Atopic dermatitis active aminoketones, their preparation and use

Abstract

The application belongs to the technical field of medicine, and discloses a kind of anti-atopic dermatitis active enaminone compound, preparation and purpose.The application specifically discloses a kind of enaminone compound 1 and compound 2 and pharmaceutically acceptable salt thereof, and pharmaceutical composition thereof, and application thereof in the preparation of treating atopic dermatitis drugs.Cell experiments prove that compound 1 and compound 2 have good treatment atopic dermatitis activity, and are expected to become the treatment drug of immune related diseases such as atopic dermatitis.

Description

Technical Field

[0001] This invention relates to a class of enaminoketone compounds and their pharmaceutically acceptable salts, methods for their preparation, pharmaceutical compositions containing such compounds, and the application of such compounds in the treatment of atopic dermatitis, belonging to the field of pharmaceutical technology. Background Technology

[0002] Atopic dermatitis (AD) is a common, chronic, relapsing, inflammatory, and itchy skin disease characterized by erythema, papules, exudation, desquamation, and itching. It often begins in childhood and can persist into adulthood. [1] The prevalence of atopic dermatitis has been rising steadily in recent years, ranging from 2.1% to 4.9% in adults and 15% to 20% in children, making it the skin disease with the highest disease burden. [2] Alzheimer's disease (AD) is heterogeneous; multiple factors, including genetics, skin barrier function, immune dysregulation, environmental factors, and microbial imbalance, can trigger the disease. Furthermore, there are significant individual differences in patients' clinical manifestations, severity, and treatment response. [3] .

[0003] Medication is an important means of treating Alzheimer's disease (AD). Topical medications are first-line therapy, including topical corticosteroids, calcineurin inhibitors, and phosphodiesterase 4 (PDE4) inhibitors. For severe cases, systemic therapy is used clinically, including antihistamines, immunosuppressants, JAK inhibitors, monoclonal antibodies, and other biologics. [1，4] Although a variety of medications are available in clinical practice, survey results show that patient satisfaction with existing medications is only 14%-42%. [5] There remains a significant need and importance in developing new drugs that are highly effective in treating Alzheimer's disease (AD).

[0004] Filaggrin (FLG) is an important protein for epidermal function, and its encoding gene is located on chromosome 1q21. FLG is first synthesized in stratum granular keratinocytes as profilaggrin, which is highly phosphorylated and insoluble in water, with a mass of approximately 400 kDa. Its N-terminus contains a calcium-binding domain and a nuclear localization element, followed by 10-12 tandem FLG monomers, and finally a C-terminus associated with degradation function. Profilaggrin is dephosphorylated and proteolytically hydrolyzed to form FLG monomers. Filaggrin plays a crucial role in epidermal barrier function, including acting as an important molecule connecting keratin fibers to form the keratin capsule, maintaining the physical barrier function of the epidermis. Furthermore, in the stratum corneum, FLG is degraded by proteases to generate natural moisturizing factors (amino acids, pyrrolidone-5-carboxylic acid, and trans-urea acid, etc.), participating in skin hydration, maintaining the acidic environment of the stratum corneum, reducing microbial residence on the skin surface, and maintaining normal skin function. [6]Studies have shown that mutations in the filaggrin gene are highly associated with the pathogenesis of Alzheimer's disease (AD). [1] More than 500 FLG mutations have been identified. [7] Meta-analyses showed that the odds ratio (OR) between AD incidence and FLG mutation was 3.1–4.8, and patients carrying FLG mutations had more severe and persistent disease. [9] .

[0005] Besides the high correlation between FLG mutations and AD incidence, approximately 50%-90% of AD patients do not carry the mutated FLG gene. However, regardless of whether they carry the mutated FLG gene, the expression levels of FLG in both lesion and non-lesion areas of all AD patients are lower than in healthy individuals.

[10] This means that atopic dermatitis is accompanied by low FLG expression. Since FLG is an important structural and functional protein for maintaining the epidermal barrier, decreased FLG expression in keratinocytes is both a significant cause and an inevitable consequence of AD. Increasing FLG expression can help rebuild the skin barrier in AD patients and can be used for wild-type (FLG-rich) skin. + / + ) and FLG monoallelic mutant genotype (FLG + / - Prevention and treatment of Alzheimer's disease (AD). Therefore, finding novel small molecule drugs that can enhance FLG expression has become one of the important directions in AD drug development.

[0006] This invention uses a filaggrin promoter model for evaluation and found that compounds 1 and 2 can significantly increase the expression of filaggrin in human keratinocytes. We believe that compounds 1 and 2 have high value for the treatment of atopic dermatitis and have application prospects.

[0007] References

[0008] [1]S. Atopic Dermatitis.N Engl J Med,2021,384,1136-1143.

[0009] [2]MRLaughter,MBCMaymone,S.Mashayekhi,BWMArents,C.Karimkhani,SMLangan,RPDellavalle,C.Flohr,The global burden of atopic dermatitis:lessons from the GBD Study-1990to 2017.British Journal of Dermatology,2021,184,304-309.

[0010] [3]SFZiegler, Thymic stromal lymphopoietin, skin barrier dysfunction, and the atopic march. Ann Allergy Asthma Immunol, 2021, 127, 306-311.

[0011] [4] Gu Heng, Zhang Jianzhong, Chinese Guidelines for the Diagnosis and Treatment of Atopic Dermatitis (2020 Edition). Chinese Journal of Dermatology, 2020, 53, 81-88.

[0012] [5]JAFOosterhaven,LSSpekhorst,J.Zhang,ANVoorberg,GLERomeijn,CMBoesjes,M.Graaf,MSBruin-Weller,MLASchuttelaar,Eczemacontrol and treatment satisfaction in atopic dermatitis patients treated with dupilumab–a cross-sectional study from the BioDay registry.J DermatologTreat,2022,33,1986-1989.

[0013] [6] CR Harding, S. Aho, CA Bosko, Filaggrin–revisited. International Journal of Cosmetic Science, 2013, 35, 412-423.

[0014] [7] J. Gupta, DJ Margolis, Filaggrin Gene Mutations with Special Reference to Atopic Dermatitis. Curr Treat Options Allergy, 2020, 7, 403-413.

[0015] [9] C. Drislane, ADIrvine, The role of filaggrin in atopic dermatitis and allergic disease. Annals of Allergy Asthma&Immunology, 2019, 124, 36-43.

[0016]

[10] JPThyssen,I.Jakasa,C.Riethmüller,MP A.Braun,M.Haftek,PGFallon,J.Wróblewski,H.Jakubowski,L.Eckhart,W.Declercq,S.Koppes,KAEngebretsen,C.Bonefeld,ADIrvine,S.Keita-Alassane,M.Simon,H.Kawasaki,A.Kubo,M.Amagai,T.Matsui,S.Kezic,Filagrin Expression and ProcessingDeficiencies Impair Corneocyte Surface Texture and Stiffness in Mice. Journal of Investigative Dermatology, 2020,140,615-623. Summary of the Invention

[0017] The technical problem solved by this invention is to provide a class of enaminoketone compounds and their pharmaceutically acceptable salts, as well as pharmaceutical compositions thereof and their use in the preparation of anti-atopic dermatitis drugs.

[0018] To solve the technical problem of this invention, the present invention provides the following technical solution:

[0019] The first aspect of the present invention is to provide compounds represented by the following group:

[0020]

[0021] The second aspect of the present invention is to provide a method for preparing the compound described in the first aspect.

[0022]

[0023] Phenylacetyl-2-phenyl-1-one 3 reacts with azacyclobutane via a 1,4-addition reaction to give compound 1. Phenylacetyl-2-phenyl-1-one 3 reacts with cis-perhydroisoindole via a 1,4-addition reaction to give compound 2.

[0024] A third aspect of the present invention is to provide a pharmaceutical composition comprising compounds as shown in structures 1 and 2 and their pharmaceutically acceptable salts, the pharmaceutical composition comprising compounds as shown in groups 1 and 2 and their pharmaceutically acceptable salts as active ingredients, and optionally comprising a pharmaceutical carrier.

[0025] Typically, the pharmaceutical compositions of this invention contain 0.1-95% by weight of the compounds of this invention.

[0026] Pharmaceutical compositions of the compounds of the present invention can be prepared according to methods known in the art. For this purpose, if desired, the compounds of the present invention can be combined with one or more solid or liquid pharmaceutical excipients and / or adjuvants to formulate suitable administration or dosage forms for use as human or veterinary medicine.

[0027] The compounds of the present invention or pharmaceutical compositions containing them can be administered in unit dose form via enteral or non-enteric routes, such as oral, intramuscular, subcutaneous, nasal, oral mucosa, skin, peritoneum or rectum, with oral administration being preferred.

[0028] The compounds of this invention or pharmaceutical compositions containing them can be administered by injection. Injection includes intravenous injection, intramuscular injection, subcutaneous injection, and intradermal injection.

[0029] Dosage forms can be liquid or solid. Liquid dosage forms include true solutions, colloids, microparticles, emulsions, and suspensions. Other dosage forms include tablets, capsules, pellets, aerosols, pills, powders, solutions, suspensions, emulsions, granules, suppositories, and lyophilized powder injections.

[0030] The extracts or compounds of this invention can be formulated into ordinary preparations, sustained-release preparations, controlled-release preparations, targeted preparations, and various microparticle delivery systems.

[0031] To formulate unit-dosage dosage forms into tablets, a wide variety of carriers known in the art can be used. Examples of carriers include diluents and absorbents such as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose, and aluminum silicate; humectants and binders such as water, glycerin, polyethylene glycol, ethanol, propanol, starch paste, dextrin, syrup, honey, glucose solution, gum arabic paste, gelatin paste, sodium carboxymethyl cellulose, shellac, methylcellulose, potassium phosphate, and polyvinylpyrrolidone; disintegrants such as dried starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfate, methylcellulose, and ethylcellulose; disintegration inhibitors such as sucrose, tristearate, cocoa butter, and hydrogenated oil; absorption promoters such as quaternary ammonium salts and sodium dodecyl sulfate; and lubricants such as talc, silica, corn starch, stearates, boric acid, liquid paraffin, and polyethylene glycol. Tablets can also be further processed into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer and multilayer tablets.

[0032] For example, various carriers known in the art can be widely used to formulate the drug delivery unit into pills. Examples of carriers include diluents and absorbents such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinylpyrrolidone, gelucire, kaolin, talc, etc.; binders such as gum arabic, tragacanth, gelatin, ethanol, honey, liquid sugar, rice paste, or flour paste, etc.; and disintegrants such as agar powder, dried starch, alginate, sodium dodecyl sulfate, methylcellulose, ethylcellulose, etc.

[0033] For example, to formulate the drug delivery unit into a capsule, the active ingredient, the extract or compound of the present invention, is mixed with the various carriers described above, and the resulting mixture is placed in a hard gelatin capsule or a soft capsule. Alternatively, the active ingredient, the compound of the present invention, can be formulated as a microcapsule, suspended in an aqueous medium to form a suspension, or filled into a hard capsule or formulated as an injectable preparation for use.

[0034] For example, the extracts or compounds of this invention can be formulated into injectable preparations, such as solutions, suspension solutions, emulsions, and lyophilized powder injections. These preparations can be aqueous or non-aqueous and may contain one or more pharmacodynamically acceptable carriers, diluents, binders, lubricants, preservatives, surfactants, or dispersants. Diluents may be selected from water, ethanol, polyethylene glycol, 1,3-propanediol, ethoxylated isostearyl alcohol, polyoxyethylene isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters, etc. Furthermore, to prepare isotonic injections, appropriate amounts of sodium chloride, glucose, or glycerol can be added to the injectable formulation. In addition, conventional solubilizers, buffers, pH adjusters, etc., can also be added. Solubilizers may be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc.; pH adjusters may be phosphates, acetates, hydrochloric acid, sodium hydroxide, etc.; osmotic pressure adjusters may be sodium chloride, mannitol, glucose, phosphates, acetates, etc. When preparing lyophilized powder injections, mannitol, glucose, etc., may also be added as support agents.

[0035] In addition, colorants, preservatives, flavorings, tasters, sweeteners or other materials may be added to pharmaceutical preparations if necessary.

[0036] To achieve the purpose of medication and enhance the therapeutic effect, the drug or drug composition of the present invention can be administered using any known method of administration.

[0037] The fourth aspect of the present invention is to provide the use of the enamine ketone compounds 1 and 2 of the present invention and their pharmaceutically acceptable salts in the preparation of drugs for the prevention or treatment of atopic dermatitis caused by immune diseases, etc.

[0038] When using the enamine ketone derivatives of the present invention and their pharmaceutically acceptable salts or the compositions of the present invention to treat the above-mentioned diseases, the dosage may refer to the dosage used when using enamine ketone derivatives for treatment; when using the indole derivatives of the present invention or the compositions of the present invention as health products, or adding them to health products, the dosage should be less than the usual therapeutic dosage.

[0039] The dosage of the compounds and pharmaceutical compositions of this invention depends on many factors, such as the nature and severity of the disease to be prevented or treated, the sex, age, weight, personality, and individual response of the patient or animal, the route of administration, frequency of administration, and therapeutic purpose. Therefore, the therapeutic dosage of this invention can vary widely. Generally speaking, the dosage of the pharmaceutical components used in this invention is well known to those skilled in the art. The actual amount of drug contained in the final formulation of the compound composition according to this invention can be appropriately adjusted to achieve the required therapeutic dose and fulfill the preventive or therapeutic purpose of this invention. The appropriate daily dosage range for the compounds of this invention is 0.001-150 mg / kg body weight, preferably 0.01-100 mg / kg body weight, more preferably 0.01-60 mg / kg body weight, and most preferably 0.1-10 mg / kg body weight. The above dosages can be administered in a single dose or divided into several doses, such as two, three, or four doses. This is limited by the clinical experience of the dispensing physician and the administration regimen, including the use of other treatment methods.

[0040] The total dose required for each treatment can be divided into multiple administrations or administered as a single dose. The compounds and compositions of the present invention can be taken alone or in combination with other therapeutic or symptomatic drugs, with dosage adjustments.

[0041] The inventors have discovered that compounds 1 and 2 of the present invention, and pharmaceutically acceptable salts thereof, have certain therapeutic effects on atopic dermatitis. Therefore, compounds 1 and 2 of the present invention, and pharmaceutically acceptable salts thereof, also relate to a method for treating and improving diseases associated with atopic dermatitis. The method comprises administering a therapeutically effective amount of compounds 1 and 2 or pharmaceutically acceptable salts thereof, or a pharmaceutical composition thereof, to a patient requiring treatment.

[0042] This invention demonstrates that compounds 1 and 2 can increase the expression of filaggrin in human keratinocytes, and have therapeutic potential for atopic dermatitis. Pharmaceutically acceptable salts of compounds 1 and 2 have not been publicly reported, nor have their therapeutic activities for atopic dermatitis been publicly reported.

[0043] Beneficial technical effects

[0044] In their research on enamine ketone compounds, the inventors of this invention synthesized new compounds 1 and 2 using chemical methods. The ability of compounds 1 and 2 to upregulate FLG gene transcription was evaluated using a classic cell model for evaluating anti-atopic dermatitis activity: the FLG gene upstream promoter dual-luciferase assay model. The results showed that compounds 1 and 2 significantly upregulated FLG gene transcription, demonstrating excellent therapeutic activity for atopic dermatitis. These are valuable new lead compounds in the development of drugs for the treatment of atopic dermatitis. Attached Figure Description

[0045] Figure 1 Results of Compound 1 and Compound 2 upregulating FLG gene transcription levels in HaCaT cells Detailed Implementation

[0046] The following examples further illustrate the invention, but do not limit the invention in any way.

[0047] Example 1: Preparation of (E)-3-(azacyclobutan-1-yl)-1-phenylprop-2-en-1-one (1)

[0048]

[0049] Weigh 390 mg of 1-phenyl-2-propyn-1-one and add it to 10.0 mL of THF solvent with stirring. Add 410 mg of aziridine. React at room temperature for 12 hours, remove the organic solvent under reduced pressure, and perform column chromatography (ethyl acetate / petroleum ether = 2:1) to give a white solid compound (E)-3-(aziridine-1-yl)-1-phenylprop-2-en-1-one 1 (336 mg, 60%). Mp: 87.7-88.5 °C; 1 H NMR (400MHz, CDCl3): δ=7.88-7.85(m,2H),7.66-7.63(m,1H),7.45-7.37(m,3H),4.12-4.10(m,1H),2.46-2.39(m,2H)ppm; 13 C NMR (100MHz, CDCl3): δ = 188.2, 151.4, 140.5, 130.9, 128.2, 127.6, 92.2, 52.4, 16.6ppm; HRMS (ESI): m / z calcd forC 12 H 13 NO[M+H] + 187.1290; found 187.1296.

[0050] Example 2: Preparation of (E)-3-((3aR,7aS)-octahydro-2H-isoindol-2-yl)-1-phenylprop-2-en-1-one (2)

[0051]

[0052] Weigh 390 mg of 1-phenyl-2-propyn-1-one and add it to 10.0 mL of THF solvent with stirring. Add 900 mg of cis-perhydroisoindole 5. React at room temperature for 12 hours, remove the organic solvent under reduced pressure, and perform column chromatography (ethyl acetate / petroleum ether = 2:1) to give a white solid compound (E)-3-((3aR, 7aS)-octahydro-2H-isoindole-2-yl)-1-phenylprop-2-en-1-one 2 (612 mg, 80%). Mp: 107.7-108.2 °C; 1 H NMR (400MHz, CDCl3): δ = 7.99 (d, J = 12.0, 1H), 7.91-7.88 (m, 2H), 7.46-7.38 (m, 3H),5.66(d,J=12.0Hz,1H),3.59(dd,J1=10.4Hz,J2=8.4Hz,1H),3.40(dd,J1= 10.4Hz, J2=5.2Hz, 1H), 3.30(dd, J1=11.2Hz, J2=8.0Hz, 1H), 3.13(dd, J1=11.2 Hz,J2=6.8Hz,1H),2.44-2.36(m,1H),2.26-2.19(m,1H),1.69-1.34(m,8H)ppm; 13 C NMR (100MHz, CDCl3): δ=188.5,151.0,140.8,130.9,128.2,127.6,93.0,56.8,50.9,37.2,37. 1,25.92,25.7,23.1,22.3ppm; IR(neat):ν=2918,1635,1533,11268,709cm-1; HRMS(ESI):m / z calcd for C 17 H 22 NO[M+H] + 256.1690; found 256.1696.

[0053] Pharmacological experiments

[0054] Experimental Example 1. Determining the effect of a compound on FLG gene transcription in HaCaT cells

[0055] (1) Principle of the dual-luciferase detection model of the upstream promoter of the FLG gene

[0056] The promoter is located upstream of the transcription start site and is a DNA sequence that is recognized and bound by transcription factors and RNA polymerase. The promoter can interact with transcription factors, thereby controlling the initiation of transcription and regulating the degree of transcription.

[0057] This detection model was constructed using the pGL4.11[luc2P] reporter gene plasmid, which uses the firefly luciferase gene as the reporter gene. The model replaced the promoter region of pGL4.11[luc2P] with the upstream -2200 / +9 promoter of the human FLG gene. When this promoter is activated, firefly luciferase is expressed, and its expression level is directly proportional to the degree of promoter activation. Simultaneously, pGL4.74[hRluc TK] was used as an internal control plasmid to correct transfection efficiency. This plasmid regulates the transcription of the Renilla luciferase gene using the HSV-TK promoter. By detecting the activity of firefly luciferase in cell lysates and correcting it with Renilla luciferase activity, the degree of FLG gene transcription can be reflected. This detection model primarily detects the degree to which compounds upregulate FLG gene transcription.

[0058] In this detection model, Ha CaT cells were co-transfected with pGL4.11-(-2200 / +9) and pGL4.74 plasmids at a ratio of 20:1. Six hours after transfection, the cells were digested and evenly seeded into 48-well plates. Twenty-four hours after seeding, the compound was added. Twenty-four hours after treatment, the activities of firefly luciferase and Renilla luciferase in the cell lysate were detected using the Luciferase Assay System (Promega) and the Renilla Luciferase Assay System (Promega), respectively. By comparing with the solvent control group, the fold increase in FLG gene transcription by the compound was calculated.

[0059] (2) Experimental methods for detecting the effect of compounds on FLG gene transcription in HaCaT cells

[0060] HaCaT cells were prepared at a density of 1.8 × 10⁶ cells per dish. 6 One cell was seeded into a 100 mm cell culture dish. After culturing for 24 hours, [the cells were then] used... HaCaT cells were co-transfected with 10 μg of pGL4.11-(-2200 / +9) plasmid and 0.5 μg of pGL4.74 internal control plasmid using transfection reagent (Polyplus). Six hours after transfection, cells in each dish were digested and divided into 3 × 10⁶ cells per well. 4Cells were seeded into 48-well plates. 24 hours after seeding, the test compound was added; benzenemod was added to the positive control group, and an equal volume of DMSO was added to the solvent control group. Three replicates were set for each group. 24 hours after treatment, the activity of firefly luciferase and Renilla luciferase in the cell lysate was measured using the Luciferase Assay System (Promega) and the Renilla Luciferase Assay System (Promega), i.e., the relative luminescence units (RLUs) in the cell lysate were determined. The fold increase in FLG gene transcription in each experimental group was calculated using formula (1). Experimental data were analyzed using GraphPad Prism software.

[0061] (1) Increase factor = (Luciferase RLUs) 受试化合物 / Renilla RLUs 受试化合物 ) / (LuciferaseRLUs 溶剂对照 / Renilla RLUs 溶剂对照 )

[0062] (3) Experimental Results

[0063] After HaCaT cells were treated with compounds 1 and 2 at a concentration of 10 μM for 24 h, the activity of firefly luciferase (Luc) and kidney luciferase (Rluc) in the cell lysate was measured, and the fold increase in FLG gene transcription by the compounds was calculated according to formula (1).

[0064] (1) Increase factor = (Luciferase RLUs) 受试化合物 / Renilla RLUs 受试化合物 ) / (LuciferaseRLUs 溶剂对照 / Renilla RLUs 溶剂对照 )

[0065] Experimental results showed that compounds 1 and 2 significantly upregulated the transcription level of the FLG gene in human keratinocytes, indicating that compounds 1 and 2 have therapeutic activity against atopic dermatitis. Specific results are shown in Table 1 and... Figure 1 .

[0066] Table 1. Compounds 1 and 2 upregulate FLG gene transcription in HaCaT cells

[0067]

Claims

1. The use of a class of compounds and their pharmaceutically acceptable salts in the preparation of drugs for the prevention or treatment of atopic dermatitis, characterized in that, The compounds are selected from the following group:

2. The application according to claim 1, characterized in that, The aforementioned atopic dermatitis is related to inflammatory skin diseases.

3. The application according to claim 2, characterized in that, The skin inflammatory diseases mentioned include eczema, seborrheic dermatitis, and neurodermatitis.

4. The use of a pharmaceutical composition in the preparation of a drug for the prevention or treatment of atopic dermatitis, characterized in that, The pharmaceutical composition contains a therapeutically effective amount of a compound of the group of claims 1, a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

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