Deuterated JAK3 inhibitor drug and application

Abstract

The invention discloses a deuterated inhibitor and pharmaceutically acceptable salts thereof, which are shown in the following formula I. The invention relates to a deuterated JAK3 inhibitor, a pharmaceutical composition and application thereof. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically relates to deuterated JAK3 inhibitor drugs and uses. Background Art

[0002] The AK / STAT (Janus protein tyrosine kinase / signal transducer and activator of transcription) signaling pathway is an important cytokine signaling pathway discovered in the 1990s when people studied interferon. It can complete signal transduction from the cytoplasm to the nucleus. The mammalian JAK family has four members, namely JAK1, JAK2, JAK3 and TYK2. This signaling pathway is related to immunity, inflammation, cell proliferation, differentiation, survival and apoptosis. Autoimmune diseases are a large class of diseases caused by tissue damage caused by the body's immune response to self-antigens, including rheumatoid arthritis (RA), psoriasis, alopecia areata and other series of diseases. Their common characteristics are unclear pathogenesis, few targets and lack of effective therapeutic drugs. JAK is a new target for the treatment of autoimmune diseases. Currently, there are 7 JAK small molecule inhibitors on the market for the treatment of different autoimmune diseases.

[0003] However, with the clinical application of JAK inhibitors, side effects such as opportunistic infections and anemia have gradually emerged. The main reason is the non-selective inhibition of different subtypes of JAK kinase. JAK kinase includes four subtypes, JAK1, JAK2, JAK3 and Tyk2. The JAK-STAT signaling pathway composed of the downstream effector molecule STAT (signal transducers and activators of transcription) regulates the inflammatory and immune signals of more than 50 cytokines in the body. This non-selective inhibition of JAK is bound to affect the physiological effects of multiple cytokines, thereby leading to corresponding side effects. Therefore, the development of highly selective JAK inhibitors is the current trend and direction of the development of such drugs. Ritlecitinib is a new oral targeted JAK3 inhibitor developed by Pfizer. Ritlecitinib is a selective JAK3 inhibitor that has no activity against JAK1, JAK2 and TYK2. In in vitro studies, it can inhibit the differentiation and function of Th1 and Th17 cells and reduce STAT5 phosphorylation caused by IL-2, IL-4, IL-7 and IL-15. In in vivo studies, Ritlecitinib reduced features of disease pathology in a murine experimental autoimmune encephalomyelitis model and in rat adjuvant arthritis.

[0004] Studies have shown that ritlecitinib can block the activity of signaling molecules and immune cells, which are believed to be the cause of alopecia areata. Compared with the first-generation pan-JAK inhibitors, ritlecitinib has an advantage in reducing toxicity. Alopecia areata, also known as alopecia areata, is a type of hair loss. It refers to the loss of hair in one or more places on the scalp. The hair loss area is round or oval, while the hair in other surrounding areas remains normal. Alopecia areata is characterized by scarless hair loss on the scalp, face or body, and has a potential immune-inflammatory pathogenesis. Alopecia areata can have a significant impact on the patient's quality of life, including psychological burdens such as anxiety or depression.

[0005] A potentially attractive strategy to improve drug metabolism properties is deuterium modification. In this method, attempts are made to slow drug metabolism or reduce the formation of undesirable metabolites by replacing one or more hydrogen atoms with deuterium atoms. Deuterium is a safe, stable, non-radioactive isotope of hydrogen. Deuterium forms a stronger bond with carbon than hydrogen. In certain cases, the bond strength agent increased by deuterium positively affects the ADME properties of the drug, thereby potentially improving the efficacy, safety and / or tolerability of the drug. At the same time, since the size and shape of deuterium are essentially the same as those of hydrogen, replacing hydrogen with deuterium is expected to have no effect on the biochemical efficacy and selectivity of the drug compared to the original chemical entity containing only hydrogen. By improving the pharmacokinetic properties of the original drug, the original defects of the drug, such as easy metabolism and large side effects, can be overcome.

[0006] Despite the beneficial activity of the current JAK3 inhibitor drug ritlecitinib, there is a continuing need for new compounds to treat the above-mentioned diseases and conditions. Summary of the invention

[0007] The deuterated JAK3 inhibitor and the pharmaceutically acceptable salt thereof provided by the present invention can improve the pharmacokinetic properties of the JAK3 inhibitor ritlecitinib, and reduce the dosage and possible toxic side effects.

[0008] In order to achieve the above object, the present invention provides a deuterated JAK3 inhibitor as shown below and a pharmaceutically acceptable salt thereof:

[0009]

[0010] The deuterated JAK3 inhibitor and its pharmaceutically acceptable salt described in the present invention are selected from methanesulfonate, maleate, hydrochloride or phosphate.

[0011] The deuterated JAK3 inhibitor and pharmaceutically acceptable salts thereof of the present invention include their use in treating autoimmune diseases such as alopecia areata.

[0012] The deuterated JAK3 inhibitor and the pharmaceutically acceptable salt thereof of the present invention can be used as a pharmaceutical composition, which comprises the deuterated JAK3 inhibitor and the pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable carrier.

[0013] The pharmaceutical composition of the deuterated JAK3 inhibitor and its pharmaceutically acceptable salt described in the present invention is selected from capsules, powders, tablets, granules, pills, injections, syrups, oral liquids, inhalants, ointments, suppositories or patches.

[0014] The term "pharmaceutically acceptable" as used herein refers to substances (such as carriers, excipients and / or adjuvants) that do not affect the biological activity or properties of the compounds of the present invention and are relatively non-toxic, that is, the substance can be administered to an individual without causing adverse biological reactions or interacting in an adverse manner with any components contained in the composition.

[0015] The term "pharmaceutical composition" as used in this application refers to a biologically active compound optionally mixed with at least one pharmaceutically acceptable chemical component, wherein the pharmaceutically acceptable chemical component includes but is not limited to carriers, excipients and / or adjuvants, such as stabilizers, diluents, dispersants, suspending agents, thickening agents, etc.

[0016] As used herein, the term "carrier" refers to relatively nontoxic chemical compounds or agents that facilitate the introduction of a compound into cells or tissues.

[0017] The term "pharmaceutically acceptable salt" as used herein refers to a salt that retains the biological effectiveness of the free acid and free base of the specified compound and has no adverse effects in biology or other aspects. The compounds of the present invention also include pharmaceutically acceptable salts. Pharmaceutically acceptable salts refer to the form of converting the base group in the parent compound into a salt. Pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of base groups such as amine (amino) groups. Pharmaceutically acceptable acid addition salts can be prepared from inorganic and organic acids. Inorganic acids derived from acid addition salts include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. Organic acids derived from acid addition salts include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.

[0018] Beneficial effects: Compared with the prior art, the present invention has the following advantages:

[0019] The present invention provides a class of deuterated JAK3 inhibitor drugs, which further improve the pharmacokinetic properties of the deuterated JAK3 inhibitors and reduce the dosage and possible toxic side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is the NMR data diagram of the compound in Example 1 of Example 1 of the present invention. DETAILED DESCRIPTION

[0021] The features and advantages of the present invention can be better understood by reference to the following specific examples. Although the application has described preferred embodiments of the present invention, these embodiments are provided only as examples. Those of ordinary skill in the art will appreciate that the variants of the embodiments of the present invention also can be used to implement the present invention, and multiple simple variants, changes and replacements of the present invention still fall within protection scope of the present invention.

[0022] All documents or portions of documents cited in this application, including but not limited to patents, patent applications, articles, books, manuals, and treatises, are incorporated herein by reference in their entirety.

[0023] The starting materials of the present invention can be obtained from commercial sources or prepared using methods well known to those skilled in the art.

[0024] Example 1

[0025] General preparation method

[0026]

[0027]

[0028] Intermediate 1-A (3 mmol) was dissolved in tert-butyl alcohol (3 mL), and then intermediate 1-B (3 mmol) and DIPEA (8 mmol) were added, and the temperature was raised to 80°C and stirred for 16 hours. After concentration, intermediate 1-C was obtained by column chromatography.

[0029] Dissolve the intermediate 1-C (3 mmol) in DMF (3 mL), then add potassium tert-butoxide (3 mmol) and 1 mL of heavy water. Heat to 100°C and stir overnight. Filter to obtain the intermediate 1-D, which is directly used in the next step.

[0030] Intermediate 1-D (1 mmol) was dissolved in 2 mL of trifluoroacetic acid, stirred at room temperature for 4 hours, and the organic phase was concentrated. 2 mL of methanol was added, and 1 mL of ethylenediamine was further added. The mixture was stirred at room temperature overnight. After the solvent was concentrated, intermediate 1-E was obtained by column chromatography.

[0031] Intermediate 1-E (0.1 mmol) was dissolved in THF (1 mL), and under nitrogen protection, 0.1 mL of triethylamine was added, and deuterated acryloyl chloride (0.05 mmol) was slowly added at 0°C, and the mixture was stirred for 20 min. The solvent was concentrated, and Example 1 (28.4 mg, yield 99.3%) was obtained by column chromatography. 1H NMR (400 MHz, DMSO) δ 11.52 (s, 1H), 8.17 – 8.06 (m, 1H), 7.37 – 7.24 (m, 1H), 7.09 (d, J = 1.7 Hz, 1H), 6.79 (td, J = 15.8,10.4 Hz, 1H), 6.10 (dd, J = 16.7, 2.4 Hz, 1H), 5.68 (dd, J = 10.5, 2.4 Hz, 1H), 4.86 – 4.51 (m, 1H), 4.39 – 3.94 (m, 2H), 2.96 – 2.60 (m, 1H), 1.87 – 1.63(m, 4H), 1.19 (dd, J = 20.4, 6.8 Hz, 3H).

[0032] Test Example 1: JAK3 enzyme activity test

[0033] Compound 001 was prepared according to patent WO 2022 / 002210A1 (see page 9 of the specification, Example 1). The test article was dissolved in dimethyl sulfoxide (DMSO) to a stock concentration of 30 mM. 11-point half-log serial dilutions were generated in DMSO, with the highest concentration being 600 μM. The test compound plate also contained a positive control well containing a known inhibitor to define 100% inhibition, and a negative control well containing DMSO to define no inhibition. The compound plate was diluted 1 to 60 so that the final maximum concentration of the measured compound was 10 μM and the DMSO concentration was 2%. The test article and the measurement control were added to a 384-well plate. The reaction mixture contained 20 mM HEPES (pH = 7.4), 10 mM magnesium chloride, 0.01% bovine serum albumin (BSA), 0.0005% Tween 20, 4 μM or 1 mM ATP. The JAK3 assay solution contains 1 μM JAKtide. The assay is started by adding 1 nM JAK3 enzyme and incubating for 75 minutes at room temperature for JAK3. The enzyme concentration and incubation time are optimized for each new enzyme preparation and slightly modified over time to ensure 20%-30% phosphorylation. The assay is stopped with a final concentration of 10 mM EDTA, 0.1% coating agent and 100 mM HEPES (pH = 7.4). The assay plate is placed on the Caliper LifeScience Lab Chip 3000 (LC3000) instrument and each well is sampled using appropriate separation conditions to measure unphosphorylated and phosphorylated peptides.

[0034]

[0035] It can be seen from the JAK3 enzyme activity data that the compound in Example 1 of the present patent shows a significant improvement in the inhibition of JAK3 enzyme activity compared with the positive drug compound 001 and ritlectinib.

[0036] Experimental Example 2: Pharmacokinetics of Compounds

[0037] 1. Experimental instruments and materials

[0038] High-speed refrigerated centrifuge, vortex oscillator (Vortex Genius3), high-speed centrifuge (Eppendorf5415D), disposable syringes, pipettes (Eppendorf), SD male rats used in the experiment were purchased from Yangzhou University, EDTA-K2 vacuum blood collection tubes, and normal saline. All rats in the oral group fasted for 12 hours before administration, and had free access to water and food during administration.

[0039] (II) Experimental steps

[0040] In Example 1, intermediate 1 and compound 001 were dissolved in DMSO / solutol / water (10 / 10 / 80) to prepare a clear solution. The intranasal administration dose of the compound was 25 mg / kg, and the tail vein administration dose of the compound was 5 mg / kg. At 2 min, 10 min, 30 min, 1 h, 2 h, 3 h, 5 h, 8 h, 12 h, 16 h, and 24 h after tail vein administration, 0.5 mL of blood was continuously collected from the venous plexus of the fundus into a heparin tube. At 5 min, 15 min, 30 min, 1 h, 2 h, 3 h, 5 h, 8 h, 12 h, 16 h, and 24 h after intranasal administration, 0.5 mL of blood was continuously collected from the venous plexus of the fundus into a heparin tube. The samples were heated at 8000r, 4 0 After centrifugation at -20 °C for 10 min, 0.15 mL of the upper plasma was collected and 0 The samples were stored at 4 °C and then analyzed by LC-MS / MS. The data were analyzed by WinNolin non-compartmental model to obtain key pharmacokinetic parameters.

[0041] (III) Experimental results

[0042] Table 1 Pharmacokinetic parameters

[0043]

[0044] Compared with ritlecitinib and compound 001, the half-life of the compound of Example 1 after oral administration is significantly improved, which can effectively improve the dosage and thus reduce the toxic side effects of high-dose administration.

[0045] Without further description, it is believed that one of ordinary skill in the art can use the foregoing description and illustrative examples to complete and utilize these compounds of the present invention and implement the claimed methods. It should be understood that the foregoing discussion and examples only present detailed descriptions of certain preferred embodiments. It should be clear to one of ordinary skill in the art that various modifications and equivalents may be made without departing from the spirit and scope of the present invention.

Claims

1. The deuterated JAK3 inhibitor and its pharmaceutically acceptable salt as shown below, 。 2. The deuterated JAK3 inhibitor and pharmaceutically acceptable salt thereof according to claim 1, characterized in that The pharmaceutically acceptable salt is selected from methanesulfonate, maleate, hydrochloride or phosphate.

3. Use of the deuterated JAK3 inhibitor according to claim 1 and a pharmaceutically acceptable salt thereof in the preparation of a drug for treating autoimmune diseases.

4. The use according to claim 3, wherein the autoimmune disease is selected from alopecia areata.

5. A pharmaceutical composition, characterized in that The invention comprises a therapeutically effective amount of the deuterated JAK3 inhibitor according to claim 1 and a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

6. The pharmaceutical composition according to claim 5, characterized in that The pharmaceutical composition is selected from capsules, powders, tablets, granules, pills, injections, syrups, oral solutions, inhalants, ointments, suppositories or patches.

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