K.sub.2CO.sub.3 solvent: DMF, RT, 45 min, 50° C., 30 min 574 [M + H].sup.+ 0.58 (A) 2.05 Int. 1.3.VII + Int. 9.2 [00224] 4 eq. K.sub.2CO.sub.3 solvent: DMF, RT 2 h 576 [M + H].sup.+ 0.66 (A) 2.06 Int. 10.4.I + Int. 9.2 [00225] 4 eq. K.sub.2CO.sub.3 solvent: DMF, RT 2 h, 50° C. 1 h 606 [M + H].sup.+ 0.53 (A) 2.07 Int. 10.4.II + Int. 9.2 [00226] 4 eq. K.sub.2CO.sub.3 solvent: DMF, RT 2 h, 50° C. 1 h 590 [M + H].sup.+ 0.54 (A) 2.08 Int. 10.4.III + Int. 9.2 [00227] 4 eq. K.sub.2CO.sub.3 solvent: DMF, RT 1 h, 50° C. 1 h 574 [M + H].sup.+ 0.51 (A) 2.09 Int. 14.4 + Int. 9.2 [00228] 4 eq. K.sub.2CO.sub.3 solvent: DMF, RT 2 h, 50° C. 4.5 h 638 [M + H].sup.+ 0.78 (A) 2.10 Int. 15.3 + Int. 9.2 [00229] 2.3 eq. K.sub.2CO.sub.3, DMF, RT 6 h 590/592 [M + H].sup.+ 0.68 (A)

Example 3.01 (General Route)

(2S,4S)-4-({5-[(35)-3-methylmorpholin-4-yl]-[3,4′-bipyridin]-6-yl}oxy)-1-[4-(trifluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.SUP.2,7.]trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]pyrrolidine-2-carboxylic acid

[0405] ##STR00230##

[0406] To a mixture of (2S,4S)-4-({5-bromo-3-[(3S)-3-methylmorpholin-4-yl]pyridin-2-yl}oxy)-1-[4-(trifluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.sup.2,7]trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]pyrrolidine-2-carboxylic acid (INTERMEDIATE 3.1.II, 50.0 mg, 0.08 mmol), (pyridin-4-yl)boronic acid (24.7 mg, 0.20 mmol), Na.sub.2CO.sub.3 solution (2.0M, 100 μL, 0.20 mmol), Xphos 3.sup.rd gen (3.40 mg) and Pd(PPh.sub.3).sub.4 (4.64 mg) was added under argon 2.00 mL dioxane. The reaction mixture was stirred for 2 h at 100° C. The reaction mixture was filtered, diluted with ACN/methanol and purified by HPLC (Xbridge, ACN/H.sub.2O/TFA).

[0407] ESI-MS: 621 [M+H].sup.+

[0408] R.sub.t (H PLC): 0.818 min (method B)

[0409] The following EXAMPLES were prepared according to the general procedure (EXAMPLE 3.01) described above. The boronates or boronic acids used are either commercially available or can be easily prepared as described in the literature (Boronic Acids: Preparation and Applications in Organic Synthesis, Medicine and Materials, 1&2, 2.sup.nd Edition, ISBN 9783527325986).

TABLE-US-00016 R.sub.t (HPLC) Reaction [min] Ex. Starting materials Structure conditions ESI-MS (method) 3.02 Int. 3.1.II +   [00231] [00232] solvent: dioxane, 2 h 100° C., RT overnight 650 [M + H].sup.+ 0.81 (B) 3.03 Int. 3.1.II +   [00233] [00234] solvent: dioxane, 2 h 100° C. 639 [M + H].sup.+ 0.56 (K) 3.04 Int. 3.1.II +   [00235] [00236] solvent: dioxane, 2 h 100° C. 671 [M + H].sup.+ 0.59 (K) 3.05 Int. 3.1.II +   [00237] [00238] solvent: dioxane, 2 h 100° C. 651 [M + H].sup.+ 1.11 (B) 3.06 Int. 3.1.II +   [00239] [00240] solvent: dioxane, 2 h 100° C., RT overnight 624 [M + H].sup.+ 0.91 (B) 3.07 Int. 3.1.II +   [00241] [00242] solvent: dioxane, 2 h 100° C., RT overnight 638 [M + H].sup.+ 0.94 (B) 3.08 Int. 3.1.I +   [00243] [00244] solvent: dioxane, 1.5 h 100° C. 674 [M + H].sup.+ 0.76 (I) 3.09 Ex. 1.10 +   [00245] [00246] solvent: dioxane, 100° C., 1.5 h 692 [M + H].sup.+ 0.72 (J) 3.10 Ex. 1.10 +   [00247] [00248] solvent: dioxane, 100° C., 1.5 h 720 [M + H].sup.+ 0.50 (K) 3.11 Ex. 1.10 +   [00249] [00250] solvent: dioxane, 100° C., 4 h 674 [M + H].sup.+ 0.92 (J) 3.12 Ex. 1.10 +   [00251] [00252] solvent: dioxane, 100° C. 2 h 691 [M + H].sup.+ 0.58 (D) 3.13 Ex. 1.10 +   [00253] [00254] solvent: dioxane, 100° C. 1 h 705 [M + H].sup.+ 0.64 (A) 3.14 Ex. 1.10 +   [00255] [00256] solvent: dioxane, 100° C. 1.5 h 693 [M + H].sup.+ 0.57 (K) 3.15 Int. 3.1.I +   [00257] [00258] solvent: dioxane, 100° C. 1.5 h 702 [M + H].sup.+ 0.69 (H) 3.16 Int. 3.1.I +   [00259] [00260] solvent: dioxane, 100° C. 1.5 h 691 [M + H].sup.+ 0.54 (D) 3.17 Ex. 1.10 +   [00261] [00262] solvent: dioxane, 100° C. 6 h 705 [M + H].sup.+ 0.83 (H) 3.18 Ex. 1.10 +   [00263] [00264] solvent: dioxane, 100° C. 1.5 h 692 [M + H].sup.+ 0.73 (H) 3.19 Int. 3.1.I +   [00265] [00266] solvent: dioxane, 100° C. overnight 673 [M + H].sup.+ 0.68 (H) 3.20 Ex. 1.10 +   [00267] [00268] solvent: dioxane, 100° C., 1.5 h 666 [M + H].sup.+ 0.69 (H) 3.21 Int. 3.1.I +   [00269] [00270] solvent: dioxane, 100° C., 2.5 h, RT overnight 670 [M + H].sup.+ 0.50 (D) 3.22 Ex. 1.10 +   [00271] [00272] solvent: dioxane, 100° C. 3 h 705 [M + H].sup.+ 0.75 (H) 3.23 Ex. 1.10 +   [00273] [00274] solvent: dioxane, 100° C. 1.5 h 688 [M + H].sup.+ 1.01 (B) 3.24 Int. 3.1.I   [00275] [00276] solvent: dioxane, 100° C. 2.5 h 687 [M + H].sup.+ 0.85 (J) 3.25 Ex. 1.10 +   [00277] [00278] solvent: dioxane, 100° C. 1.5 h, RT overnight 711 [M + H].sup.+ 0.60 (D) 3.26 Ex. 1.10 +   [00279] [00280] solvent: dioxane, 100° C. 1.5 h, RT overnight 697 [M + H].sup.+ 1.07 (J) 3.27 Ex. 1.10 +   [00281] [00282] solvent: dioxane, 100° C. 1.5 h, RT overnight 681 [M + H].sup.+ 1.04 (J) 3.28 Ex. 1.10 +   [00283] [00284] solvent: dioxane, 100° C. 1.5 h, RT overnight 688 [M + H].sup.+ 1.00 (J) 3.29 Ex. 1.10 +   [00285] [00286] solvent: dioxane, 100° C. 1.5 h 763 [M + H].sup.+ 0.59 (K) 3.30 Ex. 1.10 +   [00287] [00288] solvent: dioxane, 100° C. 2 h 720 [M + H].sup.+ 0.53 (A) 3.31 Ex. 110 +   [00289] [00290] solvent: dioxane, 100° C., 2 h 702 [M + H].sup.+ 0.55 (D) 3.32 Int. 3.1.I   [00291] [00292] solvent: dioxane, 90° C. 1 h 693 [M + H].sup.+ 1.05 (E)

Example 4.01 (General Route)

(2S,4S)-1-[(Difluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.SUP.2,7.]trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]-4-({N,N-dimethyl-5-[(9S)-9-methyl-2,5-dioxa-8-azaspiro[3.5]nonan-8-yl]-[3,4′-bipyridin]-2′-yl}oxy)pyrrolidine-2-carboxylic acid

[0410] ##STR00293##

[0411] To tert-butyl (2S,4S)-1-[4-(difluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.sup.2,7]trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]-4-({N,N-dimethyl-5-[(9S)-9-methyl-2,5-dioxa-8-azaspiro[3.5]nonan-8-yl]-[3,4′-bipyridin]-2′-yl}oxy)pyrrolidine-2-carboxylate (INTERMEDIATE 3.4.I, 13 mg, 0.020 mmol) in 2.00 mL DCM was added over a period of 1 day trifluoroacetic acid (650 μL, 8.49 mmol). The reaction mixture was stirred overnight at RT and then concentrated under reduced pressure. The crude product was purified by HPLC (Xbridge, ACN/H.sub.2O/TFA).

[0412] ESI-MS: 688 [M+H].sup.+

[0413] R.sub.t (HPLC): 0.51 min (method A)

[0414] The following EXAMPLES were prepared according to the general procedure (EXAMPLE 4.1) described above:

TABLE-US-00017 R.sub.t (HPLC) Starting Reaction [min] Ex. material Structure conditions ESI-MS (method) 4.02 Int. 3.4.II [00294] solvent: DCM, RT overnight 718 [M + H].sup.+ 0.53 (A) 4.03 Int.3.4.III [00295] solvent: DCM, RT overnight 704 [M + H].sup.+ 0.51 (A) 4.04 Int.3.4.IV [00296] solvent: DCM, RT overnight 568 [M + H].sup.+ 0.63 (A)

Example 5.01

(2S,4S)-1-[4-(Difluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.SUP.2,7.]trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]-4-({2′-methanesuIfinyl-5-[(9S)-9-methyl-2,5-dioxa-8-azaspiro[3.5]nonan-8-yl]-[3,4′-bipyridin]-6-yl}oxy)pyrrolidine-2-carboxylic acid

[0415] ##STR00297##

[0416] To a cooled solution of (2S,4S)-1-[4-(difluoromethyl)-8-oxa-3,5-diazatricyclo-[7.4.0.0.sup.2,7]trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]-4-({5-[(9S)-9-methyl-2,5-dioxa-8-azaspiro[3.5]nonan-8-yl]-2′-(methylsulfanyl)-[3,4′-bipyridin]-6-yl}oxy)-pyrrolidine-2-carboxylic acid (EXAMPLE 3.16, 25.0 mg, 0.04 mmol) in 1.00 mL DCM was added at −15° C. meta-chloroperoxybenzoic acid (7.30 mg, 0.03 mmol). The reaction mixture was stirred at −15° C. for 10 min and subsequently purified by HPLC (Xbridge, ACN/H.sub.2O/TFA).

[0417] ESI-MS: 707 [M+H].sup.+

[0418] R.sub.t (H PLC): 0.85 min (method H)

Example 6.01

(2S,4S)-4-({3-[(9S)-9-Methyl-2,5-dioxa-8-azaspiro[3.5]nonan-8-yl]-5-(prop-1-yn-1-yl)pyridin-2-yl}-oxy)-1-[4-(trifluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.SUP.2,7.]trideca-1(9),2(7),3,5,10,12hexaen-6-yl]pyrrolidine-2-carboxylic acid

[0419] ##STR00298##

[0420] To a degassed solution of (2S,4S)-4-({5-bromo-3-[(9S)-9-methyl-2,5-dioxa-8-azaspiro[3.5]nonan-8-yl]pyridin-2-yl}oxy)-1-[4-(trifluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.sup.2,7]trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]pyrrolidine-2-carboxylic acid (EXAMPLE 1.10, 150 mg, 0.230 mmol) in 5.00 mL THE was added copper(I) iodide (4.3 mg, 0.023 mmol) followed by Pd(dppf)Cl.sub.2 (33 mg, 0.050 mmol)). After stirring for a few minutes, a 1M solution of methylacetylene in THE (1.35 mL, 1.35 mmol) was added and the mixture was heated to 75° C. for 4 h. The reaction mixture was diluted with ACN, acidified with acetic acid, filtered and purified by HPLC (Xbridge, ACN/H.sub.2O/TFA).

[0421] ESI-MS: 624 [M+H].sup.+

[0422] R.sub.t (H PLC): 1.08 min (method E)

[0423] The following EXAMPLES were prepared according to the general procedure (EXAMPLE 6.01) described above:

TABLE-US-00018 R.sub.t (HPLC) Starting Reaction [min] Ex. material Structure conditions ESI-MS (method) 6.02 Ex 1.10 [00299] like Ex. 6.01 606 [M + H].sup.+ 1.01 (E) 6.03 Ex 1.33 [00300] like Ex. 6.01 608 [M + H].sup.+ 1.13 (P)

Example 7.01

(2S,4S)-4-({3-[(9S)-9-Methyl-2,5-dioxa-8-azaspiro[3.5]nonan-8-yl]-5-(pyrimidin-5-yl)pyridin-2-yl}oxy)-1-[4-(trifluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.SUP.2,7.]trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]pyrrolidine-2-carboxylic acid

[0424] ##STR00301##

[0425] To a degassed mixture of (2S,4S)-4-({3-[(9S)-9-methyl-2,5-dioxa-8-azaspiro[3.5]nonan-8-yl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl}oxy)-1-[4-(trifluoromethyl)-8-oxa-3,5-diazatricyclo-[7.4.0.02,7]-trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]pyrrolidine-2-carboxylic acid (INTERMEDIATE 16.1, 220 mg, 0.309 mmol), potassium carbonate solution (2.0 mol/L, 0.463 mL, 0.928 mmol) and 4-bromo-1-cyclopropyl-1H-pyrazole (119 mg, 618 mmol) in 5.00 mL dioxane was added Xphos 3rd gen (15 mg, 0.018 mmol). The reaction mixture was heated to 80° C. for 2 h, then cooled to RT and diluted with ethyl acetate. Saturated aqueous ammonium chloride solution was added and water. The phases were separated, and the aqueous layer extracted with ethyl acetate. The combined organic layers were washed with saturated ammonium chloride solution and dried over sodium sulfate, filtered, evaporated and purified by HPLC (Sunfire, ACN/H.sub.2O/TFA).

[0426] ESI-MS: 692 [M+H].sup.+

[0427] R.sub.t (H PLC): 1.01 min (E)

[0428] The following EXAMPLES were prepared according to the general procedure (EXAMPLE 7.01) described above:

TABLE-US-00019 R.sub.t (HPLC) Starting Reaction [min] Ex. material Structure conditions ESI-MS (method) 7.02 Ex. 1.10 +   [00302] [00303] Dioxane, K.sub.2CO.sub.3, 95° C., 30 min 699 [M + H].sup.+ 0.74 (A) 7.03 Ex. 1.10 + [00304] Dioxane, Cs.sub.2CO.sub.3, 100° C., 2 h 692 [M + H].sup.+ 0.73 (P) 7.04 Ex. 1.10 +   [00305] [00306] Dioxane, Cs.sub.2CO.sub.3, 100° C., 2 h 678 [M + H].sup.+ 1.02 (P) 7.05 Ex. 1.10 +   [00307] [00308] Dioxane, K.sub.2CO.sub.3, 100° C., 2 h 667 [M + H].sup.+ 0.72 (A)

Example 8.01

(2S,4S)-4-{[5-(1-methyl-1H-1,2,3-triazol-4-yl)-3-[(9S)-9-methyl-2,5-dioxa-8-azaspiro[3.5]nonan-8-yl]pyridin-2-yl]oxy}-1-[4-(trifluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.SUP.2,7.]trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]pyrrolidine-2-carboxylic acid

[0429] ##STR00309##

[0430] A mixture of EXAMPLE 1.12 (75 mg, 0.12 mmol), trimethylsilylmethyl azide (16 mg, 0.012 mmol), copper(II)-sulfate (20 mg, 0.12 mmol) and L-ascorbic acid sodium salt (49 mg, 0.24 mmol) in a mixture if 1.00 mL DMSO and 0.10 mL water was stirred at RT for 1 h. Then, 1M TBAF solution (240 IL) was added and the reaction mixture was stirred for 1 h at room temperature. The reaction mixture was diluted with ACN/H2O, acidified with TFA, filtered and purified by HPLC.

[0431] ESI-MS: 667 [M+H].sup.+

[0432] R.sub.t (H PLC): 0.87 min (method P)

Example 9.01

(2S,4S)-1-[4-(difluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.SUP.2,7.]trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]-4-({3-[(9S)-9-methyl-2,5-dioxa-8-azaspiro[3.5]nonan-8-yl]-5-(methylsulfanyl)pyridin-2-yl}oxy)pyrrolidine-2-carboxylic acid

[0433] ##STR00310##

[0434] A degassed mixture of (2S,4S)-4-({5-Bromo-3-[(9S)-9-methyl-2,5-dioxa-8-azaspiro[3.5]nonan-8-yl]pyridin-2-yl}oxy)-1-[4-(difluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.02,7]-trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]pyrrolidine-2-carboxylic acid (INTERMEDIATE 3.1.I (a), 160 mg, 0.25 mmol), XANTPHOS (20 mg, 0.035 mmol), sodium methanethiolate (0.25 mL, 0.59 mmol), DIPEA (0.100 mL, 0.58 mmol) and Pd.sub.2(dba).sub.3 (16 mg, 0.017 mmol) in 8.00 mL dioxane was heated at 110° C. over night. After cooling to RT, ethyl acetate and water were added to the reaction mixture. The organic layer was separated, dried, concentrated in vacuo and purified by HPLC (Sunfire C-18, H.sub.2O/ACN/TFA).

[0435] ESI-MS: 614 [M+H].sup.+

[0436] R.sub.t (HPLC): 1.01 min (method C)

Example 10.01

(2S,4S)-4-({3-[(9S)-9-methyl-2,5-dioxa-8-azaspiro[3.5]nonan-8-yl]-5-(1H-pyrazol-1-yl)pyridin-2-yl}oxy)-1-[4-(trifluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.SUP.2,7.]trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]pyrrolidine-2-carboxylic acid

[0437] ##STR00311##

[0438] To a degassed mixture of (2S,4S)-4-({5-bromo-3-[(9S)-9-methyl-2,5-dioxa-8-azaspiro[3.5]nonan-8-yl]pyridin-2-yl}oxy)-1-[4-(trifluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.sup.2,7]-trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]pyrrolidine-2-carboxylic acid (Ex. 1.10, 30 mg, 0.045 mmol), LiHMDS (1M in THF, 0.120 mL, 0.120 mmol) and pyrazole (5.00 mg, 0.073 mmol) in 1.50 mL dioxane under an argon atmosphere was added tBuBrettPhos (5.00 mg, 0.006 mmol), and the mixture was heated at 80° C. for 5 h. After cooling to RT, the mixture was diluted with methanol, concentrated in vacuo and purified by HPLC.

[0439] ESI-MS: 652 [M+H].sup.+

[0440] R.sub.t (HPLC): 1.01 min (method P)

[0441] Peparation of Prodrugs:

[0442] Prodrug P01

Methyl (2S,4S)-4-({3-[(9S)-9-methyl-2,5-dioxa-8-azaspiro[3.5]nonan-8-yl]pyridin-2-yl}oxy)-1-[4-(difluoromethyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.SUP.2,7.]trideca-1(9),2(7),3,5,10,12-hexaen-6-yl]pyrrolidine-2-carboxylate

[0443] ##STR00312##

[0444] To (2S,4S)-4-({3-[(9S)-9-methyl-2,5-dioxa-8-azaspiro[3.5]nonan-8-yl]-pyridin-2-yl}-oxy).sub.1-[4-(difluoro-methyl)-8-oxa-3,5-diazatricyclo[7.4.0.0.sup.2,7]trideca-1(13),2,4,6,9,11-hexaen-6-yl]pyrrolidine-2-carboxylic acid (EXAMPLE 4.04, 15 mg, 0.020 mmol) in 1.0 mL THE was added O-methyl-N,N′-diisopropylurea (41 μL, 0.25 mmol). The reaction mixture was stirred at RT for 60 h, then diluted with ACN/water and purified by HPLC (ACN/H2O/TFA).

[0445] ESI-MS: 582 [M+H].sup.+

[0446] R.sub.t (H PLC): 0.69 min (method A)

[0447] The following compounds were prepared according to the general procedure (PRODRUG P01) described above:

TABLE-US-00020 Pro- R.sub.t (HPLC) drug Starting Reaction [min] # material Structure /name conditions ESI-MS (method) P02 Ex 1.10 [00313] solvent: THF, RT, 3 days 678 [M + H].sup.+ 1.16 (D) P03 Ex 1.12 [00314] solvent: THF RT, 3 days 624 [M + H]+ 1.16 (H) P04 Ex 3.14 [00315] solvent: THF RT, 44 h 707 [M + H]+ 1.23 (J)

LIST OF ABBREVIATIONS

[0448] ACN acetonitrile [0449] aq. aqueous [0450] ° C. degree Celsius [0451] CH cyclohexane [0452] DBU diazabicyclo[5.4.0]undec-7-ene [0453] DCM dichloromethane [0454] DIPEA diisopropylethylamine [0455] DMA dimethylacetamide [0456] DMF N,N-dimethylformamide [0457] ds diastereoisomer [0458] ESI-MS electrospray ionisation mass spectrometry [0459] EtOAc ethyl acetate [0460] eq equivalent [0461] FA formic acid [0462] FC flash-chromatography, SiO.sub.2 is used if no further details are given [0463] h hour(s) [0464] HCl hydrogenchloride [0465] HATU [dimethylamino-(1,2,3-triazolo[4,5-b]pyridin-3-yloxy)-methylene]-dimethyl-ammonium hexafluorophosphate [0466] HPLC high performance liquid chromatography [0467] K.sub.2CO.sub.3 potassium carbonate [0468] KOH potassium hydroxide [0469] L liter [0470] LiAlH.sub.4 lithium aluminium hydride [0471] LiHMDS lithium hexamethyldisilazide [0472] MeOH methanol [0473] min minute [0474] mL milliliter [0475] M molar [0476] MS mass spectrum [0477] n.d. not determined [0478] NH.sub.4OH solution of NH.sub.3 in water [0479] Pd.sub.2(dba).sub.3 Tris(dibenzylideneacetone)dipalladium(0) [0480] Pd(dppf)Cl.sub.2 (1,1′-bis-(diphenylphosphino)-ferrocene)-dichloropalladium (II) [0481] Pd(PPh.sub.3).sub.4 tetrakis(triphenylphosphine)palladium(0) [0482] Pd(OH).sub.2/C palladium hydroxide on carbon 20% [0483] RT room temperature (about 20° C.) [0484] Sol solvent [0485] tBuBrettPhos (prop-2-en-1-yl)palladiumylium di-tert-butyl[3,6-dimethoxy-2′,4′,6′-tris(propan-2-yl)-[1,1′-biphenyl]-2-yl]phosphane trifluoromethanesulfonate [0486] TBS tert-butyl-dimethylsilyl [0487] TBAF tetrabutylammoniumfluoride [0488] TEA triethyl amine [0489] TFA trifluoroacetic acid [0490] TFAA trifluoroacetic acid anhydride [0491] THE tetrahydrofurane [0492] TIPS triisopropylsilyl [0493] TMS trimethylsilyl [0494] TosOH p-toluenesulfonic acid monohydrate [0495] TLC thin layer chromatography [0496] RP-HPLC reverse phase HPLC [0497] R.sub.f retardation factor (TLC) [0498] R.sub.t retention time in minutes [0499] Vol % volume percent [0500] Xphos 3.sup.rd gen (2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II) methanesulfonate [0501] Ziram dimethyldithiocarbamic acid zinc salt

[0502] Analytical HPLC methods:

[0503] Method A

TABLE-US-00021 Vol % water time (min) (incl. 0.1% TFA) Vol % ACN Flow [mL/min] 0.00 99 1 1.6 0.02 99 1 1.6 1.00 0 100 1.6 1.10 0 100 1.6

[0504] Analytical column: XBridge BEH C18_2.1×30 mm, 1.7 μm; column temperature: 60° C.

[0505] Method B

TABLE-US-00022 Vol % water time (min) (incl. 0.1% TFA) Vol % ACN Flow [mL/min] 0.00 97 3 2.2 0.20 97 3 2.2 1.20 0 100 2.2 1.25 0 100 3.0 1.40 0 100 3.0

[0506] Analytical column: Stable Bond (Agilent) 1.8 μm; 3.0×30 mm; column temperature: 60° C.

[0507] Method C

TABLE-US-00023 Vol % water time (min) (incl. 0.1% TFA) Vol % ACN Flow [mL/min] 0.00 97 3 2.2 0.20 97 3 2.2 1.20 0 100 2.2 1.25 0 100 3.0 1.40 0 100 3.0

[0508] Analytical column: Sunfire (Waters) 2.5.m; 3.0×30 mm; column temperature: 60° C.

[0509] Method D

TABLE-US-00024 Vol. % water time (min) (incl. 0.1% NH.sub.4OH) Vol. % ACN Flow [mL/min] 0.00 95 5 1.5 1.30 0 100 1.5 1.50 0 100 1.5 1.60 95 5 1.5

[0510] Analytical column: XBridge C18_3.0×30 mm-2.5 μm (Waters); column temperature: 60° C.

[0511] Method E

TABLE-US-00025 Vol. % water time (min) (incl. 0.1% TFA) Vol. % ACN Flow [mL/min] 0.00 95 5 1.5 1.30 0 100 1.5 1.50 0 100 1.5

[0512] Analytical column: Sunfire C18 (Waters) 2.5 μm; 3.0×30 mm; column temperature: 60° C.

[0513] Method F

TABLE-US-00026 Vol % water time (min) (incl. 0.1% NH3) Vol % ACN Flow [mL/min] 0.00 95 5 1.3 0.02 95 5 1.3 1.00 0 100 1.3 1.30 0 100 1.3

[0514] Analytical column: XBridge BEH (Waters) C18_2.1×30 mm, 2.5 μm; column temperature: 60° C.

[0515] Method G

TABLE-US-00027 Vol % water time (min) (incl. 0.1% TFA) Vol % ACN Flow [mL/min 0.00 50 50 1.6 0.02 50 50 1.6 1.00 0 100 1.6 1.10 0 100 1.6

[0516] Analytical column: XBridge BEH (Waters) C18_2.1×30 mm, 1.7 μm; column temperature: 60° C.

[0517] Method H

TABLE-US-00028 Vol. % water time (min) (incl. 0.1% TFA) Vol. % ACN Flow [mL/min] 0.00 95 5 1.5 1.30 0 100 1.5 1.50 0 100 1.5 1.60 95 5 1.5

[0518] Analytical column: Sunfire C18 (Waters) 2.5 μm; 3.0×30 mm; column temperature: 60° C.

[0519] Method I

TABLE-US-00029 Vol % water time (min) (incl. 0.1% NH3) Vol % ACN Flow [mL/min] 0.00 97 3 2.2 0.20 97 3 2.2 1.20 0 100 2.2 1.25 0 100 3.0 1.40 0 100 3.0

[0520] Analytical column: XBridge (Waters) C18_3.0×30 mm, 2.5 μm; column temperature: 60° C.

TABLE-US-00030 Vol. % water time (min) (incl. 0.1% TFA) Vol. % ACN Flow [mL/min] 0.00 95 5 1.5 1.30 0 100 1.5 1.50 0 100 1.5 1.60 95 5 1.5

[0521] Method J

[0522] Analytical column: Sunfire C18 (Waters) 2.5 μm; 3.0×30 mm; column temperature: 60° C.

[0523] Method K

TABLE-US-00031 Vol. % water time (min) (incl. 0.1% NH3) Vol. % ACN Flow [mL/min] 0.00 95 5 1.5 1.30 0 100 1.5 1.50 0 100 1.5 1.60 95 5 1.5

[0524] Analytical column: Xbridge C18 (Waters) 2.5 μm; 3.0×30 mm; column temperature: 60 mC

[0525] Method L

TABLE-US-00032 Vol. % time (min) carbon dioxide Vol. % methanol Flow [mL/min] 0.00 97 3 1.3 2.50 55 45 1.3 3.50 55 45 1.3 3.51 97 3 1.3 4.00 97 3 1.3

[0526] Analytical column: Acquity UPC2 Torus 2-PIC (Waters) 1.7 μm; 3.0×100 mm; column temperature: 30° C.

[0527] Method M

TABLE-US-00033 Vol. % water time (min) (incl. 0.1% FA) Vol. % ACN Flow [mL/min] 0.00 97 3 2.2 0.20 97 3 2.2 1.20 0 100 2.2 1.25 0 100 3.0 1.40 0 100 3.0

[0528] Analytical column: Sunfire C18 (Waters) 2.5 μm; 3.0×30 mm; column temperature: 60° C.

[0529] Method N

TABLE-US-00034 Vol. % water time (min) (incl. 0.1% TFA) Vol. % ACN Flow [mL/min] 0.00 99 1 1.5 0.20 99 1 1.5 1.00 0 100 1.5 1.10 0 100 1.5

[0530] Analytical column: XSelect HSS PEP (Waters) 1.8 μm; 2.1×30 mm; column temperature: 60° C.

[0531] Method P

TABLE-US-00035 Vol. % water time (min) (incl. 0.1% formic acid) Vol. % ACN Flow [mL/min] 0.00 80 20 0.5 0.10 80 20 0.5 1.10 0 100 0.5 2.50 80 20 0.5 3.00 80 20 0.5

[0532] Analytical column: Acquity UPLC BEH C18 (Waters) 1.7 μm; 2.1×100 mm; column temperature: 40

[0533] Method P

TABLE-US-00036 Vol. % water time (min) (incl. 0.1% TFA) Vol. % ACN Flow [mL/min] 0.00 95 5 1.5 1.30 0 100 1.5 1.50 0 100 1.5 1.60 95 5 1.5

[0534] column: Sunfire C18 (Waters) 2.5 μm; 3.0×30 mm; CT: 60° C.

EXAMPLES

5.1 Example Compounds

[0535] The following Example compounds of formula (I) or of formula (I′) as summarized in Table 1 have been synthesized and tested with respect to their pharmacological properties regarding their potency to inhibit cGAS activity.

[0536] In particular the “biochemical (in vitro) IC50-values” with regard to cGAS-inhibition (hcGAS IC50), the “IC50-value with regard to the inhibition of IFN induction in virus-stimulated THP1 cells” (THP.sub.(vir) IC50), the “IC50-value with regard to the inhibition of IFN induction in cGAMP-stimulated THP1 cells” (THP.sub.(cGAMP) IC50) and the “IC50-value with regard to inhibition of IFN induction in dsDNA-stimulated human whole blood” (hWB IC50) has been experimentally determined according to the assay methods as described in section 6 below. The results are summarized in Table 1.

[0537] The Example compounds of formula (I) or of formula (I′) as summarized in Table 1 show at the same time the following three properties: [0538] a satisfying “biochemical (in vitro) IC50-value with regard to cGAS inhibition” (with a hcGAS IC50 of ≤100 nM, preferably of ≤50 nM, in particular of ≤10 nM), [0539] a satisfying “cellular IC50-value regarding cGAS inhibition” (with a THP1.sub.(vir) IC50 of 1 μM, preferably of 500 nM, more preferably of ≤100 nM, in particular of ≤50 nM) and [0540] a satisfying selectivity for cGAS-inhibition [0541] (with a ratio THP1.sub.(CGAMP)IC50/THP1.sub.(vir) IC50 of ≥10, more preferably ≥50, more preferably ≥500, in particular ≥1000).

[0542] Additionally, the Example compounds of formula (I) or of formula (I′) also show acceptable IC50-values with regard to inhibition of IFN induction in dsDNA-stimulated human whole blood (hWB IC50).

TABLE-US-00037 TABLE 1 Pharmacological properties of the Example compounds of the invention hcGAS THP1.sub.(vir) Ratio hWB Exam. IC50 IC50 THP1.sub.(cGAMP) THP1.sub.(cGAMP) IC50/ IC50 No. Structure [nM] [nM] IC50 [nM] THP1.sub.(vir) IC50 [nM] 1.01 [00316] 8 829 24551 30 1.02 [00317] 4 697 21869 31 3439 1.03 [00318] 7 399 15136 38 8453 1.04 [00319] 6 327 10029 31 3703 1.05 [00320] 14 775 12540 16 9506 1.06 [00321] 65 951 14283 15 1.07 [00322] 5 781 18722 24 1.08 [00323] 4 831 18745 23 1.09 [00324] 2 19 22143 1169 19 1.10 [00325] 5 91 16130 178 33 1.11 [00326] 2 6 18501 3095 5 1.12 [00327] 3 7 11892 1769 15 1.13 [00328] 4 40 16406 413 102 1.14 [00329] 3 6 >16591 >2708 7 1.15 [00330] 75 332 26182 79 702 1.16 [00331] 3 7 15012 2209 23 1.17 [00332] 17 154 >16612 >108 305 1.18 [00333] 4 239 6513 27 226 1.19 [00334] 4 699 >16605 >24 667 1.20 [00335] 2 796 >16608 >21 670 1.21 [00336] 4 85 14830 175 46 1.22 [00337] 5 21 7637 355 29 1.23 [00338] 4 10 10913 1134 12 1.24 [00339] 9 848 >16597 >20 850 1.25 [00340] 4 517 >16617 >32 551 1.26 [00341] 10 583 9934 17 475 1.27 [00342] 4 516 >16604 >32 221 1.28 [00343] 3 500 >16620 >33 629 1.29 [00344] 4 296 >16623 >56 61 1.30 [00345] 3 159 22053 139 80 1.31 [00346] 3 232 >16601 >72 102 1.32 [00347] 4 487 1.33 [00348] 3 15 >10 000 677 252 1.34 [00349] 3 19 >10 000 514 51 1.35 [00350] 5 10 >10 000 >1000 47 2.01 [00351] 8 858 >16595 >19 506 2.02 [00352] 7 752 >16602 >22 1034 2.03 [00353] 3 <5 16402 3043 9 2.04 [00354] 1 3 25412 8125 5 2.05 [00355] 2 4 10808 3018 8 2.06 [00356] 4 63 11932 189 52 2.07 [00357] 3 14 14535 1005 28 2.08 [00358] 3 <5 18814 3490 6 2.09 [00359] 6 31 5177 166 37 2.10 [00360] 3 5 2746 533 24 3.01 [00361] 9 982 >16604 >17 1607 3.02 [00362] 13 306 4916 16 1538 3.03 [00363] 7 899 12838 14 2125 3.04 [00364] 12 757 7779 10 2420 3.05 [00365] 14 523 6710 13 1831 3.06 [00366] 9 525 21357 41 1988 3.07 [00367] 17 748 12339 16 1616 3.08 [00368] 5 4 4789 1179 3 3.09 [00369] 9 8 4816 585 8 3.10 [00370] 8 5 7909 1713 10 3.11 [00371] 4 5 5289 1149 11 3.12 [00372] 9 6 11138 1930 19 3.13 [00373] 9 25 8352 335 65 3.14 [00374] 11 10 7832 822 26 3.15 [00375] 10 57 19921 348 22 3.16 [00376] 15 19 11849 629 38 3.17 [00377] 20 11 16809 1542 43 3.18 [00378] 3 7 4758 645 44 3.19 [00379] 8 267 >16593 >62 47 3.20 [00380] 3 30 11521 383 50 3.21 [00381] 4 24 11689 483 55 3.22 [00382] 16 177 8912 50 70 3.23 [00383] 8 10 10027 1029 70 3.24 [00384] 6 57 19222 336 322 3.25 [00385] 9 19 9712 502 96 3.26 [00386] 11 48 9238 193 1369 3.27 [00387] 21 326 12478 38 1858 3.28 [00388] 14 712 16375 23 3.29 [00389] 66 591 6544 11 1574 3.30 [00390] 26 15 12 626 829 84 3.31 [00391] 11 3 10 899 4120 28 3.32 [00392] 9 2 12885 5194 17 4.01 [00393] 10 <5 6676 >1239 5 4.02 [00394] 12 17 12562 735 42 4.03 [00395] 8 199 7872 40 78 4.04 [00396] 2 386 >16611 >43 77 5.01 [00397] 5 579 22083 38 239 6.01 [00398] 10 <5 20480 3801 25 6.02 [00399] 6 <5 >16 000 >3000 8 6.03 [00400] 7 5 10947 2273 95 7.01 [00401] 14 5 9257 1700 50 7.02 [00402] 14 1 8320 7533 13 7.03 [00403] 21 2 >10009 4151 27 7.04 [00404] 12 2 >10011 4084 22 7.05 [00405] 9 10 13172 1292 35 8.01 [00406] 5 11 14921 1366 21 9.01 [00407] 6 16 19805 1254 38 10.01  [00408] 5 10 >10000 >1000 47

5.2 Comparison of the Example Compounds with Prior Art Compounds

5.2.1 Compounds of WO 2020/142729

[0543] In WO 2020/142729 cGAS-inhibitors with partially similar structures have been disclosed. On page 44 and 45 of WO 2020/142729 the “biochemical (in vitro) IC50-values” with regard to cGAS-inhibition (corresponding to “hcGAS IC50”) have been disclosed. Hereby compounds with a “biochemical (in vitro) IC50-value” of less than 100 nM had been designated into “group A”, compounds with a “biochemical (in vitro) IC50-value” of greater than 100 nM and less than 500 nM had been designated into “group B”, compounds with a “biochemical (in vitro) IC50-value” of greater than 500 nM and less than 1 μM had been designated into “group C”, compounds with a “biochemical (in vitro) IC50-value” of greater than 1 μM and less than 10 μM had been designated into “group D” and compounds with a “biochemical (in vitro) IC50-value” of greater than 10 μM had been designated into “group E” (see page 44 of WO 2020/142729).

[0544] On page 45 of WO 2020/142729 it is disclosed that only compound No. 25 could be designated to “group A” having a “biochemical (in vitro) IC50-value” of less than 100 nM. All other example compounds of WO 2020/142729 show “biochemical (in vitro) IC50-values” of greater than 100 nM.

5.2.2 Comparison Between the Examples of the Invention and the Examples of WO 2020/142729

[0545] Selected prior art compounds of WO 2020/142729 have been synthesized and then have been tested with respect to their pharmacological properties regarding their potency to inhibit the cGAS/STING pathway. In particular the “biochemical (in vitro) IC50-values” with regard to cGAS-inhibition (hcGAS IC50), the “cellular IC50-values with regard to inhibition of IFN induction in virus-stimulated THP1 cells” (THP1.sub.(vir) IC50), the “cellular IC50-value with regard to inhibition of IFN induction in cGAMP-stimulated THP1 cells” (THP1.sub.(CGAMP) IC50) and the “IC50-value with regard to inhibition of IFN induction in human whole blood” (hWB) have been experimentally determined for the structurally closest examples of WO 2020/142729 according to the assay methods as described in section 6 below (see Table 2).

TABLE-US-00038 TABLE 2 Pharmacological properties of a selection of Example compounds from WO 2020/142729 Example No. (as disclosed hcGAS THP1.sub.(vir) hWB in WO IC50 IC50 THP1.sub.(cGAMP) IC50 2020/142729) Structure [nM] [nM] IC50 [nM] [nM] 15 [00409] 2700 >17000 >17000 — 25 [00410] 55 >17000 >17000 >9992 28 [00411] 630 >32000 >17000 >9990 38 [00412] 3000 >17000 >17000 >9990 58 [00413] 320 21000 23000 >9982

[0546] The pharmacological properties for the Example compounds of the invention as summarized in Table 1 and the respective pharmacological properties for the compounds of WO 2020/142729 can be compared to each other, since they were experimentally determined according to the identical assay procedures as described in section 6 below.

[0547] From data as shown in Table 2 it is clear that all example compounds of WO 2020/142729 show “biochemical (in vitro) IC50-values” (=hcGAS IC50) that are significantly larger than 100 nM—with the only exception of Example No. 25 of WO 2020/142729 (in WO 2020/142729 designated in “Group A” having a “biochemical (in vitro) IC50-value” (=hcGAS IC50) of less than 100 nM). In contrast to that the Example compounds of the invention all have “biochemical (in vitro) IC50-values” (hcGAS IC50) of less than 100 nM. However, Example No. 25 of WO 2020/142729 which has a “biochemical (in vitro) IC50-value” (hcGAS IC50) of 55 nM, does not at all comply with the selection criterium of a “satisfying cellular inhibitory potency” shown by a THP1.sub.(vir) IC50 of lower than 1 μM, because THP1.sub.(vir) IC50 for Example No. 25 of WO 2020/142729 is 17 μM.

5.3 Prodrugs

[0548] It is known that esters of active agents with a carboxylic acid group may represent viable prodrugs which may i.e. show an improved oral absorption/bioavailability compared to the respective active agent. Frequently used prodrugs of active agents with a carboxylic acid group are for example methyl esters, ethyl esters, iso-propyl esters etc. (see Beaumont et al., Current Drug Metabolism, 2003, Vol. 4, Issue 6, 461-485).

[0549] Further, Nakamura et al., Bioorganic & Medicinal Chem., Vol. 15, Issue 24, p. 7720-7725 (2007), describes that also N-acylsulfonamide derivatives and N-acylsulfonylurea derivatives of a specific active agent with a free carboxylic acid group have the potential of being a viable prodrug.

[0550] Additionally, experimental hints have been found that also the methyl esters of the example compounds of formula (I) or of formula (I′) represent viable prodrugs of the cGAS inhibitors of formula (I) or of formula (I′).

[0551] Compounds P01, P02, P03 and P04 are methyl esters of the Example compounds 4.04, 1.10, 1.12 and 3.14, respectively and therefore may represent viable prodrugs of the respective Example compounds.

[0552] P01, P02, P03 and P04 have been synthesized and tested for their pharmacological properties with respect to their potency to inhibit the cGAS/STING pathway. Subsequently, the experimentally determined pharmacological properties of prodrugs P01, P02, P03 and P04 have been compared to the corresponding pharmacological properties of the respective Example compounds 4.04, 1.10, 1.12 and 3.14 as summarized in Table 3.

[0553] This comparison between the Example compound and its corresponding prodrug shows that the hcGAS IC50-values for the Example compounds are always around or even smaller than 10 nM, whereas the hcGAS IC50-values for the corresponding prodrugs are always extremely large, that means generally larger than 9000 nM. That large difference between Example compound on the one hand and its corresponding prodrug on the other hand is never observed for the respective THP1.sub.(vir) IC50-values which always stay in the same range between example compound and its corresponding prodrug (see Table 3 for Example No. 4.04 and its respective prodrug P01).

[0554] One possible explanation for that observation is that the example compounds (which represent the “drugs”) all have a free carboxyl group which seems to be crucial for inhibition of cGAS activity, whereas in all “prodrugs” the carboxyl group is masked by a carboxy-methyl ester group. Consequently, the prodrugs lose their inhibitory potency in the “in vitro human cGAS enzyme assay” (see section 6.1 below), because in this assay intracellular enzymes that cleave the carboxy-methyl ester group are absent. Therefore the prodrugs show extremely large “biochemical (in vitro) IC50-values” (=hcGAS IC50) in this “in vitro human cGAS enzyme assay”, whereas the corresponding Example compounds (which represent the drugs or active agents) show small “biochemical (in vitro) IC50-values” (=hcGAS IC50).

[0555] In the “human cGAS cell and the counter cell assay” (see section 6.2 below) endogenous cellular enzymes that cleave the carboxy-methyl ester group are present. Consequently not only the Example compounds themselves (that means the drugs or active agents themselves) show small THP1.sub.(vir) IC50-values, but also the corresponding prodrugs show relatively small “THP1.sub.(vir) IC50-values”, because in this “human cGAS cell assay” the methyl ester of the prodrugs can be cleaved by endogenous intracellular enzymes into the corresponding drug/active agent that shows inhibitory potency again.

[0556] This explanation together with the measurements as shown in Table 3 imply that methyl ester derivatives of the compounds of formula (I) or of formula (I′) really seem to represent viable prodrugs of the compounds of formula (I) or of formula (I′) which themselves have no inhibitory potency regarding the in vitro human biochemical cGAS inhibition. However, upon cleavage of the methyl ester by endogenous intracellular enzymes the compounds of formula (I) or of formula (I′) (the active agents) are formed, that exhibit again an inhibitory potency regarding the cGAS/STING pathway.

TABLE-US-00039 TABLE 3 Comparison between selected Example compound of the invention (= active agents) and their respective methyl ester prodrugs: Example No./ hcGAS THP1.sub.(vir) THP1.sub.(cGAMP) hWB Prodrug IC50 IC50 IC50 IC50 No. Structure [nM] [nM] [nM] [nM] P01 (Prodrug of Ex. 4.04) [00414] >9956 358 26058 1313 Ex. 4.04 [00415] 2 386 >16611 77 P02 (Prodrug of Ex. 1.10) [00416] >9952 444 17109 5327 Ex. 1.10 [00417] 5 91 16130 33 P03 (Prodrug of Ex. 1.12) [00418] 9617 188 22845 1088 Ex. 1.12 [00419] 3 7 11892 15 P04 (Prodrug of Ex. 3.14) [00420] >9954 67 >16621 2747 Ex. 3.14 [00421] 11 10 7832 26

6 Biological Experiments

[0557] The activity of the compounds of the invention may be demonstrated using the following in vitro cGAS enzyme and cell assays:

6.1 Method: Human cGAS Enzyme Assay (hcGAS IC50 (In Vitro))

[0558] Human cGAS enzyme was incubated in the presence of a 45 base pair double stranded DNA to activate the enzyme and GTP and ATP as substrates. Compound activity was determined by measuring the effect of compounds on the formation of the product of the enzyme reaction, cGAMP, which is measured by a mass spectrometry method.

[0559] Enzyme preparation:

[0560] Human cGAS (amino acid 1-522) with an N-terminal 6x-His-tag and SUMO-tag was expressed in E. coli BL21(DE3) pLysS (Novagen) cells for 16 h at 18° C. Cells were lysed in buffer containing 25 mM Tris (pH 8), 300 mM NaCl, 10 mM imidazole, 10% glycerol, protease inhibitor cocktail (Complete™, EDTA-free, Roche) and DNase (5 μg/mL). The cGAS protein was isolated by affinity chromatography on Ni-NTA agarose resin and further purified by size exclusion chromatography using a Superdex 200 column (GE Healthcare) equilibrated in 20 mM Tris (pH 7.5), 500 mM KCl, and 1 mM TCEP. Purified protein was concentrated to 1.7 mg/mL and stored at −80° C.

[0561] Assay Method

[0562] Compounds were delivered in 10 mM DMSO solution, serially diluted and transferred to the 384 well assay plate (Greiner #781201) using an Echo acoustic dispenser. Typically, 8 concentrations were used with the highest concentration at 10 μM in the final assay volume followed by ˜1:5 dilution steps. DMSO concentration was set to 1% in the final assay volume. The 384 well assay plate contained 22 test compounds (column 1-22), and DMSO in column 23 and 24.

[0563] After the compound transfer, 15 μL of the enzyme-DNA-working solution (12 nM cGAS, 0.32 μM 45base pair DNA in assay buffer, 10 mM Tris pH 7.5/10 mM KCl/5 mM MgCl2/1 mM DTT) were added to each well from column 1-23 via a MultiDrop Combi dispenser. In column 24, 15 μl of assay buffer without enzyme/DNA were added as a low control.

[0564] The plates were then pre-incubated for 60 min at room temperature.

[0565] Following that, 10 μL of GTP (ThermoFisher #R0461)-ATP (Promega #V915B) mix in assay buffer were added to the assay plate (columns 1-24, 30 μM final concentration each) using a Multidrop Combi.

[0566] The plates were incubated again for 90 min at room temperature.

[0567] Following the incubation, the reaction was stopped by 80 μL of 0.1% formic acid in assay buffer containing 5 nM cyclic-di-GMP (Sigma #SML1228) used as internal standard for the mass spectrometry. The total volume/well was 105 μL.

[0568] Rapidfire MS Detection

[0569] The plates were centrifuged at 4000 rpm, 4° C., for 5 min.

[0570] The RapidFire autosampler was coupled to a binary pump (Agilent 1290) and a Triple Quad 6500 (ABSciex, Toronto, Canada). This system was equipped with a 10 μL loop, C18 [12 μL bed volume] cartridge (Agilent, Part No. G9210A) containing 10 mM NH4Ac (aq) water (pH7.4) as eluent A (pump 1 at 1.5 mL/min, pump 2 at 1.25 mL/min) and 10 mM NH4Ac in v/v/v 47.5/47.5/5 ACN/MeOH/H2O (pH7.4) as eluent B (pump 3 at 1.25 mL/min). Aspiration time: 250 ms; Load time: 3000 ms; Elute time: 3000 ms; Wash volume: 500 μL.

[0571] The MS was operated in positive ion mode with HESI ion source, with a source temperature of 550° C., curtain gas=35, gas 1=65, and gas 2=80. Unit mass resolution in SRM mode. The following transitions and MS parameters (DP: declustering potential and CE: collision energy) for cGAMP and DicGMP were determined:

[0572] Analyte: cGAMP at 675.1/524, DP=130, CE=30 and

[0573] Internal standard: cyclic-di-GMP at 690.1/540, DP=130, CE=30.

[0574] The formation of cGAMP was monitored and evaluated as ratio to cyclic-di-GMP.

[0575] Data Evaluation and Calculation:

[0576] For data evaluation and calculation, the measurement of the low control was set as 0% control and the measurement of the high control was set as 100% control. The IC50 values were calculated using the standard 4 parameter logistic regression formula. Calculation: [y=(a−d)/(1+(x/c){circumflex over ( )}b)+d], a=low value, d=high value; x=conc M; c=IC50 M; b=slope

6.2 Method: Human cGAS Cell Assay and cGAMP Stimulated Counter Cell Assay (THP1.SUB.(vir) .IC50 and Thp1.SUB.(cGAMP) .Ic50)

[0577] THP1-Dual™ cells (InvivoGen #thpd-nfis) expressing IRF dependent Lucia luciferase reporter were used as basis for both assays. For the detection of cellular cGAS activity cells were stimulated by a baculovirus (pFastbac-1, Invitrogen, no coding insert) infection that delivers the cGAS enzyme stimulating double-stranded DNA (measurement of THP1.sub.(vir) IC50).

[0578] For the counter assay, cells were stimulated by cGAMP (SigmaAldrich #SML1232) to activate the identical pathway independent and directly downstream of cGAS (measurement of THP1.sub.(cGAMP) IC50). Pathway activity was monitored by measuring the Lucia luciferase activity induced by either DNA stimulated cGAS enzyme activity (measurement of THP1.sub.(vir) IC50) or by cGAMP directly (measurement of THP1.sub.(CGAMP) IC50, counter assay).

[0579] Assay Method

[0580] Compounds were delivered in 10 mM DMSO solution, serially diluted and transferred to the 384 well assay plate (Greiner #781201) using an Echo acoustic dispenser. Typically, 8 concentrations were used with the highest concentration at 10 μM in the final assay volume followed by ˜1:5 dilution steps. DMSO concentration was set to 1% in the final assay volume. The 384 well assay plate contained 21 test compounds (column 1-22), and DMSO in column 23 and 24.

[0581] Cells, cultivated according to manufacturer conditions, were harvested by centrifugation at 300 g/10 min and were 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 (Life Technologies #A11139-03)). The baculovirus solution was then added 1:200 (have varied according to virus batch) to the cells (measurement of THP1.sub.(vir) IC50). Alternatively, for the counter assay cGAMP was added to the cells at a final concentration of 10 μM (measurement of THP1.sub.(cGAMP) IC50). 30 μL of the cell/virus-mix were added to each well of the compound plate from column 1-23 via MultiDrop Combi dispenser (5000 cells/well). In column 24, 30 μl/5000 cells/well without virus were added as a low control.

[0582] The plates were then incubated for 18 h at 37° C. in a humidified incubator.

[0583] Following that, 15 μL of QuantiLuc detection reagent (InvivoGen #rep-qlcg5) were added to each well using a MultiDrop Combi. Measurement was done immediately after the addition using an EnVision reader (US-luminescence read-mode).

[0584] Data Evaluation and Calculation:

[0585] For data evaluation and calculation, the measurement of the low control was set as 0% control and the measurement of the high control was set as 100% control. The IC50 values were calculated using the standard 4 parameter logistic regression formula. Calculation: [y=(a−d)/(1+(x/c){circumflex over ( )}b)+d], a=low value, d=high value; x=conc M; c=1C50 M; b=slope

6.3 Method: Human Whole Blood Assay (Human WB IC50)

[0586] For the detection of cellular cGAS activity human whole blood was stimulated by transfection with double stranded DNA. Pathway activity was monitored by measuring the IFNα2α production.

[0587] Assay Method

[0588] Compounds were delivered as 10 mM DMSO solution and serially diluted and transferred to the 96-well cell culture plate (Corning #3595), prefilled with 20 μl OptiMEM (Gibco, #11058-021) in each well, using an Echo acoustic dispenser. Typically, 8 concentrations were used with the highest concentration at 10 μM in the final assay volume followed by ˜1:5 dilution steps. DMSO concentration was set to 0.1% in the final assay volume. The 96-well assay plate contained 10 test compounds, and DMSO in control wells.

[0589] Collection of human whole blood from 3 or more healthy donors (male or female, no medication for 7 days except contraceptive and thyroxine) as Na-Citrate blood (e.g. 3.8% in Monovettes from Sarstedt) was conducted in parallel. Whole blood was kept at room temperature for a maximum of 3 hours after collection until use in the assay.

[0590] 160 μl of the whole blood samples was transferred to each well of the 96-well assay plates filled with compound/OptiMEM. All assay plates were prepared as duplicates with blood from different donors. Blood plates were kept at room temperature for 60 minutes and continuous shaking with 450 rpm, covered with the lid, but not sealed.

[0591] DNA-Fugene mix (Herring DNA, Sigma Aldrich #D6898-1G, Fugene (5×1 mL), Promega #E2312) was prepared in OptiMEM and incubated for 10 min at RT (125 ng DNA/20 μl and Fugene ratio 9.6:1). 20 μl of the DNA Fugene mix was added to each well, resulting in 125 ng DNA/well/200 μl, and Fugene Ratio 9.6:1. 20 μl OptiMEM and 9.6:1 Fugene was added to all low control wells.

[0592] After covering assay plates with area seals and the lid, blood plates were kept at room temperature for 30 minutes and continuous shaking with 450 rpm, followed by an overnight incubation of 22 h at 37° C. in the incubator, without shaking.

[0593] For the detection of IFNα-2α in human plasma, the biotinylated capture antibody (Antibody set IFNA2, Meso Scale Diagnostics #B21VH-3, including coating and capture antibody) was diluted 1:17.5 in Diluent 100 (Meso Scale Diagnostics #R50AA-4), according to the manufacturer's directions. U-Plex MSD GOLD 96-well Small Spot Strepavidin SECTOR Plates (Meso Scale Diagnostics #L45SA-5) were coated with 25 μl diluted capture antibody. Coated plates were incubated for 60 min at room temperature under continuous shaking at 700 rpm. MSD IFNα-2a plates were washed three times with 150 μl wash buffer (1× HBSS, 0.05% Tween).

[0594] After blocking the plates with 100 μl block solution/well (1× HBSS with 0.2% Tween, 2% BSA) for 60 min at room temperature and continuous shaking at 700 rpm, plates were emptied as dry as possible by dumping just before continuing with the human plasma.

[0595] Whole Blood assay plates were centrifuged at 1600 rpm for 10 minutes. 25 μl of supernatant was transferred with a pipetting robot from each whole blood plate to the corresponding IFNα-2α plate. Plates were sealed with microplate seals and kept at room temperature again under continuous shaking at 700 rpm for two hours.

[0596] Next MSD IFNα-2α plates were washed three times with 150 μl wash buffer (1× HBSS, 0.05% Tween), before adding 25 μl MSD SULFO-TAG IFNα-2α Antibody solution (1:100 diluted in Diluent 3 (Meso Scale Diagnostics #R50AP-2) to each well of the plates.

[0597] Afterwards plates were sealed with microplate seals and kept at room temperature again under continuous shaking at 700 rpm for two hours. Finally, MSD IFNα-2α plates were washed three times with 150 μl wash buffer (1× HBSS, 0.05% Tween). 150 μl 2× Read buffer was added to each well and plates were immediately measured with the MSD Sector S600 Reader using the vendor barcode.

[0598] Data Evaluation and Calculation:

[0599] For data evaluation and calculation, % control calculation of each well was based on the mean of high (DNA stimulated control) and mean of low (unstimulated control) controls by using the following formula:

[counts(sample)−counts(low))/(counts(high)−counts(low))]*100

[0600] The IC50 values were calculated using the standard 4 parameter logistic regression formula. Calculation: [y=(a−d)/(1+(x/c){circumflex over ( )}b)+d], a=low value, d=high value; x=conc M; c=IC50 M; b=slope

7 Indications

[0601] As has been found, the compounds of formula (I) or of formula (I′) are characterized by their range of applications in the therapeutic field. Particular mention should be made of those applications for which the compounds of formula (I) or of formula (I′) according to the invention are preferably used on the basis of their pharmaceutical activity as cGAS inhibitors. While the cGAS pathway is important for host defense against invading pathogens, such as viral infection and invasion by some intracellular bacteria, cellular stress and genetic factors may also cause production of aberrant cellular dsDNA, e.g. by nuclear or mitochondrial leakage, and thereby trigger autoinflammatory responses. Consequently, cGAS inhibitors have a strong therapeutic potential to be used in the treatment of diverse autoinflammatory and autoimmune diseases.

[0602] An et al., Arthritis Rheumatol. 2017 April; 69(4):800-807, disclosed that cGAS expression in peripheral blood mononuclear cells (PBMCs) was significantly higher in patients with the autoimmune disease systemic lupus erythematosus (SLE) than in normal controls. Targeted measurement of cGAMP by tandem mass spectrometry detected cGAMP in 15% of the tested SLE patients, but none of the normal or rheumatoid arthritis controls. Disease activity was higher in SLE patients with cGAMP versus those without cGAMP. Whereas higher cGAS expression may be a consequence of exposure to type I interferon (IFN), detection of cGAMP in SLE patients with increased disease activity indicates potential involvement of the cGAS pathway in disease expression.

[0603] Park et al., Ann Rheum Dis. 2018 October; 77(10):1507-1515, also discloses the involvement of the cGAS pathway in the development of SLE.

[0604] Thim-Uam et al., iScience 2020 Sep. 4; 23(9), 101530 (doi: 10.1016/j.isci.2020.101530), discloses that the STING pathway mediates lupus via the activation of conventional dendritic cell maturation and plasmacytoid dendritic cell differentiation.

[0605] Gao et al., Proc. Natl. Acad. Sci. USA. 2015 Oct 20; 112(42):E5699-705, describes that the activation of cGAS by self-DNA leads to certain autoimmune diseases such as interferonopathies.

[0606] Tonduti et al., Expert Rev. Clin. Immunol. 2020 February; 16(2):189-198 discloses that cGAS inhibitors have particular therapeutic potential in Aicardi-Goutières syndrome which is a lupus-like severe autoinflammatory immune-mediated disorder.

[0607] In Yu et al., Cell 2020 Oct. 29; 183(3):636-649, the link between TDP-43 triggered mitochondrial DNA and the activation of the cGAS/STING pathway in amyotrophic lateral sclerosis (ALS) is described.

[0608] Ryu et al., Arthritis Rheumatol. 2020 November; 72(11):1905-1915, also shows that bioactive plasma mitochondrial DNA is associated with disease progression in specific fibrosing diseases such as systemic sclerosis (SSc) or interstitial lung deseases (ILDs), progressive fibrosing interstitial lung diseases (PF-ILDs), and idiopathic pulmonary fibrosis (IPF).

[0609] In Schuliga et al., Clin. Sci. (Lond). 2020 Apr. 17; 134(7):889-905, it is described that self-DNA perpetuates IPF lung fibroblast senescence in a cGAS-dependent manner.

[0610] Additional scientific hints linking the cause for other fibrosing diseases such as non-alcoholic steatotic hepatitis (NASH) with the cGAS/STING pathway have been described in Yu et al., J. Clin. Invest. 2019 Feb. 1; 129(2):546-555, and in Cho et al., Hepatology. 2018 October; 68(4): 1331-1346.

[0611] Nascimento et al., Sci. Rep. 2019 Oct. 16; 9(1):14848, discloses that self-DNA release and STING-dependent sensing drives inflammation due to cigarette smoke in mice hinting at a link between the cGAS-STING pathway and chronic obstructive pulmonary disease (COPD).

[0612] Ma et al., Sci. Adv. 2020 May 20; 6(21):eaaz6717, discloses that ulcerative colitis and inflammatory bowel disease (IBD) may be restrained by controlling cGAS-mediated inflammation.

[0613] Gratia et al., J. Exp. Med. 2019 May 6; 216(5):1199-1213, shows that Bloom syndrome protein restrains innate immune sensing of micronuclei by cGAS. Consequently cGAS-inhibitors have a therapeutic potential in treating Bloom's syndrome.

[0614] Kerur et al., Nat. Med. 2018 January; 24(1):50-61, describes that cGAS plays a significant role in noncanonical-inflammasome activation in age-related macular degeneration (AMD).

[0615] Further, the cGAS inhibitors of formula (I) or of formula (I′) also have a therapeutic potential in the treatment of cancer (see Hoong et al., Oncotarget. 2020 Jul. 28; 11(30):2930-2955, and Chen et al., Sci. Adv. 2020 Oct. 14; 6(42):eabb8941).

[0616] Additionally, the cGAS inhibitors of formula (I) or of formula (I′) have also a therapeutic potential in the treatment of heart failure (Hu et al., Am. J. Physiol. Heart Circ. Physiol. 2020 Jun. 1; 318(6):H1525-H1537).

[0617] Further scientific hints at a correlation between Parkinsons disease and the cGAS/STING pathway (Sliter et al., Nature. 2018 September; 561(7722):258-262) and between Sjogren's syndrome and the cGAS/STING pathway (Papinska et al., J. Dent. Res. 2018 July; 97(8):893-900) exist.

[0618] Furthermore, cGAS inhibitors of formula (I) or of formula (I′) have also a therapeutic potential in the treatment of COVID-19/SARS-CoV-2 infections as shown in Di Domizio et al., Nature. 2022 Jan. 19. doi: 10.1038/s41586-022-04421-w: “The cGAS-STING pathway drives type I IFN immunopathology in COVID-19”, and in

[0619] Neufeldt et al., Commun Biol. 2022 Jan. 12; 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”.

[0620] Additionally, cGAS inhibitors of formula (I) or of formula (I′) have a therapeutic potential in the treatment of renal inflammation and renal fibrosis as shown in Chung et al., Cell Metab. 2019 30:784-799: “Mitochondrial Damage and Activation of the STING Pathway Lead to Renal Inflammation and Fibrosis”, and in Maekawa et al., Cell Rep. 2019 29:1261-1273: “Mitochondrial Damage Causes Inflammation via cGAS-STING Signaling in Acute Kidney Injury”.

[0621] Furthermore, cGAS inhibitors of formula (I) or of formula (I′) have a therapeutic potential in the treatment of cancer as shown in Bakhoum et el., Nature. 2018 Jan. 25; 553(7689):467-472: “Chromosomal instability drives metastasis through a cytosolic DNA response”, and in Liu et al., Nature. 2018 November; 563(7729):131-136: “Nuclear cGAS suppresses DNA repair and promotes tumorigenesis”.

[0622] Additionally, cGAS inhibitors of formula (I) or of formula (I′) have a therapeutic potential in the treatment of dysmetabolism, because STING.sup.gt animals show reduced macrophage infiltration in adipose tissue upon subchronic high caloric intake (HFD) and STING.sup.gt and IRF3-deficiency leads to a decrease in blood glucose and insulin and reduced body weight (Mao et al, Arterioscler Thromb Vasc Biol, 2017; 37 (5): 920-929).

[0623] Furthermore, cGAS inhibitors of formula (I) or of formula (I′) have a therapeutic potential in the treatment of vascular diseases and leads to vascular repair/regeneration, because the release of mitochondrial DNA into the cytosol of endothelial cells results in cGAS/STING pathway activation and suppression of endothelial proliferation. Futher, knockout of the cGAS gene restores endothelial repair/regeneration in a mouse model of inflammatory lung injury (Huang et al, Immunity, 2020, March 2017; 52 (3): 475-486.e5. doi: 10.1016/j.immuni.2020,02.002).

[0624] Additionally, cGAS inhibitors of formula (I) or of formula (I′) have a therapeutic potential in the treatment of age-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).

[0625] Consequently the compounds of formula (I) or of formula (I′) as cGAS inhibitors can be used in the therapy of autoinflammatory and autoimmune diseases such as systemic lupus erythematosus (SLE), interferonopathies, Aicardi-Goutieres syndrome, age-related macular degeneration (AMD), amyotrophic lateral sclerosis (ALS), inflammatory bowel disease (IBD), chronic obstructive pulmonary disease (COPD), Bloom's syndrome, Sjogren's syndrome and Parkinson disease.

[0626] Additionally the compounds of formula (I) or of formula (I′) as cGAS inhibitors can be used in the therapy of fibrosing disease such as systemic sclerosis (SSc), interferonopathies, non-alcoholic steatotic hepatitis (NASH), interstitial lung disease (ILD), preferably progressive fibrosing interstitial lung disease (PF-ILD), in particular idiopathic pulmonary fibrosis (IPF).

[0627] Further, the compounds of formula (I) or of formula (I′) as cGAS inhibitors can be used in the therapy of age-related macular degeneration (AMD), heart failure, COVID-19/SARS-CoV-2 infection, renal inflammation, renal fibrosis, dysmetabolism, vascular diseases, cardiovascular diseases and cancer.

8 Combinations

[0628] The compounds of formula (I) or of formula (I′) may be administered to the patient alone or in combination with one or more other pharmacologically active agents.

[0629] In a preferred embodiment of the invention the compounds of formula (I) or of formula (I′) may be combined with one or more pharmacologically active agents selected from the group of anti-inflammatory agents, anti-fibrotic agents, anti-allergic agents/anti-histamines, bronchodilators, beta 2 agonists/betamimetics, adrenergic agonists, anticholinergic agents, methotrexate, mycophenolate mofetil, leukotriene modulators, JAK inhibitiors, anti-interleukin antibodies, non-specific immunotherapeutics such as interferones or other cytokines/chemokines, cytokine/chemokine receptor modulators (i.e. cytokine receptor agonists or antagonists), Toll-like receptor agonists (=TLR agonists), immune checkpoint regulators, anti-TNF antibodies (Humira™), and anti-BAFF agents (Belimumab and Etanercept).

[0630] Anti-fibrotic agents are preferably selected from Pirfenidone and tyrosine kinase inhibitors such as Nintedanib, wherein Nintedanib is preferred in particular.

[0631] Preferred examples of anti-inflammatory agents are NSAIDs and corticosteroids.

[0632] NSAIDs are preferably selected from ibuprofen, naproxen, diclofenac, meloxicam, celecoxib, acetylsalicylic acid (Aspirin™), indomethacin, mefenamic acid and etoricoxib.

[0633] Corticosteroids are preferably selected from Flunisolide, Beclomethasone, Triamcinolone, Budesonide, Fluticasone, Mometasone, Ciclesonide, Rofleponide and Dexametasone.

[0634] Antiallergic agents/anti-histamines are preferably selected from Epinastine, Cetirizine, Azelastine, Fexofenadine, Levocabastine, Loratadine, Ebastine, Desloratidine and Mizolastine.

[0635] Beta 2 agonists/betamimetics may be either long acting beta 2 Agonists (LABAs) or short acting beta agonists (SABAs). Particularly preferred beta 2 agonists/betamimetics are selected from Bambuterol, Bitolterol, Carbuterol, Clenbuterol, Fenoterol, Formoterol, Hexoprenalin, Ibuterol, Pirbuterol, Procaterol, Reproterol, Salmeterol, Sulfonterol, Terbutalin, Tolubuterol, Olodaterol, and Salbutamol, in particular Olodaterol.

[0636] Anticholinergic agents are preferably selected from ipratropium salts, tiotropium salts, glycopyrronium salts, and theophylline, wherein tiotropium bromide is preferred in particular.

[0637] Leukotriene modulators are preferably selected from Montelukast, Pranlukast, Zafirlukast, Ibudilast and Zileuton.

[0638] JAK inhibitors are preferably selected from Baricitinib, Cerdulatinib, Fedratinib, Filgotinib, Gandotinib, Lestaurtinib, Momelotinib, Pacritinib, Peficitinib, Ruxolitinib, Tofacitinib, and Upadacitinib.

[0639] Anti-interleukin antibodies are 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™, anti-IL-12 antibodies, anti-IL-15 antibodies.

9 Formulations

[0640] The compounds of the invention may be administered by any suitable route of administration, including both systemic administration and topical administration. Systemic administration includes oral administration, parenteral administration, transdermal administration, rectal administration, and administration by inhalation. Parenteral administration refers to routes of administration other than enteral, transdermal, or by inhalation, and is typically by injection or infusion. Parenteral administration includes intravenous, intramuscular, intrasternal, and subcutaneous injection or infusion. Inhalation refers to administration into the patient's lungs whether inhaled through the mouth or through the nasal passages. Topical administration includes application to the skin. The compounds of the invention may be administered via eye drops to treat Sjogren's syndrome.

[0641] Suitable forms for administration are for example tablets, capsules, solutions, syrups, emulsions or inhalable powders or aerosols. The content of the pharmaceutically effective compound(s) in each case should be in the range from 0.1 to 90 wt. %, preferably 0.5 to 50 wt. % of the total composition, i.e. in amounts which are sufficient to achieve the dosage range specified hereinafter.

[0642] The preparations may be administered orally in the form of a tablet, as a powder, as a powder in a capsule (e.g. a hard gelatin capsule), as a solution or suspension. When administered by inhalation the active substance combination may be given as a powder, as an aqueous or aqueous-ethanolic solution or using a propellant gas formulation.

[0643] Preferably, therefore, pharmaceutical formulations are characterized by the content of one or more compounds of formula (I) or of formula (I′) according to the preferred embodiments above.

[0644] It is particularly preferable if the compounds of formula (I) or of formula (I′) are administered orally, and it is also particularly preferable if they are administered once or twice a day. Suitable tablets may be obtained, for example, by mixing the active substance(s) with known excipients, for example inert diluents such as calcium carbonate, calcium phosphate or lactose, disintegrants such as corn starch or alginic acid, binders such as starch or gelatine, lubricants such as magnesium stearate or talc and/or agents for delaying release, such as carboxymethyl cellulose, cellulose acetate phthalate, or polyvinyl acetate. The tablets may also comprise several layers.

[0645] Coated tablets may be prepared accordingly by coating cores produced analogously to the tablets with substances normally used for tablet coatings, for example kollidone or shellac, gum arabic, talc, titanium dioxide or sugar. To achieve delayed release or prevent incompatibilities the core may also consist of a number of layers. Similarly, the tablet coating may consist of a number of layers to achieve delayed release, possibly using the excipients mentioned above for the tablets.

[0646] Syrups containing the active substances or combinations thereof according to the invention may additionally contain a sweetener such as saccharine, cyclamate, glycerol or sugar and a flavor enhancer, e.g. a flavoring such as vanillin or orange extract. They may also contain suspension adjuvants or thickeners such as sodium carboxymethyl cellulose, wetting agents such as, for example, condensation products of fatty alcohols with ethylene oxide, or preservatives such as p-hydroxybenzoates.

[0647] Capsules containing one or more active substances or combinations of active substances may for example be prepared by mixing the active substances with inert carriers such as lactose or sorbitol and packing them into gelatin capsules. Suitable suppositories may be made for example by mixing with carriers provided for this purpose, such as neutral fats or polyethylene glycol or the derivatives thereof.

[0648] Excipients which may be used include, for example, water, pharmaceutically acceptable organic solvents such as paraffins (e.g. petroleum fractions), vegetable oils (e.g. groundnut or sesame oil), mono- or polyfunctional alcohols (e.g. ethanol or glycerol), carriers such as e.g. natural mineral powders (e.g. kaolins, clays, talc, chalk), synthetic mineral powders (e.g. highly dispersed silicic acid and silicates), sugars (e.g. cane sugar, lactose and glucose), emulsifiers (e.g. lignin, spent sulphite liquors, methylcellulose, starch and polyvinylpyrrolidone) and lubricants (e.g. magnesium stearate, talc, stearic acid and sodium lauryl sulphate).

[0649] For oral administration the tablets may, of course, contain, apart from the abovementioned carriers, additives such as sodium citrate, calcium carbonate and dicalcium phosphate together with various additives such as starch, preferably potato starch, gelatin and the like. Moreover, lubricants such as magnesium stearate, sodium lauryl sulphate and talc may be used at the same time for the tableting process. In the case of aqueous suspensions, the active substances may be combined with various flavor enhancers or colorings in addition to the excipients mentioned above.