Morpholine derivative, pharmaceutical composition and application of morpholine derivative in medicine

Abstract

The invention provides a compound shown in a general formula (I), or all stereoisomers, pharmaceutically acceptable salts or deuterated substances thereof, and application of the compound in preparation of drugs for autoimmune diseases.

Description

Technical Field

[0001] This invention relates to a morpholine derivative, a pharmaceutical composition, and its application in medicine. Background Technology

[0002] Toll-like receptors (TLRs) are a class of molecular pattern recognition receptors that are widely distributed in different tissues. They monitor and recognize different pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs), playing an important role in both innate and adaptive immunity.

[0003] TLRs belong to type I transmembrane proteins. To date, 13 members of the TLR family have been discovered, 10 of which exist in humans. TLR1, TLR2, TLR4, TLR5, TLR6, TLR10, and TLR11 are located on the cell membrane and can recognize lipids, lipoproteins, and other substances in microorganisms. TLR3, TLR7, TLR8, and TLR9 are located in intracellular vesicle structures (such as lysosomes, endosomes, and endoplasmic reticulum) and recognize nucleic acids in microorganisms.

[0004] TLR7 and TLR8 are most similar in sequence and function. Numerous studies have shown that activation of TLR7 / 8 can trigger type I interferon responses and various inflammatory reactions. In autoimmune disorders such as systemic lupus erythematosus (SLE), abnormal and persistent activation of TLR7 / 8 leads to the exacerbation of the disease state. Therefore, developing compounds with selective and potent inhibitory activity to antagonize TLR7 / 8 and inhibit overactivated immune responses holds promise as a new approach to treating autoimmune diseases. Summary of the Invention

[0005] The purpose of this invention is to provide a novel morpholinane derivative or stereoisomer, its pharmaceutical composition, and its application in the preparation of drugs for autoimmune diseases.

[0006] One or more embodiments of the present invention provide a compound of general formula (I), or all its stereoisomers, pharmaceutically acceptable salts, or deuterated derivatives:

[0007]

[0008] R1 is selected from -CN, -CF3, -OCH3 or -OCF3;

[0009] A is selected from A was further replaced or not replaced;

[0010] B is selected from

[0011] R2 is selected from 4-8 membered heterocyclic alkyl groups, and R2 may be further substituted or unsubstituted.

[0012] One or more embodiments of the present invention provide a compound of general formula (I), or all its stereoisomers, pharmaceutically acceptable salts, or deuterated derivatives, wherein R2 is further selected from one or more of C 1-3 Alkyl, halogen, C 1-3 Substitution with haloalkyl, 3-6 membered cycloalkyl or 3-6 membered heterocycloalkyl groups.

[0013] One or more embodiments of the present invention provide a compound of general formula (I), or all its stereoisomers, pharmaceutically acceptable salts, or deuterated derivatives, wherein A is further selected from one or more of C. 1-3 Alkyl, halogen, C 1-3 Substitution of alkyl halogens, 3-6 membered cycloalkyl groups or 3-6 membered heterocycloalkyl groups

[0014] One or more embodiments of the present invention provide a compound of general formula (I), or all its stereoisomers, pharmaceutically acceptable salts, or deuterated derivatives, said compound being selected from one of the following structures:

[0015]

[0016]

[0017] One or more embodiments of the present invention provide a pharmaceutical composition comprising:

[0018] Compounds of general formula (I) or their stereoisomers, pharmaceutically acceptable salts or deuterated derivatives;

[0019] One or more other active ingredients may be selected; and

[0020] Pharmaceutically acceptable carriers and / or excipients.

[0021] One or more embodiments of the present invention provide the use of the above-described pharmaceutical compositions, compounds of general formula (I) or their stereoisomers, pharmaceutically acceptable salts or deuterated derivatives in the preparation of medicaments for autoimmune diseases.

[0022] One or more embodiments of the present invention provide the use of the above-described compound or its stereoisomers, pharmaceutically acceptable salts or deuterated derivatives, or the above-described pharmaceutical compositions as TLR7 / 8 inhibitors. Detailed Implementation

[0023] The following embodiments illustrate the technical solutions of the present invention in detail, but the scope of protection of the present invention includes, but is not limited to, these embodiments.

[0024] Example 1

[0025] 5-((2R,6R)-2-methyl-6-(5-(1-methylpiperidin-4-yl)-1,3,4-oxadiazol-2-yl)morpholino)quinoline-8-nitrile compound 1

[0026] 5-((2R,6R)-2-methyl-6-(5-(1-methylpiperidin-4-yl)-1,3,4-oxadiazol-2-yl)morpholino)qui noline-8-carbonitrile

[0027]

[0028]

[0029] first step:

[0030] (2R,6R)-2-methyl-6-(5-(1-methylpiperidin-4-yl)-1,3,4-oxadiazol-2-yl)morpholine-4-carboxylic acid tert-butyl ester compounds 1-3

[0031] tert-butyl(2R,6R)-2-methyl-6-(5-(1-methylpiperidin-4-yl)-1,3,4-oxadiazol-2-yl)morpholine-4-carbox ylate

[0032] Under a nitrogen atmosphere, ((2R,6R)-4-(tert-butyloxycarbonyl)-6-methylmorpholine-2-carboxylic acid 1-1 (900 mg, 3.67 mmol) was dissolved in DCM (20.0 mL), followed by the sequential addition of DIPEA (1.18 g, 9.18 mmol) and HATU (2.09 g, 5.50 mmol). After activation at 40 °C for 30 min, 1-methyl-4-piperidincarbamoylhydrazide 1-2 (692 mg, 4.40 mmol) was added. The atmosphere was purged with nitrogen three times, and the reaction was carried out at room temperature for 2 h. Burgess reagent (2.63 g, 11.0 mmol) was added, and the reaction was carried out overnight at room temperature. The solvent was evaporated to dryness, and the mixture was extracted with water and DCM. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated to dryness. The mixture was then purified by reverse phase (0.1% TFAin). (water) yields the target product (2R,6R)-2-methyl-6-(5-(1-methylpiperidin-4-yl)-1,3,4-oxadiazol-2-yl)morpholine-4-carboxylic acid tert-butyl ester compound 1-3 (yellow solid, 800 mg, yield 53.6%).

[0033] LC-MSm / z(ESI) = 367.2[M+1].

[0034] Step Two:

[0035] (2R,6R)-2-methyl-6-(5-(1-methylpiperidin-4-yl)-1,3,4-oxadiazol-2-yl)morpholine compounds 1-4

[0036] (2R,6R)-2-methyl-6-(5-(1-methylpiperidin-4-yl)-1,3,4-oxadiazol-2-yl)morpholine

[0037] Under a nitrogen atmosphere, (2R,6R)-2-methyl-6-(5-(1-methylpiperidin-4-yl)-1,3,4-oxadiazol-2-yl)morpholine-4-carboxylic acid tert-butyl ester 1-3 (800 mg, 2.18 mmol) was dissolved in DCM (20.0 mL), followed by the addition of TFA (5 mL) N2 to purge the gas three times, and the reaction was carried out at room temperature for 1 h. The solvent was then evaporated to obtain the target crude product (2R,6R)-2-methyl-6-(5-(1-methylpiperidin-4-yl)-1,3,4-oxadiazol-2-yl)morpholine compound 1-4 (yellow oil, 581 mg, 100% yield).

[0038] LC-MSm / z(ESI) = 267.2[M+1].

[0039] Step 3:

[0040] 5-((2R,6R)-2-methyl-6-(5-(1-methylpiperidin-4-yl)-1,3,4-oxadiazol-2-yl)morpholino)quinoline-8-nitrile compound 1

[0041] 5-((2R,6R)-2-methyl-6-(5-(1-methylpiperidin-4-yl)-1,3,4-oxadiazol-2-yl)morpholino)qui noline-8-carbonitrile

[0042] Under a nitrogen atmosphere, (2S,6S)-2-methyl-6-(5-(1-methylpiperidin-4-yl)-1,3,4-oxadiazol-2-yl)morpholine 1-4 (280 mg, 0.73 mmol) was dissolved in 1,4-dioxane (6.00 mL), followed by the sequential addition of 5-bromoquinoline-8-carbamate 1-5 (292 mg, 1.26 mmol), cesium carbonate (2.10 g, 5.50 mmol), RuPhosPdG3 (42 mg, 0.055 mmol), and water (0.60 mL). The nitrogen atmosphere was purged three times, and the mixture was heated to 80 °C and reacted for 2 h. The solvent was evaporated, the mixture was extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, the solvent was evaporated, the reaction solution was directly concentrated, and purified by reversed-phase C18 column chromatography (acid method) to obtain 450 mg of crude product. TLC (DCM:MeOH = 10:1) yielded the target product 5-((2R,6R)-2-methyl-6-(5-(1-methylpiperidin-4-yl)-1,3,4-oxadiazol-2-yl)morpholino)quinoline-8-nitrile compound 1 (yellowish-white solid, 280 mg).

[0043] LC-MSm / z(ESI) = 419.10[M+1].

[0044] 1 H NMR(400MHz,Chloroform-d)δ9.10(dd,1H),8.49(dd,1H),8.08(d,1H),7.55(dd,1H),7.18(d,1H),5.23(dd,1H),4.23(dd t,1H),3.67(dt,1H),3.42–3.29(m,2H),2.97(d,3H),2.83(dd,1H),2.37(s,3H),2.17(d,4H),,2.04(t,2H),1.36(d,3H).

[0045] Biological testing

[0046] HEK-Blue-hTLR7 / 8 / 9 cell inhibition assay

[0047] 1. HEK-Blue-hTLR7 / 8 cells (1×10⁸) 4 (cells / well), HEK-Blue-hTLR9 cells (1.5×10⁻⁶) 4 (30 μL per well) cells were seeded into 384-well cell culture plates. The plates were incubated at 37°C in a 5% CO2 incubator for 4 hours.

[0048] 2. The compounds were prepared using DMSO and diluted in DMEM medium to 10 concentrations (1:3 dilution), with final concentrations of 10000, 3333.3, 1111.1, 370.4, 123.5, 41.2, 13.7, 4.6, 1.5, and 0.5 nM, respectively; R848 was prepared using DMSO and diluted in DMEM medium to final concentrations of 0.8 μM (HEK-Blue-hTLR7) and 3 μM (HEK-Blue-hTLR8); ODN2006 was prepared using endotoxin-free water and diluted in DMEM medium to final concentration of 1 μM.

[0049] 3. Cells treated with DMSO served as the blank control group. Simultaneously, cells treated with Resiquimod (R848) alone served as the positive control group for HEK-Blue-hTLR7 / 8, and cells treated with ODN2006 alone served as the positive control group for HEK-Blue-hTLR9. Cells treated with the test compound and either R848 or ODN2006 served as the test group. Each group had two parallel wells, which were incubated at 37°C in a 5% CO2 incubator.

[0050] 4. After 4 hours of incubation, remove the 384-well plates of HEK-Blue-hTLR7 / 8 from the incubator. Add R848 to each well individually, or add R848 and the diluted compound simultaneously. Incubate at 37°C in a 5% CO2 incubator for 16 hours. Then, remove the 384-well plates of HEK-Blue-hTLR9 from the incubator. Add ODN2006 to each well individually, or add ODN2006 and the diluted compound simultaneously. Incubate at 37°C in a 5% CO2 incubator for 16 hours.

[0051] 5. After culturing for 16 hours, remove the 384-well plate from the incubator, centrifuge at 1000 rpm for 1 minute, and use a multi-mode microplate reader to read the optical density value of each well at 620 nm.

[0052] 6. Calculate the cell inhibition rate = (1 - (OD620 test group - OD620 blank group) / (OD620 positive group - OD620 blank group)) × 100%, and calculate the half maximal inhibitory concentration (IC50) through curve fitting. 50 ).

[0053] Table 1: Activity of the compounds of the present invention in HEK-Blue-hTLR7 / 8 / 9 cell assays

[0054] compound <![CDATA[TLR7IC 50 (nM)]]> <![CDATA[TLR8IC 50 (nM)]]> <![CDATA[TLR9IC 50 (nM)]]> Compound 1 ＜10 ＜10 >5000

[0055] Conclusion: The compounds of this invention exhibit superior antagonistic activity against HEK-Blue-hTLR7 / 8 cells, showing significant antagonistic effects, while having no significant antagonistic effect against HEK-Blue-hTLR9 cells.

[0056] This invention specification provides a detailed description of specific embodiments. Those skilled in the art should recognize that the above embodiments are exemplary and should not be construed as limiting the invention. For those skilled in the art, various improvements and modifications can be made to the invention without departing from its principles, and the resulting technical solutions also fall within the scope of protection of the claims of this invention.

Claims

1. A compound represented by general formula (I), or all its stereoisomers, pharmaceutically acceptable salts, or deuterated derivatives: in, R1 is selected from -CN, -CF3, -OCH3, or -OCF3; A is selected from A was further replaced or not replaced; B is selected from R2 is selected from 4-8 membered heterocyclic alkyl groups, and R2 may be further substituted or unsubstituted.

2. The compound according to claim 1, or all its stereoisomers, pharmaceutically acceptable salts, or deuterated derivatives, wherein, R2 is further selected from C by one or more factors. 1-3 Alkyl, halogen, C 1-3 Substitution with haloalkyl, 3-6 membered cycloalkyl or 3-6 membered heterocycloalkyl groups.

3. The compound according to claim 1, or all its stereoisomers, pharmaceutically acceptable salts, or deuterated derivatives, wherein, A is further selected from one or more of C. 1-3 Alkyl, halogen, C 1-3 Substitution with haloalkyl, 3-6 membered cycloalkyl or 3-6 membered heterocycloalkyl groups.

4. The compound according to any one of claims 1-3, or all its stereoisomers, pharmaceutically acceptable salts, or deuterated derivatives, wherein the compound is selected from one of the following structures:

5. A pharmaceutical composition comprising: The compound or its stereoisomer, pharmaceutically acceptable salt or deuterated derivative as described in any one of claims 1-4; One or more other active ingredients may be selected; as well as Pharmaceutically acceptable carriers and / or excipients.

6. Use of the compound or stereoisomer of any one of claims 1-4, a pharmaceutically acceptable salt or deuterated product thereof, or the pharmaceutical composition of claim 5 in the preparation of a medicament for autoimmune diseases.

7. Use of the compound or stereoisomer of any one of claims 1-4, a pharmaceutically acceptable salt or deuterated product thereof, or the pharmaceutical composition of claim 5 as a TLR7 / 8 inhibitor.

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