阅读说明：本技术 用于治疗发炎性病症的方法 (Methods for treating inflammatory disorders ) 是由 詹维康 于 2020-01-03 设计创作，主要内容包括：本发明提供一种用于治疗发炎性病症的方法,尤其是用于治疗需要此类治疗的个体的与发炎性细胞因子过度表达相关的发炎性病症的方法,其包括向所述个体投与具有(经取代苯基)-丙烯醛部分的化合物。(The present invention provides a method for the treatment of inflammatory disorders, in particular inflammatory disorders associated with the overexpression of inflammatory cytokines, in a subject in need of such treatment, comprising administering to the subject a compound having a (substituted phenyl) -propenal moiety.)

1. A method for treating an inflammatory disorder associated with overexpression of an inflammatory cytokine in a subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising an effective amount of a compound according to formula V or VIII, and optionally a pharmaceutically acceptable carrier or excipient;

wherein in formula V:

each' n₅"is independently 1, 2 or 3;

R₃₅、R₄₅、R₃₅' and R₄₅' is independently selected from the group consisting of: -H, -OH and-OCH₃；

L₅-Z₅Does not storeAt least one of the following steps; or

L₅-Z₅Is present and when Z is₅When not present, L₅Selected from the group consisting of: C0-C8 alkylene, unsaturated alkenylene, and alkynyl;

Z₅selected from the group consisting of: -H, -OH, aromatic ring, cycloalkyl, -COR₁₅、-CO₂R₁₅、-CONR₁₅R₂₅、-NR₁₅R₂₅、-C(X₅)₃；

R₁₅And R₂₅Independently selected from the group consisting of: -H, -CH₃and-C₂H₅(ii) a And is

X₅Is a halogen atom selected from the group consisting of-F, -Cl and-Br;

R₁₈and R₂₈Is a mono or di-substituted group and is independently selected from the group consisting of: methoxy, hydroxy and alkylsulfonyl;

R₃₈selected from the group consisting of:

(CH₂)₃CH₃、and is

R₄₈Selected from the group consisting of: CH (CH)₃H, F and Cl; and is

n₈Is 1 or 2;

wherein the inflammatory cytokine is selected from the group consisting of INF- α, I L-1 β, I L-17A, I L-22, and I L-31.

2. The method of claim 1, wherein the compound is formula V, and "n₅"is 1; r₃₅、R₄₅、R₃₅' and R₄₅' is-OCH₃And L₅-Z₅The side chain is absent.

3. The method of claim 1, wherein the compound is of formula VIII, R₁₈And R₂₈Is disubstituted methoxy, R₃₈Is composed ofR₄₈Is H, and n₈Is 1.

4. The method of claim 1, wherein the pharmaceutical composition is a topical pharmaceutical composition.

5. The method of claim 4, wherein the pharmaceutical composition further comprises an emollient, a skin penetrating agent, an emulsifier, a neutralizer, a gelling agent, a preservative, or a chelating agent.

6. The method of claim 1, wherein the inflammatory cytokine is synthesized by a dendritic cell or a macrophage.

7. The method of claim 1, wherein the disorder is an epidermal disorder.

8. The method of claim 1, wherein the disorder is a dermal disorder.

9. The method of claim 1, wherein the disorder is an inflammatory skin disorder.

10. The method of claim 9, wherein the disorder is actinic keratosis, atopic dermatitis, chickenpox, cold sores, contact dermatitis, eczema, urticaria, impetigo, follicular keratosis, latex hypersensitivity, measles, psoriasis, tinea, rosacea, keloids, hypertrophic scars, pruritus or seborrheic eczema.

11. The method of claim 10, wherein the disorder is psoriasis.

12. The method of claim 10, wherein the condition is pruritus.

13. The method of claim 1, wherein the disorder is an autoimmune disease.

14. The method of claim 13, wherein the autoimmune disease is edison's disease, celiac disease, graft-versus-host disease GvHD, graves' disease, hashimoto's thyroiditis, inflammatory bowel disease, multiple sclerosis, myasthenia gravis, pernicious anemia, psoriatic arthritis, rheumatoid arthritis RA, sjogren's syndrome, systemic lupus erythematosus (lupus), type 1 diabetes, or vasculitis.

15. The method of claim 14, wherein the inflammatory bowel disease is ulcerative colitis or crohn's disease.

Technical Field

The present invention relates to a method of treatment, more precisely, the invention provides a method of treating inflammatory disorders with compounds having at least one (substituted phenyl) -propenal moiety.

Background

It is well known that certain natural products can have therapeutic effects, which have led to their use in many cultures for the treatment and prevention of human diseases (e.g. chinese herbs and many other folk medicines). The effectiveness of such treatments has led the pharmaceutical industry to seek and isolate active compounds from these natural products and to develop active ingredients as therapeutic or prophylactic agents for the treatment and prevention of various diseases or medical conditions. Thus, many commonly used drugs have been developed or produced from natural products. However, compounds isolated from natural products are known to exert certain physiological functions in their natural hosts; and their therapeutic effect against human diseases is not readily apparent. Historically, the therapeutic treatment was only deduced by cumulative experience or "trial and error" in humans. Furthermore, because the compounds are not originally produced for human use, the compounds in their native form are often not structurally and functionally optimal for the treatment of human diseases. Today, however, modern chemical techniques, including analytical and synthetic chemical methods, as well as advances in pharmaceutical biology have made it possible to dissect the chemical structure and locate the "pharmacophore" (the core structure essential for therapeutic activity) within a compound, such as one isolated from a natural product; furthermore, these novel techniques allow the synthesis of novel compounds with optimal or even better therapeutic efficacy based on the structure of the pharmacophore.

The compound curcumin (present as the major pigment in turmeric plants) and many of its analogs have been reported to possess a number of biological activities in vitro, such as antioxidant, anti-inflammatory, anti-tumor and anti-angiogenic activities; however, none of curcumin or its analogs have been developed as therapeutic agents for the treatment of human diseases. This suggests that curcumin in its native form may not be the optimal molecule for development into a therapeutic drug.

Psoriasis is an immune-related chronic inflammatory skin disease affecting the quality of life in 2% to 3% of the global population psoriasis is typically associated with redness, scales, raised plaques, and significant thickening of the epidermis, leukocyte infiltration in the epidermis and dermis induced by inflammation and enhanced keratinocyte proliferation this disease may be caused by a variety of external and internal factors including microbial infection, skin damage, environmental, weather, stress, immune disorders and heredity the leukocyte infiltration in psoriatic lesions contains mainly dendritic cells, macrophages, neutrophils and T cells the dendritic cells produce a variety of pro-inflammatory cytokines that promote the development of psoriasis, including TNF- α, I L-1 β, I L-6 and I L-23. TNF- α are potent pro-inflammatory stimulators produced by I L-23 in activated DCs the I L-1 β may promote I L-17 secretion of I L-6 from cells 17 and I L-6 protect the T cells from T cells and promote a potent pro-inflammatory response to T cell responses in the immune response of T cells induced by the primary immune response of T cells in the T cells induced by T cells.

Disclosure of Invention

The present invention provides a method for treating an inflammatory disorder associated with overexpression of an inflammatory cytokine in a subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising an effective amount of a compound according to formula V or VIII, and optionally a pharmaceutically acceptable carrier or excipient;

wherein in formula V:

each' n₅"is independently 1, 2 or 3;

R₃₅、R₄₅、R₃₅' and R₄₅' is independently selected from the group consisting of: -H, -OH and-OCH₃；

L₅-Z₅Is absent; or

L₅-Z₅Is present and when Z is₅When not present, L₅Selected from the group consisting of: C0-C8 alkylene, unsaturated alkenylene, and alkynyl;

Z₅selected from the group consisting of: -H, -OH, aromatic ring, cycloalkyl, -COR₁₅、-CO₂R₁₅、-CONR₁₅R₂₅、-NR₁₅R₂₅、-C(X₅)₃；

R₁₅And R₂₅Independently selected from the group consisting of: -H, -CH₃and-C₂H₅(ii) a And is

X₅Is a halogen atom selected from the group consisting of-F, -Cl and-Br;

R₁₈and R₂₈Is a mono or di-substituted group and is independently selected from the group consisting of: methoxy, hydroxy and alkylsulfonyl;

R₃₈selected from the group consisting of:

(CH₂)₃CH₃、and is

R₄₈Selected from the group consisting of: CH (CH)₃H, F and Cl; and is

n₈Is 1 or 2;

wherein the inflammatory cytokine is selected from the group consisting of INF- α, I L-1 β, I L-17A, I L-22, and I L-31.

The present invention is described in detail in the following sections. Other features, objects, and advantages of the invention will be apparent from the description and from the claims.

Drawings

FIG. 1 shows the results of cytotoxicity of ASC-J9(J9) and ASC-JM17(JM17) on mouse dendritic cells and macrophages. Cells were treated with different concentrations of ASC-J9 and ASC-JM17 for 24 hours. The viability of the cells was measured by MTS assay. *: p < 0.05.

FIG. 2 shows the effect of ASC-J9(J9) and ASC-JM17(JM17) on inflammatory cytokine-induced cytokine expression in dendritic cells were pretreated with different concentrations of ASC-J9 and ASC-JM17 for 1 hour and then inflammatory cytokines TNF- α or I L-1 β 24 were added for 24 hours.

FIG. 3 shows the effect of ASC-J9(J9) and ASC-JM17(JM17) on R848-induced cytokine expression in mouse splenocytes mice splenocytes pretreatment of mouse splenocytes with 1 μ M ASC-J9 or ASC-JM17 for 1 hour, followed by addition of T L R ligand, R8486 hours expression of mouse cytokine genes was measured by RT-qPCR.

FIG. 4 shows the effect of ASC-J9(J9) and ASC-JM17(JM17) on R848-induced cytokine expression in mouse bone marrow-derived macrophages the macrophages were pretreated with 1 μ M ASC-J9 or ASC-JM17 for 1 hour and then T L R ligand was added, R8486 hours.

Figures 5A to 5D show the evaluation of the anti-inflammatory effect of ASC-J9(J9) on IMQ-induced psoriatic inflammation in vivo. Balb/c mice were treated topically with Imiquimod (IMQ) and with control cream, ASC-J9 or clobetasol (clobetasol) for 10 consecutive days. (A) An exemplary schedule is shown. Photographic observations in each group on day 0 (B), day 5 (C) and day 10 (D) are shown.

Figures 6A to 6D show the evaluation of the anti-inflammatory effect of ASC-J9(J9) on IMQ-induced psoriatic inflammation in vivo. Balb/c mice were treated topically with Imiquimod (IMQ) and with control cream, ASC-J9, or clobetasol for 10 consecutive days. (A) The Severity of the inflammatory response on the skin was assessed based on the Psoriasis Area Severity Index (Psoriasis Area Severity Index). (B) On the tenth day after treatment, mice were sacrificed and their skin thickness was measured with a vernier caliper to assess the severity of the psoriatic response. (C) Cytokine gene expression in mouse skin measured by RT-qPCR after 10 days treatment. (F) H & E staining of the diseased skin after 10 days of treatment. *: p < 0.05; **: p < 0.05; ns: not significant.

Figures 7A to 7C show the evaluation of the anti-inflammatory effect of ASC-J9(J9) on IMQ-induced psoriatic inflammation in vivo. Balb/c mice were treated topically with Imiquimod (IMQ) and with control cream, different doses of ASC-J9 for 5 consecutive days. (A) An exemplary schedule is shown. Photographic observations in each group on day 0 (B), day 5 (C) are shown.

Figures 8A to 8C show the evaluation of the anti-inflammatory effect of ASC-J9(J9) on IMQ-induced psoriatic inflammation in vivo. Balb/c mice were treated topically with Imiquimod (IMQ) and with control cream, different doses of ASC-J9 for 5 consecutive days. (A) The severity of the inflammatory response on the skin was assessed based on the psoriasis area severity index. (B) On the fifth day post-treatment, mice were sacrificed and their skin thickness was measured with a vernier caliper to assess the severity of the psoriatic response. (C) Cytokine gene expression in mouse skin measured by RT-qPCR after 10 days treatment. *: p < 0.05; **: p < 0.05; ns: not significant.

Fig. 9 shows ASC-J9 inhibits itch markers in keloid patient-derived fibroblasts.

Detailed Description

The present invention may be understood more readily by reference to the following detailed description of various embodiments of the invention, the examples and the chemical diagrams and tables having associated descriptions. It is to be understood that, unless otherwise specifically indicated by the claims, the invention is not limited to a particular method of manufacture, carrier, or formulation, or to a particular mode of formulating the compounds of the invention into products or compositions intended for topical, oral, or parenteral administration, as such may, of course, vary, as is well understood by those of ordinary skill in the relevant art. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

The present invention provides a method for treating an inflammatory disorder associated with overexpression of an inflammatory cytokine in a subject in need of such treatment, comprising administering to the subject a pharmaceutical composition comprising an effective amount of a compound according to formula V or VIII, and optionally a pharmaceutically acceptable carrier or excipient;

wherein in formula V:

each' n₅"is independently 1, 2 or 3;

R₃₅、R₄₅、R₃₅' and R₄₅' is independently selected from the group consisting of: -H, -OH and-OCH₃；

L₅-Z₅Is absent; or

L₅-Z₅Is present and when Z is₅When not present, L₅Selected from the group consisting of: C0-C8 alkylene, unsaturated alkenyleneAnd an alkynyl group;

Z₅selected from the group consisting of: -H, -OH, aromatic ring, cycloalkyl, -COR₁₅、-CO₂R₁₅、-CONR₁₅R₂₅、-NR₁₅R₂₅、-C(X₅)₃；

R₁₅And R₂₅Independently selected from the group consisting of: -H, -CH₃and-C₂H₅(ii) a And is

X₅Is a halogen atom selected from the group consisting of-F, -Cl and-Br;

R₁₈and R₂₈Is a mono or di-substituted group and is independently selected from the group consisting of: methoxy, hydroxy and alkylsulfonyl;

R₃₈selected from the group consisting of:

(CH₂)₃CH₃、and is

R₄₈Selected from the group consisting of: CH (CH)₃H, F and Cl; and is

n₈Is 1 or 2;

wherein the inflammatory cytokine is selected from the group consisting of INF- α, I L-1 β, I L-17A, I L-22, and I L-31.

As used herein, the term "(substituted phenyl) -acrolein moiety" refers to a composition comprising a phenyl group having an acrolein moiety (when m equals 1) and an alkoxy or hydroxyl moiety, or an alkyl or substituted alkyl moiety attached thereto. The substitution may be positioned meta or para or ortho with respect to the acrolein moiety as used herein, and refers to the general formulaWherein q may be any number of 1, 2, 3 or 4; and m can be any number of 1, 2, 3,4, or more.

As used herein, the term "alkyl" refers to a straight or branched hydrocarbon chain group consisting of only carbon and hydrogen atoms, containing no unsaturated groups, having from one to ten carbon atoms, and attached to the rest of the molecule by a single bond, such as methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1-dimethylethyl (tert-butyl), and the like.

As used herein, the term "alkenyl" refers to a straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, containing at least one double bond, having two to ten carbon atoms, and attached to the remainder of the molecule by a single or double bond, such as vinyl, prop-1-enyl, pent-1, 4-dienyl, and the like.

The term "alkenylene" as used herein refers to a compound containing a carbon-carbon double bond and represented by the formula C_pH_2p-2Straight or branched hydrocarbon chains are represented wherein the hydrogen may be replaced by additional carbon-carbon double bonds or monovalent substituents (e.g., ethenylene, prop-1-enylene, etc.).

The term "alkoxy" as used herein refers to a group having the formula-OR, wherein R is alkyl, haloalkyl, OR cycloalkyl. "optionally substituted alkoxy" refers to a group having the formula-OR ', wherein R' is optionally substituted alkyl as described herein.

The term "alkynyl" as used herein refers to a straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, containing at least one triple bond, having two to ten carbon atoms, and attached to the remainder of the molecule by a single or triple bond, e.g., ethynyl, prop-1-ynyl, but-1-ynyl, pent-3-ynyl, and the like.

As used herein, the term "aryl" refers to a group of a carbocyclic ring system in which at least one of the rings is aromatic. The aryl group may be fully aromatic or may contain a combination of aromatic and non-aromatic rings. A "biaryl system" is a compound comprising at least two aryl groups.

As used herein, the term "cycloalkyl" refers to a stable monovalent monocyclic or bicyclic hydrocarbon group consisting only of carbon and hydrogen atoms, having three to ten carbon atoms, and being saturated and attached to the rest of the molecule by single bonds, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.

As used herein, the term "diketone bridge" or "keto-enol bridge" refers to a linear or branched hydrocarbon chain containing two ketones or an enol positioned in close proximity to a ketone, respectively. A "diketone bridge" or "keto-enol bridge" is located between at least two aryl moieties.

As used herein, the term "hydroxyalkyl" refers to a straight or branched hydroxy-substituted hydrocarbon chain radical having from one to ten carbon atoms, e.g., -CH₂OH、-(CH₂)₂OH and the like.

In some embodiments, compounds according to formula V are provided according to formula V-1 or V-2:

in a preferred embodiment of the invention, the compound according to formula VIII comprises a 4, 4-disubstituted 1, 7-bis- (3, 4-dimethoxyphenyl) -hepta-1, 6-diene-3, 5-dione and a 6, 6-disubstituted 1, 11-bis (substituted phenyl) -undec-1, 3,8, 10-tetraene-5, 7-dione framework as shown in table 1.

Table 1:

in another preferred embodiment of the invention, the compound is of formula V, and "n₅"is 1; r₃₅、R₄₅、R₃₅' and R₄₅' is-OCH₃And L₅-Z₅The side chain is absent and is designated ASC-J9.

In another preferred embodiment of the invention, the compound is of formula VIII, R₁₈And R₂₈Is disubstituted methoxy, R₃₈Is composed ofR₄₈Is H, and n₈Is 1 and is called ASC-JM 17.

The present invention also provides a composition comprising a compound having a (substituted phenyl) -propenal moiety. The composition according to the invention is preferably a pharmaceutical, food or cosmetic composition.

Preferably, the pharmaceutical composition comprises a compound having a (substituted phenyl) -propenal moiety and optionally a pharmaceutically acceptable carrier or excipient.

Preferably, the pharmaceutical composition comprises an effective amount of the compound.

Typically, ranges are expressed herein as "about" one particular value, and/or to "about" another particular value. When such a range is expressed, an embodiment includes one particular value and/or a range to another particular value. Similarly, when values are expressed as approximations, by use of the word "about," it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint.

"optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not. For example, the phrase "optionally including an agent" means that the agent may or may not be present.

It must be noted that, as used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, unless the context requires otherwise, singular terms shall include the plural and plural terms shall include the singular.

As used herein, the term "individual" means any animal, preferably a mammal, and more preferably a human. Examples of subjects include humans, non-human primates, rodents, guinea pigs, rabbits, sheep, pigs, goats, cows, horses, dogs, and cats.

The term "effective amount" of an active ingredient as provided herein means a sufficient amount of the ingredient to be adjusted to provide the desired function. As will be noted below, the exact amount required will vary from individual to individual depending on the disease state, physical condition, age, sex, species and weight of the individual, the particular characteristics and formulation of the composition, and the like. The dosing regimen may be adjusted to induce an optimal therapeutic response. For example, several divided doses may be administered daily, or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. Therefore, it is not possible to specify an exact "effective amount". However, an appropriate effective amount can be determined by one of ordinary skill in the art using only routine experimentation.

As used herein, the term "treating" or "treatment" means reversing, alleviating, inhibiting the progression of, or improving: the term applies to a disorder, disease, or condition, or one or more symptoms of the disorder, disease, or condition.

As used herein, the term "carrier" or "excipient" refers to any substance that is not a therapeutic agent by itself, that acts as a carrier and/or diluent and/or adjuvant, or vehicle, for delivering a therapeutic agent to an individual or adding to a formulation to improve its handling or storage characteristics, or to permit or facilitate the formation of dosage units of the composition into discrete articles suitable for oral administration, such as capsules or tablets. Suitable carriers or excipients are well known to those of ordinary skill in the art of making pharmaceutical formulations or food products. The carrier or excipient may include, by way of illustration and not limitation, buffers, diluents, disintegrants, binders, adhesion agents, wetting agents, polymers, lubricants, glidants, substances added to mask or counteract an unpleasant taste or odor, flavoring agents, dyes, fragrances, and substances added to improve the appearance of the composition. Acceptable carriers or excipients include citrate buffers, phosphate buffers, acetate buffers, bicarbonate buffers, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acid, magnesium carbonate, talc, gelatin, acacia, sodium alginate, pectin, dextrin, mannitol, sorbitol, lactose, sucrose, starch, gelatin, cellulosic materials (e.g., cellulose esters and cellulose alkyl esters of alkanoic acids), low melting wax cocoa butter, amino acids, urea, alcohols, ascorbic acid, phospholipids, proteins (e.g., serum albumin), Ethylene Diamine Tetraacetic Acid (EDTA), dimethyl sulfoxide (DMSO), sodium chloride or other salts, liposomes, mannitol, sorbitol, glycerin or powders, polymers (e.g., polyvinyl-pyrrolidone, polyvinyl alcohol, and polyethylene glycol), and other pharmaceutically acceptable materials. The carrier should not destroy the pharmacological activity of the therapeutic agent and should be non-toxic when administered at a dose sufficient to deliver a therapeutic amount of the agent.

The pharmaceutical compositions according to the present invention are preferably administered locally or systemically by any method known in the art, including, but not limited to, intramuscular, intradermal, intravenous, subcutaneous, intraperitoneal, intranasal, oral, mucosal or external routes. The appropriate route, formulation and dosing schedule can be determined by one of skill in the art. In the present invention, the pharmaceutical composition may be formulated in various ways according to the respective routes of administration, such as liquid solutions, suspensions, emulsions, syrups, tablets, pills, capsules, sustained-release formulations, powders, granules, ampoules, injections, infusions, kits, ointments, emulsions, liniments, creams or combinations thereof. If necessary, it can be sterilized or mixed with any pharmaceutically acceptable carrier or excipient, many of which are known to those of ordinary skill in the art.

As used herein, external routes are also referred to as topical administration, including, but not limited to, administration by insufflation and inhalation. Examples of various types of formulations for topical administration include ointments, creams, gels, foams, formulations for delivery by transdermal patches, powders, sprays, aerosols, capsules or cartridges for inhalers or insufflators or drops (e.g., eye drops or nasal drops), solutions/suspensions for nebulization, suppositories, pessaries, retention enemas, and chewable or suckable tablets or granules or liposomes or microencapsulated formulations.

Ointments, creams and gels may, for example, be formulated with an aqueous or oily base with the addition of suitable thickening and/or gelling agents and/or solvents. The base may thus for example comprise water and/or an oil, such as liquid paraffin or a vegetable oil, such as peanut oil or castor oil, or a solvent, such as polyethylene glycol. Thickeners and gelling agents which may be used depending on the nature of the base include soft paraffin, aluminium stearate, cetostearyl alcohol, polyethylene glycol, lanolin, beeswax, carboxypolymethylene and cellulose derivatives and/or glyceryl monostearate and/or nonionic emulsifiers.

Emulsions may be formulated with an aqueous or oily base and will generally also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents or thickening agents.

Powders for external application may be formed with the aid of any suitable powder base, such as talc, lactose or starch. Drops may be formulated with an aqueous or non-aqueous base also including one or more dispersing agents, solubilizing agents, suspending agents or preservatives.

The spray compositions may be formulated, for example, as aqueous solutions or suspensions, or as aerosols delivered from pressurized packs (e.g., metered dose inhalers), using a suitable liquefied propellant. Aerosol compositions suitable for inhalation may be suspensions or solutions. The aerosol composition may optionally contain additional formulation excipients well known in the art, such as surfactants, for example oleic acid or lecithin, and co-solvents, for example ethanol.

The topical formulation may be administered to the affected area by one or more applications per day; an occlusive dressing may be advantageously used on the skin area. Continuous or long term delivery can be achieved by a bonded reservoir system.

The cosmetic composition according to the invention may be an aqueous phase formulation consisting essentially of water; it may also include a mixture of water and a water-miscible (miscibility in water greater than 50 wt% at 25 ℃) solvent, for example, a lower monoalcohol containing from 1 to 5 carbon atoms, such as ethanol or isopropanol; diols having 2 to 8 carbon atoms, such as propylene glycol, ethylene glycol, 1, 3-butanediol or dipropylene glycol, C3-C4 ketones and C2-C4 aldehydes; and glycerol. The aqueous formulation is preferably in the form of an aqueous gel or hydrogel formulation. The hydrogel formulation includes a thickener to thicken the liquid solution. Examples of thickeners include, but are not limited to, carbomers, cellulosic based materials, gums, algins, agar, pectins, carrageenans, gelatin, mineral or modified mineral thickeners, polyethylene glycols and polyols, polyacrylamides, and other polymeric thickeners. Thickeners are preferably used which provide stability and optimal flow characteristics of the composition.

The cosmetic composition according to the invention may be in the form of an emulsion or cream formulation. It may contain an emulsifying surfactant. These surfactants may be selected from anionic and nonionic surfactants. With regard to the definition of the properties and function (emulsification) of surfactants, reference may be made to the encyclopedia of chemical Technology, Kock-Othmer, 22, 333-432, 3 rd edition, 1979, Weili (Wiley), in particular, the reference at 347-377 relates to anionic and nonionic surfactants.

Preferably the surfactants used in the cosmetic compositions according to the invention are selected from: nonionic surfactant: fatty acids, fatty alcohols, polyethoxylated or polyglycerolated fatty alcohols (such as polyethoxylated stearyl alcohol or cetylstearyl alcohol), fatty acid esters of sucrose, alkyl glucose esters (specifically, polyoxyethylated fatty esters of C1-C6 alkyl glucose), and mixtures thereof; anionic surfactant: a C16-C30 fatty acid neutralized with an amine, aqueous ammonia or a basic salt, and mixtures thereof. Preferably surfactants are used which make it possible to obtain oil-in-water or wax-in-water emulsions.

The cosmetic composition according to the present invention may further comprise an effective amount of a physiologically acceptable antioxidant selected from the group consisting of: butylated p-cresol, butylated hydroquinone monomethyl ether and tocopherol.

The cosmetic composition according to the present invention may further include natural or modified amino acids, natural or modified sterol compounds, natural or modified collagen, silk protein or soybean protein.

The cosmetic composition according to the invention is preferably formulated for topical application to keratin materials such as the skin, hair, eyelashes or nails. It may be in any presentation form commonly used for this type of application, in particular in the form of an aqueous or oily solution, an oil-in-water or water-in-oil emulsion, a silicone emulsion, a microemulsion or nanoemulsion, an aqueous or oily gel or a liquid, paste-like or solid anhydrous product.

The cosmetic composition according to the invention may be nearly fluid and may have the appearance of a white or colored cream, ointment, milk, lotion, serum, paste, mousse or gel. It may optionally be topically applied to the skin in the form of an aerosol, patch or powder. It may also be in solid form, for example in the form of a stick. It can be used as a skin care product and/or as a cosmetic product for the skin. Alternatively, it can be formulated into shampoo or hair conditioner.

The cosmetic compositions according to the invention may also contain, in a known manner, additives and adjuvants commonly found in cosmetics, such as hydrophilic or lipophilic gelling agents, preservatives, antioxidants, solvents, fragrances, fillers, pigments, odor absorbers and dyes.

The compound may be added to a conventional food composition (i.e., an edible food or beverage or precursor thereof) during the manufacture of the food composition. Almost all food compositions can be supplemented with the compounds of the present invention. Food compositions that can be supplemented with the compounds of the present invention include, but are not limited to, confections, baked goods, ice creams, dairy products, sweet and flavored snacks, snack bars, meal replacement products, snack foods, soups, pasta, noodles, canned foods, frozen foods, dried foods, refrigerated foods, oils and fats, baby foods, or soft foods coated on bread, or mixtures thereof.

In a preferred embodiment of the invention, the pharmaceutical composition is a topical pharmaceutical composition.

In a preferred embodiment of the present invention, the pharmaceutical composition further comprises an emollient, a skin penetrating agent, an emulsifier, a neutralizing agent, a gelling agent, a preservative or a chelating agent.

The present invention also provides a method for treating an inflammatory disorder associated with the overexpression of an inflammatory cytokine in a subject in need of such treatment, comprising administering to the subject a pharmaceutical composition as mentioned above.

While not wishing to be bound by theory, applicants believe that the method according to the invention treats the condition by inhibiting inflammatory cytokines synthesized by activated dendritic cells, macrophages or T cells.

Preferably, the inflammatory cytokines include, but are not limited to, INF- α, I L-1 β, I L-17A, I L-22, and I L-31.

In a preferred embodiment of the invention, the disorder is an epidermal disorder.

In another preferred embodiment of the invention, the disorder is a dermal disorder.

In another aspect, the disorder is preferably an inflammatory skin disorder.

Preferably, the inflammatory skin condition comprises, but is not limited to, actinic keratosis, atopic dermatitis, chickenpox, cold sores, contact dermatitis, eczema, urticaria, impetigo, follicular keratosis, latex hypersensitivity, measles, psoriasis, tinea, rosacea, keloids, hypertrophic scars, pruritus or seborrheic eczema.

The present invention also provides a method for treating an autoimmune disease in a subject in need of such treatment, comprising administering to the subject a pharmaceutical composition as mentioned above.

In a preferred embodiment of the invention, the autoimmune disease comprises, but is not limited to, Addison' sdisease), celiac disease, graft-versus-host disease (GvHD), Graves ' disease, Hashimoto's thyroiditis, inflammatory bowel disease, multiple sclerosis, myasthenia gravis, pernicious anemia, psoriatic arthritis, Rheumatoid Arthritis (RA), sjogren's syndrome (syndrome), systemic lupus erythematosus (lupus), type 1 diabetes, or vasculitis.

Preferably, the inflammatory bowel disease comprises, but is not limited to, ulcerative colitis or Crohn's disease.

In one embodiment of the invention, we have established an Imiquimod (IMQ) -induced psoriatic model in which imiquimod induces psoriatic inflammation on the skin of mice similar to the psoriatic response of humans.We evaluated ASC-J9 and related compounds for the treatment of psoriatic inflammation using immune cells comprising dendritic cells and macrophages and this IMQ-induced psoriatic mouse model in an in vitro study, we found that ASC-J9 and ASC-JM17 can inhibit inflammatory cytokines or T L R ligand-induced cytokine expression in mouse splenocytes, mouse bone marrow-derived dendritic cells, and bone marrow-derived macrophages.in addition, we observed that the in vivo ASC-J9 cream reduces IMQ-induced psoriatic inflammation.J 2 also reduces IMQ-induced psoriatic inflammatory inflammation, which comprises reducing erythema, reducing scaling and thickness of the lesioned skin, and inhibiting inflammatory gene expression in psoriatic skin.

In a preferred embodiment of the invention, ASC-J9 inhibits the itch marker in keloid patient-derived fibroblasts. While not wishing to be bound by theory, applicants believe that ASC-J9 inhibits pruritus by down-regulating Attominin (ARTN) expression and Transient Receptor Potential (TRP) channels in patient-derived keloid fibroblasts. Overexpression of ARTN was shown to enhance expression of TRPV1 and TRPA1 in cutaneous sensory neurons that cause pruritus (molecular Pain (Mol Pain) 2015 3,8 days; 11: 8; pharmaceutical (basel) s (pharmaceuticals (basel) 2018, 12 months; 11(4): 100).

The following examples are provided to assist those skilled in the art in practicing the invention.

Examples of the invention

Example 1: psoriasis vulgaris

Materials and methods

Storage conditions of the test article

ASC-J9(J9) and ASC-JM17(JM17) powders for in vitro studies were dissolved in DMSO and stored at-20 ℃.

The J9 control cream and the J9 cream, which contained 0.05% J9, 0.1% J9, and 0.5% J9, were stored at room temperature.

Dose preparation

J9 and JM17 powders were dissolved in DMSO as stock solutions to 10mg/m L, and then serially diluted to various concentrations for studies.

The test system comprises:

MTS assay for cell viability: the cytotoxicity of the test substance was measured by MTS assay. The MTS assay is based on the reduction of MTS tetrazolium compounds by viable cells to produce colored formazan dyes that are soluble in cell culture medium. This conversion is believed to be carried out by NAD (P) H-dependent dehydrogenases in metabolically active cells. The formazan dye was quantified by measuring absorbance at 490-500 nm. A non-toxic dose will be administered for further analysis.

Cell culture mouse bone marrow-derived dendritic cells (BMDCs) and bone marrow-derived macrophages (BMDMs) were produced from mouse bone marrow cells briefly, dendritic cells can be produced by two induction procedures.bone marrow cells can be induced into bone marrow DCs called GMDCs using GM-CSF and I L-4 for 10 days, while mixed type DCs containing bone marrow and plasma cell DCs called Fit3DC can be produced by induction with Fit 3-ligand for 10 days.to produce BMDM, bone marrow cells are induced into BMDM using 30% L929 modified medium for 7 days.

Analysis of gene expression: according to the manufacturer's protocol, use of Isol-RNA dissolving reagent (De)Total RNA was purified from cells by the national Hilden 5PRIME company (5PRIME GmbH, Hilden, Germany)). Use ofReverse transcription kit (QIAGEN)^TM) Reverse transcription with oligo-dT primer for the first cDNA Synthesis Using L ightCycler480 detection System (Roche Diagnostics GmbH, Penzberg, Germany) andSYBR Green RT-PCR kit, quantitative PCR with gene specific primers (see appendix) for gene expression analysis. mRNA expression levels were normalized to the expression level of GAPDH.

Animal models of psoriatic inflammation: 30 or 40mg of 5% IMQ gel (Aldara) per day^TM) The severity of inflammatory responses of the skin was assessed on the basis of the Psoriasis Area Severity Index (PASI) as described, briefly, three parameters of the psoriasis response (erythema, scaling and skin thickness) were independently scored on a scale of 0 to 4, 0: none, 1: mild, 2: moderate, 3: significant, and 4: very significant, the severity of the response was measured using an accumulated score of 0 to 12 by adding the scores from these three parameters, at the end of the experiment, the mice were sacrificed to measure skin thickness with a vernier caliper, the grouping and dosage of the animal studies are described as follows, two animal experiments are listed in tables 2 and 3:

table 2: animal experiment No. 1:

table 3: animal experiment No. 2:

statistical analysis: all data are presented as mean ± SD. Statistical analysis of data from three independent experiments was performed using the schraden t-test (Student's t-test). P values <0.05 were considered statistically significant differences in the experimental groups.

Data analysis

In vitro anti-inflammatory Activity of J9 and JM17 on macrophages and dendritic cells

In vitro toxicity testing was performed by MTS assay.

The anti-inflammatory effects of J9 and JM17 on inflammatory cytokines induced by T L R ligand, including TNF- α, I L-1 β, I L-17A, I L-22, I L-23, and I L-31, in various immune cells, including mouse splenocytes, mouse bone marrow-derived dendritic cells, and bone marrow-derived macrophages, were analyzed by quantitative PCR (RT-qPCR).

Inhibition of J9 and JM17 in animal models with Imiquimod (IMQ) -induced psoriatic inflammation.

Mice were (1) photographed for evaluation, and analyzed for (2) Psoriasis Area Severity Index (PASI), (3) skin thickness measurement We selected samples with better inhibition in the experiment for further analysis by taking (1) H & E staining, and (2) inflammatory gene expression comprising TNF- α, I L-1 β, I L-17A, I L-22, I L-23, and I L-31 in diseased tissues as measured by quantitative PCR (RT-qPCR).

Results

In vitro anti-inflammatory Activity of J9 and JM17 on macrophages and dendritic cells

The objective of this example was to evaluate the anti-inflammatory capacity of J9 and JM17 against psoriatic inflammation in vitro and in vivo. To find safe doses of J9 and JM17 for in vitro studies, the cytotoxicity of J9 and JM17 was measured by MTS assay. Given that dendritic cells and macrophages are important cells for eliciting psoriatic inflammation, these two cells were selected for cytotoxicity testing. These cells were treated with different concentrations of J9 and JM17 from 0. mu.M to 10. mu.M. As shown in fig. 1, GMDC, Fit3DC and BMDM were up to 10 μ M, 10 μ M and 1 μ M resistant to J9 (non-toxic dose), respectively. Furthermore, resistance of GMDC, Fit3DC and BMDM to JM17 (non-toxic dose) was up to 5. mu.M, 3. mu.M and 1. mu.M, respectively.

To evaluate the anti-inflammatory effects of J and JM in vitro, GMDC was pretreated with J and JM for 1 hour and stimulated with inflammatory cytokines, TNF-or I-1 for 24 hours the expression of cytokines including TNF-0, I1-1 5, I4-17 7-22, I9-23 and I0-31 was measured by RT-qPCR in FIG. 2, J and JM significantly inhibited TNF-3 or I2-1 8 induced TNF-6 and I4-1 expression in GMDC, but J and JM did not attenuate TNF-1 or I5-1 3 induced I-22 expression in GMDC, I-17A expression was not induced by TNF-or I-1, moreover, I-23 and I-31 expression was not detectable in dendritic cells in the experimental group.

We also evaluated the anti-inflammatory effects of J9 and JM17 on mouse splenocytes and bone marrow-derived macrophages J9 and JM17 pre-treated mouse splenocytes for 1 hour, and then added T L R ligand R8486 hours RT-qPCR analysis revealed that J9 and JM17 significantly inhibited R848-induced TNF- α, I α 1-1 α 0, I α 2-17A, and I α 3-22 expression in mouse splenocytes, but the expression of I α 5-23 and I α 7-31 in mouse splenocytes was undetectable (fig. 3.) in BMDM, we found that R848-induced expression of TNF- α 4, I L-1 α 6, I L-17A, I L-22, and I L-31 was inhibited by J9, and the expression of R848-induced TNF- α, I L-1 β, and I5L-17A was inhibited by JM17 (fig. 3).

Inhibition of J9 and JM17 in animal models with Imiquimod (IMQ) -induced psoriatic inflammation.

We further evaluated the in vivo, meso-inflammatory effects of J9 on psoriatic inflammation. First, we compared the effects of the J9 control cream, the J9 cream, and clobetasol on inhibiting IMQ-induced psoriatic inflammation over the 10-day experimental schedule (fig. 5A). Mice of each group were photographed for evaluation on day 0, day 5 and day 10 (fig. 5B, 5C and 5D). After 5 days of treatment, the diseased skin of the IMQ-treated group was observed to have severe erythema, flaking and increased skin thickness (fig. 5B). In the control cream group, no inhibition was observed (fig. 5B). J9(2 mg/cm)²And 10mg/cm²) Treatment significantly reduced erythema, scaling and thickness of the affected skin (fig. 5B). In addition, treatment with the corticosteroid drug clobetasol significantly inhibited IMQ-induced erythema, scaling, and skin thickness increase in lesional skinPlus (fig. 5B). However, clobetasol treatment significantly reduced skin thickness (fig. 5B and 6B). In this experiment, we found that these parameters did not increase at day 10 compared to the severity of erythema, scaling and increased skin thickness at day 5. These phenomena reached the most severe on day 5 (fig. 5 and 6A). The Psoriasis Area Severity Index (PASI) scores of these groups were compared and we found that 2mg/cm was passed from day 3 to day 10²The PASI score was significantly reduced in the group of J9-treated mice. From day 4 to day 6, at a rate of 10mg/cm²The group of J9-treated mice was significantly reduced (fig. 6A). Skin thickness was measured on day 10. J9(2 mg/cm)²And 10mg/cm²) Treatment significantly inhibited IMQ-induced thickness increase (fig. 6B) we also found that clobetasol treatment reduced skin thickness compared to sham-treated controls (fig. 6B) expression of cytokine genes including TNF- α, I L-1 β, I L-17A, I L-22, I L-23, and I L-31 in diseased skin was also analyzed, among these cytokines we observed that J9 attenuated IMQ-induced I L-1 β expression on day 10 (fig. 6C)&The E staining results revealed that J9 treatment reduced the thickness of epidermis and dermis, and keratinocyte proliferation and inflammation in the epidermis (fig. 6D).

To further understand the therapeutic effect of the different concentrations of the J9 cream, 0.5% J9(2 mg/cm) was used²And 6mg/cm²)、0.1％J9(10mg/cm²) And 0.05% J9 cream (2 mg/cm)²) The therapeutic effect on IMQ-induced psoriatic inflammation was evaluated in a five day experimental schedule (fig. 7A). Mice of each group were photographed for evaluation on day 0 and day 5 (fig. 7B and 7C). After 5 days of treatment, we found 0.5% J9(2 mg/cm)²And 6mg/cm²) And 0.1% J9(10 mg/cm)²) IMQ-induced inflammation was inhibited in treated mice, but with 0.05% J9 cream (2 mg/cm)²) The treated mice were not inhibited (fig. 7C). PASI scores, skin thickness and gene expression of diseased skin were also measured. With 0.5% J9(2 mg/cm)²And 6mg/cm²) And 0.1% J9(10 mg/cm)²) PASI scores of treated mice decreased significantly from day 4 to day 5 (fig. 8A). Similar results were also observed in skin thickness (fig. 8B). After the reaction with 0.5% J9(2 mg/cm)²And 6mg/cm²) And 0.1% J9(10 mg/cm)²) IMQ-induced expression of TNF- α and I L-1 β was also inhibited in the lesional skin in treated mice (fig. 8C) we also analyzed the induction and inhibition of I L-17A, I L-22, I L-23 and I L-31 in lesional skin by IMQ and J9 in this experiment, but no significant results were obtained due to the lower expression levels of these cytokines.

Example 2: itching (pruritus)

Two keloid patient-derived fibroblasts, designated 256K and 268K, were treated with ASC-J9 for 24 hours. Cells were then harvested and mRNA expression of ARTN and GAPDH was determined by qPCR.

As shown in fig. 9, ASC-J9 inhibited the itch marker in keloid patient-derived fibroblasts. ASC-J9 suppresses pruritus by down-regulating attritor factor (ARTN) expression in patient-derived keloid fibroblasts.

While the present invention has been described in conjunction with the specific embodiments set forth above, many alternatives, modifications, and variations thereof will be apparent to those of ordinary skill in the art. All such alternatives, modifications and variations are considered to be within the scope of the present invention.