Tricyclic Janus kinase (JAK) inhibitors and their use in treating autoimmune diseases

Abstract

The present invention relates to compounds of formula (1) or (2), wherein R 1 and R 3 is a purine or a purine analog, and R 2 and R 4 is a small functional group. The present invention also relates to intermediates for the synthesis of compounds of formula 1 or 2. Compounds of formula 1 or 2 are Janus kinase inhibitors and are therefore useful for the treatment of diseases, in particular autoimmune diseases, cancer, Alzheimer's disease or for the prevention of rejection of allografts or xenografts.

Description

Background Art

[0001] The Janus kinase (JAK) family consists of four members, JAK1, JAK2, JAK3, and TYK2, and they are involved in cell growth, survival, development, and differentiation through cytokine-activated cell signaling. Therefore, Janus kinase inhibitors have a wide range of applications ranging from autoimmune diseases to cancer to Alzheimer's disease. Known JAK inhibitors include Tofacitinib, Delgocitinib, PF-06651600, Baricitinib, Upadacitinib, and Filgotinib.

[0002] Tofacitinib is an FDA-approved pan-JAK kinase inhibitor (mainly JAK1 and 3) for the treatment of rheumatoid arthritis and ulcerative colitis. It is undergoing clinical trials for psoriatic arthritis (Phase 3), psoriasis (Phase 3), Crohn's disease (Phase 2), and renal transplantation (Phase 2).

[0003] Digatinib (JTE-052) is another pan-JAK kinase inhibitor, a Phase 2 compound for the treatment of atopic dermatitis and chronic hand eczema.

[0004] PF-06651600 is a selective JAK3 inhibitor currently in Phase 3 for alopecia areata and Phase 2 for ulcerative colitis, Crohn's disease, rheumatoid arthritis, and non-segmental vitiligo.

[0005] Baricitinib is an approved JAK1 and JAK2 inhibitor for the treatment of rheumatoid arthritis.

[0006] Upadacitinib is a selective JAK1 inhibitor in clinical trials for rheumatoid arthritis (Phase 3), Crohn's disease (Morbus Crohn) (Phase 2), ulcerative colitis (Phase 2), atopic dermatitis (Phase 2), psoriatic arthritis (Phase 3), and axial spondyloarthritis (Phase 2).

[0007] Filgotinib is also a selective JAK1 inhibitor. It is in stage 3 for rheumatoid arthritis, Crohn's disease, and ulcerative colitis. It also targets small intestinal CD, fistulizing CD, Sjögren's syndrome, Phase 2 studies are underway in the treatment of leukopenia, ankylosing spondylitis, cutaneous lupus, lupus nephropathy, and uveitis.

[0008] Based on the above prior art, the object of the present invention is to provide means and methods for providing additional JAK inhibitors having a novel tricyclic skeleton ( Figure 1 ). This object is achieved by the subject matter of the independent claims of this description.

[0009] describe

[0010] Terms and Definitions

[0011] The term Janus kinase refers to a non-receptor tyrosine kinase that transduces cytokine-mediated signals through the JAK-STAT pathway. Janus kinases may be abbreviated as "JAKs". Four JAK family members are known: Janus kinase 1 (JAK1), Janus kinase 2 (JAK2), Jansu kinase 3 (JAK3), and tyrosine kinase 2 (TYK2). Human Janus kinase 1 is encoded by the gene JAK1, human Janus kinase 2 is encoded by the gene JAK2, human Janus kinase 3 is encoded by the gene JAK3, and human tyrosine kinase 2 is encoded by the gene TYK2.

[0012] The term allograft relates to an organ, tissue or cell transplanted from a donor to a recipient of the same species. Such an organ is also called an allogeneic organ or allograft, for example an allogeneic kidney.

[0013] The term xenograft refers to an organ, tissue or cells transplanted from a donor to a recipient of a different species.

[0014] As used herein, the term treating or treatment of any disease or illness (e.g., cancer) refers in one embodiment to alleviating the disease or illness (e.g., slowing or preventing or reducing the development of the disease or at least one of its clinical symptoms). In another embodiment, "treating" or "treatment" refers to alleviating or reducing at least one physical parameter, including those that the patient may not be able to discern. In yet another embodiment, "treating" or "treatment" refers to regulating the disease or illness physically (e.g., stabilization of discernible symptoms), physiologically (e.g., stabilization of physical parameters), or both. Unless specifically described herein, methods for assessing the treatment and / or prevention of a disease are generally known in the art. DETAILED DESCRIPTION

[0015] The first aspect of the present invention relates to a compound of Formula 1 or Formula 2 or a salt thereof,

[0016] in,

[0017] R1 and R 3 Selected from

[0018]

[0019] R 2 and R 4 Selected from

[0020]

[0021] z is 1, 2, 3, or 4,

[0022] x and p are 1 or 2, and

[0023] y and q are 1 or 2.

[0024] The compounds of formula 1 or 2 are potent Janus kinase inhibitors characterized by a tricyclic or polycyclic skeleton. The skeleton includes 1 or R 3 Purine or purine analogs at R 2 or R 4 Purine or purine analogs and small functional groups can interact with Janus kinases to inhibit Janus kinases, where it is assumed that purine or purine analogs bind to the hinge region of Janus kinases. In particular, compared with known bicyclic JAK inhibitors such as digaotinib, the JAK inhibitors according to the present invention, such as KMC420, show enhanced activity and increased selectivity in different Janus kinases. Enhanced activity can be achieved in particular by compounds of formula 1, while increased selectivity can be achieved by compounds of formula 2.

[0025] In particular, tricyclic JAK inhibitors (Formula 1) are more soluble in water compared to polycyclic JAK inhibitors (Formula 2).

[0026] In certain embodiments, the compound is a compound of Formula 1.

[0027] In certain embodiments, z is 1 or 2.

[0028] In certain embodiments, z, x, y, p, and q are 1.

[0029] In certain embodiments, R 1 and R 3 Selected from

[0030] in particular More specifically

[0031] In certain embodiments, R 2 and R 4 Selected from

[0032] In certain embodiments, R 2 and R 4 Selected from

[0033] The compounds according to the invention can be synthesized as shown in Schemes 2 to 4 (see the "Examples" section). The second, third and fourth aspects of the invention relate to intermediates for the synthesis of compounds according to the first aspect of the invention.

[0034] The second aspect of the present invention relates to an intermediate of formula 3 or 4 or a salt thereof,

[0035] in,

[0036] R 5 , R 6 , R 7 and R 8 Selected from

[0037] -H,

[0038] - a protecting group cleavable under acidic conditions,

[0039] - a protecting group cleavable under alkaline conditions,

[0040] - a protecting group cleavable under reducing conditions,

[0041] - a hydrogenolytically cleavable protecting group,

[0042] - a protecting group cleavable using a metal catalyst,

[0043] z, x, y, p and q are defined as described above.

[0044] The compounds according to the invention comprise at position R 1 (Formula 1) or R 3 (Formula 2) at position R 2 (Formula 1) or R 4 (Formula 2) is a small functional group at the position. In order to allow the synthesis of compounds according to the present invention, the N atom in the intermediate of Formula 3 or Formula 4 can be protected. Suitable protecting groups are cleavable under acidic conditions, such as Boc or Ts, cleavable under alkaline conditions, such as Fmoc, Ac or Ac-CF3, cleavable under reducing conditions, such as Ts, cleavable by hydrogenolysis, such as Bn or Cbz, and cleavable using a metal catalyst, such as Alloc or Troc.

[0045] In certain embodiments, R 5 , R 6 , R7 and R 8 Selected from

[0046] -H,

[0047]

[0048] z, x, y, p and q are defined as described above.

[0049] In particular, orthogonal protection facilitates the synthesis of compounds according to Formula 1 and 2. Orthogonally protected intermediates include two protecting groups that can be cleaved by reaction conditions that are different from each other.

[0050] In certain embodiments,

[0051] -R 5 and R 6 Both and R 7 and R 8 Both are –H, or

[0052] -R 5 and R 7 One or R 6 and R 8 one of them is –H and the other R is a protecting group, or

[0053] -R 5 and R 7 Both and R 6 and R 8 Both are protecting groups, where R 5 and R 7 The cleavage of the protecting group at R 6 and R 8 Cleavage of protecting groups.

[0054] In certain embodiments,

[0055] -R 5 and R 7 One or R 6 and R 8 one of them is –H and the other R is a protecting group, or

[0056] -R 5 and R 7 Both and R 6 and R 8 Both are protecting groups, where R 5 and R 7 The cleavage of the protecting group at R 6 and R 8 Cleavage of protecting groups.

[0057] In certain embodiments, R 5 and R 7 Both and R 6 and R 8 Both are protecting groups, where R 5 and R 7 The cleavage of the protecting group at R 6 and R 8 Cleavage of protecting groups.

[0058] The third aspect of the present invention relates to an intermediate according to formula 3 or 4 or a salt thereof, wherein

[0059] R 5 and R 7 One or R 6 and R 8 One of the selected and

[0060] Another R is selected from

[0061] -H,

[0062] - protective groups cleavable under acidic conditions, in particular Boc, Ts,

[0063] - protective groups cleavable under alkaline conditions, in particular Fmoc, Ac, Ac-CF3,

[0064] - a protecting group cleavable under reducing conditions, in particular Ts,

[0065] - hydrogenolytically cleavable protecting groups, in particular Bn, Cbz,

[0066] - protective groups cleavable using metal catalysts, in particular Alloc, Troc,

[0067] z, x, y, p and q are defined as described above.

[0068] As mentioned above, the orthogonal protection strategy allows the stepwise synthesis of 1 or R 3 Purine or purine analogs at R 2 or R 4 If a purine or purine analogue is attached first, an intermediate according to the third aspect of the invention is obtained.

[0069] In certain embodiments, R 5 and R 7 One or R 6 and R 8 One of the selected

[0070] in particular More specifically and

[0071] Another R is selected from

[0072] -H,

[0073] - protective groups cleavable under acidic conditions, in particular Boc, Ts,

[0074] - protective groups cleavable under alkaline conditions, in particular Fmoc, Ac, Ac-CF3,

[0075] - a protecting group cleavable under reducing conditions, in particular Ts,

[0076] - hydrogenolytically cleavable protecting groups, in particular Bn, Cbz,

[0077] - protective groups cleavable using metal catalysts, in particular Alloc, Troc,

[0078] z, x, y, p and q are defined as described above.

[0079] The fourth aspect of the present invention relates to an intermediate of formula 3 or 4 or a salt thereof, wherein R 5 and R 7 One or R 6 and R 8 One of the selected and

[0080] Another R is selected from

[0081] -H,

[0082] - protective groups cleavable under acidic conditions, in particular Boc, Ts,

[0083] - protective groups cleavable under alkaline conditions, in particular Fmoc, Ac, Ac-CF3,

[0084] - a protecting group cleavable under reducing conditions, in particular Ts,

[0085] - hydrogenolytically cleavable protecting groups, in particular Bn, Cbz,

[0086] - protective groups cleavable using metal catalysts, in particular Alloc, Troc,

[0087] z, x, y, p and q are defined as described above.

[0088] As mentioned above, the orthogonal protection strategy allows the stepwise synthesis of 1 or R 3Purine or purine analogs at R 2 or R 4 If the small functional group is attached first, an intermediate according to the fourth aspect of the invention is obtained.

[0089] In certain embodiments, R 5 and R 7 One or R 6 and R 8 One of the selected

[0090] Especially selected from and

[0091] Another R is selected from

[0092] -H,

[0093] - protective groups cleavable under acidic conditions, in particular Boc, Ts,

[0094] - protective groups cleavable under alkaline conditions, in particular Fmoc, Ac, Ac-CF3,

[0095] - a protecting group cleavable under reducing conditions, in particular Ts,

[0096] - hydrogenolytically cleavable protecting groups, in particular Bn, Cbz,

[0097] - protective groups cleavable using metal catalysts, in particular Alloc, Troc,

[0098] z, x, y, p and q are defined as described above.

[0099] A fifth aspect of the present invention relates to an intermediate of formula 5 or 6,

[0100] in,

[0101] R 9 and R 10 Selected from

[0102] -H,

[0103] - protective groups cleavable under acidic conditions, in particular Boc, Ts,

[0104] - protective groups cleavable under alkaline conditions, in particular Fmoc, Ac, Ac-CF3,

[0105] - a protecting group cleavable under reducing conditions, in particular Ts,

[0106] - hydrogenolytically cleavable protecting groups, in particular Bn, Cbz,

[0107] - protective groups cleavable using metal catalysts, in particular Alloc, Troc,

[0108] z, y and q are defined as described above.

[0109] In certain embodiments of the first, second, third, fourth or fifth aspect of the invention, the compound or the intermediate is an enantiomer.

[0110] The compounds and intermediates disclosed herein include one or more chiral C atoms. Therefore, the compounds and intermediates may be obtained as racemates. The activity and selectivity for Janus kinases may be increased by using only one enantiomer.

[0111] According to a sixth aspect of the present invention, there is provided a compound according to the first aspect of the present invention, for use in treating a disease.

[0112] As shown in Table 1 (see the "Examples" section), the compounds according to the invention inhibit all four members of the JAK family, in particular JAK 1. Various diseases are associated with the activity of the kinases of the JAK family.

[0113] JAK is related to cytokine receptors. After the ligand binds to the cytokine receptor, the kinase of the JAK family is activated by phosphorylation. Phosphorylated JAK phosphorylates the STAT protein downstream of the JAK-STAT signal transduction pathway. Phosphorylated STAT protein acts as a transcription factor in the nucleus. The JAK-STAT signal transduction pathway is related to the expression of cytokines, interferons or interleukins. Therefore, diseases (such as autoimmune diseases) particularly involving cytokines, interferons or interleukins can be treated by inhibiting JAK.

[0114] In certain embodiments of the sixth aspect of the invention, enantiomers or racemates, in particular enantiomers, are used.

[0115] According to a seventh aspect of the present invention, there is provided a compound according to the first aspect of the present invention for use in treating autoimmune diseases, cancer, Alzheimer's disease, asthma or for preventing rejection of allogeneic transplants or xenografts.

[0116] Particularly in the treatment of cancer, the compounds of the present invention may also be used to support anticancer therapy by altering the tumor microenvironment, thereby allowing antitumor drugs (eg antibodies or antibody-drug conjugates) to better access malignant cells.

[0117] In certain embodiments, the compounds according to the first aspect are used to treat rheumatoid arthritis, ulcerative colitis, psoriatic arthritis, psoriasis, Crohn's disease, atopic dermatitis, chronic hand eczema, non-segmental vitiligo, axial spondyloarthritis, small intestinal CD, fistulizing CD, Sjögren's syndrome, ankylosing spondylitis, cutaneous lupus, lupus nephropathy, uveitis, myelofibrosis and Alzheimer's disease, asthma, or for preventing rejection of an allogeneic transplant, particularly an allogeneic kidney.

[0118] In certain embodiments, the compounds according to the first aspect are used to treat rheumatoid arthritis, ulcerative colitis, psoriatic arthritis, psoriasis, Crohn's disease, atopic dermatitis, chronic hand eczema, non-segmental vitiligo, axial spondyloarthritis, small bowel CD, fistulizing CD, Sjögren's syndrome, ankylosing spondylitis, cutaneous lupus, lupus nephropathy, uveitis, asthma and Alzheimer's disease.

[0119] In certain embodiments, the compounds according to the first aspect are used to treat rheumatoid arthritis, ulcerative colitis, psoriatic arthritis, psoriasis, Crohn's disease, atopic dermatitis, chronic hand eczema, non-segmental vitiligo, axial spondyloarthritis, small intestinal CD, fistulizing CD, Sjögren's syndrome, ankylosing spondylitis, cutaneous lupus, lupus nephropathy, uveitis and Alzheimer's disease.

[0120] In certain embodiments, the compounds according to the first aspect are used to treat rheumatoid arthritis, ulcerative colitis, psoriatic arthritis, psoriasis, Crohn's disease, atopic dermatitis, chronic hand eczema, non-segmental vitiligo, axial spondyloarthritis, small intestinal CD, fistulizing CD, Sjögren's syndrome, ankylosing spondylitis, cutaneous lupus, lupus nephropathy and uveitis.

[0121] In certain embodiments of the seventh aspect of the invention, enantiomers or racemates, in particular enantiomers, are used.

[0122] In another embodiment, the present invention is directed to a pharmaceutical composition comprising at least one compound of the present invention or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier, diluent or excipient.

[0123] As used herein, the term pharmaceutical composition refers to a compound of the invention or a pharmaceutically acceptable salt thereof, together with at least one pharmaceutically acceptable carrier. In certain embodiments, the pharmaceutical composition according to the invention is provided in a form suitable for topical, parenteral or injectable administration.

[0124] As used herein, the term pharmaceutically acceptable carrier includes any solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial agents, antifungal agents), isotonic agents, absorption delaying agents, salts, preservatives, drugs, drug stabilizers, binders, excipients, disintegrants, lubricants, sweeteners, flavoring agents, dyes, etc. and combinations thereof, as would be known to those skilled in the art (see, e.g., Remington: the Science and Practice of Pharmacy, ISBN 0857110624).

[0125] The skilled artisan will appreciate that any specifically mentioned drug may be present as a pharmaceutically acceptable salt of that drug.A pharmaceutically acceptable salt comprises an ionized drug and an oppositely charged counter ion. Non-limiting examples of pharmaceutically acceptable anionic salt forms include acetate, benzoate, benzenesulfonate, bitatrate, bromide, carbonate, chloride, citrate, edetate, edisylate, embonate, estolate, fumarate, gluceptate, gluconate, hydrobromide, hydrochloride, iodide, lactate, lactobionate, malate, maleate, mandelate, methanesulfonate, methyl bromide, methylsulfate, mucate, naphthenate, nitrate, pamoate, phosphate, diphosphate, salicylate, disalicylate, stearate, succinate, sulfate, tartrate, toluenesulfonate, triethiodide, and valerate. Non-limiting examples of pharmaceutically acceptable cationic salt forms include aluminum, benzathine, calcium, ethylenediamine, lysine, magnesium, meglumine, potassium, procaine, sodium, tromethamine, and zinc.

[0126] A further aspect of the invention relates to a process for preparing a compound of formula 1 or 2.

[0127] The synthesis of the compound of formula 1 or 2 starts from a diketone. The diketone is an extract for preparing a bicyclic or tricyclic intermediate of formula 5 or 6 as described in the fifth aspect of the present invention.

[0128] In certain embodiments, the method for preparing the intermediate of formula 5 or 6 comprises the following steps:

[0129] - providing a diketone of formula 6 or 7,

[0130] where z is 1 or 2, especially 1,

[0131] - using a compound of formula 8 and a diketone of formula 6 or 7 for condensation reaction, followed by reduction, to obtain an intermediate of formula 9 or 10,

[0132] in,

[0133] B is a protecting group, especially Boc,

[0134] y is 1 or 2, especially 1,

[0135] q is 1 or 2, especially 1,

[0136] z is 1 or 2, especially 1,

[0137] - allylation, in particular by using allyl bromide and potassium carbonate as base, to obtain an intermediate of formula 11 or 12,

[0138] in,

[0139] B is a protecting group, especially Boc,

[0140] y is 1 or 2, especially 1,

[0141] q is 1 or 2, especially 1,

[0142] z is 1 or 2, especially 1,

[0143] - Removal of the protecting groups and intramolecular reductive amination, in particular by using NaBH3CN, to obtain intermediates of formula 5 or 6.

[0144] For the intermediates of formula 5 or 6, see the fifth aspect of the invention.

[0145] Starting from the intermediate of formula 5 or 6, the intermediate of formula 3 or 4 as described in the second aspect of the present invention can be obtained.

[0146] In certain embodiments, the method for preparing the intermediate of Formula 3 or 4 comprises the following steps:

[0147] - providing an intermediate of formula 5 or 6,

[0148] - Ozonolysis followed by two reductive aminations, in particular using benzylamine and NaBH3CN, to obtain intermediates of formula 3 or 4.

[0149] In order to allow purine or purine analogs (R 1 and R 3 ) and for R 2 and R 4 For the coupling of small functional groups, the intermediate of formula 3 or 4 may include orthogonal protecting groups.

[0150] For the intermediates of formula 3 or 4, see the second aspect of the invention.

[0151] In particular, ozonolysis is performed followed by two reductive aminations to obtain intermediates of formula 3 or 4, wherein x and p are 1.

[0152] Intermediates of formula 3 or 4, wherein x and p are 2, can be obtained by hydroazidation or ozonolysis of olefins followed by selective aldehyde reduction, tosylation, and displacement with sodium cyanide as described in Li, et al. J. Am. Chem. Soc. 2019141239415-9421.

[0153] Intermediates of formula 3 or 4 as described in the third and fourth aspects of the invention may be obtained by standard methods.

[0154] In particular, the intermediate of formula 3 or 4 as described in the third aspect of the present invention can be obtained by selective deprotection followed by nucleophilic substitution with chloropurine or its analogue.

[0155] Finally, the compound of formula 1 or 2 according to the first aspect of the present invention is deprotected and then coupled by forming an amide bond as for R 2 and R 4 It is obtained by the small functional groups described above. BRIEF DESCRIPTION OF THE DRAWINGS

[0156] Figure 1 Shown are the known JAK inhibitors Tofacitinib, JTE-052 (Diagotinib) or PF-066051600 compared to the JAK inhibitors according to the invention (KMC420 and KMC423).

[0157] Example

[0158] Synthesis of diamine 1

[0159] Diamine 1 was synthesized as shown in Scheme 1. Diallyl-1,3-cyclopentanedione 3 was obtained by palladium-catalyzed allylation of 1,3-cyclopentanedione 2 (Schwartz, CE; Curran, DP, AJ Am. Chem. Soc. 1990, 112(25), 9272–9284). Simultaneous ring closure was achieved by a one-pot tandem ozonolysis and reductive amination with benzylamine and NaBH(OAc)3 (Kyasa, S.; Fisher, T.; Dussault, P., Synthesis 2011, 2011(21), 3475–3481) to afford benzyl-protected diamine 4. Diamine 1 was then deprotected with palladium on carbon and hydrogen. Its structure was confirmed by crystallization of the HCl salt.

[0160]

[0161] Scheme 1: Synthesis of diamine 1.

[0162] Synthesis of orthogonally protected diamine 9

[0163] The synthesis of tricyclic diamine 9 with two different protecting groups is shown in Scheme 2. Proline-catalyzed Knoevenagel condensation of 1,3-cyclopentanedione 2 with Boc-protected aminoacetaldehyde followed by in situ reduction with Hantzsch ester afforded monoalkylated diketone 5 (Ramachary, DB; Kishor, M., Org. Biomol. Chem. 2008, 6(22), 4176), which was then subjected to palladium-catalyzed allylation to afford an equilibrium mixture of C- and O-allylated products 6 and 7. Boc removal followed by intramolecular reductive amination with NaBH3CN and Boc reprotection afforded bicyclic ketone 8. Finally, ozonolysis followed by two reductive aminations with benzylamine and NaBH3CN afforded 9, a useful derivative of 1 bearing two orthogonally protected secondary amines.

[0164]

[0165] Scheme 2: Synthesis of orthogonally protected diamine 9.

[0166] Synthesis of JAK inhibitors KMC420 and KMC423 (inhibitors of Formula 1)

[0167] KMC420 and KMC423 were obtained from intermediate 9 (see also Figure 1 ), as shown in Scheme 3. Selective deprotection of the benzyl protecting group followed by nucleophilic substitution with 6-chloro-7-deazapurine afforded compound 10. Boc-deprotection followed by amide bond formation with cyanoacetic acid or acrylic acid, respectively, afforded two JAK inhibitors, KMC420 and KMC423.

[0168]

[0169] Scheme 3: Synthesis of new JAK inhibitors KMC420 and KMC423.

[0170] Synthesis of JAK inhibitors according to Formula 2

[0171] The JAK inhibitor of Formula 2 is synthesized according to the reaction scheme shown in Scheme 4.

[0172] Similar to the synthesis of 9, tetracyclic orthogonally protected diamine 15 can be obtained according to Scheme 4. Proline-catalyzed Knoevenagel condensation of 1,3-indadione 11 with Boc-protected aminoacetaldehyde followed by in situ reduction with Hantzsch ester afforded the monoalkylated diketone 12 (Ramachary, DB; Kishor, M., Org. Biomol. Chem. 2008, 6(22), 4176). Allylation with allyl bromide and potassium carbonate as base afforded product 13. Boc removal followed by intramolecular reductive amination with NaBH3CN and Boc reprotection afforded tricyclic ketone 14. Finally, ozonolysis followed by two reductive aminations with benzylamine and NaBH3CN afforded 15, a useful derivative of 16 with two orthogonally protected secondary amines.

[0173]

[0174] Scheme 4: Synthesis of phenylenediamine analog 15 and its kinase inhibitor.

[0175] JAK inhibition

[0176] Activity assays showed that KMC420 and KMC423 were potent kinase inhibitors (Table 1).

[0177] Table 1: IC 50 Kinase inhibitory activity is given in nM.

[0178]

[0179] In the presence of ATP (K m Enzyme assays were performed with 5% paraformaldehyde (100 nM) and Ulight-CAGAGAIETDKEYYTVKD (100 nM) as described by Zhou, YJ. et al. (1997), Proc. Natl. Acad. Sci. USA, 94: 13850-13855 for JAK1, Brizzi, MF et al. (1996), J. Biol. Chem., 271: 3562-3567 for JAK2, Yamaoka, K. et al. (2004), Gen. Biol., 5: 253 for JAK3 and Ide, H. et al. (2008), Biochem. Biophys. Res. Commun., 369: 292-296 for TYK2.

[0180] Detailed description of the synthesis of the compounds according to the invention

[0181] Tert-butyl (2-(2-hydroxy-5-oxyylidenecyclopent-1-en-1-yl)ethyl)carbamate (5)

[0182]

[0183] To a suspension of 1,3-cyclopentanedione (3.32 g, 33.8 mmol, 1.0 eq), Hantzsch ester (11.1 g, 43.8 mmol, 1.3 eq) and crude N-Boc-2-aminoacetaldehyde (6.98 g, 43.8 mmol, 1.3 eq) in non-anhydrous DCM (110 ml), proline (1.01 g, 8.8 mmol, 0.3 eq) was added and the mixture was stirred at 22° C. for 25 h. After completion of the reaction, the solvent was evaporated under reduced pressure and purified using flash column chromatography (SiO 2: Hexane / EtOAc 2:8 to pure EtOAc) to give the desired title product 5 (7.15 g, 29.6 mmol, 88%) as a white powder.

[0184] R f =0.35 (9:1 DCM / MeOH);

[0185] mp: 170–171°C;

[0186] 1 H-NMR (400MHz, CDCl3): δ = 3.11 (t, J = 7.1Hz, 2H), 2.52 (s, 4H), 2.42 (t, J = 7.1Hz, 2H), 1.45 (s, 9H);

[0187] 13 C-NMR (100MHz, CDCl3): δ = 157.9, 115.3, 80.9, 40.0, 30.4, 28.4, 21.5;

[0188] HRMS(ESI):C 12 H 20 O4N + [M+H] + The calculated m / z value is 242.1387, the found value is 242.1386.

[0189] tert-Butyl 3a-allyl-4-oxyylidenehexahydrocyclopenta[b]pyrrole-1(2H)-carboxylate (8)

[0190]

[0191] To a suspension of compound 5 (4.82 g, 20.0 mmol, 1.0 eq.) and allyl acetate (2.2 ml, 20.4 mmol, 1.0 eq.) in anhydrous THF (40 ml), Pd(PPh3)4 (0.23 g, 0.2 mmol, 0.01 eq.) was added under argon atmosphere and stirred at 22 ° C for 4.5 hours. The solvent was evaporated and purified by column chromatography (SiO2: 8: 2 hexane / EtOAc) to give the C- and O-alkylated intermediates as an inseparable mixture. The mixture was then redissolved in DCM (40 ml) and TFA (10 ml) was slowly added at 0 ° C, and the solution was then stirred at 22 ° C for 12 hours. The volatiles were evaporated under reduced pressure, and excess TFA was removed by coevaporation with toluene (3 x 30 ml) to give the imine intermediate as a brown oil. The residue was dissolved in MeOH (200ml) and NaBH3CN (1.26g, 20.0mmol, 1.0 equivalent) was added at 0°C, the solution was warmed to 22°C, and then stirred at this temperature for 24 hours. The reaction was then quenched with an aqueous NaOH solution (3M, 20ml), and the solvent was reduced to about a quarter. The aqueous phase was extracted with diethyl ether (3x 150ml), dried over Na2SO4, filtered and dried under reduced pressure. The intermediate was dissolved in DCM (200ml), triethylamine (4.2ml, 30.1mmol, 1.5 equivalent) and Boc2O (4.80g, 22.0mmol, 1.1 equivalent) were added at 0°C, and the solution was stirred at 22°C for 16 hours. The solvent was evaporated and then purified by column chromatography (SiO2: 9:1 to 8:2 hexanes / EtOAc) to give the title compound 8 (2.44 g, 9.2 mmol, 46%) as a colorless crystalline compound.

[0192] R f =0.30 (hexane / EtOAc: 9:1);

[0193] mp: 46-47℃;

[0194] 1 H-NMR (400MHz, CDCl3): δ=5.75–5.65(m,1H),5.12–5.08(m,2H),4.18–4.09(m,1H),3.60(b r,1H),3.11(br,1H),2.40–2.21(m,5H),2.14–2.01(m,2H),1.82–1.74(m,1H),1.48(s,9H);

[0195] 13C-NMR (100MHz, CDCl3): δ=221.6,221.0,154.0,132.8,119.1,79.7,63.7,60.4,59.7,45.9,38.2,36.8,32.9,32.5,28.5,26.4,25.3;

[0196] HR-MS(ESI):C 15 H 23 O3N + [M+Na] + The calculated m / z value is 288.1570, the found value is 288.1572.

[0197] Due to the presence of rotamers in NMR measurements, an analytical aliquot was deprotected using a mixture of DCM and TFA (1:1, 1.0 ml).

[0198] 1 H-NMR (400MHz, CDCl3): δ = 10.11 (br, 1H; NH2 + ),9.71(br,1H;NH2 + ),5.67–5.57(m,1H),5.18–5.14(m,2H),4.12(br,1H),3.32(br,1H),3.16(br,1 H),2.67–2.57(m,1H),2.52–2.36(m,2H),2.32–2.20(m,4H),2.06–1.99(m,1H);

[0199] 13 C-NMR (100MHz, CDCl3): δ=217.4,131.4,120.6,64.1,59.5,45.3,38.5,36.5,33.9,23.1;

[0200] 6-Benzyloctahydrocyclopenta[2,1-b:5,1-b']dipyrrole-3(3aH)-carboxylic acid tert-butyl ester (9)

[0201]

[0202] At -78 ° C, O3 / O2 (≈3g O3 / h) flow was introduced into a solution of compound 8 (1.06g, 4.00mmol, 1.0 equivalent) in DCM and MeOH (1:1, 40mL) for 30 minutes. The unique blue color indicates the end of ozonolysis, and excess ozone is removed with the O2 flow. Dimethyl sulfide (3.00mL, 41.0mmol, 10 equivalents) was added and the cooling bath was removed. The reaction mixture was allowed to reach 22 ° C and stirred at this temperature for 8 hours. The solvent was then evaporated and the residue was redissolved in MeOH (40mL). Benzylamine (440μL, 4.03mmol, 1.0 equivalents) was added and stirred at 22 ° C for one hour, and then acetic acid (230μL, 4.02mmol, 1.0 equivalents) and sodium cyanoborohydride (251mg, 3.99mmol, 1.0 equivalents) were added in sequence. The mixture was stirred at 22 ° C for 16.5 hours, and then quenched with the NaOH aqueous solution (2M, 20mL). The reaction volume was reduced to about 1 / 3rd of the original volume, and the crude product was extracted with diethyl ether (3x 100ml). The organic phase merged was dried over Na2SO4, filtered and concentrated under reduced pressure. The crude product was purified by column chromatography (SiO2: 8: 2 to 1: 1 hexane / EtOAc+0.1% triethylamine), title compound 9 (956mg, 2.79mmol, 70%) was obtained, which is a colorless oil.

[0203] R f =0.13–0.38 (SiO2; DCM / MeOH: 50:1+NEt3 drops);

[0204] 1 H-NMR (400MHz, CD2Cl2): δ=7.32–7.21(m,5H),3.89(d,J=13.1Hz,1H),3.70–3.63(m,2H),3.29(d,J=13.1Hz,1H),3.08(dt,J=11.1,6.0Hz,1H ),2.79(t,J=7.8Hz,1H),2.60-2.56(m,1H),2.24–2.18(m,1H),2.05–1 .94(m,3H),1.90–1.72(m,3H),1.63–1.57(m,1H),1.55–1.44(m,10H);

[0205] 13C-NMR (100MHz, CD2Cl2): δ=154.3(br),154.1(br),140.4,129.1,128.5,127.1,78.9,77.5,69.3,61.1(br),6 0.3(br),59.5,54.1,47.7(br),47.2(br),37.5(br),37.0(br),36.8,32.0(br),30.9(br),30.6,28.7,28.5;

[0206] HR-MS(ESI):C 21 H 31 O2N2 + [M+H] + m / z calculated value 343.2380, found value 343.2371

[0207] Due to the presence of rotamers in NMR measurements, an analytical aliquot was deprotected using a mixture of DCM and TFA (1:1, 1.0 ml).

[0208] 1 H-NMR (400MHz, CD3OD): δ=7.57–7.47(m,5H),4.48–4.41(m,2H),3.97(br,2H),3.64–3.58(m,1H),3.50– 3.41(m,2H),3.35–3.27(m,1H),2.50–2.43(m,1H),2.41–2.18(m,4H),2.07(br,2H),1.85(br,1H),ppm;

[0209] 13 C-NMR (100MHz, CD3OD): δ = 163.1 (q, J = 34Hz, COOCF3 - ),131.9,131.5,131.2,130.4,118.3(q,J=294Hz,COOCF3 - ),78.7(br),70.1,62.6(br),59.5(br),56.0(br),47.3(br),37.7,35.2(br),30.0,29.5(br),ppm;

[0210] 6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)octahydrocyclopenta[2,1-b:5,1-b']dipyrrole-3(3aH)-carboxylic acid tert-butyl ester (10)

[0211]

[0212] To a solution of compound 9 (829 mg, 2.42 mmol, 1.0 eq.) in methanol (24.0 mL, 0.10 M) was added acetic acid (280 μL, 4.90 mmol, 2.0 eq.) and Pd / C (10% Pd; 83 mg, 10 m / m%). The flask was placed under vacuum and flushed with hydrogen three times. The reaction mixture was then stirred at 22 °C for 3 days under a hydrogen atmosphere (1 atm, balloon) until the starting material was completely consumed. The reaction mixture was filtered through celite and the solvent was evaporated to dryness. The intermediate was redissolved in anhydrous NMP (5.0 mL, 0.48 M), then triethylamine (1.0 mL, 7.17 mmol, 3.0 eq.) and 6-chloro-7-deazapurine (409 mg, 2.66 mmol, 1.1 eq.) were added under argon, heated to 110 °C and stirred at this temperature for 14 hours. The solution was diluted with deionized water (50 mL) and extracted with DCM (3 x 30 mL). The combined organic phases were dried over Na2SO4, filtered and the solvent was reduced. Flash column chromatography (SiO2: DCM / MeOH 50: 1 to 20: 1) gave the title compound 10 (603 mg, 1.63 mmol, 67%) as a light brown powder.

[0213] R f =0.23(SiO2;50:1DCM / MeOH);

[0214] mp: 219-222℃;

[0215] 1 H-NMR (400MHz, CD2Cl2): δ = 11.24 (br, 1H), 8.27 (s, 1H), 7.09 (d, J = 3.7Hz, 1H), 6.55 (d,, J = 3.7Hz, 1H), 4.48–4.46 (m, 1H), 4. 25–4.20(m,1H),3.96–3.91(m,1H),3.71–3.70(m,2H),3.34–3.27(m,1H),2.25–2.23(m,1H),2.14-1.74(m,8H),1.45(s,9H);

[0216] 13 C-NMR (100MHz, CD2Cl2): δ=155.1,154.1,151.8,151.6,120.5,103.4,101.9, 79.3,70.2,69.1,61.4,49.2,47.5,47.1,36.3,35.9,33.9,33.4,32.9,28.7;

[0217] HR-MS(ESI):C 20 H28 O2N5 + [M+H] + m / z calculated value 370.2238, found value 370.2245

[0218] 3-(6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)octahydrocyclopenta[2,1-b:5,1-b']dipyrrol-3(3aH)-yl)-3-oxyylidenepropionitrile (KMC420)

[0219]

[0220] Compound 10 (66.5 mg, 0.18 mmol, 1.0 eq.) was dissolved in a mixture of DCM and TFA (1:1, 2 mL) and the solution was stirred for 2 hours at 22° C. After the deprotection was completed, the volatile compounds were evaporated under reduced pressure.

[0221] The dried residue was suspended in DCM (2 mL) and N,N-diisopropylethylamine (156 μ L, 0.89 mmol, 4.9 equiv) was added to form a clear solution. Then cyanoacetic acid (30.8 mg, 0.36 mmol, 2.0 equiv), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (as HCl salt) (69.0 mg, 0.36 mmol, 2.0 equiv) and oxime (51.0 mg, 0.36 mmol, 2.0 equiv) were subsequently added and the reaction was stirred at 22 ° C for 21 hours. The organic phase was diluted with EtOAc (30 mL) and washed with saturated NaHCO (2 x 30 mL). The combined organic phase was dried over Na SO, filtered and reduced. RP-HPLC (gradient: 0–20% D in 40 min) yielded the final compound KMC420 as a TFA salt (18.0 mg, 0.04 mmol, 23%; purity >98%)

[0222] UPLC: t R =2.17min;

[0223] RP-HPLC: t R =19–28min;

[0224] 1 H-NMR (400MHz, D2O): δ = 8.22 (s, 1H), 7.41 (s, 1H), 6.92 (s, 1H), 4.45 (br, 1H), 4. 28–4.27(m,1H),4.05–3.75(m,4.5H),3.62–3.42(br,1.5H),2.46–1.95(m,8H);

[0225] 13C-NMR (100MHz, D2O): δ=163.1,162.7,147.6,142.0,124.2,115.7,104.6 ,71.3,70.4,69.0,48.4,47.2,34.5,33.8,33.3,32.9,31.6,26.0,25.2;

[0226] Due to the obstructed rotation, some 13 The C signal was split, and the quaternary carbon signal could not be resolved.

[0227] HR-MS(ESI):C 18 H 21 N6O + [M+H] + The calculated m / z value is 337.1771, the found value is 337.1768.

[0228] 1-(6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)octahydrocyclopenta[2,1-b:5,1-b']dipyrrol-3(3aH)-yl)prop-2-en-1-one (KMC423)

[0229]

[0230] Compound 10 (49.9 mg, 0.14 mmol, 1.0 equivalent) was dissolved in a mixture of DCM and TFA (1:1, 1 mL) and the solution was stirred at 22 ° C for 2 hours. The volatiles were evaporated under reduced pressure and the dried residue was suspended in DCM (1 mL). DIPEA (23 μL, 0.13 mmol, 1.0 equivalent) was added to form a clear solution, followed by the addition of acrylic acid (175 μL, 2.55 mmol, 19 equivalents), EDCl (58.8 mg, 0.31 mmol, 2.3 equivalents) and DMAP (2.8 mg, 0.02 mmol, 0.2 equivalents) and the reaction was stirred at 22 ° C for 19.5 hours. The organic phase was diluted with DCM (5 mL), washed with saturated Na2CO3 (3x 5 mL), and then the aqueous phase was extracted with DCM (3x 5 mL). The combined organic phase was dried over Na2SO4, filtered and reduced. RP-HPLC (gradient: 0-50% D in 40 min) yielded the final compound KMC423 as a TFA salt (19.0 mg, 0.04 mmol, 31%; purity >99%).

[0231] UPLC: t R =2.40min;

[0232] RP-HPLC: t R =20-23min;

[0233] 1 H-NMR (400MHz, CD3OD): δ = 8.25 (d, J = 2.5Hz, 1H), 7.39 (d, J = 3.4Hz, 1H), 6.97 (d, J = 3.6Hz, 1H), 6.70–6.61 (m, 1H), 6.29 (dt, J = 16.8, 1.8Hz ,1H),5.78(ddd,J=10.4,6.1,1.8Hz,1H),4.43(br,1H),4.32(br,1H),4.14–3.96(m,2H),3.69(br,1H),3.48(br,1H),2.46–1.95(m,8H);

[0234] 13 C-NMR (100MHz, CD3OD): δ = 166.3, 166.3, 162.3 (q, J = 34.7Hz, COOCF3 - ),143.2,130.1,129.6,128.8,128.5,1225.3,105.7,72.3,69.9,69.3,48.9,47.7,36.4,36.0,35.0,33.4;

[0235] Due to the obstructed rotation, some 13 The C signal was split, and the quaternary carbon signal could not be resolved.

[0236] HR-MS(ESI):C 18 H 22 N5O + [M+H] + The calculated m / z value is 324.1819, the found value is 324.1820.

[0237] Tert-butyl (2-(1,3-dioxy-2,3-dihydro-1H-inden-2-yl)ethyl)carbamate (12)

[0238]

[0239] To a suspension of 1,3-indanedione (2.93 g, 20.0 mmol, 1.0 eq), Hantzsch ester (5.07 g, 20.0 mmol, 1.0 eq) and crude N-Boc-2-aminoacetaldehyde (3.18 g, 20.0 mmol, 1.0 eq) in DCM (65 ml) was added proline (460 mg, 4 mmol, 0.2 eq) and the mixture was stirred at 22° C. for 19.5 h. After completion of the reaction, the solvent was evaporated under reduced pressure and the product was purified by flash column chromatography (SiO 2 : hexane / EtOAc 9:1 to pure EtOAc) and then recrystallized from hot hexane / EtOAc mixture (8 / 2, 100 ml) to give the desired title compound 12 (2.99 g, 10.3 mmol, 52%) as an off-white powder.

[0240] R f =0.18 (hexane / EtOAc: 8 / 2);

[0241] mp: 97-99℃;

[0242] 1 H-NMR (400MHz, CDCl3): δ = 7.94–7.98 (m, 2H), 7.80–7.85 (m, 2H), 4.9 (br, 1H; NH), 3.36(q,J=6.2Hz,2H),3.06(t,J=6.4Hz,1H),2.11(q,J=6.5Hz,2H),1.35(s,9H);

[0243] 13 C-NMR (100MHz, CDCl3): δ=200.3,157.9,142.1,135.7,123.3,79.2,50.1,38.3,28.3,26.9;

[0244] HR-MS(ESI):C 16 H 19 O4n + [M+Na] + The calculated m / z value is 312.1206, the found value is 312.1190.

[0245] Tert-butyl (2-(2-allyl-1,3-dioxy-2,3-dihydro-1H-inden-2-yl)ethyl)carbamate (13)

[0246]

[0247] To compound 12 (1.46 g, 5.0 mmol, 1 eq.), KCO (1.39 g, 10.1 mmol, 2 eq.) and tetrabutylammonium hydrogen sulfate (TBAHS) (0.26 g, 0.75 mmol, 0.15 eq.) in anhydrous acetonitrile (35 ml) and in a yellow suspension under an argon atmosphere, allyl bromide (0.9 ml, 10.4 mmol, 2.1 eq.) was added dropwise. The mixture was stirred at 22 °C for 19 hours, and after the reaction was complete, excess KCO was filtered out and the solvent was reduced. The residue was distributed between deionized water and DCM, and the aqueous layer was extracted with DCM (3 x 25 ml). The combined organic phase was dried over MgSO, filtered and reduced. Purification by MPLC (24 g SiO2: 1:0 to 1:1 hexanes / EtOAc) gave the title compound 13 (1.48 g, 4.49 mmol, 90%) as a light yellow powder.

[0248] R f =0.31 (hexane / EtOAc: 8 / 2);

[0249] mp: 63-65℃;

[0250] 1 H-NMR (400MHz, CDCl3): δ=7.98–7.94(m,2H),7.85–7.80(m,2H),5.50–5.40(m,1H),5.04–4.87(m,2 H),4.27(br,1H;NH),3.04–2.99(m,2H),2.52(d,J=7.5Hz,1H),2.06(t,J=7.2Hz,2H),1.29(s,9H);

[0251] 13 C-NMR (100MHz, CDCl3): δ=203.3,155.2,142.0,135.8,131.1,123.2,119.7,79.2,56.8,28.3,39.6,36.7,34.2;

[0252] HR-MS(ESI):C 19 H 23 O4n + [M+Na] + m / z = calculated value 352.1519, found value 352.1509.

[0253] 3a-Allyl-4-oxydeoxy-3,3a,4,8b-tetrahydroindeno[1,2-b]pyrrole-1(2H)-carboxylic acid tert-butyl ester (14)

[0254]

[0255] Compound 13 (1.47g, 4.46mmol, 1.0 equivalent) is stirred at 22 ° C for 18 hours in a MeOH solution of HCl (7.2mL, ≈1.25M, 2.0 equivalent). After the reaction is complete, the volatiles are evaporated and the residue is dissolved in MeOH (45mL). NaOAc (0.38g, 4.48mmol, 1.0 equivalent) and NaBH3CN (0.28g, 4.48mmol, 1.0 equivalent) are added and stirred at 22 ° C for 16 hours. The reaction is quenched with an aqueous NaOH solution (2M, 10ml) and the volume is reduced to about a quarter. The aqueous phase is extracted with EtOAc (3x 50mL), the organic phase merged is washed with brine, dried over Na2SO4, filtered and the solvent is evaporated. The residue was then dissolved in DCM (10 mL), and NEt (0.65 mL, 4.50 mmol, 1.0 eq.) and Boc O (1.18 g, 5.41 mmol, 1.2 eq.) were added at 0° C. The solution was stirred at 22° C. for 20 hours, and after the reaction was complete, the solvent was evaporated and the product was purified by column chromatography (SiO : hexane / EtOAc 1:0 to 1:1) to give the title compound 14 (0.74 g, 2.36 mmol, 53%) as a colorless oil.

[0256] R f =0.35 (hexane / EtOAc: 9 / 1);

[0257] 1 H-NMR (400MHz, CDCl3): δ = 8.02–7.81 (m, 1H), 7.71–7.63 (m, 2H), 7.47–7. 45(m,1H),5.71–5.59(m,1H),5.30–5.18(m,1H),5.15–5.10(m,1H),5.05– 5.00(m,1H),3.81–3.62(m,1H),3.07–2.96(m,1H),2.67–2.62(m,1H),2. 49–2.43(m,1H),2.15–2.11(m,1H),1.96–1.88(m,1H),1.61–1.48(m,9H);

[0258] 13C-NMR (100MHz, CDCl3): δ=207.7,207.2,154.8,154.1,152.9,152.3,136.1,135.9,133.3,133.1,129.5,129.4,12 8.5,127.2,123.7,123.4,119.2,80.6,80.1,63.9,63.8,61.1,60.1,45.8,45.2,39.4,39.2,34.2,33.7,28.9,28.6

[0259] Due to the obstructed rotation, some 1 H- and 13 C-signal splitting.

[0260] HR-MS(ESI):C 19 H 23 O3N + [M+Na] + The calculated m / z value is 336.1570, the found value is 336.1564.

[0261] Due to the presence of rotamers in NMR measurements, an analytical aliquot was deprotected using a mixture of DCM and TFA (1:1, 1.0 ml).

[0262] 1 H-NMR (400MHz, D2O): δ=7.95–7.91(m,1H),7.86–7.84(m,2H),7.75–7.71(m,1H),5.67–5.56(m,1H),5.32(s,1H),5.20 –5.04(m,2H),3.53–3.48(m,1H),3.0(dt,J=11.7,6.4Hz,1H),2.75–2.54(m,2H),2.43–2.37(m,1H),2.24–2.16(m,1H);

[0263] 13 C-NMR (100MHz, D2O): δ = 208.8, 162.8 (q, J = 36Hz, COOCF3 - ),145.5,137.8,136.8,131.8,131.8,127.5,124.4,120.1,116.3(q,J=292Hz,COOCF3 - ),64.0,59.9,45.0,38.4,32.8

[0264] 6-Benzyl-2,3,5,6,6a,10b-hexahydroindeno[1,2-b:3,2-b']dipyrrole-1(4H)-carboxylic acid tert-butyl ester (15)

[0265]

[0266] At -78 ° C, an O3 / O2 (≈3 g O3 / h) stream was introduced into a solution of compound 14 (683 mg, 2.18 mmol, 1.0 equiv) in DCM and MeOH (1:1, 22 mL) for 30 minutes. The characteristic blue color indicated the end of ozonolysis, and excess ozone was removed with the O2 stream. Dimethyl sulfide (1.6 mL, 21.6 mmol, 9.9 equiv) was added and the cooling bath was removed. The reaction mixture was allowed to reach 22 ° C and stirred at this temperature for 5 hours. The solvent was then evaporated and then co-evaporated with toluene. The residue was dissolved in anhydrous MeOH (22 mL), then benzylamine (238 μL, 2.18 mmol, 1.0 equiv), acetic acid (125 μL, 2.19 mmol, 1.0 equiv) were added, and stirred at 22 ° C for one hour. Then sodium cyanoborohydride (274mg, 4.36mmol, 2.0 equivalents) was added and the solution was stirred at 22 ° C for 19.5 hours. After the reaction was completed, it was quenched using the NaOH aqueous solution (2M, 20mL). The reaction volume was reduced to about one-third of the original volume, and the crude product was extracted with diethyl ether (3x 100ml). The organic phase merged was dried over Na2SO4, filtered and concentrated under reduced pressure. The product was purified by column chromatography (SiO2: hexane / EtOAc 8:2+0.1% triethylamine) to obtain title compound 15 (687mg, 1.76mmol, 81%), which is colorless oil.

[0267] R f =0.34 (hexane / EtOAc: 8 / 2+NEt 3 drops);

[0268] 1 H-NMR (400MHz, CD2Cl2): δ=7.73–7.62(m,1H),7.36–7.22(m,8H),5.09–5.04(m,1H),4.16–4.12(m,1H),3.93–3.90(m,1H),3. 67–3.62(m,2H),3.28–3.22(m,1H),2.86(br,1H),2.61–2.55(m,1H),2.14–2.07(m,1H),2.01–1.88(m,3H)1.57–1.48(m,9H);

[0269] 13C-NMR (100MHz, CDCl3): δ=155.2,154.8,145.4,144.9,143.4,143.2,140.0,129.2,128.8,128.7,128.6,128.4,127.8,127.6,127. 3,127.2,126.8,125.9,125.7,79.9,79.4,77.7,77.6,72.4,62.3,61.1,59.2,54.3,46.9,46.5,37.0,36.7,36.5,36.2,28.8,28.7;

[0270] HR-MS(ESI):C 25 H 31 O2N2 + [M+H] + The calculated m / z value is 391.2380, the found value is 391.2365.

[0271] Due to the presence of rotamers in NMR measurements, an analytical aliquot was deprotected using a mixture of DCM and TFA (1:1, 1.0 ml).

[0272] 1 H-NMR (400MHz, D2O): δ=7.63–7.54(m,8H),7.06–7.04(m,1H),5.30(s,1H),5.24(s,1H),4.83–4.79(m ,1H),4.63–4.60(m,1H),3.66–3.51(m,3H),3.23–3.16(m,1H),2.63–2.54(m,2H),2.49–2.39(m,2H);

[0273] 13 C-NMR (100MHz, D2O): δ = 162.8 (q, J = 36Hz, COOCF3 - ),136.7,136.2,132.2,131.8,130.9,130.5,129.7,126.9,126.1,116.3(q,J=292Hz,COOCF3 - ),79.6,71.6,59.9,58.9,54.4,45.8,35.7,34.0;

[0274] 1,2,3,4,5,6,6a,10b-Octahydroindeno[1,2-b:3,2-b']dipyrrole-1,6-diolium dichloride (16)

[0275]

[0276] Pd / C (10wt.%) (32mg, 10wt.%) was added to a solution of compound 15 (324.6mg, 0.83mmol, 1.0 equivalent) and acetic acid (95μL, 1.66mmol, 2.0 equivalent) in MeOH (10mL). The mixture was stirred for 20 hours at 22°C under H2 atmosphere (1atm, balloon), and then filtered through diatomaceous earth and concentrated under reduced pressure. The residue was absorbed in DCM (10ml) and TFA (5ml) and stirred for 2 hours at 22°C. After the reaction was completed, the solvent was evaporated, alkalized using NaOH aqueous solution (3M, 10mL) and extracted with EtOAc (3x 100mL). The combined organic phase was dried over Na2SO4, filtered and concentrated. Final compound 16 (206mg, 0.75mmol, 91%) was precipitated from diethyl ether with HCl (1.25M, in MeOH) as a colorless crystalline solid.

[0277] 1 H-NMR (400MHz, D2O): δ=7.68–7.62(m,4H),5.36(s,2H),3.62–3.56(m,2H),3.20–3.13(m,2H),2.59–2.53(m,2H),2.44–2.36(m,2H);

[0278] 13 C-NMR (100MHz, D2O): δ=137.1,131.8,126.5,72.0,59.2,45.5,35.1;

[0279] HR-MS(ESI):C 13 H 17 N2 + [M+H] + The calculated m / z value is 201.1386, the found value is 201.1392.

[0280] References

[0281] Schwartz, CE; Curran, DP, AJAm.Chem.Soc.1990,112(25),9272–9284

[0282] Kyasa,S.;Fisher,T.;Dussault,P.,Synthesis 2011,2011(21),3475–3481

[0283] Ramachary,DB;Kishor,M.,Org.Biomol.Chem.2008,6(22),4176

[0284] Zhou,YJ.et al.(1997),Proc.Natl.Acad.Sci.USA,94:13850-13855

[0285] Brizzi,MFet al.(1996),J.Biol.Chem.,271:3562-3567

[0286] Yamaoka,K.et al.(2004),Gen.Biol.,5:253

[0287] Ide,H.et al.(2008),Biochem.Biophys.Res.Commun.,369:292-296

[0288] Li,et al.J.Am.Chem.Soc.2019141239415-9421。

Claims

1. A compound of formula 1 or a salt thereof, in, R 1 Selected from R 2 Selected from z, x, and y are 1.

2. The compound according to claim 1, wherein R 1 Selected from 3. The compound according to claim 2, wherein R 1 Selected from 4. The compound according to claim 2, wherein R 1 yes 5. The compound according to any one of claims 1 to 4, wherein R 2 Selected from 6. The compound according to claim 5, wherein R 2 Selected from 7. An intermediate of formula 3 or a salt thereof, in, R 5 and R 6 Selected from -H， - a protecting group cleavable under acidic conditions, - a protecting group cleavable under alkaline conditions, - a protecting group cleavable under reducing conditions, - a hydrogenolytically cleavable protecting group, - a protecting group cleavable using a metal catalyst, z, x, and y are 1.

8. The intermediate according to claim 7, wherein the protecting group cleavable under acidic conditions is 9. The intermediate according to claim 7, wherein the protecting group cleavable under alkaline conditions is 10. The intermediate according to claim 7, wherein the protecting group cleavable under reducing conditions is 11. The intermediate according to claim 7, wherein the hydrogenolytically cleavable protecting group is 12. The intermediate according to claim 7, wherein the protective group cleavable using a metal catalyst is 13. The intermediate according to claim 7, wherein -R 5 and R 6 Both are –H, or -R 5 or R 6 is –H, and the other R is a protecting group, or -R 5 and R 6 are all protecting groups, among which R 5 The cleavage of the protecting group at R 6 Cleavage of the protecting group.

14. An intermediate according to formula 3 or a salt thereof, in, R 5 or R 6 Selected from and Another R is selected from -H， - a protecting group cleavable under acidic conditions, - a protecting group cleavable under alkaline conditions, - a protecting group cleavable under reducing conditions, - a hydrogenolytically cleavable protecting group, - a protecting group cleavable using a metal catalyst, z, x, and y are 1.

15. The intermediate according to claim 14, wherein the protecting group cleavable under acidic conditions is Boc or Ts.

16. The intermediate according to claim 14, wherein the protecting group cleavable under basic conditions is Fmoc, Ac or Ac-CF3.

17. The intermediate according to claim 14, wherein the protecting group cleavable under reducing conditions is Ts.

18. The intermediate according to claim 14, wherein the hydrogenolytically cleavable protecting group is Bn or Cbz.

19. The intermediate according to claim 14, wherein the protecting group cleavable using a metal catalyst is Alloc or Troc.

20. An intermediate according to formula 3 or a salt thereof, in, R 5 or R 6 Selected from and Another R is selected from -H， - a protecting group cleavable under acidic conditions, - a protecting group cleavable under alkaline conditions, - a protecting group cleavable under reducing conditions, - a hydrogenolytically cleavable protecting group, - a protecting group cleavable using a metal catalyst, z, x, and y are 1.

21. The intermediate according to claim 20, wherein the protecting group cleavable under acidic conditions is Boc or Ts.

22. The intermediate according to claim 20, wherein the protecting group cleavable under basic conditions is Fmoc, Ac or Ac-CF3.

23. The intermediate according to claim 20, wherein the protecting group cleavable under reducing conditions is Ts.

24. The intermediate of claim 20, wherein the hydrogenolytically cleavable protecting group is Bn or Cbz.

25. The intermediate according to claim 20, wherein the protecting group cleavable using a metal catalyst is Alloc or Troc.

26. An intermediate of formula 5, in, R 9 Selected from -H， - a protecting group cleavable under acidic conditions, - a protecting group cleavable under alkaline conditions, - a protecting group cleavable under reducing conditions, - a hydrogenolytically cleavable protecting group, - a protecting group cleavable using a metal catalyst, z and y are 1.

27. The intermediate according to claim 26, wherein the protecting group cleavable under acidic conditions is Boc or Ts.

28. The intermediate according to claim 26, wherein the protecting group cleavable under basic conditions is Fmoc, Ac or Ac-CF3.

29. The intermediate according to claim 26, wherein the protecting group cleavable under reducing conditions is Ts.

30. The intermediate of claim 26, wherein the hydrogenolytically cleavable protecting group is Bn or Cbz.

31. The intermediate according to claim 26, wherein the protecting group cleavable using a metal catalyst is Alloc or Troc.

32. A compound according to any one of claims 1 to 6, an intermediate according to any one of claims 7 to 13, an intermediate according to any one of claims 14 to 19, an intermediate according to any one of claims 20 to 25, or an intermediate according to any one of claims 26 to 31, wherein The compound or the intermediate is an enantiomer.

33. Use of a compound according to any one of claims 1 to 6 for the preparation of a medicament for the treatment of a disease associated with the activity of a kinase of the JAK family.

34. The use according to claim 33, wherein Enantiomers or racemates are used.

35. The use according to claim 33, wherein Enantiomers were used.

36. Use of a compound according to any one of claims 1 to 6 for the preparation of a medicament for the treatment of autoimmune diseases associated with the activity of kinases of the JAK family, cancer, Alzheimer's disease, asthma or for the prevention of rejection of allografts or xenografts.

37. The use according to claim 36, wherein The drug is used to treat rheumatoid arthritis, ulcerative colitis, psoriatic arthritis, psoriasis, Crohn's disease, atopic dermatitis, chronic hand eczema, non-segmental vitiligo, axial spondyloarthritis, small intestinal CD, fistulizing CD, Sjögren's syndrome, ankylosing spondylitis, cutaneous lupus, lupus nephropathy, uveitis, myelofibrosis and Alzheimer's disease or asthma.

38. The use according to claim 36, wherein The medicament is used to prevent rejection of allografts.

39. The use according to claim 38, wherein The medicament is used to prevent rejection of an allogeneic kidney.

40. The use according to any one of claims 36 to 39, wherein Enantiomers or racemates are used.

41. The use according to any one of claims 36 to 39, wherein Enantiomers were used.

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