WO2023232123A1 - 一类倍半萜聚酮化合物作为免疫调节剂在防治免疫性疾病的用途 - Google Patents

一类倍半萜聚酮化合物作为免疫调节剂在防治免疫性疾病的用途 Download PDF

Info

Publication number
WO2023232123A1
WO2023232123A1 PCT/CN2023/097904 CN2023097904W WO2023232123A1 WO 2023232123 A1 WO2023232123 A1 WO 2023232123A1 CN 2023097904 W CN2023097904 W CN 2023097904W WO 2023232123 A1 WO2023232123 A1 WO 2023232123A1
Authority
WO
WIPO (PCT)
Prior art keywords
formula
compound
salt
sesquiterpene
disease
Prior art date
Application number
PCT/CN2023/097904
Other languages
English (en)
French (fr)
Inventor
尹芝南
高昊
唐欣
王传喜
杨恒文
姚新生
李振华
胡丹
任菲菲
邝润桥
韩雪
Original Assignee
暨南大学
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 暨南大学 filed Critical 暨南大学
Publication of WO2023232123A1 publication Critical patent/WO2023232123A1/zh

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
    • C07D209/44—Iso-indoles; Hydrogenated iso-indoles
    • C07D209/46—Iso-indoles; Hydrogenated iso-indoles with an oxygen atom in position 1
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/11—Aldehydes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
    • A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
    • A61K31/222—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin with compounds having aromatic groups, e.g. dipivefrine, ibopamine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/404—Indoles, e.g. pindolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/06—Antiasthmatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/10—Anti-acne agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/14—Drugs for disorders of the endocrine system of the thyroid hormones, e.g. T3, T4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C47/00—Compounds having —CHO groups
    • C07C47/52—Compounds having —CHO groups bound to carbon atoms of six—membered aromatic rings
    • C07C47/56—Compounds having —CHO groups bound to carbon atoms of six—membered aromatic rings containing hydroxy groups
    • C07C47/57—Compounds having —CHO groups bound to carbon atoms of six—membered aromatic rings containing hydroxy groups polycyclic
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C47/00—Compounds having —CHO groups
    • C07C47/52—Compounds having —CHO groups bound to carbon atoms of six—membered aromatic rings
    • C07C47/575—Compounds having —CHO groups bound to carbon atoms of six—membered aromatic rings containing ether groups, groups, groups, or groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
    • C07C69/02—Esters of acyclic saturated monocarboxylic acids having the carboxyl group bound to an acyclic carbon atom or to hydrogen
    • C07C69/12—Acetic acid esters
    • C07C69/21—Acetic acid esters of hydroxy compounds with more than three hydroxy groups
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
    • C12P17/10—Nitrogen as only ring hetero atom
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/62—Carboxylic acid esters
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/07—Optical isomers
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2602/00—Systems containing two condensed rings
    • C07C2602/02—Systems containing two condensed rings the rings having only two atoms in common
    • C07C2602/04—One of the condensed rings being a six-membered aromatic ring
    • C07C2602/10—One of the condensed rings being a six-membered aromatic ring the other ring being six-membered, e.g. tetraline
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the invention belongs to the field of natural medicine, and specifically relates to the use of a class of sesquiterpene polyketide compounds as immunomodulators in the preparation of medicines for preventing or treating immune-related diseases.
  • autoimmune diseases include inflammatory bowel disease (IBD), multiple sclerosis (MS) and rheumatoid arthritis (RA). ), including more than 100 kinds of diseases, have severe morbidity, high incidence rate, long onset period, no cure, and seriously endanger human health.
  • Immunomodulators can be used to treat immune diseases caused by immune dysfunction by regulating the body's immune response (enhancement, suppression or bidirectional regulation).
  • modulators that enhance the body's immunity can be used to intervene.
  • immunosuppressants are currently mainly used to improve disease symptoms and prognosis.
  • CD4 + T cells differentiate into Th17 cells with the participation of TGF ⁇ and IL6.
  • IL17 is a strong pro-inflammatory cytokine mainly secreted by Th17 cells, which mediates a variety of inflammatory and autoimmune diseases [2] .
  • IL17 plays a role in inflammatory bowel disease, type 1 diabetes, multiple sclerosis, rheumatoid arthritis, psoriasis, systemic lupus erythematosus, asthma, Sjogren's syndrome and ankylosing spondylitis.
  • IL17 inhibitors have been used clinically for therapeutic intervention in various immune diseases such as psoriasis, arthritis, and multiple sclerosis, and have shown certain efficacy [3] .
  • IL10 is a class of cytokines with immunosuppressive properties and broad anti-inflammatory properties. IL10 can regulate the activity of a variety of myeloid or lymphoid cells, inhibit their production of pro-inflammatory factors, and thus play a role in inhibiting immune rejection, allergies, inflammation and autoimmune diseases in organ transplantation [4] .
  • Type I regulatory T cells (Tr1) secrete a large amount of IL10 and are a type of cell with immunosuppressive effects.
  • Tr1 secrete a large amount of IL10 and are a type of cell with immunosuppressive effects.
  • Tr1 secrete a large amount of IL10 and are a type of cell with immunosuppressive effects.
  • Currently, clinical trials of transferring Tr1 cells to prevent and treat autoimmune diseases or protect immune rejection of organ transplants have shown efficacy [5] .
  • compounds that inhibit IL17 and/or promote IL10 can be used as immunomodulators to prevent and treat organ transplant immune rejection, allergies, inflammation and autoimmune diseases (inflammatory bowel disease, type 1 diabetes, multiple sclerosis, rheumatoid arthritis, silver Sclerosis, systemic lupus erythematosus, asthma, Sjogren's syndrome and ankylosing spondylitis, uveitis, autoimmune nephritis, autoimmune thyroiditis, Behcet's disease, lichen planus, various respiratory diseases, various liver diseases and various cardiovascular diseases, etc.).
  • organ transplant immune rejection inflammatory bowel disease, type 1 diabetes, multiple sclerosis, rheumatoid arthritis, silver Sclerosis, systemic lupus erythematosus, asthma, Sjogren's syndrome and ankylosing spondylitis, uveitis, autoimmune nephritis, autoimmune thyroiditis, Behcet's disease, lichen planus, various respiratory diseases, various
  • compounds that inhibit IL17 and/or promote IL10 can also be used as immunomodulators for the treatment of inflammation-related diseases such as acne, disc herniation, and celiac disease.
  • IL17 is also closely related to the occurrence of tumors, especially inflammation-related tumors.
  • IL10 has a bidirectional regulatory effect on the occurrence and development of tumors.
  • IL10 can exert anti-tumor effects by inhibiting the production of inflammatory factors that promote tumor development and activating tumor-specific CD8 + T [4] . Therefore, compounds that inhibit IL17 and/or promote IL10 can also be used as immunomodulators for tumor treatment.
  • DC Dendritic cells
  • BMDC mouse bone marrow-derived dendritic cells
  • Th2 cells mainly secrete the cytokine IL4.
  • IL4 has a variety of biological functions and plays an important role in humoral immunity and acquired immunity [7] .
  • IL4 can induce the activation of B cells and T cells and the differentiation of B cells into plasma cells.
  • IL4 can also inhibit lymphocyte apoptosis, promote the antigen presentation ability of macrophages, and improve the function of killing tumor cells. Therefore, compounds that promote IL4 can be used as immunomodulators for diseases related to anti-tumor and anti-infection (especially anti-parasitic infection).
  • the object of the present invention is to provide a class of sesquiterpene polyketide compounds, pharmaceutical compositions and their uses. Specifically, the inventor discovered and isolated and identified a type of sesquiterpene polyketide compound from a strain of Rhizoma fungus. Experiments have proven that it has immunomodulatory activity and can be used to prevent and treat immune diseases, especially inflammatory bowel disease, Autoimmune diseases such as multiple sclerosis, psoriasis and sepsis.
  • a first aspect of the present invention provides the use of a class of sesquiterpene polyketides or pharmaceutically acceptable salts thereof as immunomodulators.
  • the sesquiterpene polyketides are represented by formula (I):
  • R 2 is selected from: methyl, hydroxymethyl, acetylhydroxymethyl, aldehyde, or dimethoxymethyl;
  • R 3 is selected from: methyl, hydroxymethyl, acetylhydroxymethyl, or aldehyde;
  • R 2 , R 3 and benzene ring form a 5-membered lactam ring or lactone ring;
  • R 4 , R 5 , R 6 and R 7 are carbon atoms, and any adjacent group is selected from double bonds, and the remaining groups are selected from single bonds.
  • the pharmaceutically acceptable salt is a salt formed by a compound of formula (I) and an organic base or an inorganic base.
  • the salt formed is sodium salt, potassium salt, calcium salt, iron salt, magnesium salt, zinc salt, aluminum salt, barium salt or ammonium salt.
  • the sesquiterpene polyketide compound is selected from compounds of formula II to formula XII, and the specific structural formula is as follows:
  • a second aspect of the present invention provides a method for preparing the above-mentioned compound, which includes: fermenting a microorganism that produces a sesquiterpene polyketide compound represented by formula (I) and then separating it by chromatography.
  • microorganism is a fungus of the genus Lactobacillus.
  • the fungus of the genus Lactobacillus is ZLW0801-19, and its preservation number is CGMCC No. 19039.
  • the third aspect of the present invention provides a pharmaceutical composition for immunomodulation, including the above-mentioned sesquiterpene polyketide compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.
  • the salt is any one of sodium salt, potassium salt, calcium salt, iron salt, magnesium salt, zinc salt, aluminum salt, barium salt and ammonium salt.
  • the amount of the active ingredient (i.e., the compound of the present invention) contained in the pharmaceutical composition can be specifically applied according to the patient's condition and the situation diagnosed by the doctor.
  • the amount or concentration of the active compound is within a relatively small range. Adjusted within a wide range, the content of the compound of formula (I) or a pharmaceutically acceptable salt thereof is 1-90% by weight of the composition.
  • pharmaceutically acceptable carriers include diluents, lubricants, binders, disintegrants, stabilizers, solvents, etc.
  • diluents of the present invention include but are not limited to starch, microcrystalline cellulose, sucrose, dextrin, lactose, powdered sugar, glucose, etc.
  • lubricants include but are not limited to magnesium stearate, stearic acid, and sodium chloride.
  • the binders include but are not limited to water, ethanol, starch slurry, syrup, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, seaweed sodium bicarbonate, polyvinylpyrrolidone, etc.
  • the disintegrants include but are not limited to starch effervescent mixture, namely sodium bicarbonate and citric acid, tartaric acid, low-substituted hydroxypropyl cellulose, etc.
  • the stabilizers include but are not limited to polysaccharides Such as acacia gum, agar, alginic acid, cellulose ether, carboxymethyl chitosan, etc.
  • the solvent includes but is not limited to water, balanced salt solution, etc.
  • the pharmaceutical composition is an oral preparation or injection; preferably, the oral preparation includes but is not limited to ordinary tablets, dispersible tablets, enteric-coated tablets, granules, capsules, dropping pills, powders, oral liquids or Any one of emulsions; preferably, the injection is selected from any one of small water injections, infusions or freeze-dried powder injections.
  • the fourth aspect of the present invention also provides the above-mentioned compound of formula (I) or a pharmaceutically acceptable salt thereof for preparing an immunomodulator.
  • the immunomodulatory agent is used for the prevention and/or treatment of immune-related diseases.
  • the immunomodulator is used to prevent and treat diseases or conditions for which inhibiting IL17 and/or promoting IL10 is beneficial.
  • the disease or indication is selected from the group consisting of psoriasis, inflammatory bowel disease, type I diabetes, multiple sclerosis, rheumatoid arthritis, systemic lupus erythematosus, asthma, Sjogren's syndrome, ankylosing spondylitis , immune rejection of organ transplantation, allergies, sepsis, uveitis, autoimmune nephritis, autoimmune thyroiditis, Behcet's disease, autoimmune liver disease, lichen planus, acne, disc herniation, and celiac disease.
  • the immunomodulatory agent is used for diseases or indications in which promoting IL4 is beneficial.
  • the disease or indication is related to parasitic infection.
  • the immunomodulatory agent is used to prevent and treat diseases or indications for which suppression of DC immune response is beneficial.
  • the disease or indication is inflammatory and autoimmune diseases, sepsis, and organ transplant rejection.
  • the sesquiterpene polyketide compound of the present invention is derived from microbial fermentation and is convenient for large-scale fermentation and industrial preparation; through cell and animal activity tests, it was first discovered that this type of compound has significant immunomodulatory activity.
  • Figure 1 shows that the compound of formula (III) significantly inhibits the expression of human IL17.
  • Figure 2 shows that the compound of formula (III) inhibits the ratio of IL12/IL23p40 + , IL6 + and TNF ⁇ + cells in LPS-stimulated BMDC.
  • Figure 3 shows that the compound of formula (III) significantly inhibited the weight loss (Figure 3A), increased DAI score (Figure 3B), and colon shortening (Figure 3C) of mice with enteritis; colon histopathological analysis showed that formula (III ) compounds significantly inhibited the extent of colon inflammation, epithelial damage, and intestinal structural lesions ( Figures 3D and 3E).
  • Figure 4 shows: Compound of formula (III) significantly reduced the EAE score of EAE model mice (Figure 4A); spinal cord tissue pathological analysis showed that treatment with compound of formula (III) significantly inhibited inflammatory cell infiltration and demyelination in the spinal cord. extent ( Figure 4B).
  • Figure 5 shows: Compared with the model group, treatment with the compound of formula (III) significantly inhibited the ear thickening of psoriasis-like mice ( Figures 5A and 5B); the results of ear histopathological analysis showed that treatment with the compound of formula (III) Significantly inhibited the pathological damage of parakeratosis, hyperkeratosis, Munro body formation, acanthosis, inflammatory cell infiltration, and dermal blood vessel dilation and congestion in the ear skin of psoriasis-like mice ( Figures 5C and 5D).
  • Figure 6 shows that the compound of formula (III) significantly improves the survival rate of mice with LPS-induced sepsis.
  • Figure 7 shows that the compound of formula (III) significantly inhibits the level of ALT in the serum of mice with liver injury.
  • the mass spectrometer is an LCQ-Advantage mass spectrometer produced by Finnigan Company in the United States.
  • the superconducting NMR instrument was BrukerAV-400.
  • Silica gel GF254 for thin layer chromatography and silica gel for column chromatography (200-300 mesh) are both products of Qingdao Ocean Chemical Factory.
  • the reversed-phase ODS filler 50 ⁇ m is a product of Japan YMC Company.
  • the medium and low pressure liquid chromatograph is a product of Shanghai Lisui Electronic Technology Co., Ltd.
  • the chromatographic column used for liquid phase separation is Phenomenex Gemini C18 column (10.0 ⁇ 250mm, 5 ⁇ m). Methanol and acetonitrile used for liquid chromatography were of chromatographic grade, water was double distilled water, and other reagents were of analytical grade.
  • the in vitro cell level experimental data shown in the examples are expressed as mean ⁇ standard deviation (Mean ⁇ SD), and the animal experimental data are expressed as mean ⁇ standard error (Mean ⁇ SEM).
  • Two-tailed Student's t test was used for analysis; to compare the differences between more than 2 groups of data, One-wayAnova was used for analysis; for each subject at different time points, For repeated measurement data (such as weight measurement, ear thickness measurement), repeated measures analysis of variance in Two-wayAnova was used to compare differences between groups.
  • Statistical differences are marked as follows: * represents P ⁇ 0.05, ** represents P ⁇ 0.01, *** represents P ⁇ 0.001, **** represents P ⁇ 0.0001, ns represents not significant difference.
  • the acquired data were analyzed using Graphpad Prism 7.00 and FlowJo V10 (for flow cytometry), and all pathology sections were evaluated by pathologists.
  • Example 1 Large-scale fermentation of the fungus ZLW0801-19 and its sample pretreatment method
  • the fungal strain ZLW0801-19 of the genus ZLW0801-19 was cultured on potato dextrose agar (PDA) slant at 25°C for 5 days. After activation on the PDA slope, inoculate into four Erlenmeyer flasks (250 mL) containing potato dextrose (PDB) medium to prepare seed liquid. Each Erlenmeyer flask contains 100 mL of PDB medium. The rotation speed is 200 rpm and cultured at 25°C for 5 days to prepare the seed liquid. . Fermentation was carried out in 24 Erlenmeyer flasks (500mL), each containing 70g of rice.
  • PDA potato dextrose agar
  • Distilled water (105mL) was first added to each Erlenmeyer flask, the rice was soaked overnight and then autoclaved at 120°C. 30 minutes. After cooling to room temperature, 5.0 mL of seed solution was inoculated into each Erlenmeyer flask and cultured at room temperature in the dark for 51 days.
  • Example 1 the crude extract of ethyl acetate (81.7g) was subjected to silica gel column chromatography, using cyclohexane-ethyl acetate (100:0, 98:2, 95:5, 90:10, 80:20, 70:30, 50:50, 0:100, v/v) and methanol were eluted, and each gradient elution volume was 6L to obtain 7 fraction samples (F1-F7).
  • Fraction F5 was subjected to medium and low pressure liquid phase ODS column chromatography, and was eluted sequentially with methanol-water (70:30, 80:20, 90:10, and 100:0, v/v), and each gradient elution volume was 2.5L.
  • Fraction F5.1-F5.9 9 fraction samples (F5.1-F5.9) were obtained.
  • Fraction F5.2 was subjected to medium and low pressure liquid phase ODS column chromatography, and was eluted sequentially with methanol-water (60:40, 70:30, 80:20, 90:10, v/v), with each gradient elution volume of 0.7 L, 5 fraction samples (F5.2.1-F5.2.5) were obtained.
  • Fraction F5.2.4 (387 mg) was prepared by reverse-phase preparative HPLC (Cosmosil Packed C18 column), using acetonitrile-water (50:50, v/v) with a flow rate of 3 mL/min for elution to obtain compound of formula (X) ( t R :53.6min, 26.2mg).
  • Fraction F5.2.5 (365 mg) was prepared by reverse-phase preparative HPLC (Phenomenex, Packed C18 column), and acetonitrile-water (55:45, v/v) with a flow rate of 8 mL/min was used for elution to obtain formula (XII) Compound (t R :86.4min, 7.0mg).
  • Fraction F5.3 was subjected to gel column LH-20 column chromatography and eluted with methanol to obtain 7 fractions (F5.3.1–F5.3.7).
  • Fraction F5.3.7 (144mg) was prepared by reversed-phase preparative HPLC (Cosmosil Packed C18 column), using methanol-water (75:25, v/v) with a flow rate of 3mL/min for elution, to obtain compound of formula (II) ( t R : 63.7 min, 17.0 mg) and the compound of formula (V) ( t R : 69.9 min, 17.5 mg).
  • Fraction F5.5 (1550 mg) was prepared by reverse-phase preparative HPLC (Phenomenex, Packed C18 column), and methanol-water (75:25, v/v) with a flow rate of 8 mL/min was used for elution to obtain formula (IV) Compound (t R :73.5min, 507.0mg).
  • Fraction F5.7 was subjected to silica gel column chromatography, using cyclohexane-ethyl acetate (90:10, 85:15, 80:20, 75:25, 70:30, 60:40, 50:50, 0: 100, v/v), each gradient elution volume was 1.0L, and 8 fraction samples (F5.7.1-F5.7.8) were obtained.
  • Fraction F5.7.4 (62.1 mg) was prepared by reverse-phase preparative HPLC (Cosmosil Packed C18 column), using methanol-water (82:18, v/v) with a flow rate of 3 mL/min for elution, to obtain the compound of formula (VI) (t R :21.3min,12.1mg).
  • Fraction F6 was subjected to medium and low pressure liquid phase ODS column chromatography, and was eluted with methanol-water (60:40, 70:30, 80:20, 90:10, 100:0, v/v) in sequence, with each gradient elution volume 2.5 L, 7 fraction samples (F6.1-F6.7) were obtained.
  • Fraction F6.3 was subjected to silica gel column chromatography, and eluted with cyclohexane-ethyl acetate (75:25, 70:30, 60:40, 50:50, 0:100v/v) in sequence, each gradient elution The volume is 0.3L, and 6 fraction samples (F6.3.1-F6.3.6) are obtained.
  • Fraction F6.3.6 (349.7 mg) was prepared by reverse-phase preparative HPLC (Cosmosil Packed C18 column), and acetonitrile-water (40:60, v/v) with a flow rate of 3 mL/min was used for elution to obtain the compound of formula (IX) (t R :17.5min,7.7mg).
  • Fraction F6.4 was subjected to medium and low pressure liquid phase ODS column chromatography, using methanol-water (70:30, v/v) to elute, with an elution volume of 0.7L, and 5 fraction samples (F6.4.1-F6.4.5) were obtained. .
  • Fraction F6.4.2 (1630 mg) was prepared by reverse-phase preparative HPLC (Phenomenex, Packed C18 column), and methanol-water (70:30, v/v) with a flow rate of 8 mL/min was used for elution to obtain formula (III) Compound (t R : 35.2 min, 820 mg) and compound of formula (XI) (t R : 25.4 min, 4.0 mg).
  • Fraction F6.4.3 (212.9 mg) was prepared by reversed-phase preparative HPLC (Cosmosil Packed C18 column), using acetonitrile-water (55:45, v/v) with a flow rate of 3 mL/min for elution, to obtain the compound of formula (VII) (t R : 63.3 min, 10.3 mg) and the compound of formula (VIII) (t R : 25.0 min, 10.1 mg).
  • T cells produce specific cytokines after differentiation, such as Th17 producing the cytokine IL17, Th2 producing the cytokine IL4, and Tr1 producing the cytokine IL10.
  • IL17-GFP mice, IL4-eGFP mice and IL10-eGFP mice are transgenic mice with a green fluorescent reporter gene (GFP), which produces specific cytokines after T cell differentiation.
  • the gene downstream is connected to the GFP fluorescent protein coding sequence. Therefore, a fluorescence detection system can be used to detect the expression of GFP to measure the expression level of cytokines.
  • Anti-mouse CD3Ab (antibody, antibody) coating plate Add anti-mouse CD3Ab diluted in IMDM medium to a final concentration of 10 ⁇ g/mL in a 96-well plate, and incubate at 37°C for 2 hours.
  • IL17-GFP mice raised in a specific pathogen-free (SPF) environment were sacrificed by cervical dislocation.
  • the spleens were removed and placed in IMDM culture medium.
  • the spleens were ground in a clean bench using sterilized pathology slides.
  • the ground cell suspension was filtered into a centrifuge tube using a 40 ⁇ m cell strainer filter; centrifuge (4°C, 1400 rpm, 7 minutes), discard the supernatant, add red blood cell lysate, mix well, let stand at room temperature for 5 minutes, and then add IMDM terminates the lysis of red blood cells; filter through a 40 ⁇ m cell strainer filter and then centrifuge (4°C, 1400 rpm, 7 minutes), discard the supernatant, and resuspend the cells in IMDM containing 10% FBS; dilute the cells 100 times, and count the cells under a microscope. Count the results and prepare a cell suspension of 1 ⁇ 10 6 cells/mL (final concentration).
  • Th17 cell differentiation inducing factors anti-mouse CD28Ab (1 ⁇ g/mL), rhTGFb1.2 (0.25ng/mL), rmIL6 (40ng/mL), anti-mouse IL4 (5 ⁇ g/mL), anti-mouse IFN ⁇ (5 ⁇ g /mL) (cytokines here are final concentrations).
  • Dilution of the sample to be tested Dilute the compound to 10 ⁇ M (final concentration) with IMDM containing 10% FBS; dilute the positive compound SR2211 that inhibits IL17 to 1 ⁇ M (final concentration).
  • Plating Aspirate the antibodies on the plate, wash them with IMDM, add 100 ⁇ L cell suspension and 100 ⁇ L final concentration sample solution to each well for co-culture (such as 100 ⁇ L cell suspension with a concentration of 4 ⁇ 10 6 cells/mL and 100 ⁇ L 20 ⁇ M compound co-culture , the final concentration of cells in the culture system was 2 ⁇ 10 6 cells/mL, and the compound concentration was 10 ⁇ M).
  • Cell culture The sample solution and cells were cultured in a humidified incubator at 37°C, 5% CO2 for a total of 72 hours.
  • Cell surface staining and flow cytometry detection Collect the cultured cells into a flow tube, add 3 mL of pre-cooled PBS, centrifuge (4°C, 1400 rpm, 7 minutes), discard the supernatant, and add 0.25 ⁇ L of LAPc anti to 50 ⁇ L of cell suspension. -Mouse CD4 antibody surface staining, place at 4°C in the dark for 20 minutes. Then resuspend the cells in 1 mL of pre-cooled PBS, add 2 mL of PBS, centrifuge again (4°C, 1400 rpm, 7 minutes), discard the supernatant, add 200 ⁇ L of PBS to each tube, and analyze the ratio of GFP + cells to CD4 + cells by flow cytometry.
  • IMDM medium and fetal bovine serum (FBS) were purchased from Gibco; anti-mouse CD28Ab, anti-mouse CD3Ab, anti-mouse IL4, anti-mouse IFN ⁇ , APC anti-mouse CD4 were purchased from Surgie; rmIL6 was purchased from Peprotech company; rhTGFb1.2 was purchased from R&D Company; red blood cell lysate was purchased from Tiangen Company; PBS was purchased from Solarbio Company.
  • the relative inhibitory activity (Relative Inhibition Activity) of the compound against IL17 was calculated using the formula: (ratio of GFP + in the DMSO group to CD4 + - ratio of GFP + in the compound group to CD4 + )/ratio of GFP + in the DMSO group ⁇ 100% ).
  • Th2 differentiation conditions Primary spleen lymphocytes from fluorescent reporter mice IL4-eGFP mice were extracted, co-cultured with compounds under Th2 differentiation conditions, and IL4-GFP levels were detected by flow cytometry to evaluate the effects of compounds on IL4 expression.
  • the culture medium used for Th2 is RPMI-1640; the cell density is 2 ⁇ 10 6 cells/mL; the differentiation conditions used are: anti-mouse CD28Ab (1 ⁇ g/mL), rmIL2 (2ng/mL), rmIL4 (40ng/mL) , Anti-mouse IFN ⁇ (10 ⁇ g/mL).
  • Other experimental steps for Th2 differentiation are the same as those for Th17 differentiation in Example 3.
  • rmIL4 and rmIL2 were purchased from Peprotech Company; RPMI-1640 culture medium was purchased from Sigma Company.
  • the formula is used: the ratio of GFP + to CD4 + in the compound group/the ratio of GFP + to CD4 + in the DMSO group ⁇ 100% to calculate the relative activity (RelativeActivity) of the compound on IL4.
  • Tr1 differentiation conditions Primary spleen lymphocytes from fluorescent reporter IL10-eGFP mice were extracted and sorted, co-cultured with compounds under Tr1 differentiation conditions, and the effects of compounds on IL10 expression were evaluated by detecting IL10-eGFP levels by flow cytometry.
  • the culture medium used for Tr1 is RPMI-1640; the cell density is 1 ⁇ 10 6 cells/mL; the differentiation conditions used are: anti-mouse CD28Ab (1 ⁇ g/mL), rmIL27 (50ng/mL), rhTGFb1.2 (0.5ng /mL).
  • Other experimental steps for Tr1 differentiation are the same as those for Th17 differentiation in Example 3.
  • rmIL27 was purchased from Biolegend; RPMI-1640 culture medium was purchased from Sigma.
  • the formula is used: the ratio of GFP + in the compound group to CD4 + / the ratio of GFP + in the DMSO group to CD4 + ⁇ 100% to calculate the relative activity (RelativeActivity) of the compound on IL10.
  • the compounds of the present invention are formula (II), formula (III), formula (IV), formula (VI), formula (VII), and formula (VIII).
  • the relative activity of formula (IX), formula (X), formula (XI) and formula (XII) on IL10 is significantly enhanced. This indicates that the above compounds have the effect of promoting the expression of IL10 and can exert broad-spectrum anti-inflammatory effects and be used for anti-inflammatory and autoimmune diseases.
  • sesquiterpene polyketides have the activity of inhibiting IL17, promoting IL4 and promoting the expression of IL10, and can be used for the prevention and treatment of immune diseases.
  • Example 6 Compound of formula (III) inhibits the expression of human IL17
  • PBMC Human peripheral blood mononuclear cells
  • human Th17 After human Th17 is differentiated, it is stimulated with 50ng/mL PMA, 1 ⁇ g/mL Ionomycin and Golgi-Plug for 5 hours. Add flow cytometry antibodies BV421anti-human IL17A and BV500anti-human CD4, and stain according to the experimental steps of the intracellular staining kit.
  • anti-human CD3, anti-human CD28, anti-human IFN ⁇ , and anti-human IL4 were purchased from Tonbo Biosciences; rhIL6 was purchased from Peprotech; BV421 anti-human IL17A was purchased from BioLegend, and BV500 anti-human CD4 was purchased from BD.
  • Example 7 Compounds of formula (III) inhibit BMDC from secreting inflammatory cytokines
  • Bone marrow-derived cells were extracted from the leg bones of C57BL/6 mice and cultured in bone marrow-derived dendritic cell (BMDC) culture medium (1640 medium containing 50ng/mL GM-CSF, 20ng/mL IL4 pre-warmed 10% FBS ) for 2 days, transfer the supernatant to a new dish and add BMDC culture medium to continue culturing for 2 days, half-change the medium and continue culturing for 2 days; collect cells, enrich BMDC with magnetic beads, and dilute the cells to 2 ⁇ 10 5 / mL, 1 mL in each tube was inoculated into the flow tube and cultured overnight; DMSO and 5 ⁇ M compound of formula (III) were added to the BMDC cultured overnight, cultured under LPS (100ng/mL) stimulation for 18 hours, and then Golgi-Plug was added for treatment 6 hours; add PurifiedRatAnti-Mouse CD16/CD32 to block for 15 minutes, add flow cytometry antibodies
  • Percepcy5.5 anti-mouse CD11c, FITC anti-mouse TNF ⁇ , PE anti-mouse IL6 and APC anti-mouse IL12/IL23p40 were purchased from Biolegend; eF450 anti-mouse MHCII was purchased from Invitrogen; PurifiedRatAnti-Mouse CD16/CD32 was purchased from BD Company.
  • Example 8 Compound of formula (III) has the effect of inhibiting DSS-induced acute enteritis
  • Dextran sulfate sodium (DSS)-induced enteritis in mice is a classic model of inflammatory bowel disease (IBD).
  • the modeling steps are as follows: From day 0 to day 4, C57BL/6 mice drink 3% DSS (purchased from MP Company) freely; on day 5, the 3% DSS is removed and replaced with water, and the mice are fed for another 3 days.
  • mice Ten-week-old male C57BL/6 mice were divided into 4 groups, with 8 mice in each group. Each group was treated as follows: blank control group (Control+Vehicle): drink pure water freely and inject solvent into the abdominal cavity twice a day; DSS model group ( DSS+Vehicle): Drink 3% DSS freely, and intraperitoneally inject the solvent twice a day; High-dose group (DSS+Formula III(H)): Drink 3% DSS freely, and inject 40 mg/kg formula (III) compound intraperitoneally twice a day. ; Low-dose group (DSS+Formula III(L)): drink 3% DSS freely and intraperitoneally inject 20 mg/kg compound of formula (III) twice a day. Vehicle refers to the solvent that dissolves the compound of formula (III), which is composed of 5% DMSO + 5% Solutol HS 15 (polyethylene glycol-15 hydroxystearate, purchased from BASF) + 90% water for injection.
  • Vehicle refers to the solvent that
  • DAI Clinical disease activity index
  • mice On the 8th day of the experiment, the mice were sacrificed, and the complete colons were removed and the length of the mouse colons was measured. The colon was rinsed with PBS, and the distal colon was dissected for pathological analysis.
  • Example 9 Compound of formula (III) has the effect of treating EAE in mice
  • MOG35-55-induced experimental autoimmune meningitis is a classic mouse model that mimics human multiple sclerosis (MS).
  • the modeling steps are as follows: take MOG35-55 (Shanghai Gill Biochemical Company), adjust the concentration to 2 mg/mL; take incomplete Freund's adjuvant (Sigma Company), add inactivated Mycobacterium tuberculosis H37RA (DIFCO Company), and adjust the concentration to 5mg/mL; mix the MOG35-55 solution and Freund's adjuvant according to the volume ratio of 1:1, and emulsify on ice for 2 hours to a water-in-oil state; after the mice are anesthetized, do it subcutaneously on the back of the mice's ears and thighs.
  • mice immunized in this experiment were 10-week-old female C57BL/6 mice.
  • the EAE score is an indicator used to evaluate the degree of EAE disease activity. It can be scored from 0 to 5 points according to the severity of the disease: 0 points - no obvious abnormality; 0.5 points - tail tip weakness, no abnormality in motor ability; 1 point - no abnormality in movement ability; The tail is weak, and there is no abnormality in the movement ability; 1.5 points - the whole tail is weak, the two hind limbs can be separated, and the movement is occasionally missed; 2 points - the whole tail is weak, the two hind limbs are brought together, and the movement is often missed; 2.5 points - dragging one hind limb; 3 points - dragging two hind limbs, and can turn over on its own after being placed on its back; 3.5 points - dragging two hind limbs, and cannot turn over on its own after being placed on its back, and its forelimbs can normally grasp the metal bars of the cage frame; 4 points - the mouse drags its two hind limbs and cannot turn over on its own after being placed
  • mice The EAE score of the mice was scored every day during the experiment. On the 5th day of intervention with the compound of formula (III), the mice were sacrificed, and tissue from the spinal cord enlargement area was collected for pathological analysis.
  • Example 10 Compound of formula (III) has the effect of treating psoriasis in mice
  • IMQ Topically applied imiquimod
  • Ear thickness was measured daily during the experiment. On the 21st day of the experiment, the mice were sacrificed, the ears of the mice were photographed, and the ear tissues were taken for pathological analysis.
  • Example 11 Compound of formula (III) has a protective effect on LPS-induced sepsis in mice
  • LPS lipopolysaccharide
  • model group LPS+Vehicle
  • formula (III) compound treatment group LPS+Formula III
  • Example 12 Compound of formula (III) has protective effect on APAP-induced liver injury
  • Acetaminophen (APAP)-induced liver injury mimics clinical drug-induced liver injury.
  • the mice were not allowed to eat or drink water the day before the experiment.
  • drug-induced liver injury was induced in mice by intraperitoneal injection of 250 mg/kg APAP (purchased from Sigma Company).
  • model group APAP+Vehicle
  • formula (III) compound treatment group APAP+Formula III
  • ALT alanine aminotransferase

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Immunology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Diabetes (AREA)
  • Rheumatology (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Dermatology (AREA)
  • Epidemiology (AREA)
  • Pulmonology (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • General Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Endocrinology (AREA)
  • Emergency Medicine (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Ophthalmology & Optometry (AREA)
  • Urology & Nephrology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Transplantation (AREA)
  • Hematology (AREA)
  • Pain & Pain Management (AREA)
  • Obesity (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

一类倍半萜聚酮化合物、药物组合物及其作为免疫调节剂和防治免疫性疾病的用途。生物活性实验显示涉及的倍半萜聚酮类化合物，可作为免疫调节剂用于免疫相关疾病的预防和/或治疗。

Description

一类倍半萜聚酮化合物作为免疫调节剂在防治免疫性疾病的用途
相关申请的交叉引用
本申请要求2022年06月02日提交的中国申请号2022106262021的权益。所述申请号2022106262021据此全文以引用方式并入本文。
技术领域
本发明属于天然药物领域，具体涉及一类倍半萜聚酮化合物作为免疫调节剂在制备预防或治疗免疫相关疾病的药物中的用途。
背景技术
免疫系统平衡对身体健康至关重要，当其被打破时容易患免疫性疾病。肿瘤和感染与机体免疫低下相关。器官移植免疫排斥、过敏和自身免疫病与机体免疫反应过强相关[1]，其中，自身免疫病包括炎症性肠病(IBD)、多发性硬化症(MS)和类风湿性关节炎(RA)在内的100余种疾病，发病重，发病率高，发病周期长，无治愈方法，严重危害人类健康。免疫调节剂可通过调节机体的免疫反应(增强、抑制或双向调节作用)，用于治疗免疫功能紊乱所引起的免疫性疾病。对于肿瘤和感染等与免疫力低下相关的疾病，可采用增强机体免疫力的调节剂进行干预。而对于器官移植免疫排斥、过敏和自身免疫病等与机体免疫反应过强相关的疾病，目前主要使用免疫抑制剂来改善疾病症状和预后。
皮质激素、甲氨蝶呤、环孢素A等化学药物属于非特异性免疫抑制剂，长期使用毒副作用大；生物制剂价格昂贵，治疗效果个体差异大，且具有增加病人感染与患肿瘤的风险。因此，开发新型免疫抑制剂来防治与免疫反应过强相关的疾病仍受到全世界的重视。近年来，我们对免疫性疾病的致病机制的研究取得了重要进展。抑制致炎因子IL17表达、促进抑炎因子IL10表达以及抑制DC细胞产生促炎因子均已成为抑制与免疫反应过强相关的免疫性疾病的有效策略，这为研发新型免疫抑制剂提供了新思路。
CD4+T细胞在TGFβ、IL6的参与下分化成Th17细胞。IL17是由Th17细胞主要分泌的强致炎性细胞因子，介导了多种炎症和自身免疫病[2]。越来越多的证据表明，IL17在炎症性肠病、Ⅰ型糖尿病、多发性硬化症、类风湿性关节炎、银屑病、系统性红斑狼疮、哮喘、干燥综合症和强直性脊柱炎等发病中起着关键性的作用，并且与葡萄膜炎、自身免疫性肾炎、自身免疫甲状腺炎、白塞氏病、扁平苔藓、呼吸道疾病(包括非嗜酸粒细胞性哮喘、类固醇抵抗型哮喘和囊性纤维化)、肝脏疾病(包括药物性肝损伤、酒精性肝病、非酒精性脂肪肝、病毒性肝炎、肝细胞癌、自身免疫性肝炎，原发性胆汁性肝硬化和原发性硬化性胆管炎)以及心血管疾病(包括动脉粥样硬化、心肌梗死、动脉瘤和缺血性脑卒中)相关联。临床使用IL17的抑制剂对银屑病、关节炎和多发性硬化症等多种免疫性疾病进行治疗干预，均显示出一定的疗效[3]。
IL10是一类具有免疫抑制特性和广泛抗炎特性的细胞因子。IL10能调节多种髓系或淋系来源细胞的活性，抑制其产生促炎因子，从而发挥抑制器官移植免疫排斥、过敏、炎症以及自身免疫病的作用[4]。I型调节性T细胞(Tr1)分泌大量的IL10，是一类具有免疫抑制作用的细胞，目前，转输Tr1细胞防治自身免疫性疾病或保护器官移植免疫排斥的临床实验已现疗效[5]。
因此抑制IL17和/或促进IL10的化合物可作为免疫调节剂防治器官移植免疫排斥、过敏、炎症和自身免疫病(炎症性肠病、Ⅰ型糖尿病、多发性硬化症、类风湿性关节炎、银屑病、系统性红斑狼疮、哮喘、干燥综合症和强直性脊柱炎、葡萄膜炎、自身免疫性肾炎、自身免疫甲状腺炎、白塞氏病、扁平苔藓、多种呼吸道疾病、多种肝脏疾病以及多种心血管疾病等)。此外，抑制IL17和/或促进IL10的化合物还可作为免疫调节剂用于痤疮、椎间盘突出、腹腔疾病等与炎症相关的疾病的治疗。IL17还与肿瘤的发生密切相关，尤其是炎症相关的肿瘤。IL10对肿瘤的发生发展具有双向调节的作用。IL10可通过抑制促进肿瘤发生发展的炎性因子的产生以及通过活化肿瘤特异性CD8+T来发挥抗肿瘤作用[4]。因此抑制IL17和/或促进IL10的化合物还可作为免疫调节剂用于肿瘤治疗。
树突状细胞(Dendritic cell，DC)是一种抗原提呈细胞，DC细胞可将抗原呈递到T细胞为T细胞的活化提供第二信号，也可通过分泌细胞因子IL6、TNFα、IL12诱导T细胞的极化。然而，当DC细胞介导的免疫反应过度激活时，可促进炎症因子风暴甚至脓毒症的发生、也会介导器官移植排斥反应、炎症和自身免疫病等免疫性疾病[6]。因此，在抗原刺激下，抑制DC细胞分泌炎症因子如IL6、TNFα、IL12等的化合物可用于炎症和自身免疫病、脓毒症、器官移植排斥反应的治疗。对于抑制DC免疫反应药效学实验，常采用小鼠骨髓来源的树突状细胞(Bone Marrow-Derived Dendritic Cells，BMDC)。
Th2细胞主要分泌细胞因子IL4。IL4具有多种生物学功能，在体液免疫和获得性免疫中发挥重要作用[7]。IL4能诱导B细胞、T细胞活化以及B细胞向浆细胞分化。此外，IL4还能抑制淋巴细胞凋亡、促进巨噬细胞的抗原提呈能力和提高杀伤肿瘤细胞的功能。因此，促进IL4的化合物可作为免疫调节剂用于抗肿瘤、抗感染(尤其是抗寄生虫感染)相关的疾病。
发明内容
本发明的目的在于提供一类倍半萜聚酮化合物、药物组合物及其用途。具体地，发明人从一株漆斑属真菌中发现并分离鉴定了一类倍半萜聚酮化合物，经实验证明其具有免疫调节活性，可用于防治免疫性疾病，尤其是炎症性肠病、多发性硬化症、银屑病和脓毒症等自身免疫性疾病。
本发明的第一方面提供了一类倍半萜聚酮化合物或其药学上可接受的盐作为免疫调节剂的用途，所述倍半萜聚酮类化合物如式(I)所示：
其中，R1选自：羟基，乙酰氧基，或＝O；
R2选自：甲基，羟甲基，乙酰羟甲基，醛基，或二甲氧基甲基；
R3选自：甲基，羟甲基，乙酰羟甲基，或醛基；
或者R2，R3与苯环组成5元内酰胺环或内酯环；
R4，R5，R6，R7为碳原子，并且任一相邻基团间选自双键，其余基团间选自单键。
进一步的，所述药学上可接受的盐为式(I)化合物与有机碱或无机碱形成的盐。
更进一步的，所述形成的盐为钠盐、钾盐、钙盐、铁盐、镁盐、锌盐、铝盐、钡盐或铵盐。
优选的，所述倍半萜聚酮类化合物选自式Ⅱ-式Ⅻ的化合物，具体结构式如下：
本发明的第二方面提供了一种上述化合物的制备方法，包括：由产生如式(I)所示的倍半萜聚酮类化合物的微生物发酵后，再用色谱法分离后得到。
进一步的，所述微生物为漆斑属真菌。
进一步可选的，所述漆斑属真菌为ZLW0801-19，其保藏号为CGMCC No.19039。
本发明的第三方面提供了一种用于免疫调节的药物组合物，包括上述倍半萜聚酮类化合物或其药学上可接受的盐以及药学上可接受的载体。
进一步可选的，所述盐为钠盐、钾盐、钙盐、铁盐、镁盐、锌盐、铝盐、钡盐、铵盐中的任意一种。
进一步可选的，在一个实施方式中药物组合物中含有的活性成份(即本发明化合物)的量可以根据患者的病情、医生诊断的情况特定的加以应用，活性化合物的量或浓度在一个较宽的范围内调节，所述式(I)化合物或其药学上可接受的盐的含量为组合物重量的1-90％。
进一步可选的，药学上可接受的载体包括稀释剂、润滑剂、粘合剂、崩解剂、稳定剂、溶剂等。本发明所述稀释剂包括但不限于淀粉、微晶纤维素、蔗糖、糊精、乳糖、糖粉、葡萄糖等；所述润滑剂包括但不限于硬脂酸镁、硬脂酸、氯化钠、油酸钠、月桂醇硫酸钠、泊洛沙母等；所述粘合剂包括但不限于水、乙醇、淀粉浆、糖浆、羟丙基甲基纤维素、羧甲基纤维素钠、海藻酸钠、聚乙烯吡咯烷酮等；所述崩解剂包括但不限于淀粉泡腾混合物即碳酸氢钠和枸橼酸、酒石酸、低取代羟丙基纤维素等；所述稳定剂包括但不限于多糖如金合欢胶、琼脂、藻酸、纤维素醚和羧甲基甲壳酯等；所述溶剂包括但不限于水、平衡的盐溶液等。
进一步可选的，所述药物组合物为口服制剂或注射剂；优选的，所述口服制剂包括但不限于普通片剂、分散片、肠溶片、颗粒、胶囊、滴丸、散剂、口服液或乳剂中的任意一种；优选的，所述注射剂选自小水针剂、输液剂或冻干粉针剂中的任意一种。
本发明的第四方面还提供了制备免疫调节剂的上述式(I)化合物或其药学上可接受的盐。
进一步的，所述免疫调节剂用于免疫相关疾病的预防和/或治疗。
更进一步的，所述免疫调节剂用于防治抑制IL17和/或促进IL10是有益的疾病或适应症。
优选的，所述疾病或适应症选自银屑病、炎症性肠病、I型糖尿病、多发性硬化症、类风湿性关节炎、系统性红斑狼疮、哮喘、干燥综合症、强直性脊柱炎、器官移植免疫排斥、过敏、脓毒症、葡萄膜炎、自身免疫性肾炎、自身免疫甲状腺炎、白塞氏病、自身免疫性肝病、扁平苔藓、痤疮、椎间盘突出、腹腔疾病。
更进一步的，所述免疫调节剂用于促进IL4是有益的疾病或适应症。
优选的，所述疾病或适应症为寄生虫感染相关。
更进一步的，所述免疫调节剂用于防治抑制DC免疫反应是有益的疾病或适应症。
优选的，所述疾病或适应症为炎症和自身免疫病、脓毒症、器官移植排斥反应。
有益效果
本发明所述的倍半萜聚酮类化合物是从微生物发酵来源，便于大量发酵和工业化制备；通过细胞和动物活性测试，首次发现该类化合物具有显著的免疫调节活性。
附图说明
图1所示为式(III)化合物显著抑制人IL17的表达。
图2所示为式(III)化合物抑制了LPS刺激的BMDC中IL12/IL23p40+、IL6+和TNFα+细胞的比例。
图3所示为：式(III)化合物显著抑制了肠炎小鼠体重下降(图3A)、DAI评分增高(图3B)、结肠缩短(图3C)的现象；结肠组织病理分析显示，式(III)化合物显著抑制了结肠炎症、上皮损伤以及肠道结构病变的程度(图3D和3E)。
图4所示为：式(III)化合物显著降低了EAE模型小鼠的EAE评分(图4A)；脊髓组织病理分析显示，式(III)化合物处理显著抑制了脊髓炎性细胞浸润以及脱髓鞘程度(图4B)。
图5所示为：相对于模型组，式(III)化合物处理显著抑制类银屑病小鼠耳朵增厚的现象(图5A和5B)；耳朵组织病理分析结果显示，式(III)化合物处理显著抑制了类银屑病小鼠耳朵皮肤角化不全、角化过度、Munro小体形成、棘层肥厚、炎性细胞浸润和真皮血管扩张充血的病理损伤(图5C和5D)。
图6所示为式(III)化合物显著提高了LPS诱导的脓毒症小鼠的存活率。
图7所示为式(III)化合物显著抑制了肝损伤小鼠血清中ALT的水平。
具体实施方式
下面将进一步的来举例说明本发明。需要指出的是，所述实施例说明了一些制备或使用方法，然而，要理解的是，这些实施例不限制本发明。本发明的保护范围以所附权利要求书记载的内容为准。
下列实施例中，质谱仪为美国Finnigan公司生产的LCQ-Advantage质谱仪。超导核磁共振仪为BrukerAV-400。薄层色谱用硅胶GF254和柱色谱硅胶(200-300目)均为青岛海洋化工厂产品。反相ODS填料50μm为日本YMC公司产品。中低压液相色谱仪为上海利穗电子科技有限公司产品。液相分离所使用色谱柱为Phenomenex Gemini C18column(10.0×250mm, 5μm)。液相色谱用甲醇和乙腈为色谱纯，水为双重蒸馏水，其他试剂均为分析纯。
实施例中显示的体外细胞水平的实验数据以平均值±标准差(Mean±SD)表示，动物实验数据以平均值±标准误差(Mean±SEM)表示。对于比较两组数据间的差异，采用Two-tailed Student’s t test进行分析；比较多于2组的数据间的差异，采用One-wayAnova进行分析；对于每个受试对象在不同的时间点进行了重复测量的数据(如体重测量、耳朵厚度测量)，采用Two-wayAnova中的重复测量方差分析来比较组间差异。统计学差异采用以下标示：*代表P<0.05，**代表P<0.01，***代表P<0.001，****代表P<0.0001，ns代表差异不显著。采用Graphpad Prism 7.00和FlowJo V10(用于流式)分析获取的数据，所有病理切片由病理医生进行评估。
实施例1漆斑属真菌ZLW0801-19大量发酵及其样品前处理方法
(1)漆斑属真菌ZLW0801-19真菌菌株在25℃的马铃薯葡萄糖琼脂(PDA)斜面上培养5天。经PDA斜面活化后接种至4个含马铃薯葡萄糖(PDB)培养基的锥形瓶(250mL)中制备种子液，每个锥形瓶含有100mLPDB培养基，转速200rpm在25℃培养5天制备种子液。在24个锥形瓶(500mL)中进行发酵，每个锥形瓶中含有70g大米，首先向每个锥形瓶中加入蒸馏水(105mL)，将大米浸泡过夜，然后在120℃下高压灭菌30分钟。冷却至室温后，向每个锥形瓶接种5.0mL种子液，并在室温避光培养51天。
(2)将发酵物加入乙酸乙酯进行浸泡提取3次，将提取液减压浓缩至干，得到粗提物(81.7g)。
实施例2式(II)—式(XII)化合物的制备
实施例1中乙酸乙酯粗提物(81.7g)利用硅胶柱层析，依次采用环己烷-乙酸乙酯(100:0,98:2,95:5,90:10,80:20,70:30,50:50,0:100,v/v)、甲醇进行洗脱，每个梯度洗脱体积6L，得到7个馏分样品(F1-F7)。馏分F5经中低压液相ODS柱层析，依次采用甲醇-水(70:30,80:20,90:10,and 100:0,v/v)洗脱，每个梯度洗脱体积2.5L，得到9个馏分样品(F5.1-F5.9)。馏分F5.2经中低压液相ODS柱层析，依次采用甲醇-水(60:40,70:30,80:20,90:10,v/v)洗脱，每个梯度洗脱体积0.7L，得到5个馏分样品(F5.2.1-F5.2.5)。馏分F5.2.4(387mg)经反相制备级HPLC制备(Cosmosil Packed C18column)，使用流速为3mL/min的乙腈-水(50:50,v/v)进行洗脱，得到式(X)化合物(tR:53.6min,26.2mg)。将馏分F5.2.5(365mg)经反相制备级HPLC制备(Phenomenex,Packed C18column)，使用流速为8mL/min的乙腈-水(55:45,v/v)进行洗脱，得到式(XII)化合物(tR:86.4min,7.0mg)。馏分F5.3经凝胶柱LH-20柱层析，采用甲醇洗脱得到7个馏分(F5.3.1–F5.3.7)。馏分F5.3.7(144mg)经反相制备级HPLC制备(Cosmosil Packed C18column)，使用流速为3mL/min的甲醇-水(75:25,v/v)进行洗脱，得到式(II)化合物(tR:63.7min,17.0mg)和式(V)化合物(tR:69.9min,17.5mg)。将馏分F5.5(1550mg)经反相制备级HPLC制备(Phenomenex,Packed C18column)，使用流速为8mL/min的甲醇-水(75:25,v/v)进行洗脱，得到式(IV)化合物(tR:73.5min,507.0mg)。馏分F5.7经硅胶柱层析，依次采用环己烷-乙酸乙酯(90:10,85:15,80:20,75:25,70:30,60:40,50:50,0:100,v/v)洗脱，每个梯度洗脱体积1.0L，得到8个馏分样品(F5.7.1-F5.7.8)。馏分F5.7.4(62.1mg)经反相制备级HPLC制备(Cosmosil Packed C18column)，使用流速为3mL/min的甲醇-水(82:18,v/v)进行洗脱，得到式(VI)化合物(tR:21.3min,12.1mg)。馏分F6经中低压液相ODS柱层析，依次采用甲醇-水(60:40,70:30,80:20,90:10,100:0,v/v)洗脱，每个梯度洗脱体积2.5L，得到7个馏分样品(F6.1-F6.7)。馏分F6.3经硅胶柱层析，依次采用环己烷-乙酸乙酯(75:25,70:30,60:40,50:50,0:100v/v)洗脱，每个梯度洗脱体积0.3L，得到6个馏分样品(F6.3.1-F6.3.6)。馏分F6.3.6(349.7mg)经反相制备级HPLC制备(Cosmosil Packed C18column)，使用流速为3mL/min的乙腈-水(40:60,v/v)进行洗脱，得到式(IX)化合物(tR:17.5min,7.7mg)。馏分F6.4经中低压液相ODS柱层析，采用甲醇-水(70:30,v/v)洗脱，洗脱体积0.7L，得到5个馏分样品(F6.4.1-F6.4.5)。将馏分F6.4.2(1630mg)经反相制备级HPLC制备(Phenomenex,Packed C18column)，使用流速为8mL/min的甲醇-水(70:30,v/v)进行洗脱，得到式(III)化合物(tR:35.2min,820mg)和式(XI)化合物(tR:25.4min,4.0mg)。馏分F6.4.3(212.9mg)经反相制备级HPLC制备(Cosmosil Packed C18column)，使用流速为3mL/min的乙腈-水(55:45,v/v)进行洗脱，得到式(VII)化合物(tR:63.3min,10.3mg)和式(VIII)化合物(tR:25.0min,10.1mg)。
理化常数如下：
式(II)化合物：无色片状晶体；(c 1.0,CH3OH)；UV(CH3OH)λmax(logε)205(4.31),225(4.28),221(4.10),300(4.27)nm；ECD(c 3.4×10–4M CH3OH)λmax(Δε)221(+1.86),245(+1.71),334(–0.55),306(–0.74)；IR(KBr)νmax 3399,3137,2974,2929,2864,1616,1430,1293,1262,1117,1045cm–1；ESI-MS(positive)m/z 373[M+H]+,395[M+Na]+；ESI-MS(negative)m/z 371[M–H]–,743[2M–H]–；HR-ESI-MS(positive)m/z 373.2379[M+H]+(calcd.for C23H33O4,373.2379)，确定化合物分子式为C23H32O4；1H和13C NMR见表1。
式(III)化合物：淡黄色固体；(c 1.0,CH3OH)；UV(CH3OH)λmax(logε)206(4.43),225(4.35),240(4.18),299(4.31)nm；IR(KBr)νmax 3146,2969,2929,2881,1621,1625,1448,1258cm-1；ESI-MS(positive)m/z 389[M+H]+,411[M+Na]+；ESI-MS(negative)m/z 387[M–H]–,775[2M–H]–；HR-ESI-MS(positive)m/z 389.2331[M+H]+(calcd.for C23H33O5,389.2328)，确定化合物分子式为C23H32O5；1H和13C NMR见表1。
式(IV)化合物：淡黄色针状晶体；(c 1.0,CH3OH)；(CH3OH)λmax(logε)205(4.27),223(4.23),240(4.09),299(4.27)nm；IR(KBr)νmax 3399,3160,2973,2838,2868,1708,1619,1435,1268,1002cm–1；ESI-MS(positive)m/z 453[M+Na]+,883[2M+Na]+；ESI-MS(negative)m/z 429[M–H]–,859[2M–H]–；HR-ESI-MS(positive)m/z 453.2261[M+Na]+(calcd.for C25H34O6Na,453.2253)，确定化合物分子式为C25H34O6；1H和13C NMR见表1。
式(V)化合物：淡黄色固体；(c 1.0,CH3OH)；UV(CH3OH)λmax(logε)205(4.47),226(4.43),242(4.31),300(4.44)nm；IR(KBr)νmax 3430,2961,2931,2868,1719,1622,1435,1265,1039cm–1；ESI-MS(positive)m/z 453[M+Na]+,883[2M+Na]+；ESI-MS(negative)m/z429[M–H]–,859[2M–H]–；HR-ESI-MS(positive)m/z 431.2433[M+H]+(calcd.for C25H35O6,431.2434)，确定化合物分子式为C25H34O6；1H和13C NMR见表1。
式(VI)化合物：淡黄色油状；(c 1.0,CH3OH)；UV(CH3OH)λmax(logε)205(4.52),226(4.47),242(4.31),298(4.45)nm；IR(KBr)νmax 3279,2934,2867,1708,1618,1434,1267,1032cm–1；ESI-MS(positive)m/z 495[M+Na]+,967[2M+Na]+；ESI-MS(negative)m/z 471[M–H]–,943[2M–H]–；HR-ESI-MS(positive)m/z 495.2368[M+Na]+(calcd.for C27H36O7Na,495.2359)，确定化合物分子式为C27H36O7；1H和13C NMR见表2。
式(VII)化合物：淡黄色油状；(c 1.0,CH3OH)；UV(CH3OH)λmax(logε)205(4.37),225(4.35),239(4.18),300(4.35)nm；ECD(c 3.2×10–4M CH3OH)λmax(Δε)201(+1.34),235(+2.04),274(–0.80),324(+0.46)；IR(KBr)νmax 3411,2929,2858,1618,1434,1386,1267,1240,1125,1050cm–1；ESI-MS(positive)m/z 387[M+H]+,409[M+Na]+；ESI-MS(negative)m/z385[M–H]–,771[2M–H]–；HR-ESI-MS(positive)m/z 409.2009[M+Na]+(calcd.for C23H30O5Na,409.2015)，确定化合物分子式为C23H30O5；1H和13C NMR见表2。
式(VIII)化合物：淡黄色固体；(c 1.0,CH3OH)；UV(CH3OH)λmax(logε)206(4.36),226(4.30),240(4.16),299(4.33)nm；ECD(c 5.8×10–4M CH3OH)λmax(Δε)224(+0.99),249(+3.46),302(–0.76),361(–1.02)；IR(KBr)νmax 3399,2934,2850,1617,1386,1267,1245,1125,1050,988cm–1；ESI-MS(positive)m/z 455[M+Na]+,895[2M+Na]+；ESI-MS(negative)m/z 431[M–H]–,863[2M–H]–；HR-ESI-MS(positive)m/z 455.2415[M+Na]+(calcd.for C25H36O6Na,455.2410)，确定化合物分子式为C25H36O6；1H和13C NMR见表2。
式(IX)化合物：淡绿色固体；(c 1.0,CH3OH)；UV(CH3OH)λmax(logε)215(4.57),262(4.06),305(3.27)nm；IR(KBr)νmax 3385,2929,2863,1675,1452,1386,1262cm–1； ESI-MS(positive)m/z 386[M+H]+,793[2M+Na]+；ESI-MS(negative)m/z 769[2M–H]–；HR-ESI-MS(positive)m/z 386.2340[M+H]+(calcd.for C23H32NO4,386.2331)，确定化合物分子式为C23H31NO4；1H和13C NMR见表2。
式(X)化合物：淡绿色固本；(c 1.0,CH3OH)；UV(CH3OH)λmax(logε)206(4.44),223(4.46),240(4.42),298(4.33)nm；ECD(c 6.0×10–4M CH3OH)λmax(Δε)204(+7.00),247(+18.5),330(+1.81)；IR(KBr)νmax 3283,2970,2925,2867,1621,1618,1430,1267,1005cm–1；ESI-MS(positive)m/z 409[M+Na]+,795[2M+Na]+；ESI-MS(negative)m/z 385[M–H]–,771[2M–H]–；HR-ESI-MS(positive)m/z 387.2176[M+H]+(calcd.for C23H31O5,387.2176)，确定化合物分子式为C23H30O5；1H和13C NMR见表3。
式(XI)化合物：白色固体；(c 1.0,CH3OH)；UV(CH3OH)λmax(logε)206(4.39),226(4.06),289(3.69)nm；ECD(c 6.4×10–4M CH3OH)λmax(Δε)220(+3.81),236(+3.09),246(+3.74),323(–0.54)；IR(KBr)νmax3301,2961,2872,1628,1462,1284,1125,1076cm–1；ESI-MS(positive)m/z 389[M+H]+,411[M+Na]+；ESI-MS(negative)m/z 387[M–H]–,775[2M–H]–；HR-ESI-MS(positive)m/z 389.2329[M+H]+(calcd.for C23H33O5,389.2328)，确定化合物分子式为C23H32O5；1H和13C NMR见表3。
式(XII)化合物：白色固体；(c 1.0,CH3OH)；UV(CH3OH)λmax(logε)206(4.41),227(4.09),290(3.76)nm；ECD(c 2.9×10–4M CH3OH)λmax(Δε)220(+5.47),235(+4.19),246(+5.58),320(–1.30)；IR(KBr)νmax 3479,29743,2881,1740,1623,1489,1270,1045cm–1；ESI-MS(positive)m/z 453[M+Na]+；ESI-MS(negative)m/z 429[M–H]–,HR-ESI-MS(positive)m/z 453.2246[M+Na]+(calcd.for C25H34O6Na,453.2353)，确定化合物分子式为C25H34O6；1H和13C NMR见表3。
表1式(II)—式(V)化合物的13C NMR及1HNMR数据和归属

a The data recorded in DMSO-d6(1H NMR for 300MHz,13C NMR for 75MHz)b The data recorded in DMSO-d6(1H NMR for 400MHz,13C NMR for 100MHz)
表2式(VI)—式(IX)化合物的13C NMR及1H NMR数据和归属

a The data recorded in DMSO-d6(1H NMR for 300MHz,13C NMR for 75MHz)b The data recorded in DMSO-d6(1H NMR for 400MHz,13C NMR for 100MHz)
表3式(X)—式(XII)化合物的13C NMR及1H NMR数据和归属


a The data recorded in DMSO-d6(1H NMR for 300MHz,13C NMR for 75MHz)
实施例3倍半萜聚酮类化合物式(II)—式(XII)对IL17表达的影响
T细胞分化后产生特定细胞因子，如Th17产生细胞因子IL17，Th2产生细胞因子IL4，Tr1产生细胞因子IL10。IL17-GFP小鼠、IL4-eGFP小鼠和IL10-eGFP小鼠(均引自JAX公司)是带有绿色荧光报告基因(GFP)的转基因小鼠，其在T细胞分化后产生的特定细胞因子的基因下游连接上GFP荧光蛋白编码序列。因此可用荧光探测系统检测GFP的表达量来衡量细胞因子的表达水平。
提取荧光报告小鼠IL17-GFP小鼠的原代脾脏淋巴细胞，在Th17分化条件下与化合物共培养，通过流式细胞术检测IL17-GFP水平来评价化合物对IL17表达的影响(此方法使用原代淋巴细胞更好模拟复杂的生理过程；使用GFP小鼠和流式细胞术使实验简便，重复性好且结果可靠)。具体实验步骤如下：
Anti-mouse CD3Ab(antibody，抗体)包板：在96孔板中加入IMDM培养基稀释的终浓度为10μg/mL的anti-mouse CD3Ab，37℃孵育2小时。
制备单细胞悬液：无特定病原体(SPF)环境下饲养的IL17-GFP小鼠，颈椎脱臼处死，取脾脏置于IMDM培养基中，用灭菌的病理玻片在超净台中磨碎脾脏，磨碎后的细胞悬液用40μm cell strainer滤网过滤至离心管；后离心(4℃，1400rpm，7分钟)，弃上清，加红细胞裂解液，混匀，室温静置5分钟，后加IMDM终止裂解红细胞；40μm cell strainer滤网过滤后再离心(4℃，1400rpm，7分钟)，弃上清，用含10％FBS的IMDM重悬细胞；细胞稀释100倍，显微镜下细胞计数，根据计数结果，配成1×106个/mL(终浓度)的细胞悬液。
加入Th17细胞分化诱导因子：anti-mouse CD28Ab(1μg/mL)，rhTGFb1.2(0.25ng/mL)，rmIL6(40ng/mL)，anti-mouse IL4(5μg/mL)，anti-mouse IFNγ(5μg/mL)(此处细胞因子均为终浓度)。
待检测样本稀释：用含10％FBS的IMDM将化合物稀释成10μM(终浓度)；将抑制IL17的阳性化合物SR2211稀释成1μM(终浓度)。
铺板：将包板的抗体吸去，用IMDM洗一遍，每孔加入100μL细胞悬液和100μL终浓度样本溶液共培养(如100μL4×106个/mL浓度的细胞悬液和100μL 20μM化合物共培养，培养体系中细胞终浓度为2×106个/mL，化合物浓度为10μM)。
细胞培养：样本溶液和细胞置于37℃，5％的CO2的湿润培养箱中共培养72小时。
细胞表面染色和流式检测：收取培养的细胞于流式管，加入3mL预冷的PBS，离心(4℃，1400rpm，7分钟)，弃上清，后在50μL细胞悬液中加入0.25μLAPC anti-mouse CD4抗体表面染色，4℃避光放置20分钟。后用1mL预冷的PBS重悬细胞，补2mLPBS，再离心(4℃，1400rpm，7分钟)，弃上清，每管加200μLPBS，流式检测分析GFP+的细胞占CD4+细胞的比率。
其中IMDM培养基和胎牛血清(FBS)购自Gibco公司；anti-mouse CD28Ab，anti-mouse CD3Ab，anti-mouse IL4，anti-mouse IFNγ，APC anti-mouse CD4购自Sungene公司；rmIL6购自Peprotech公司；rhTGFb1.2购自R&D公司；红细胞裂解液购自Tiangen公司；PBS购自Solarbio公司。
采用公式：(DMSO组GFP+占CD4+的比率-化合物组GFP+占CD4+的比率)/DMSO组GFP+占CD4+的比率×100％来计算化合物对IL17的相对抑制活性(Relative Inhibition Activity)。
结果如表4所示：脾脏淋巴细胞向Th17细胞分化条件下，本发明化合物式(II)、式(III)、式(V)、式(XI)和式(XII)对IL17的相对抑制活性显著增强。这表明，本发明化合物式(II)、式(III)、式(V)、式(XI)和式(XII)具有抑制IL17表达的效果，可用于防治免疫相关疾病尤其是IL17介导的免疫性疾病。
表4式(II)—式(XII)化合物对IL17表达影响(10μM)

实施例4倍半萜聚酮类化合物式(II)—式(XII)对IL4表达的影响
提取荧光报告小鼠IL4-eGFP小鼠的原代脾脏淋巴细胞，在Th2分化条件下与化合物共培养，通过流式细胞术检测IL4-GFP水平来评价化合物对IL4表达的影响。Th2使用的培养基为RPMI-1640；细胞密度为2×106个/mL；使用的分化条件为：anti-mouse CD28Ab(1μg/mL)，rmIL2(2ng/mL)，rmIL4(40ng/mL)，Anti-mouse IFNγ(10μg/mL)。其他Th2分化的实验步骤同实施例3中的Th17分化一致。
其中rmIL4，rmIL2购自Peprotech公司；RPMI-1640培养基购自Sigma公司。
在评价化合物对IL4表达的影响时，采用公式：化合物组GFP+占CD4+的比率/DMSO组GFP+占CD4+的比率×100％来计算化合物对IL4的相对活性(RelativeActivity)。
具体结果如表5所示：脾脏淋巴细胞向Th2细胞分化条件下，本发明化合物式(II)、式(III)、式(X)、式(XI)和式(XII)对IL4的相对活性显著增强。这表明本发明化合物式(II)、式(III)、式(X)、式(XI)和式(XII)具有促进IL4表达的效果，可用于抗寄生虫感染。
表5式(II)—式(XII)化合物对IL4表达影响(10μM)

实施例5倍半萜聚酮类化合物式(II)—式(XII)对IL10表达的影响
提取分选荧光报告小鼠IL10-eGFP小鼠的原代脾脏淋巴细胞，在Tr1分化条件下与化合物共培养，通过流式细胞术检测IL10-eGFP水平来评价化合物对IL10表达的影响。Tr1使用的培养基为RPMI-1640；细胞密度为1×106个/mL；使用的分化条件为：anti-mouse CD28Ab(1μg/mL)，rmIL27(50ng/mL)，rhTGFb1.2(0.5ng/mL)。其他Tr1分化的实验步骤同实施例3中的Th17分化一致。
其中rmIL27购自Biolegend公司；RPMI-1640培养基购自Sigma公司。
在评价化合物对IL10的影响时，采用公式：化合物组GFP+占CD4+的比率/DMSO组GFP+占CD4+的比率×100％来计算化合物对IL10的相对活性(RelativeActivity)。
具体结果如表6所示：脾脏淋巴细胞向Tr1细胞分化条件下，本发明化合物式(II)、式(III)、式(IV)、式(VI)、式(VII)、式(VIII)、式(IX)、式(X)、式(XI)和式(XII)对IL10的相对活性显著增强。这表明上述化合物具有促进IL10表达的效果，可发挥广谱的抗炎作用，用于抗炎和自身免疫疾病。
表6式(II)—式(XII)化合物对IL10表达影响(20μM)
综合上述实施例3、4和5的结果可知：倍半萜聚酮类化合物具有抑制IL17、促进IL4和促进IL10表达的活性，可用于免疫性疾病的防治。
实施例6式(III)化合物抑制人IL17表达
从健康人外周血液样本中分离出人外周血单核细胞(PBMC)，采用EasySepTMHumanCD4+T Cell Isolation Kit II试剂盒(购自Stem cell)从人的PBMC中分离出人的naive CD4+T细胞，用于后续人的Th17分化。
用含10％FBS的IMDM将分选的人的naive CD4+T细胞稀释到5×105个/mL，加入anti-human CD28Ab(1μg/mL)；rhIL6(40ng/mL)；rhTGFβ1.2(0.5ng/mL)；anti-human IL4(10μg/mL)；anti-human IFNγ(10μg/mL)；在上述细胞悬液中分别加入DMSO和2.5μM式(III)化合物，接种于anti-human CD3Ab(5μg/mL)包板2小时的培养板，放入细胞培养箱中培养；在第三天补加rhIL6，第五天洗去anti-human CD23Ab和anti-human CD28Ab，用含rhIL6，rhTGFβ1.2，anti-human IL4，anti-human IFNγ的新的培养基转孔，继续培养2天；用含rhIL6，rhTGFβ1.2，anti-human IL4，anti-human IFNγ的新的培养基分孔，再培养2天。
人的Th17分化好后，用50ng/mLPMA、1μg/mL Ionomycin和Golgi-Plug刺激5小时。加入流式抗体BV421anti-human IL17A和BV500anti-human CD4，按照胞内染色试剂盒的实验步骤进行染色。
其中anti-human CD3，anti-human CD28，anti-human IFNγ，anti-human IL4购自Tonbo Biosciences公司；rhIL6购自Peprotech公司；BV421anti-human IL17A购自BioLegend公司，BV500anti-human CD4购自BD公司。
实验结果如图1所示：人的naive CD4+T细胞向人的Th17分化的条件下，式(III)化合物显著抑制了人的CD4+T细胞中IL17+细胞的比例。这说明，式(III)化合物具有抑制人的IL17表达的效果。
实施例7式(III)化合物抑制BMDC分泌炎性细胞因子
从C57BL/6小鼠腿骨提取骨髓来源的细胞，在骨髓来源的树突状细胞(BMDC)培养液(含50ng/mL GM-CSF、20ng/mL IL4预温的10％FBS的1640培养基)中培养2天，转移上清到新的皿并补加BMDC培养液继续培养2天，半换液继续培养2天；收集细胞，磁珠富集BMDC后将细胞稀释到2×105/mL，每管1mL接种于流式管，培养过夜；在培养过夜的BMDC中分别加入DMSO和5μM式(III)化合物，在LPS(100ng/mL)刺激下培养18小时后加入Golgi-Plug处理6小时；加入PurifiedRatAnti-Mouse CD16/CD32封闭15分钟，加入流式抗体Percepcy5.5anti-mouse CD11c、eF450anti-mouse MHCⅡ、FITC anti-mouse TNFα、PE anti-mouse IL6和APC anti-mouse IL12/IL23p40，按照胞内染色试剂盒的实验步骤进行染色。
其中Percepcy5.5 anti-mouse CD11c、FITC anti-mouse TNFα、PE anti-mouse IL6和APC anti-mouse IL12/IL23p40购自Biolegend公司；eF450anti-mouse MHCⅡ购自Invitrogen公司；PurifiedRatAnti-Mouse CD16/CD32购自BD公司。
实验结果如图2所示：式(III)化合物抑制了LPS刺激的BMDC(CD11c+MHCII+细胞)中IL12/IL23p40+、IL6+和TNFα+细胞的比例。这说明式(III)化合物具有抑制BMDC产生炎性细胞因子IL12/IL23、IL6和TNFα的效果。
实施例8式(III)化合物具有抑制DSS诱导的急性肠炎的效果
硫酸葡聚糖钠(DSS)诱导的小鼠肠炎是经典的炎性肠病(IBD)模型。建模步骤如下：第0到第4天，C57BL/6小鼠自由饮用3％DSS(购自MP公司)；第5天将3％DSS撤离换成水，继续饲养3天。
将10周龄雄性C57BL/6小鼠分为4组，每组8只，各组处理如下：空白对照组(Control+Vehicle)：自由饮用纯水，每天腹腔注射溶剂2次；DSS模型组(DSS+Vehicle)：自由饮用3％DSS，每天腹腔注射溶剂2次；高剂量组(DSS+Formula III(H))：自由饮用3％DSS，每天腹腔注射40mg/kg式(III)化合物2次；低剂量组(DSS+Formula III(L))：自由饮用3％DSS，每天腹腔注射20mg/kg式(III)化合物2次。Vehicle指溶解式(III)化合物的溶剂，由5％DMSO+5％Solutol HS 15(聚乙二醇-15羟基硬脂酸酯，购自巴斯夫公司)+90％注射用水组成。
实验期间每天称量小鼠体重，监测小鼠粪便成型程度以及便隐血情况，按照发病严重程度打分(每项0-4分)。临床疾病活动指数(DAI)是临床上用于评价疾病活动程度的一项指标，DAI分数＝(体重下降评分+大便成型评分+便血评分)/3。
实验第8天处死小鼠，取出完整结肠测量小鼠结肠长度。PBS冲洗结肠，解剖远端结肠用于病理分析。
实验结果发现：式(III)化合物显著抑制了肠炎小鼠体重下降(图3A)、DAI评分增高(图3B)、结肠缩短(图3C)的现象(n＝8)；病理分析的结果显示式(III)化合物显著抑制了结肠炎症、上皮损伤以及肠道结构病变的程度(图3D和3E)(n＝8)。这些结果说明式(III)化合物具有抑制DSS诱导的急性肠炎的效果，可用作炎症性肠病治疗。
实施例9式(III)化合物具有治疗小鼠EAE的效果
MOG35-55诱导的实验性自身免疫性脑膜炎(EAE)是经典的模拟人类多发性硬化症(MS)的小鼠模型。建模步骤如下：取MOG35-55(上海吉尔生化公司)，调整浓度为2mg/mL；取不完全弗氏佐剂(Sigma公司)，加入灭活的结核杆菌H37RA(DIFCO公司)，调整浓度为5mg/mL；按照体积比1:1将MOG35-55溶液与弗氏佐剂混合，冰上乳化2小时至油包水状态；小鼠麻醉后，在小鼠双耳后侧及股侧皮下进行免疫，每个部位注射50μL，共200μL/只，此时设为第0天；在免疫的第0天及第2天，每只小鼠每次腹腔注射300ng百日咳毒素(List labs公司)。该实验免疫小鼠为10周龄雌性C57BL/6小鼠。
EAE评分是用于评价EAE疾病活动程度的一项指标，按照疾病严重程度可评分0-5分：0分-无明显异常；0.5分-尾尖无力，运动能力未见异常；1分-整条尾巴无力，运动能力未见异常；1.5分-整条尾巴无力，两条后肢可分开，运动偶有踩空；2分-整条尾巴无力，两条后肢合在一起，运动常见踩空；2.5分-一条后肢拖行；3分-两条后肢拖行，仰面放置后可自行翻身；3.5分-两条后肢拖行，仰面放置后不能自行翻身，前肢可正常抓取笼架金属条；4分-小鼠两条后肢拖行，仰面放置后不能自行翻身，一条前肢无力，不能正常抓取笼架金属条；4.5分-小鼠四肢瘫痪；5分-小鼠死亡或连续2天超过4分。
建模10天后，将23只已初步发病(EAE评分0.5-1分)的小鼠随机分为2组，各组处理如下：EAE模型组(EAE+Vehicle)：每天腹腔注射溶剂2次，连续5天，n＝11；式(III)化合物处理组(EAE+Formula III)：每天腹腔注射40mg/kg式(III)化合物2次，连续5天，n＝12。
实验期间每天对小鼠的EAE评分进行打分。在式(III)化合物进行干预的第5天，处死小鼠，取脊髓膨大区组织用于病理分析。
实验结果发现：式(III)化合物显著降低了EAE模型小鼠的EAE评分(图4A)；脊髓病理分析结果显示，式(III)化合物处理显著抑制了脊髓炎性细胞浸润以及脱髓鞘程度(图4B)。这些结果说明式(III)化合物具有治疗小鼠EAE的效果，可用作多发性硬化症的治疗。
实施例10式(III)化合物具有治疗小鼠类银屑病的效果
局部涂抹咪喹莫特(IMQ)诱导的小鼠类银屑病在表型和组织学特征上与人类银屑病非常相似。建模步骤如下：实验第0天到第7天小鼠耳朵每天涂抹25mg IMQ(四川明欣利迪公司)，第7天发病达高峰期；之后间隔一天涂抹IMQ维持病情。
将8周龄雄性Babl/c小鼠分为3组，各组处理如下：空白对照组(Control+Vehicle)：实验全程不涂IMQ，第7天后每天腹腔注射溶剂2次，n＝6；模型组(IMQ+Vehicle)：连续7天涂抹IMQ，之后间隔一天涂抹IMQ维持病情，第7天后每天腹腔注射溶剂2次，n＝6；式(III)化合物处理组(EAE+Formula III)：连续7天涂抹IMQ，之后间隔一天涂抹IMQ维持病情，第7天后每天腹腔注射20mg/kg式(III)化合物，n＝8。
实验期间每天测量耳朵厚度。实验第21天处死小鼠，对小鼠耳朵拍照，取耳朵组织用于病理分析。
实验结果发现：相对于模型组，式(III)化合物处理显著抑制类银屑病小鼠耳朵增厚的现象(图5A和5B)；耳朵病理切片H&E染色评分显示，式(III)化合物处理显著抑制了类银屑病小鼠耳朵皮肤角化不全、角化过度(鳞状上皮增生)、Munro小体形成、棘层肥厚、炎性细胞浸润和真皮血管扩张充血(图5C和5D)。这些结果说明式(III)化合物具有治疗小鼠类银屑病的效果，可用作银屑病的治疗。
实施例11式(III)化合物对LPS诱导的小鼠脓毒症具有保护作用
脂多糖(LPS)诱导的小鼠脓毒症模型是经典的脓毒症动物模型。实验采用腹腔注射致死剂量的35mg/kg LPS(购自Sigma公司)诱导脓毒症。
将雄性C57BL/6小鼠分为2组，各组处理如下：模型组(LPS+Vehicle)：腹腔单次注射溶剂，n＝11；式(III)化合物处理组(LPS+Formula III)：腹腔单次注射20mg/kg式(III)化合物，n＝13。小鼠给与溶剂或式(III)化合物预处理2小时后，腹腔注射致死剂量的35mg/kg LPS诱导脓毒症。
观察小鼠存活情况，同时记录小鼠死亡时间，直到第6天。
使用Graphpad 7.0绘制生存曲线，进行显著性分析。具体结果如图6所示：式(III)化合物显著提高了小鼠的存活率。这些结果说明式(III)化合物对LPS诱导的小鼠脓毒症具有保护作用，可用于脓毒症的防治。
实施例12式(III)化合物对APAP诱导的肝损伤具有保护作用
对乙酰氨基酚(APAP)诱导的肝损伤模拟了临床的药物性肝损伤。实验前一天小鼠禁食不禁水，实验采用腹腔注射250mg/kgAPAP(购自sigma公司)诱导小鼠药物性肝损伤。
将雄性C57BL/6小鼠分为2组，各组处理如下：模型组(APAP+Vehicle)：腹腔单次注射溶剂，n＝6；式(III)化合物处理组(APAP+Formula III)：腹腔单次注射20mg/kg式(III)化合物，n＝6。小鼠给与溶剂或式(III)化合物预处理0.5小时后，腹腔注射250mg/kgAPAP诱导小鼠药物性肝损伤。
分别于APAP注射前以及注射后3小时，6小时，12小时，24小时从小鼠眼框采集血样，肝素钠抗凝。血液样本采集后使用ALT试剂盒(购自上海荣盛生物药业有限公司)检测小鼠血清中谷丙转氨酶(ALT)水平。
具体结果如图7所示：式(III)化合物显著抑制了肝损伤小鼠血清中ALT的水平。这些结果说明式(III)化合物对APAP诱导的肝损伤具有保护作用，可用于药物性肝损伤的防治。
本发明内容仅仅举例说明了要求保护的一些具体实施方案，其中一个或更多个技术方案中所记载的技术特征可以与任意的一个或多个技术方案相组合，这些经组合而得到的技术方案也在本申请保护范围内，就像这些经组合而得到的技术方案已经在本发明公开内容中具体记载一样。
参考文献：
[1]Horwitz D A,Fahmy T M,Piccirillo C A,et al.Rebalancing immune homeostasis to treat autoimmune diseases.Trends Immunol,2019,40(10):888-908.
[2]Maddur M S,Miossec P,Kaveri S V,et al.Th17 cells.The American Journal of Pathology,2012,181(1):8-18.
[3]Patel D D,Kuchroo V K.Th17 cell pathway in human immunity:Lessons from genetics and therapeutic interventions.Immunity,2015,43(6):1040-1051.
[4]Bedke T,Muscate F,Soukou S,et al.Il-10-producing t cells and their dual functions.Semin Immunol,2019,44:101335.
[5]Roncarolo M G,Gregori S,Bacchetta R,et al.The biology of t regulatory type 1cells and their therapeutic application in immune-mediated diseases.Immunity,2018,49(6):1004-1019.
[6]Reizis B.Plasmacytoid dendritic cells:development,regulation,and function.Immunity,2019,50(1):37-50.
[7]Ho I C,Miaw S C.Regulation of il-4 expression in immunity and diseases.Advances in Experimental Medicine and Biology,2016,941:31-77.

Claims (18)

  1. 一类倍半萜聚酮化合物或其药学上可接受的盐作为免疫调节剂的用途，其特征在于，所述倍半萜聚酮化合物如式(I)所示：
    其中，R1选自：羟基，乙酰氧基，或＝O；
    R2选自：甲基，羟甲基，乙酰羟甲基，醛基，或二甲氧基甲基；
    R3选自：甲基，羟甲基，乙酰羟甲基，或醛基；
    或者R2，R3与苯环组成5元内酰胺环或内酯环；
    R4，R5，R6，R7为碳原子，并且任一相邻基团间选自双键，其余基团间选自单键。
  2. 根据权利要求1所述的用途，其特征在于，所述药学上可接受的盐为所述倍半萜聚酮化合物与有机碱或无机碱形成的盐。
  3. 根据权利要求2所述的倍半萜聚酮类化合物或其药学上可接受的盐，其特征在于，所述形成的盐为钠盐、钾盐、钙盐、铁盐、镁盐、锌盐、铝盐、钡盐或铵盐。
  4. 一类倍半萜聚酮化合物或其药学上可接受的盐，其特征在于所述化合物为：
  5. 根据权利要求1-4中任一项所述倍半萜聚酮化合物或其药学上可接受的盐的制备方法，其特征在于，包括：由产生如式(I)所示的倍半萜聚酮类化合物的微生物，经发酵后再用色谱法分离后得到。
  6. 根据权利要求5所述的制备方法，其特征在于，所述微生物为漆斑属真菌。
  7. 根据权利要求6所述的制备方法，其特征在于，所述漆斑属真菌为ZLW0801-19，其保藏号为CGMCC No.19039。
  8. 根据权利要求1所述的用途，其特征在于，所述免疫调节剂用于免疫相关疾病的预防和/或治疗。
  9. 根据权利要求8所述的用途，其特征在于，所述的免疫相关疾病的预防和/或治疗是防治抑制IL17和/或促进IL10是有益的疾病或适应症。
  10. 根据权利要求9所述的用途，其特征在于，所述疾病或适应症选自银屑病、炎症性肠病、I型糖尿病、多发性硬化症、类风湿性关节炎、系统性红斑狼疮、哮喘、干燥综合症、强直性脊柱炎、器官移植免疫排斥、过敏、脓毒症、葡萄膜炎、自身免疫性肾炎、自身免疫甲状腺炎、白塞氏病、自身免疫性肝病、药物性肝损伤、扁平苔藓、痤疮、椎间盘突出、腹腔疾病。
  11. 根据权利要求8所述的用途，其特征在于，所述的免疫相关疾病的预防和/或治疗是防治促进IL4是有益的疾病或适应症。
  12. 根据权利要求11所述的用途，其特征在于，所述疾病或适应症为寄生虫感染相关。
  13. 根据权利要求8所述的用途，其特征在于，所述的免疫相关疾病的预防和/或治疗是防治抑制DC免疫反应是有益的疾病或适应症。
  14. 根据权利要求13所述的用途，其特征在于，所述疾病或适应症为炎症和自身免疫病、脓毒症、器官移植排斥反应。
  15. 一种用于免疫调节的药物组合物，其特征在于，包括权利要求1-4中任意一项所述倍半萜聚酮化合物或其药学上可接受的盐以及药学上可接受的载体。
  16. 根据权利要求15所述的药物组合物，其特征在于，所述倍半萜聚酮化合物或其药学上可接受的盐的含量为组合物重量的1-90％。
  17. 根据权利要求15-16所述的药物组合物，其特征在于，所述药物组合物为口服制剂或注射剂。
  18. 根据权利要求17所述的药物组合物，其特征在于，所述口服制剂选自普通片剂、分散片、肠溶片、颗粒、胶囊、滴丸、散剂、口服液或乳剂，所述注射剂为小水针剂、输液剂或冻干粉针。
PCT/CN2023/097904 2022-06-02 2023-06-01 一类倍半萜聚酮化合物作为免疫调节剂在防治免疫性疾病的用途 WO2023232123A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202210626202.1A CN115040507A (zh) 2022-06-02 2022-06-02 一类倍半萜聚酮化合物作为免疫调节剂在防治免疫性疾病的用途
CN202210626202.1 2022-06-02

Publications (1)

Publication Number Publication Date
WO2023232123A1 true WO2023232123A1 (zh) 2023-12-07

Family

ID=83158988

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2023/097904 WO2023232123A1 (zh) 2022-06-02 2023-06-01 一类倍半萜聚酮化合物作为免疫调节剂在防治免疫性疾病的用途

Country Status (2)

Country Link
CN (1) CN115040507A (zh)
WO (1) WO2023232123A1 (zh)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115040507A (zh) * 2022-06-02 2022-09-13 暨南大学 一类倍半萜聚酮化合物作为免疫调节剂在防治免疫性疾病的用途
CN114956974B (zh) * 2022-06-02 2023-10-31 暨南大学 一类倍半萜聚酮化合物、药物组合物及其用途
CN114796171A (zh) * 2022-06-02 2022-07-29 暨南大学 一个倍半萜聚酮化合物用于防治肺动脉高压的用途

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114796171A (zh) * 2022-06-02 2022-07-29 暨南大学 一个倍半萜聚酮化合物用于防治肺动脉高压的用途
CN114956974A (zh) * 2022-06-02 2022-08-30 暨南大学 一类倍半萜聚酮化合物、药物组合物及其用途
CN115040507A (zh) * 2022-06-02 2022-09-13 暨南大学 一类倍半萜聚酮化合物作为免疫调节剂在防治免疫性疾病的用途

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114796171A (zh) * 2022-06-02 2022-07-29 暨南大学 一个倍半萜聚酮化合物用于防治肺动脉高压的用途
CN114956974A (zh) * 2022-06-02 2022-08-30 暨南大学 一类倍半萜聚酮化合物、药物组合物及其用途
CN115040507A (zh) * 2022-06-02 2022-09-13 暨南大学 一类倍半萜聚酮化合物作为免疫调节剂在防治免疫性疾病的用途

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
KUANG, RUNQIAO: "A Study of Secondary Metabolites of Wetland-derived Myrothecium gramineum", MASTER THESIS, JINAN UNIVERSITY, CN, 18 December 2018 (2018-12-18), CN, XP009551320 *
LAUBE, T. ; BERNET, A. ; DAHSE, H.M. ; JACOBSEN, I.D. ; SEIFERT, K.: "Synthesis and pharmacological activities of some sesquiterpene quinones and hydroquinones", BIOORGANIC & MEDICINAL CHEMISTRY, ELSEVIER, AMSTERDAM, NL, vol. 17, no. 4, 15 February 2009 (2009-02-15), AMSTERDAM, NL, pages 1422 - 1427, XP025949546, ISSN: 0968-0896, DOI: 10.1016/j.bmc.2009.01.028 *
LIU, Q. ; SHU, X. ; SUN, A. ; SUN, Q. ; ZHANG, C. ; AN, H. ; LIU, J. ; CAO, X.: "Plant-derived small molecule albaconol suppresses LPS-triggered proinflammatory cytokine production and antigen presentation of dendritic cells by impairing NF-@kB activation", INTERNATIONAL IMMUNOPHARMACOLOGY, ELSEVIER, AMSTERDAM, NL, vol. 8, no. 8, 1 August 2008 (2008-08-01), NL , pages 1103 - 1111, XP022710377, ISSN: 1567-5769, DOI: 10.1016/j.intimp.2008.04.001 *
TANG XIN, WANG CHUANXI, WANG LEI, REN FEIFEI, KUANG RUNQIAO, LI ZHENHUA, HAN XUE, CHEN YIMING, CHEN GUODONG, WU XIUQING, LIU JIE, : "Aureane-type sesquiterpene tetraketides as a novel class of immunomodulators with interleukin-17A inhibitory activity", ACTA PHARMACEUTICA SINICA B, vol. 13, no. 9, 1 September 2023 (2023-09-01), pages 3930 - 3944, XP093115352, ISSN: 2211-3835, DOI: 10.1016/j.apsb.2023.03.017 *

Also Published As

Publication number Publication date
CN115040507A (zh) 2022-09-13

Similar Documents

Publication Publication Date Title
WO2023232123A1 (zh) 一类倍半萜聚酮化合物作为免疫调节剂在防治免疫性疾病的用途
Plevy et al. A role for TNF-alpha and mucosal T helper-1 cytokines in the pathogenesis of Crohn's disease.
Zlotta et al. What are the immunologically active components of Bacille Calmette‐Guérin in therapy of superficial bladder cancer?
WO2023232125A1 (zh) 一类倍半萜聚酮化合物、药物组合物及其用途
US20040208863A1 (en) Anti-inflammatory activity from lactic acid bacteria
Li et al. Aconitine: a potential novel treatment for systemic lupus erythematosus
KR101599982B1 (ko) 노노무라에아 종으로부터의 고리형 펩티드, 그에 대한 생산 프로세스 및 이를 포함하는 마이코박테리아 관련 질병의 치료 혹은 예방을 위한 약학적 조성물
KR101446179B1 (ko) 바우미 상황버섯 유래의 폴리페놀 추출물을 유효성분으로 함유하는 염증질환의 예방 및 치료용 조성물
JP2011505349A (ja) 肥満細胞活性化の阻害用組成物を製造するためのエル・カゼイ株の使用
Aldinucci et al. Modulating dendritic cells (DC) from immunogenic to tolerogenic responses: a novel mechanism of AZA/6-MP
Madouri et al. Protein kinase Cθ controls type 2 innate lymphoid cell and TH2 responses to house dust mite allergen
Zhou et al. The effect of triptolide on CD4+ and CD8+ cells in Peyer's patch of SD rats with collagen induced arthritis
JPWO2018135556A1 (ja) Ｈｔｌｖ−１関連脊髄症を治療することに用いるための医薬組成物
Ma et al. Dysfunction of IL-10-producing type 1 regulatory T cells and CD4+ CD25+ regulatory T cells in a mimic model of human multiple sclerosis in Cynomolgus monkeys
KR102031706B1 (ko) 나노크기의 산화 그래핀을 포함하는 항염증용 조성물
KR100913406B1 (ko) Ｔｈ１－매개 면역 질환의 예방 또는 치료용 조성물
TW201249454A (en) Non-polysaccharide compound and usage thereof, and extraction method from Dendrobium genus plant
JPWO2003051382A1 (ja) アポトーシスを誘導する方法およびそのための組成物
US5336666A (en) Immunostimulant drug based on polar glyopeptidolipids of mycobacterium chelonae
Kansa et al. Phenotypic identification of suppressor‐effector, suppressor‐amplifier and suppressor‐inducer T cells of B cell differentiation in man
US20200297680A1 (en) Immunotherapeutic agent
CN102584924A (zh) 碳-21甾体化合物的用途
CN106916226B (zh) 以ctla-4为靶点的免疫调节药物治疗结核病的应用
CN116064391A (zh) 诱导Treg细胞的酵母衍生多糖的结构和功能特征
CN111481565A (zh) 戈氏副拟杆菌的脂多醣用于抑制发炎反应的医药组合物的用途

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23815300

Country of ref document: EP

Kind code of ref document: A1