WO2019129249A1 - 含三萜类化合物的药物组合物及其用途 - Google Patents

含三萜类化合物的药物组合物及其用途 Download PDF

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WO2019129249A1
WO2019129249A1 PCT/CN2018/125075 CN2018125075W WO2019129249A1 WO 2019129249 A1 WO2019129249 A1 WO 2019129249A1 CN 2018125075 W CN2018125075 W CN 2018125075W WO 2019129249 A1 WO2019129249 A1 WO 2019129249A1
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substituted
group
compound
unsubstituted
formula
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PCT/CN2018/125075
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English (en)
French (fr)
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郑钦元
朱敬杰
曲婧
罗煜
丁时澄
秦丽军
周想燕
孙传民
赵芬琴
张存清
季中伟
郭丽
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上海蓝木化工有限公司
郑钦元
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Priority claimed from CN201711480630.3A external-priority patent/CN109985050A/zh
Priority claimed from CN201711480659.1A external-priority patent/CN109985051A/zh
Priority claimed from CN201711484482.2A external-priority patent/CN109985071A/zh
Priority claimed from CN201711489391.8A external-priority patent/CN109985052A/zh
Application filed by 上海蓝木化工有限公司, 郑钦元 filed Critical 上海蓝木化工有限公司
Priority to CN201880084560.6A priority Critical patent/CN111526878A/zh
Priority to US16/959,084 priority patent/US20200384003A1/en
Publication of WO2019129249A1 publication Critical patent/WO2019129249A1/zh

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    • A—HUMAN NECESSITIES
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    • A61K9/00—Medicinal preparations characterised by special physical form
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    • A61K31/575—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
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    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
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    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
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    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
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    • 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
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A61P27/10—Ophthalmic agents for accommodation disorders, e.g. myopia
    • 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
    • A61P27/12—Ophthalmic agents for cataracts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
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    • A61K9/0051—Ocular inserts, ocular implants
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    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J17/00—Normal steroids containing carbon, hydrogen, halogen or oxygen, having an oxygen-containing hetero ring not condensed with the cyclopenta(a)hydrophenanthrene skeleton
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    • C07J—STEROIDS
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    • C07J—STEROIDS
    • C07J41/00—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring
    • C07J41/0033—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005
    • C07J41/0055—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring not covered by C07J41/0005 the 17-beta position being substituted by an uninterrupted chain of at least three carbon atoms which may or may not be branched, e.g. cholane or cholestane derivatives, optionally cyclised, e.g. 17-beta-phenyl or 17-beta-furyl derivatives
    • C—CHEMISTRY; METALLURGY
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    • C07J—STEROIDS
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    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J71/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton is condensed with a heterocyclic ring
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    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J9/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J9/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane
    • C07J9/005—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane containing a carboxylic function directly attached or attached by a chain containing only carbon atoms to the cyclopenta[a]hydrophenanthrene skeleton

Definitions

  • the present invention relates to the field of medicine, and in particular, the present invention relates to a novel use of a triterpenoid compound or an extract of Inonotus obliquus.
  • the lens of a normal human eye is composed of a plurality of aligned lens proteins. Once these lens proteins are arranged incorrectly or misfolded, protein aggregates are formed, which affects the normal transparency and refractive index of the lens. Among them, cataract is a major form of manifestation, and it is also the world's highest blinding disease. Therefore, the treatment and prevention of lens diseases, especially cataracts, is important. Unfortunately, the current treatment of cataracts is still the main treatment for surgery and replacement of intraocular lenses.
  • lanosterol can be used to treat cataracts in animals.
  • the structural formula of lanosterol is as follows:
  • the sustained-release lanosterol drug has a cataract treatment effect on dogs only when combined with high-frequency intravitreal injection.
  • different researchers have also pointed out that the eyeball or crystal is directly administered. Wool sterol does not restore or improve the transparency of cataract crystals in primates.
  • a first aspect of the invention provides a compound of formula I, or an optical isomer thereof, or a racemate thereof, or a solvate thereof, or a pharmaceutically acceptable salt thereof, or a prodrug thereof, or a hydrazine thereof Use of the compounds for the preparation of a pharmaceutical composition or formulation for the prevention and/or treatment of ocular diseases caused by lens lesions;
  • q 0, 1 or 2;
  • R1a, R1b, R2a, R2b, R3a and R3b are each independently selected from: hydrogen, substituted or unsubstituted C1 - C20 alkyl, substituted or unsubstituted C2 - C20 alkenyl, substituted or unsubstituted C2 - C20 alkyne , substituted or unsubstituted C3 ⁇ C10 cycloalkyl, ⁇ OH, substituted or unsubstituted C1-C10 alkoxy, ⁇ COOH, ⁇ CHO, substituted or unsubstituted C1 ⁇ C10 ester, ⁇ SH, substituted Or unsubstituted C1 ⁇ C10 alkylthio, ⁇ A ⁇ B,
  • A is a non-divalent linking group
  • B is H, ⁇ OH, ⁇ SH, C1-C3 alkoxy, C1-C3 alkylthio, ⁇ CHO, ⁇ COOH, C1-C4 ester, C3 ⁇ C10 cycloalkyl, aryl, C3-C10 5- to 6-membered heteroaryl, or benzyl;
  • R1a, R2a, R3a, R1b, R2b and R3b is a group containing O or S;
  • Z is selected from the group consisting of H, substituted or unsubstituted C1 - C20 alkyl, substituted or unsubstituted C2 - C20 alkenyl, substituted or unsubstituted C2 - C20 alkynyl, substituted or unsubstituted C3 - C10 ring Alkyl, substituted or unsubstituted aryl, substituted or unsubstituted 5-8-membered heteroaryl, substituted or unsubstituted ⁇ (C1-C6 alkylene)-aryl, substituted or unsubstituted ⁇ (C1 ⁇ C6 alkylene) ⁇ 5 ⁇ 8 membered heteroaryl, ⁇ OH, substituted or unsubstituted C1-C20 alkoxy, ⁇ SH, substituted or unsubstituted C1-C20 alkylthio, substituted or unsubstituted a C1-C10 ester group, a substituted or un
  • R4 is hydrogen, substituted or unsubstituted C1-C4 alkyl
  • R7, R12 and R15 are each independently selected from: none, hydrogen, substituted or unsubstituted C1-C4 alkyl;
  • R5, R6, R8, R9a, R9b, R14, R16, R17a and R17b are each independently selected from the group consisting of hydrogen, OH, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkoxy, halogen.
  • the dotted line The total number of double bonds represented is 0, 1, 2, 3 or 4.
  • the double bond is located between the following positions: a and b, b and c, c and d, e and f, c and f, f and g, g and h, h and a , b and i, i and j, a and k, and/or k and l.
  • the double bond is located between the following positions: a and b.
  • the double bond is located between the following positions: h and a, and/or b and i.
  • the valence state of each C conforms to the requirements of the chemically stable structure (ie, C is tetravalent).
  • the divalent linking group has 1 to 10 linking units selected from the group consisting of -CRaRb-; -C(OH)Ra-, -NRa-, -O-, -CO
  • Ra and Rb are each independently H, C1 - C6 alkyl, C1 - C6 haloalkyl, C3 - C6 cycloalkyl, or benzyl.
  • A is independently a substituted or unsubstituted C1-C4 alkylene group
  • Y is a group having a hydroxyl group or a SH substituent.
  • the R4 is hydrogen or methyl.
  • R7, R12 and R15 are each independently selected from the group consisting of: none, hydrogen, methyl.
  • R13a and R13b are -OH, or -SH.
  • the R10a, R10b are each independently selected from the group consisting of hydrogen, OH, ⁇ SH, ⁇ OSO 3 H, —OPO 3 H, —COOH, —CHO; or R 10a and R 10b constitute a carbonyl group.
  • R5, R6, R8, R9a, R9b, R14, R16, R17a and R17b are each independently selected from the group consisting of hydrogen and methyl.
  • the R9a, R9b are each independently selected from the group consisting of hydrogen, methyl and halogen.
  • the R9a is halogen (F, Cl, or Br) and R9b is selected from hydrogen or methyl.
  • At least one of R1a and R1b is a group containing O or S.
  • At least one of R2a and R2b is a group containing O or S.
  • At least one of R3a and R3b is a group containing O or S.
  • the Z is not ⁇ CH 2 C(CH 3 ) 3 ;
  • Z is ⁇ CH 2 C(CH 3 ) 2 —OH.
  • q is 1 or 2.
  • the compound of formula I is a compound of formula 1-1:
  • R1a, R1b, R2a, R2b, R3a, R3b, R4, R7, R10a, R10b, R11a, R11b, R12, R13a, R13b, R15 and Z are as defined above.
  • R11a and/or R11b are methyl.
  • the compound of formula I is a compound of formula I-2:
  • R1a, R1b, R2a, R2b, R3a, R3b, R4, R7, R11a, R11b, R13a, R13b, R15 and Z are as defined above.
  • the compound of formula I is selected from the group consisting of the following compounds:
  • R1a, R1b, R2a, R2b and Z are as defined above.
  • the compound of formula I is selected from the group consisting of the following compounds:
  • R1a, R1b, R2a, and R2b are as defined above.
  • the compound of formula I is of the formula shown in formula I-7:
  • R1a, R1b, R2a, R2b are as defined above.
  • the compound of formula I is of the formula shown in formula I-8:
  • R1a, R1b, R2a, R2b are as defined above.
  • the compound of formula I is selected from the group consisting of: Table A:
  • the compound of formula I is selected from the group consisting of:
  • the compound of formula I is selected from the group consisting of
  • the ocular disease caused by the lens lesion is selected from the group consisting of cataract, presbyopia, myopia, cortical opacity, presbyopia, and ocular complications caused by diabetes.
  • the ocular disease is selected from the group consisting of a congenital cataract, an acquired cataract.
  • the ocular disease is selected from the group consisting of mature cataracts and immature cataracts.
  • the cataract is selected from the group consisting of a traumatic cataract, a metabolic cataract, an elderly cataract, a congenital cataract, a self-style cataract, a concurrent cataract, or a combination thereof.
  • the metabolic cataract comprises a diabetic metabolizing cataract.
  • the traumatic cataract includes a surgically related cataract.
  • the self-styled cataract includes an ageing spontaneous cataract.
  • the pharmaceutical composition or formulation comprises: (a) a therapeutically effective amount of a compound of formula I as a first active ingredient, or an optical isomer thereof or a racemate thereof, Or a solvate thereof, or a pharmaceutically acceptable salt thereof, or a prodrug thereof; (b) a pharmaceutically acceptable carrier;
  • the content of the first active ingredient is from 0.001 to 99% by weight, preferably from 0.01 to 70% by weight, more preferably from 0.05 to 40% by weight, based on the total weight of the composition.
  • the pharmaceutical composition or formulation comprises: (a) a therapeutically effective amount of a compound of formula I as a first active ingredient, or an optical isomer thereof or a racemate thereof, Or a solvate thereof, or a pharmaceutically acceptable salt thereof, or a prodrug thereof, or a deuterated compound thereof; (b) a pharmaceutically acceptable carrier;
  • the content of the first active ingredient is from 0.001 to 99% by weight, preferably from 0.01 to 70% by weight, more preferably from 0.05 to 40% by weight, based on the total weight of the composition.
  • the concentration of the first active ingredient is preferably from 1 to 500 mM or from 10 to 200 mM, preferably from 15 to 150 mM, more preferably from 20 to 50 mM; most preferably from 20 to 30 mM.
  • the pharmaceutical composition or formulation is selected from the group consisting of an injection, an external preparation, and an oral preparation.
  • the pharmaceutical composition or preparation is in the form of an ophthalmic preparation, and the ophthalmic preparation is an eye drop, an emulsion, a gel, an ophthalmic ointment, a sustained release microsphere, and an intraocular lens. Released tablets, medicinal sustained release film.
  • the pharmaceutical composition or formulation further comprises: (c) a second active ingredient, wherein the second active ingredient is selected from the group consisting of lanolin compounds, lanosterol, all rust Any of the compounds (especially steroid compounds, terpenoids), azole compounds, glucocorticoids, antibiotics, or a combination thereof contained in the fungus are contained in the genus Pseudomonas or the polyporus.
  • the second active ingredient is selected from the group consisting of lanolin compounds, lanosterol, all R. grisea or polypores, and any compound contained in the fungus (especially Steroids, terpenoids, azoles, amyloid modulators, glucocorticoids, antibiotics, or combinations thereof.
  • the second active ingredient is present in an amount of from 0.01 to 20% by weight, preferably from 5 to 15% by weight, based on the total weight of the composition.
  • the concentration thereof is preferably 10 to 200 mM, preferably 15 to 150 mM, more preferably 20 to 50 mM; Good ground 20 ⁇ 30mM.
  • the lanolin compound is selected from the group consisting of lanosterol, dihydro lanosterol, 25-hydroxycholesterol, or a combination thereof.
  • amyloid modulator is selected from the group consisting of TPPB (CAS. 497259 ⁇ 23 ⁇ 1), rosmarinic acid, and doxycycline.
  • the glucocorticoid compound is selected from the group consisting of dexamethasone, hydrocortisone, or a combination thereof.
  • the antibiotic is selected from the group consisting of tobramycin, gentamicin sulfate, chlortetracycline, chloramphenicol, or a combination thereof.
  • the azole compound is selected from the group consisting of econazole, isoconazole, bifonazole, clotrimazole, aripiprazole, ketoconazole, fluconazole, phenylimidazole, and imidium.
  • the azole compound is selected from the group consisting of econazole, fluconazole, isoconazole, tebuconazole, propiconazole, or a combination thereof.
  • the mass ratio of the compound of formula I to the azole compound is from 50:1 to 1:50, preferably from 10:1 to 1:10.
  • the compound of formula I further comprises a deuterated compound of a compound of formula I.
  • the pharmaceutical composition or formulation is also used to (b) inhibit, reverse (dissolve or depolymerize) lens protein aggregation.
  • the pharmaceutical composition or formulation is also used to (c) prevent and/or treat a disease associated with lens protein aggregation.
  • the lens protein comprises an ⁇ B crystallin protein.
  • the disease associated with lens protein aggregation is selected from the group consisting of cataract, presbyopia, myopia, cortical opacity, presbyopia, and ocular complications caused by diabetes.
  • a non-therapeutic and/or non-diagnostic method for improving or maintaining lens transparency in vitro comprising: crystallizing a compound of formula I, or an optical isomer thereof or Contacting the racemate, or a solvate thereof, or a pharmaceutically acceptable salt thereof, or a prodrug thereof, or a deuterated compound thereof, thereby improving (e.g., increasing) or maintaining lens clarity, wherein the compound of formula I is as above Said.
  • a method for preventing and/or treating an ocular disease caused by a lens lesion comprising the steps of: administering a compound of the formula I, or an optical isomer thereof, or a substance thereof Racemic, or a solvate thereof, or a pharmaceutically acceptable salt thereof, or a prodrug thereof, or a deuterated compound thereof, wherein the compound of formula I is as defined above.
  • the subject is a human and a non-human mammal.
  • the non-human mammals include, but are not limited to, pets (such as dogs and cats), livestock (such as cattle, sheep, horses, pigs), various zoo animals (pandas, elephants, etc.) .
  • the subject further includes other animals than human and non-human mammals, such as non-mammals.
  • a novel compound of formula II or an optical isomer thereof, or a racemate thereof, or a solvate thereof, or a pharmaceutically acceptable salt thereof, or a prodrug thereof, Or its deuterated compound,
  • P is 0, 1 or 2;
  • R18a, R18b are each independently hydrogen, substituted or unsubstituted C1-C4 alkyl, or -COOH;
  • R19a, R19b are each independently hydrogen, substituted or unsubstituted C1-C4 alkyl, or halogen;
  • R20 is a substituted or unsubstituted C1-C6 acyl group, a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted saccharyl-O-(C1-C6 alkyl)-, a substituted or unsubstituted fluorenyl group, a substituted or unsubstituted C3-C6 heterocycloalkyl group, or a substituted or unsubstituted C2-C6 alkenyl group;
  • R21a, R21b are each independently hydrogen, hydroxy, decyl, or substituted or unsubstituted C1-C4 alkyl;
  • R22a, R22b are each independently hydrogen, or a substituted or unsubstituted C1-C4 alkyl group
  • R23a, R23b are each independently hydrogen, hydroxy, thiol, glycosyl, or substituted or unsubstituted C1-C alkoxy;
  • R24a, R24b are each independently hydrogen, hydroxy, or halogen
  • the glycosyl group is a monosaccharide (such as pentose or hexose), or a disaccharide.
  • the dotted line The total number of double keys represented is 0, 1, or 2.
  • the double bond is located between m and n.
  • the double bond is between m and l, and/or between o and n.
  • R22a and/or R22b are methyl.
  • p is 1.
  • p is 2.
  • R23a is hydrogen and R23b is hydroxy, thiol or saccharide.
  • R20 is a glycosyl-O-(C1-C6 alkyl)-.
  • the glycosyl group is a pentose or hexose group, preferably selected from the group consisting of glucosyl, fructosyl, mannosyl, arabinose, ribosyl, or combinations thereof.
  • the heterocycloalkyl group contains from 1 to 3 heteroatoms selected from the group consisting of N, O and S.
  • the heterocycloalkyl group is a C3 to C6 heterocycloalkyl group having 1 O hetero atom.
  • the compound of formula II has one or more characteristics selected from the group consisting of:
  • R18a, R18b are each independently -COOH, or methyl
  • R19a, R19b are each independently hydrogen or halogen
  • R21a, R21b are each independently hydrogen, hydroxy, or fluorenyl
  • R22a, R22b are each independently methyl
  • R23a, R23b are each independently hydrogen, hydroxy, thiol or hexavalent monosaccharide
  • R24a is hydrogen and R24b is fluorine.
  • two or more hydroxyl groups are not contained on the same carbon atom.
  • two or more fluorenyl groups are not contained on the same carbon atom.
  • the six-membered monosaccharide is glucose
  • the compound of formula II is selected from the group consisting of: Table B:
  • an extract of Inonotus obliquus for preparing a pharmaceutical composition or preparation for: (a) preventing and/or treating a lens disease caused by a disease Eye disease.
  • the extract comprises a fat-soluble extract.
  • the extract comprises an alcohol extract.
  • the extract comprises an extract of a non-aqueous solvent.
  • the extract is a water-insoluble extract or a poorly water-soluble extract.
  • the extract contains the following components: terpenoids, terpenoids, or a combination thereof.
  • the extract comprises a steroid extract.
  • the extract comprises a triterpenoid, preferably a tetracyclic triterpenoid extract.
  • the extract contains one or more compounds selected from the group C below:
  • the extract contains a compound selected from the group consisting of:
  • the extract contains a compound selected from the group consisting of:
  • the weight percentage of the inoculum is 0.01-99.99% by weight, preferably 1-99% by weight.
  • the weight percentage of the tylosic acid in the extract is from 0.01 to 99.99% by weight, preferably from 1 to 99% by weight.
  • the inoculum and the oleic acid account for 5 to 100% by weight, preferably 10 to 100% by weight, more preferably 20 to 100% by weight of the tetracyclic triterpenoid content. More preferably, it is 30-100% by weight, more preferably 70-100% by weight, and most preferably 80-100% by weight.
  • the extract is not pure lanosterol.
  • the weight ratio of Inotodiol to lanosterol in the extract is ⁇ 2:1, preferably ⁇ 5:1, more preferably ⁇ 10:1.
  • the extract is an extract of C1-C6 alcohol of Inonotus obliquus.
  • the C1-C6 alcohol comprises methanol, ethanol, propanol or a combination thereof.
  • the extract is further purified after alcohol extraction.
  • the further purification comprises: extracting with petroleum ether and/or an ester solvent.
  • the extract of Inonotus obliquus comprises a fat-soluble extract or a water-soluble extract of a fruiting body of Inonotus obliquus.
  • the ocular disease is selected from the group consisting of cataract, presbyopia, myopia, cortical opacity, presbyopia, and ocular complications caused by diabetes.
  • the ocular disease is selected from the group consisting of a congenital cataract, an acquired cataract.
  • the ocular disease is selected from the group consisting of mature cataracts and immature cataracts.
  • the cataract is selected from the group consisting of a traumatic cataract, a metabolic cataract, a cataract cataract, an elderly cataract, a self-style cataract, or a combination thereof.
  • the traumatic cataract includes a surgically related cataract.
  • the self-styled cataract includes an ageing spontaneous cataract.
  • the metabolic cataract comprises a diabetic metabolizing cataract.
  • the extract of Inonotus obliquus is prepared by the following method:
  • step (3) the extract of Inonotus obliquus is subjected to silica gel column chromatography, and the eluate is collected, and the purified extract of Inonotus obliquus is isolated.
  • the extract of Inonotus obliquus is prepared by the following method:
  • the preparation method further comprises the step (3) of performing the silica gel column chromatography on the extract of the Inonotus obliquus, collecting the eluate, and separating the purified birch. Inoculillus extract.
  • the weight ratio of the coarse powder of the B. obliqu. fruit body to the alcohol solvent is 1:30-100, preferably 1:60-90.
  • the alcohol solvent comprises ethanol.
  • the alcohol solvent comprises a 90-100% (v/v) aqueous solution of ethanol.
  • the fruiting body is a dry fruiting body.
  • the pharmaceutical composition or formulation comprises: (a) a therapeutically effective amount of an extract of Inonotus obliquus as the first active ingredient; (b) a pharmaceutically acceptable carrier.
  • the content of the first active ingredient is from 0.001 to 99% by weight, preferably from 0.01 to 70% by weight, more preferably from 0.05 to 40% by weight, based on the total weight of the composition or formulation.
  • the pharmaceutical composition or formulation is selected from the group consisting of an oral preparation, an injection, and an external preparation.
  • the pharmaceutical composition or preparation is in the form of an ophthalmic preparation, and the ophthalmic preparation is an eye drop, an emulsion, a gel, an ophthalmic ointment, a sustained release microsphere, and an intraocular lens. Released tablets, medicinal sustained release film.
  • the pharmaceutical composition or formulation further comprises: (c) a second active ingredient, wherein the second active ingredient is selected from the group consisting of lanosterol compounds, azole compounds, sugars Corticosteroids, antibiotics, or a combination thereof.
  • the pharmaceutical composition or formulation further comprises: (c) a second active ingredient, wherein the second active ingredient is selected from the group consisting of lanosterol compounds, azole compounds, sugars Corticosteroids, amyloid modulators, antibiotics, or a combination thereof.
  • the second active ingredient is present in an amount of from 0.01 to 20% by weight, preferably from 5 to 15% by weight, based on the total weight of the composition.
  • the concentration thereof is preferably 10 to 200 mM, preferably 15 to 150 mM, more preferably 20 to 50 mM; most preferably 20 to ⁇ 30 mM.
  • the lanosterol compound is selected from the group consisting of: lanosterol.
  • the glucocorticoid compound is selected from the group consisting of dexamethasone, hydrocortisone, or a combination thereof.
  • amyloid modulator is selected from the group consisting of TPPB (CAS. 497259 ⁇ 23 ⁇ 1), rosmarinic acid, and doxycycline.
  • the antibiotic is selected from the group consisting of tobramycin, gentamicin sulfate, chlortetracycline, chloramphenicol, or a combination thereof.
  • the azole compound is selected from the group consisting of econazole, isoconazole, bifonazole, clotrimazole, aripiprazole, ketoconazole, fluconazole, phenylimidazole, and imidium.
  • the mass ratio of the extract of Inonotus obliquus to the azole compound is from 50:1 to 1:50, preferably from 10:1 to 1:10.
  • the pharmaceutical composition or formulation is further for: (b) inhibiting, reversing lens protein aggregation; and/or (c) preventing and/or treating a disease associated with lens protein aggregation.
  • the lens protein comprises an ⁇ B crystallin protein.
  • the disease associated with lens protein aggregation is selected from the group consisting of cataract, presbyopia, myopia, cortical opacity, presbyopia, and ocular complications caused by diabetes.
  • a method for improving or maintaining lens transparency in vitro, non-therapeutic and/or non-diagnostic comprising the steps of: contacting a lens with an extract of Inonotus obliquus to improve (or enhance) Or maintaining lens transparency, wherein the inonotus extract is as described in the fourth aspect of the invention.
  • a method for preventing and/or treating an ocular disease caused by a lens lesion comprising the steps of: administering a desired object, an extract of Inonotus obliquus, wherein the extract of Inonotus obliquus
  • the object is as described in the sixth aspect of the invention.
  • the subject is a human and a non-human mammal.
  • the non-human mammals include, but are not limited to, pets (such as dogs and cats), livestock (such as cattle, sheep, horses, pigs), various zoo animals (pandas, elephants, etc.) .
  • the subject further includes other animals than human and non-human mammals, such as non-mammals.
  • a pharmaceutical composition comprising:
  • a second active ingredient wherein the second active ingredient is selected from the group consisting of an azole compound, an amyloid modulator, or a combination thereof.
  • the content of the first active ingredient is from 0.001 to 99% by weight, preferably from 0.01 to 70% by weight, more preferably from 0.05 to 40% by weight, based on the total weight of the composition.
  • the concentration of the first active ingredient is preferably from 1 to 500 mM or from 10 to 200 mM, preferably from 15 to 150 mM, more preferably from 20 to 50 mM; most preferably from 20 to 30 mM.
  • the second active ingredient is present in an amount of from 0.01 to 20% by weight, preferably from 5 to 15% by weight, based on the total weight of the composition.
  • the concentration thereof is preferably 10 to 200 mM, preferably 15 to 150 mM, more preferably 20 to 50 mM; most preferably 20 to 30 mM. .
  • the azole compound is selected from the group consisting of econazole, isoconazole, bifonazole, clotrimazole, aripiprazole, ketoconazole, fluconazole, phenylimidazole, and imidium.
  • the azole compound is selected from the group consisting of econazole, fluconazole, isoconazole, tebuconazole, propiconazole, or a combination thereof.
  • amyloid modulator is selected from the group consisting of: TPPB (CAS. 497259 ⁇ 23 ⁇ 1), rosmarinic acid, doxycycline, or a combination thereof.
  • the mass ratio of the compound of formula I (or the compound of formula II) to the azole compound is from 50:1 to 1:50, preferably from 10:1 to 1:10.
  • the compound of formula I is selected from the group consisting of: inonotc, alginic acid, or a combination thereof.
  • the pharmaceutical composition or formulation is selected from the group consisting of an injection, an external preparation, and an oral preparation.
  • the pharmaceutical composition or preparation is in the form of an ophthalmic preparation, and the ophthalmic preparation is an eye drop, an emulsion, a gel, an ophthalmic ointment, a sustained release microsphere, and an intraocular lens. Released tablets, medicinal sustained release film.
  • FIG. 1 is a view showing the morphology of an upper light source of a rat lens after successful cataract extraction in the first embodiment of the present invention.
  • FIG. 2 is a view showing the form of a lower light source of a rat lens after successful cataract extraction in the first embodiment of the present invention.
  • Fig. 3 is a view showing the morphology of the upper light source of the rat lens after successful cataract extraction in the first group of the present invention.
  • Fig. 4 is a view showing the morphology of a lower light source of a rat lens after successful cataract extraction in the first group of the present invention.
  • Figure 5 is a view showing the morphology of the upper light source of the rat lens after the end of the experiment selected in the first embodiment of the present invention.
  • Fig. 6 is a view showing the form of a lower light source of a rat lens after the end of the experiment selected in the first embodiment of the present invention.
  • Figure 7 is a view showing the morphology of the upper light source of the rat lens after the end of the experiment selected in the first group of the present invention.
  • Figure 8 is a view showing the morphology of the lower light source of the rat lens after the end of the experiment selected in the first group of the present invention.
  • Fig. 9 is a view showing the morphology of the upper light source of the rat lens after successful cataract extraction in the second embodiment of the present invention.
  • Fig. 10 is a view showing the form of a lower light source of a rat lens after successful cataract extraction in the second embodiment of the present invention.
  • Figure 11 is a view showing the morphology of the upper light source of the rat lens after successful cataract extraction in the second group of the present invention.
  • Figure 12 is a view showing the morphology of the lower light source of the rat lens after successful cataract extraction in the second group of the present invention.
  • Figure 13 is a view showing the morphology of the upper light source of the rat lens after the end of the experiment selected in the second embodiment of the present invention.
  • Figure 14 is a view showing the morphology of the lower light source of the rat lens after the end of the experiment selected in the second embodiment of the present invention.
  • Figure 15 is a view showing the morphology of the upper light source of the rat lens after the end of the experiment selected in the second group of the present invention.
  • Figure 16 is a view showing the morphology of the lower light source of the rat lens after the end of the experiment selected in the second group of the present invention.
  • Figure 17 is a view showing the morphology of an eyeball before and after treatment of a cynomolgus monkey congenital cataract (left eye) in Example 8 of the present invention, wherein 17A is before administration and 17B is after administration.
  • 18 is a view showing the morphology of an eyeball before and after treatment of a cynomolgus monkey congenital cataract (right eye) in Example 8 of the present invention, wherein 18A is before administration and 18B is after administration.
  • Figure 19 is a view showing the morphology of the eyeball before and after treatment of cynomolgus monkey (No. 013321) self-style cataract (left eye) in Example 9 of the present invention, wherein 19A is before administration, and 19B is after administration. .
  • Figure 20 is a view showing the morphology of the eyeball before and after treatment of cynomolgus monkey (No. 990447) self-style cataract (left eye) in Example 9 of the present invention, wherein 20A is before administration and 20B is after administration. .
  • Figure 21 is a view showing the morphology of the eyeball before and after treatment of cynomolgus monkey (No. 993747) self-style cataract (left eye) in Example 9 of the present invention, wherein 21A is before administration and 21B is after administration.
  • Figure 22 is a view showing the morphology of an eyeball before and after treatment of a traumatic cataract in a rat (the right eye of a female rat is selected as a photograph) in Example 10 of the present invention, wherein 22A is before administration. 22B is after administration.
  • Figure 23 is a view showing the morphology of an eyeball before and after treatment of a traumatic cataract in a rat with a traumatic cataract according to Example 10 of the present invention, wherein 23A is before administration, 23B. For after administration.
  • Figure 24 is a view showing the morphology of the eyeball before and after treatment of the vaginal eye ointment of the tenth embodiment of the present invention (selecting the left eye of a female mouse as a photograph), wherein 24A is before administration and 24B is given After the medicine.
  • Figure 25 is a view showing the morphology of an eyeball before and after treatment of a metabolite cataract of diabetic rats (selecting the right eye of a male rat as a photograph) in Example 11 of the present invention, wherein 25A is administered Before, 25B is after administration.
  • Figure 26 is a view showing the morphology of the eyeball before and after treatment of the cataract cataract of diabetic rats (selecting the right eye of a male rat as a photograph) in Example 11 of the present invention, wherein 26A is before administration. 26B is after administration.
  • Figure 27 is a view showing the morphology of the eyeball before and after treatment of the lanosterol eye ointment in the diabetic rat (the right eye of a male rat is selected as a photograph) in Example 11 of the present invention, wherein 27A is before administration, 27B For after administration.
  • 28 is a view showing the morphology of an eyeball before and after treatment of a blank eye ointment for a diabetic rat with metabolite cataract in a right eye of a male rat according to Example 11 of the present invention, wherein 28A is before administration, and 28B is After administration.
  • Figure 29 is a view showing the morphology of the eyeball before and after the right eye treatment of the metabotropic cataract of a diabetic rat in the positive control group of Example 11 of the present invention, wherein Figure 29A is before administration, and Figure 29B is administered. Rear.
  • Figure 30 is a nuclear magnetic resonance (NMR) chart of the extract of Inonotus obliquus prepared in Example 14 of the present invention.
  • Figure 31 is a mass spectrum (MS) chart of the extract of Inonotus obliquus prepared in Example 14 of the present invention.
  • Figure 32 is a high performance liquid chromatography (HPLC) chart of the extract of Inonotus obliquus prepared in Example 14 of the present invention.
  • Figure 33 is a graph showing the results of detecting the ThT fluorescence signal of lens protein aggregates significantly by Inophyllum sinensis in Example 22 of the present invention.
  • a compound of formula I (including pures, mixtures or corresponding extracts) can significantly prevent and/or treat ocular diseases caused by lens lesions.
  • the experimental results of the present invention show that the compound of the formula I can quickly achieve the effect of curing and preventing ocular diseases caused by lens lesions in a simple administration manner. The present invention has been completed on this basis.
  • R1 As used herein, “R1”, “R 1 ", and “R 1” have the same meaning. Other similar definitions have the same meaning.
  • C1-C20 alkyl or “C1-C10 alkyl” or “C1-C8 alkyl” refers to a straight chain having from 1 to 20 or from 1 to 10 or from 1 to 8 carbon atoms.
  • Branched alkyl groups such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, heptyl, octyl, or the like.
  • C1-C8 alkoxy or “C1-C4 alkoxy” refers to a straight or branched alkoxy group having from 1 to 8 and from 1 to 4 carbon atoms, such as methoxy. , ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, hexyloxy, heptyloxy, octyloxy, or Similar group.
  • C1-C8 alkylthio or “C1-C4 alkylthio” refers to a straight or branched alkylthio group having from 1 to 8 and from 1 to 4 carbon atoms, such as methyl sulfide.
  • Base ethylthio, propylthio, isopropylthio, butylthio, isobutylthio, sec-butylthio, tert-butylthio, pentylthio, hexylthio, heptylthio, octylthio, Or a similar group.
  • C3C10 cycloalkyl refers to a cycloalkyl group having from 3 to 10 carbon atoms, such as cyclopropyl, cyclobutyl, methylcyclobutyl, cyclopentyl, or the like.
  • C1 - C8 alkylene or "C1 - C7 alkylene” or “C1 - C6 alkylene” refers to having 1-8, 1-7, and 1-6 carbons.
  • a carbon atom of a branched or linear alkyl molecule of an atom removes a group of -CH2- formed by two hydrogen atoms, such as a methylene group (-CH2-), or the like.
  • C2-C10 alkenyl refers to a hydrocarbon radical formed by a linear or branched olefin molecule of 2 to 10 carbon atoms having one or more double bonds and having one hydrogen atom bonded to the double bond.
  • C(CH3) 2 CH-
  • C2-C8 alkynyl refers to a hydrocarbon radical formed by a straight or branched alkynyl molecule of 2-8 carbon atoms having one or more triple bonds, with one hydrogen atom bonded to the triple bond.
  • ethynyl CH tri-CH-
  • H3C-C tri-CH- or the like.
  • C1-C6 haloalkyl refers to a substituent of a straight or branched alkyl-halogen having from 1 to 6 carbon atoms, such as ⁇ CH 2 Cl, —CH 2 CH 2 Br, CH 2 . CHCH 2 Cl, or a similar group.
  • halogen refers to F, Cl, Br, and I.
  • aryl refers to a C6-C20 aryl group, including monocyclic or bicyclic or tricyclic aryl groups such as phenyl, naphthyl, anthracenyl.
  • C6-C10 member aryl refers to a cyclic group having an aromatic structure, such as phenyl, naphthyl.
  • the term "5-8 membered heteroaryl” has a 5- to 8-membered monocyclic or fused polycyclic ring and has a heteroaryl group of N, O or S on the ring system, such as pyrrolyl, pyridyl, furan. Base, or a similar group.
  • cyanate group has the formula: -O-C ⁇ N.
  • R 1 and R 2 represent a linked group, respectively, and R 1 and R 2 may be the same or different.
  • glycosyl refers to a monovalent substituent formed by the removal of a hemiacetal hydroxyl group from a cyclic form of a monosaccharide (or disaccharide).
  • Representative monosaccharides include pentoses and hexoses.
  • a preferred glycosyl group is a monosaccharide substituted one or more OH groups of the compound of formula I to form a ⁇ O-glycosyl group.
  • the terms “comprising,” “comprising,” and “comprising” are used interchangeably and encompasses not only a closed definition but also a semi-closed, and open-ended definition. In other words, the terms include “consisting of” and “consisting essentially of.”
  • ⁇ (D) generation means that one or more hydrogens in a compound or group are replaced by deuterium. Deuterated can be monosubstituted, disubstituted, polysubstituted or fully substituted.
  • the cerium isotope content of cerium at the cerium substitution site is greater than the natural strontium isotope content (0.015%), more preferably greater than 50%, more preferably greater than 75%, and even more preferably greater than 95%, more preferably The ground is greater than 97%, more preferably greater than 99%, and even more preferably greater than 99.5%.
  • D has an isotope content of ⁇ 95%, more preferably ⁇ 99% at the position of the hydrogen atom.
  • presbyopia also referred to as “presbyopia” refers to the visual state in which the lens of the eye loses its flexibility, making it difficult to focus on close objects.
  • the ocular diseases caused by the lens lesions do not include conjunctivitis, eye infections caused by pathogens (such as bacteria, viruses, etc.) or ocular inflammation.
  • prevention refers to the administration of a therapeutically effective amount of a compound of formula I (including neats, mixtures) and/or an extract of Inonotus obliquus, or an optical isomer thereof or a racemate thereof, prior to an ocular disease.
  • treating means administering a therapeutically effective amount of a compound of formula I, or an optical isomer thereof, or a racemate thereof, or a solvate thereof, or a pharmaceutically acceptable salt thereof, or a prodrug thereof Thereby reducing, alleviating or slowing the progression or progression of an ocular disease, disorder or condition.
  • “treating” refers to alleviating, slowing the progression of an ocular disease, disorder or condition, or ameliorating one or more signs or symptoms of an ocular disease, disorder or condition.
  • “Prophylaxis” and “treatment” as used in the present invention include delaying and terminating the progression of the disease, or eliminating ocular diseases caused by lens lesions, without requiring 100% inhibition, elimination and reversal.
  • the compound of Formula I and/or the Inonotus obliquus of the present invention and its composition or formulation are absent (eg, without being exposed to the compound of Formula I of the present invention and Extraction of the compound of formula I and/or B. obliquus of the present invention compared to the level observed in a control subject or specimen biologically matched to its composition or formulation.
  • composition or formulation thereof that alleviates, prevents, inhibits, and/or reverses, for example, at least about 1%, at least about 10%, at least about 30%, at least about 50%, of an ocular disorder caused by a lens lesion, such as a cataract. Or at least about 80%.
  • the term "cataract” refers to a disease or condition that causes turbidity (turbidity) or opacity, or induces swelling of the lens, including congenital cataracts and acquired cataracts, on the surface and/or inside of the lens.
  • the cataract includes, but is not limited to, age-related cataract, diabetic cataract, surgery-related cataract, cataract caused by radiation exposure, cataract caused by hereditary disease, cataract caused by infection, or drug Caused by cataracts.
  • the present invention has for the first time discovered the use of a compound of formula I for the preparation of a pharmaceutical composition or formulation for the prevention and/or treatment of a lens disease caused by the treatment of a lens disease associated ophthalmopathy. Eye disease.
  • compound of the invention refers to a compound of formula I, or an optical isomer thereof, or a racemate thereof, or a solvate thereof, or pharmaceutically acceptable thereof Accepted salt, or its prodrug.
  • the term also includes the corresponding deuterated compounds. It will be understood that the term also includes mixtures of the above components.
  • the chiral carbon atom may be in the R configuration, in the S configuration, or a mixture of the two.
  • the dotted line The total number of double bonds represented is 0, 1, 2, 3 or 4.
  • the double bond is located between the following positions: a and b, b and c, c and d, e and f, c and f, f and g, g and h, h and a , b and i, i and j, a and k, and/or k and l.
  • the valence state of each C conforms to the requirements of the chemically stable structure (ie, C is tetravalent).
  • A is independently a substituted or unsubstituted C1-C4 alkylene group
  • Y is a group having a hydroxyl group or a SH substituent.
  • the R4 is hydrogen or methyl.
  • R7, R12 and R15 are each independently selected from the group consisting of: none, hydrogen, methyl.
  • the R10a, R10b are each independently selected from: hydrogen, -OH, -SH, -0SO 3 H , -OPO 3 H, -COOH, -CHO, or R10a and R10b constituting a carbonyl group.
  • R5, R6, R8, R9a, R9b, R14, R16, R17a and R17b are each independently selected from the group consisting of hydrogen and methyl.
  • said Z is not ⁇ CH 2 C(CH 3 ) 3 .
  • said Z is -CH 2 C(CH 3 ) 2 -OH.
  • the compound of formula I is any one of formulas I-1, I-2, I-3, I-4, I-5, I-6, I-7, and I-8. .
  • the compound of formula I is selected from the group consisting of
  • the term "pharmaceutically acceptable salt” means a salt suitable for use as a medicament formed by the compound of the present invention and an acid or a base.
  • Pharmaceutically acceptable salts include inorganic and organic salts.
  • a preferred class of salts are the salts of the compounds of the invention with acids.
  • Suitable acids for forming salts include, but are not limited to, mineral acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, Organic acids such as maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzoic acid, and benzenesulfonic acid; and acidic amino acids such as aspartic acid and glutamic acid.
  • a preferred class of salts are the salts of the compounds of the invention with bases.
  • Suitable acids for forming salts include, but are not limited to, inorganic bases such as sodium hydroxide, potassium hydroxide, sodium carbonate, sodium hydrogencarbonate, sodium phosphate, and the like, and organic bases such as ammonia, triethylamine, and diethylamine.
  • inorganic bases such as sodium hydroxide, potassium hydroxide, sodium carbonate, sodium hydrogencarbonate, sodium phosphate, and the like
  • organic bases such as ammonia, triethylamine, and diethylamine.
  • prodrug is also referred to as a prodrug, a prodrug, a prodrug, etc., and refers to a compound which has a pharmacological action after being transformed in vivo.
  • the prodrug itself has no biological activity or low activity, and becomes an active substance after metabolism in the body. The purpose of this process is to increase the bioavailability of the drug, enhance the targeting, and reduce the toxicity and side effects of the drug.
  • a prodrug of a compound of formula A can be metabolized to a compound of formula I (representatively, such as, for example, inoculum).
  • a preferred prodrug is an ester formed by esterification of a compound of the formula I of the present invention with a compound containing a hydroxyl group or a compound having a carboxyl group.
  • the hydroxyl group-containing compound includes a C1-C6 lower alcohol (e.g., ethanol, propanol, etc.), a lower sugar (e.g., glucose, lactose, etc.).
  • the carboxyl group-containing compound includes a C1-C6 lower acid.
  • the C1-C6 lower acid includes an organic or inorganic acid.
  • the C1-C6 lower acid is hydrochloric acid, sulfuric acid, acetic acid, propionic acid, oxalic acid, fumaric acid, maleic acid, malic acid, tartaric acid, and the like.
  • Another preferred class of prodrugs are glycosylated compounds, i.e., one or more OH groups in the compound of Formula I (e.g., R10a, R10b, Z or other positions in Formula 1) are substituted with a glycosyl group to form The product of O-glycosylation.
  • the compound of the formula I of the present invention can be produced by a method well known to those skilled in the art in the prior art, and the reaction parameters of the respective steps are not particularly limited. Furthermore, the compounds of the invention are also commercially available. Typically, the compound of formula I of the present invention is prepared by extraction, isolation and purification from Inonotus obliquus.
  • the inventors conducted pharmacokinetic studies on different compounds of formula I, and as a result, found that at least one of R1a, R2a, R3a, R1b, R2b and R3b is a group containing O or S in the structure of the compound of formula I. (such as ⁇ OH or ⁇ SH), can significantly extend the half-life of the compound of formula I in vivo, especially to increase the concentration of concentration in the eye, prolong the residence time in aqueous humor, and improve the therapeutic effect on eye diseases.
  • the pharmaceutical composition or formulation is further used to: (b) inhibit, reverse (solvent or depolymerize) lens protein aggregation; and/or (c) prevent and/or treat and crystallize protein aggregation Related diseases.
  • the ocular diseases include, but are not limited to, cataracts, presbyopia, myopia, cortical opacity, presbyopia, and ocular complications caused by diabetes.
  • the present invention finds for the first time the use of an extract of Inonotus obliquus in the treatment of lens disease-related eye diseases for preparing a pharmaceutical composition or preparation for preventing and/or treating an eye caused by a lens lesion. disease.
  • extract or “effective portion” includes water-soluble and/or fat-soluble extracts.
  • the term also includes alcohol extracts or aqueous extracts, especially water-insoluble, or water-insoluble extracts or fat-soluble extracts.
  • an effective site group that is, an extract containing a fat-soluble effective site and a water-soluble effective site, or a mixture thereof.
  • the chemical component contained in the effective part of the present invention includes at least a substance selected from the group consisting of a steroid, a steroid or a combination thereof.
  • the method which can be used for preparing the extract of Inonotus obliquus of the present invention is not particularly limited.
  • a water-soluble and/or fat-soluble extract can be obtained by a conventional method using Borian hominis as a raw material.
  • the preparation of the active site is carried out by solvent extraction, extraction, and/or chromatography.
  • the solvent used for the solvent extraction method is not particularly limited, and representative examples include, but are not limited to, one of water, ethanol, methanol, acetone, ethyl acetate or several solvents. Mixed solvent.
  • the number of extractions can be one or more times.
  • the solvent used for the solvent extraction method is not particularly limited, and representative examples include, but are not limited to, n-butanol, ethyl acetate, dichloromethane, chloroform, cyclohexane, petroleum ether. a mixed solvent of one or several solvents in the medium. The number of extractions may be one or more times;
  • column chromatography is not particularly limited, and representative examples include, but are not limited to, one of activated carbon, silica gel, reverse phase silica gel, macroporous resin, and dextran gel. Combination of species or several.
  • the extract comprises a terpenoid component, such as a triterpenoid component.
  • the extract comprises a tetracyclic triterpenoid component.
  • the extract comprises a mixture of two or more extracts obtained by using different extraction methods for Inonotus obliquus.
  • the extract contains a compound selected from the group consisting of:
  • the weight percentage of the inoculum is 0.01-99.99% by weight in the extract. In another preferred embodiment, the weight percentage of the tylosic acid in the extract is 0.01-99.99% by weight.
  • the preparation method further comprises the step (3) of performing the silica gel column chromatography on the extract of the Inonotus obliquus, collecting the eluate, and separating the purified birch. Inoculillus extract.
  • the crude powder of the dried fruit body of Inonotus obliquus was refluxed with ethanol, and the extract was evaporated under reduced pressure to obtain an ethanol extract.
  • the ethanol extract was further suspended with water, extracted with petroleum ether and ethyl acetate in that order, and the solvent was distilled away under reduced pressure to obtain petroleum ether extract and ethyl acetate extract.
  • the petroleum ether extract was subjected to silica gel column chromatography and eluted with a petroleum ether-ethyl acetate gradient (for example, 15:1 - 2:1), and the eluate was collected each time according to the results of thin layer chromatography, and the same fraction was combined.
  • the single component was recrystallized after distilling off the solvent under reduced pressure.
  • Elliptic acid was obtained by elution with petroleum ether-ethyl acetate (eg, about 5:1); or the ethyl acetate extract was subjected to silica gel column chromatography, first eluting with a gradient of chloroform-methanol (eg, 100:1 - 10:1), eluted in chloroform-methanol (eg 70:1) to give mixture I, and eluted with petroleum ether-ethyl acetate gradient (eg 30:1 -1:1) in petroleum ether-acetic acid When ester (such as 8:1) is eluted, the inoculum is obtained.
  • the crude powder of the dried fruit body of Inonotus obliquus was refluxed with ethanol, and the extract was evaporated under reduced pressure to obtain an ethanol extract.
  • the ethanol extract was further suspended with water, extracted with petroleum ether and ethyl acetate in that order, and the solvent was distilled away under reduced pressure to obtain petroleum ether extract and ethyl acetate extract.
  • the ethyl acetate extract is subjected to silica gel column chromatography, eluting with a gradient of chloroform-methanol (e.g., 100:1 - 10:1) and eluting with chloroform-methanol (e.g., 50:1) to give mixture II.
  • the mixture is subjected to silica gel column chromatography, eluting with a petroleum ether-ethyl acetate gradient (e.g., 20:1 - 1:2), eluting with petroleum ether-ethyl acetate (e.g., 3:1) to give a mixture III.
  • the mixture III is subjected to silica gel column chromatography, eluting with a petroleum ether-ethyl acetate gradient (e.g., 5:1 - 1:2), eluting with petroleum ether-ethyl acetate (e.g., 2:1) .
  • the present invention provides for the first time the extract of Inonotus obliquus for the preparation of a pharmaceutical composition or preparation for preventing and/or treating an ocular disease caused by a lens lesion.
  • the pharmaceutical composition or formulation is further for: (b) inhibiting, reversing lens protein aggregation; and/or (c) preventing and/or treating a disease associated with lens protein aggregation.
  • the ocular disease is (but not limited to): cataract, presbyopia, myopia, cortical opacity, presbyopia sclerotherapy, diabetes-induced ocular complications.
  • compositions or formulation and method of administration are provided.
  • the pharmaceutical composition or formulation comprises: a compound of the formula I of the invention and/or an extract of Inonotus obliquus; and a pharmaceutically acceptable carrier.
  • the content of the first active ingredient is from 0.001 to 99% by weight, preferably from 0.01 to 70% by weight, more preferably from 0.05 to 40% by weight, based on the total weight of the composition.
  • the pharmaceutical composition or formulation may further comprise: (c) a second active ingredient, wherein the second active ingredient is selected from the group consisting of lanolin compounds, lanosterol, all Any of the compounds contained in the fungus, the azole compound, the amyloid modulator, the glucocorticoid compound, the antibiotic, or a combination thereof may be contained in the bacterium or the polyporus.
  • a second active ingredient selected from the group consisting of lanolin compounds, lanosterol, all Any of the compounds contained in the fungus, the azole compound, the amyloid modulator, the glucocorticoid compound, the antibiotic, or a combination thereof may be contained in the bacterium or the polyporus.
  • the second active ingredient is present in an amount of from 0.01 to 20% by weight, preferably from 5 to 15% by weight, based on the total weight of the composition.
  • the concentration thereof is preferably 10 to 200 mM, preferably 15 to 150 mM, more preferably 20 to 50 mM; most preferably 20 to ⁇ 30 mM.
  • the lanolin compound is selected from the group consisting of: lanosterol.
  • the glucocorticoid compound is selected from the group consisting of dexamethasone, hydrocortisone, or a combination thereof.
  • the antibiotic is selected from the group consisting of tobramycin, gentamicin sulfate, chlortetracycline, chloramphenicol, or a combination thereof.
  • the azole compound is selected from the group consisting of econazole, isoconazole, bifonazole, clotrimazole, aripiprazole, ketoconazole, fluconazole, phenylimidazole, and imidium.
  • amyloid modulator also known as amyloid modulator, amyloid inhibitor, amyloid inhibitor, specifically refers to the regulation or inhibition of protein accumulation, adhesion, precipitation, affecting proteins A class of molecules or compounds that form fibrillar protein aggregates.
  • the term "effective therapeutic amount” refers to an amount that produces a function or activity to a human and/or animal and that is acceptable to humans and/or animals. It will be understood by one of ordinary skill in the art that the "effective amount” or “effective amount” may vary depending on the form of the pharmaceutical composition, the route of administration, the excipient of the drug used, the severity of the disease, and the combination with other drugs. The situation varies.
  • pharmaceutically acceptable carrier means: one or more compatible solid, semi-solid, liquid or gel fillers which are suitable for use in humans or animals and which must be of sufficient purity and sufficiently low toxicity.
  • Cosmetic refers to the ingredients of the pharmaceutical composition and the active ingredients of the drug and their intermixing without significantly reducing the efficacy.
  • the carrier is not particularly limited and may be selected from materials commonly used in the art, or obtained by a conventional method, or commercially available.
  • Examples of pharmaceutically acceptable carriers are cellulose and its derivatives (such as methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, etc.), gelatin, talc, solid lubricants. (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oil (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyol (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifier (such as spit Warm), wetting agent (such as sodium lauryl sulfate), buffer, chelating agent, thickener, pH adjuster, transdermal enhancer, colorant, flavoring agent, stabilizer, antioxidant, preservative, Bacteriostatic agents, pyrogen-free water, liposomes, and the like.
  • solid lubricants such as stearic acid, magnesium stearate), calcium sulfate, vegetable oil (such as
  • the liquid dosage form may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or a mixture of these substances.
  • inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or a mixture of these
  • compositions may also contain adjuvants such as wetting agents, emulsifying and suspending agents, and the like.
  • the dosage form of the pharmaceutical composition includes, but is not limited to, an oral preparation, an injection, and an external preparation.
  • Representative examples include, but are not limited to, tablets, injections, infusions, ointments, gels, solutions, microspheres, films.
  • a preferred dosage form is an ophthalmic formulation.
  • the ophthalmic preparations are eye drops, emulsions, gels, ophthalmic ointments, sustained release microspheres, intraocular sustained release grafts, and medicinal sustained release drug films.
  • the ophthalmic formulation comprises a pharmaceutically acceptable pharmaceutical carrier, including but not limited to: a solvent or diluent, a surfactant, a thickener, an osmotic pressure adjusting agent, a pH adjusting agent, a bacteriostatic agent, a chelate mixture.
  • a pharmaceutically acceptable pharmaceutical carrier including but not limited to: a solvent or diluent, a surfactant, a thickener, an osmotic pressure adjusting agent, a pH adjusting agent, a bacteriostatic agent, a chelate mixture.
  • ophthalmic preparations such as eye drops, eye drops
  • a solvent or diluent including an aqueous solvent or a non-aqueous solvent.
  • the aqueous solvent includes distilled water, physiological saline, water for injection, and the like;
  • the nonaqueous solvent used includes ethanol, propylene glycol, glycerin, vegetable oil (such as olive oil, castor oil, corn oil, soybean oil for injection) and the like.
  • the surfactant is selected from the group consisting of anionic surfactants, cationic surfactants, nonionic surfactants, chaotropic surfactants, or combinations thereof.
  • the nonionic surfactant is selected from the group consisting of: Tween, Span, fatty acid glycerides, polyoxyethylenes, polyoxyethylene-polyoxypropylene copolymers, or combinations thereof.
  • the amount (or content) of the surfactant is generally from 0 to 2% by weight, more preferably from 0.1 to 1% by weight.
  • thickeners can be used to increase the viscosity of the system, keep the system in a stable and stable suspension state or opaque state, increase the retention time of the drug in the eye by adding an appropriate amount of thickener, thereby increasing the eye The absorption of effective small ingredients.
  • the thickener is preferably chitosan, hydroxypropylmethylcellulose (HPMC), methylcellulose (MC), and povidone (PVP), gelatin, sodium carboxymethylcellulose ( CMC-Na) and so on.
  • HPMC hydroxypropylmethylcellulose
  • MC methylcellulose
  • PVP povidone
  • CMC-Na sodium carboxymethylcellulose
  • the thickener is used in an amount (or content) of from 0 to 6 wt%, preferably from 0.1 to 5 wt%.
  • the osmotic pressure adjusting agent is added to the ocular preparation to make the osmotic pressure of the ophthalmic preparation similar to the osmotic pressure of the human eye environment, thereby reducing the irritation to the eye.
  • Representative, commonly used osmotic pressure adjusting agents include, but are not limited to, acetic acid, sodium acetate, sodium bicarbonate.
  • the amount (or amount) of the osmotic pressure adjusting agent maintains the ophthalmic formulation in an isotonic or isotonic environment.
  • pH adjusting agent is added to the ocular preparation to maintain the pH of the ophthalmic preparation in an appropriate range, similar to the pH of the human eye environment, thereby reducing irritation to the eye.
  • pH adjusting agents include, but are not limited to, sodium chloride, potassium chloride, and glucose.
  • the amount (or content) of the pH adjusting agent maintains the pH of the ophthalmic preparation at 5-9.
  • bacteriostatic agents can kill or inhibit the growth of bacteria in the cream, prevent the bacteria from breeding too much, and endanger human health.
  • the bacteriostatic agent is not particularly limited and may be one or a combination of one of paraben or paraben.
  • the bacteriostatic agent of the present invention is selected from the group consisting of methylparaben, ethylparaben, propylparaben, or a combination thereof.
  • a certain amount of a chelating agent such as EDTA may be appropriately added to increase the stability of the preparation.
  • concentration of the chelating agent ranges from 0 to 0.05% by weight.
  • the manner of administration of the composition or formulation of the present invention is not particularly limited, and representative modes of administration include, but are not limited to, topical administration, oral administration, injection, and the like.
  • a preferred mode of administration is the topical administration of the composition or formulation to the eye including, but not limited to, conjunctiva, retrobulbar, periocular, retinal, choroidal or intraocular administration, etc., representative of Drop eye drops, intraocular injection, ocular mucosal injection, ocular mucosal coating, etc.
  • the pharmaceutical preparation should be matched to the mode of administration.
  • the agents of the invention may also be used with other synergistic therapeutic agents (including before, during or after).
  • a pharmaceutical composition or formulation is used, a safe and effective amount of the drug is administered to a subject, such as a human or non-human mammal, typically at least about 10 micrograms per kilogram of body weight, and in most cases. Not more than about 8 mg/kg of body weight, preferably the dose is from about 10 micrograms per kilogram of body weight to about 1 milligram per kilogram of body weight.
  • specific doses should also consider factors such as the route of administration, the health of the patient, etc., which are within the skill of the skilled physician.
  • the present invention provides a method for non-therapeutic improvement or maintenance of lens transparency in vitro, comprising crystallizing a compound of formula I, or an optical isomer thereof, or a racemate thereof, or a solvate thereof, or a pharmaceutically acceptable thereof
  • the accepted salt, or a prodrug thereof, or a deuterated compound thereof and/or an extract of Inonotus obliquus is contacted to inhibit lens lesions.
  • the compounds of formula I are as described herein.
  • lens lesions include opacity or oxidation, etc., inhibiting lens lesions including improving or maintaining lens transparency.
  • the present invention also provides a method for preventing and/or treating an ocular disease, the method comprising the steps of administering to a subject in need thereof a compound of the formula I according to the present invention, or an optical isomer thereof or a racemate thereof, or a solvate thereof, or a pharmaceutically acceptable salt thereof, or a prodrug thereof, or a deuterated compound thereof and/or an extract of Inonotus obliquus.
  • the subject is a human and a non-human mammal.
  • the non-human mammals include, but are not limited to, pets (such as dogs, cats), livestock (such as cattle, sheep, horses, pigs), various zoo animals (pandas, elephants, tigers). )Wait.
  • the subject further includes other animals than human and non-human mammals, such as non-mammals.
  • the present invention has found for the first time that the compound of the formula I and the extract of Inonotus obliquus can prevent and/or treat ocular diseases such as cataract prevention and treatment, and have a remarkable therapeutic effect.
  • the oleic acid eye drops formula:
  • Example 3 the therapeutic effects of the two active ingredients of Example 1 and Comparative Example 1 on lens lesions (cataracts) were compared.
  • a rat cataract model was established. Wistar rats were selected 10-13 days after birth, male or female, modeled with sodium selenite, and each rat was injected subcutaneously in the back and neck according to 20 ⁇ mol/kg body weight. Sodium selenite is prepared by using physiological saline, and the prepared solution is sterilized by a 0.22 ⁇ m membrane and stored at room temperature and protected from light. Significant nuclear cataract in the eye can be observed after the eye is rubbed in the rat.
  • Example 1 Eight cataract rat models were equally divided into two groups according to the left and right eyes of each rat, and each group of 8 corresponding intraocular lenses were respectively formulated with the formulation of Example 1 (Example 1) and the formulation of Comparative Example 1 ( In the first group, the immersion treatment was carried out. Before the start of the experiment, the morphology of the lens of the eyeballs of the two groups was observed and photographed. The upper light source and the lower eye of a rat lens were selected after successful cataract modeling in the first group. The morphology of the light source is shown in Fig. 1 and Fig. 2 respectively; the morphology of the upper and lower light sources of a rat lens of the lens selected after successful cataract modeling in the first group is shown in Fig. 3 and Fig. 4, respectively. All the rats' lens was taken out and the score was 5 points.
  • the left and right eye lenses of the rats in the group 1 and the control group 1 were respectively immersed in the eye drops of the inonotus and the same concentration of the structural analog lanosterol eye drops, and the effect was observed and scored two weeks later, wherein the examples
  • the morphology of the upper and lower light sources of a rat lens of the eye lens selected after the end of one group of experiments is shown in Fig. 5 and Fig. 6 respectively; the upper and lower light sources of a rat lens of the lens selected after the end of the experiment in the first group of the comparison group
  • the shapes are shown in Figures 7 and 8, respectively; the scoring criteria are:
  • the compound of the formula I of the present invention (a compound having an O or S group at a specific position) represented by the inoculum, is a simple administration even with respect to the structural analog lanosterol.
  • the method also has excellent therapeutic effect on cataract, which can quickly relieve, cure and prevent animal cataract.
  • Example 4 the therapeutic effects of the two active ingredients of Example 2 and Comparative Example 2 on lens lesions (cataracts) were compared.
  • a rat cataract model was established. Wistar rats were selected from 10 to 13 days after birth, male or female, modeled with sodium selenite, and each rat was injected subcutaneously in the back and neck according to 20 ⁇ mol/kg body weight. Sodium selenite is prepared by using physiological saline, and the prepared solution is sterilized by a 0.22 ⁇ m membrane and stored at room temperature and protected from light. Significant nuclear cataract in the eye can be observed after the eye is rubbed in the rat.
  • Example 2 Eight cataract rat models were divided into two groups according to the left and right eyes of each rat, and each group of 8 corresponding intraocular lenses was used in the formulation of Example 2 (Example 2) and the formulation of Comparative Example 2 ( In the comparison group 2), before the start of the experiment, the morphology of the lens of the eyeballs of the two groups of rats was observed and photographed, and the upper and lower light sources of a rat lens of the eyeball selected in the cataract model in the second group of Example 2 were taken. The morphology is shown in Fig. 9 and Fig. 10 respectively; the morphology of the upper and lower light sources of a rat lens of the eye selected after successful cataract modeling in the group 2 of Fig. 2 is shown in Fig. 11 and Fig. 12, respectively. The photographs of all the rats were taken out and scored 5 points.
  • the left and right eye lenses of each of the rats of Example 2 and Comparative Example 2 were each immersed in oleic acid eye drops and the same concentration of structural analog lanosterol eye drops, and the effect was observed and scored on the 9th day (Table 4).
  • Table 4 the morphology of the upper and lower light sources of a rat lens of the eye selected after the end of the experiment in Example 2 are shown in Fig. 13 and Fig. 14 respectively;
  • the morphology of the upper and lower light sources of the mouse eye lens are as shown in Figures 15 and 16, respectively;
  • the scoring standard is the same as in Example 3.
  • the compound of the formula I of the present invention (compound having an O or S group at a specific position) represented by oleic acid is relatively simple to administer with respect to lanosterol.
  • the method also has an excellent effect of treating cataracts (the cataractous lens after the treatment of the catalyzed eye drops, the opacity area at the center position is significantly reduced), which can quickly alleviate, cure and prevent animal cataracts.
  • the concentration of lanosterol decreased by 82% ⁇ 14.7%, while the concentration of Inophyllum sinensis decreased by 11% ⁇ 2.3%, and the concentration of sputum acid only decreased by 8% ⁇ 1.5%.
  • Component content Inoculum 1.25g White Vaseline 75g Liquid paraffin 12.5g
  • Blank eye ointment formula and preparation method thereof :
  • Example 8 the primate cynomolgus monkey was used as an animal model, and the therapeutic effect of the inoculum ophthalmic ointment of Example 6 on congenital cataract was examined.
  • Fig. 17 and Fig. 18 The morphological comparisons of the left and right eyes of cynomolgus monkeys before and after administration of Inoculum sinensis ophthalmic ointment are shown in Fig. 17 and Fig. 18, respectively. It can be seen from Fig. 17 and Fig. 18 that the treatment with inoculum Then, using the same slit lamp inspection condition to shoot the contrast picture, the cataract opacity of the left and right eye lens of the cynomolgus monkey was significantly reduced, and the turbid area before partial administration was completely turbid after treatment with the inoculum. Disappeared, the turbid area continued to decrease, and no new turbid area appeared in the whole crystal.
  • Example 9 the effect of the treatment of self-style cataract by the inoculum of the genus Inoculum in Example 6 was examined using an aging primate cynomolgus monkey as an animal model, and Comparative Example 4 was used as a control group.
  • the administration method is the inoculum of the inoculum of the inoculum of the inoculum of the inoculum in the sixth embodiment, and is directly applied to the corneal surface of the left eye of the monkey on the left side of the numbers 013321 and 990447, and the monkey face is upward when the medicine is applied, and the animal is fixed after the medicine is applied. 3-5 minutes. After 42 days of continuous administration of the left eye of the two monkeys, their eyeballs of the left eye were examined again in the same manner and photographed. At the same time as the experimental group, a 19-year-old monkey with self-style cataract (No.
  • the opacity area of the entire crystalline cataract region was also significantly reduced, and the lens cataract level of the drug treatment appeared to be transparent ( Normal) level of reversal.
  • the therapeutic effect of lanosterol in the control group is shown in Figure 21 (the left eye of the same subject). It can be clearly observed from the comparison pictures before and after administration that lanosterol does not slow the progression of self-style cataract in elderly primates, and Compared with before treatment, the turbid area inside the crystal increased by nearly 50% after lanosterol treatment, and the light transmittance of the whole crystal further decreased. The above results indicate that inonotc have an obvious and unique therapeutic effect on self-style cataract in elderly primates.
  • the rat skin model was used to examine the therapeutic effects of the inoculum eye cream and the oleic acid eye ointment of Example 6 and Example 7 on the traumatic cataract.
  • the blank eye ointment was used as a blank control, and the lanosterol eye ointment of Comparative Example 4 was used as a positive control group.
  • Each group contained 2 male and female SD rats (the numbers of each group were A, B, C and D, respectively). Wherein A and B are male rats, and C and D are female rats).
  • a and B are male rats
  • C and D are female rats.
  • the cornea and the lens capsule were pierced with an injection needle, and the needle was repeatedly slid in the crystal cortex under microscope observation until obvious turbidity appeared in the crystal.
  • the rats were re-anesthetized and expanded, photographed, and the degree of traumatic cataract and its degree were evaluated.
  • the lens cortex was observed with a slit lamp.
  • the score was evaluated according to the Lens opacities classification system II (LOCS II) standard. Because it is a local cataract caused by trauma, the turbid area is ignored when the first score is scored. Only the degree of cortical turbidity is examined, and the depth of cortical opacity is later. The extent of the change to assess the efficacy.
  • the first photograph was started every other day, and four groups of SD rats were administered with Example 6 (experimental group 1), Example 7 (experimental group 2), comparative example 3 (solvent control group 3), and birch brown in Comparative Example 4, respectively.
  • the size of the drug is applied directly to the rat cornea. After three months, the rats were anesthetized again and expanded, photographed (except solvent control group 3), and rat traumatic cataract and its degree were evaluated.
  • Example 6 The birch in Example 6, Example 7, Comparative Example 3 and Comparative Example 4
  • the results of the treatment of cataract cataracts with melanobacteria ophthalmic ointment, oleic acid eye ointment, blank eye ointment and lanosterol eye ointment are shown in Table 7 ⁇ 10 and Figure 22 ⁇ 24.
  • the rat model was used as an animal model to examine the therapeutic effect of the inoculum anophylin ointment and the oleic acid eye ointment of Example 6 and Example 7 on the metabolic cataract in the diabetic animal model.
  • the blank eye ointment in Comparative Example 3 was used as a blank control, and the lanosterol eye ointment in Comparative Example 4 was used as a positive control. Since the commercial pirenoxine sodium eye drops (white end stop) is an ophthalmic drug for the treatment of mild diabetic cataract or complicated cataract, it is also used as a positive control.
  • the inoculum eye cream, the oleic acid eye ointment, the blank eye ointment and the lanosterol eye ointment were continuously administered on weekdays, not administered on non-working days, and the daily dosing frequency was once a day.
  • the dosage is half a mung bean size, which is directly applied to the cornea of the right eye of the rat, and the left eye is not treated; the other right eye of the pirenoxine sodium eye drops group is administered according to the commercial specification and dosage. (Not administered on non-working days), the left eye is not administered. All animal models were anesthetized again after 2 months of treatment.
  • Example 6 The blood samples were taken from the rats to determine the blood glucose level (both at 18.6 mmol/L or more) and photographed to evaluate the degree of cataract in the right lens and cortex of the rat.
  • Example 6 The results of the treatment of diabetic metabolite cataract in the treatment of diabetic metabolite cataract in Inoculum 3 and Comparative Example 4 are shown in Table 11 ⁇ 14 and Figure 25 ⁇ 28.
  • the results of the right eye treatment in the pirenoxine sodium eye drops group are shown in Figure 29.
  • Table 11 The therapeutic effect of the inoculum ophthalmic ointment of Inoculum in Example 6 on diabetic metabolite cataract in rats
  • mice B rat C mouse Before administration CII CII Ctr After administration
  • Solvent control group 3 A mouse B rat C mouse Before administration CI Ctr Ctr After administration CIII CII CII
  • the turbid area in the lens of the rat increased after the administration of lanosterol, and the area where the cataract was not originally formed also showed turbidity, and the light transmittance of the whole crystal continued to decrease, and the condition deteriorated.
  • the lanosterol-induced metabolic cataract caused by diabetes does not reverse the effect of restoring crystal transparency.
  • Example 12 the therapeutic effect of the inoculum in the Example 6 on the senile cataract was examined using a canine animal model.
  • LOCS II Lens opacities classification system II
  • the left and right eyes were determined to be old-type cataracts, and the grades of cortical cataracts were CIV and CIII.
  • the inoculum eye cream of Example 2 was administered once a day, twice a day, each time two soybeans were administered in a dose, and the ointment was directly applied to the cornea, and the animal head was fixed after the medicine was applied. In the 5th minute, the drug utilization rate was increased.
  • the lens was examined again in the same way and the degree of cortical cataract was evaluated. The left and right cataracts were found to be reduced to CI and CI, respectively.
  • Porcine alcohol has a significant therapeutic effect on senile cataract.
  • Inonotus obliquus extract 1 1 kg of crude powder of dried fruit body of Inonotus obliquus, refluxing with 95% ethanol at a ratio of material to liquid 1:75 (g: ml), and extracting the solvent under reduced pressure to obtain 40 Gram ethanol extract.
  • the whole ethanol extract was suspended with water, extracted with petroleum ether, and the extract was separately distilled under reduced pressure to obtain 1.25 g of the extract of Inonotus obliquus.
  • Inonotus obliquus extract 2 The extract of Inonotus obliquus prepared in Example 13 was subjected to silica gel column chromatography and eluted with a petroleum ether-ethyl acetate gradient (15:1 - 2:1), each time collected. The eluate was 50 mL and the same fraction was combined according to the results of thin layer chromatography. The solvent was distilled off under reduced pressure, and the single component was recrystallized, and the extract was obtained by eluting with petroleum ether-ethyl acetate 5:1. 2.
  • the Inotodiol extract 2 is a single inotodiol component with a purity of 98.36%.
  • the structural formula of the inonotol is as follows:
  • the extract 1 of Example 13 was used as an active ingredient, and an ophthalmic ointment was prepared in a usual manner.
  • the recipe is as follows:
  • Extract 1 was weighed, the mass was 1.25 g, white petrolatum 75 g, liquid paraffin 12.5 g, placed in a container, and sonicated in a 70 ° C water bath until all the compounds were uniformly dispersed. Transfer quickly to a clean, sterile eye tube, chill in an ice bath until the ointment is solidified and stored at 4 °C.
  • the extract of Inonotus obliquus contains a large number of animal-safe ingredients that can be used to treat cataract in animals. These ingredients can be used to prepare treatment, prevent, stop the progression of the disease and / Or drugs that delay the progression of the disease against cataracts.
  • Example 17 the eye drug safety of the extracts of Inoculum, Trametes, and Inonotus obliquus 1 in Example 6, Example 7, and Example 15 was examined using rabbit as an animal model. At the same time, the blank eye ointment in Comparative Example 3 was used as a blank control.
  • the first group of left eyes was given the antibacterial ointment of Inonotus obliquus in Example 6.
  • the right eye was given 0.9% sodium chloride injection
  • the second group of left eyes was given the thrombisic acid eye ointment of Example 7
  • the right eye was given 0.9% sodium chloride injection
  • the third group of left eye was given the example 15
  • the administration volume was about soybean-sized eye ointment/eye, and the 0.9% sodium chloride injection was administered in a volume of 0.1 ml/eye.
  • Fluorescein sodium was examined in both eyes of each rabbit before the test. Animals with eye irritation, corneal defects, and conjunctival injury were not used for the test, and qualified rabbits were used for the test. The eyes were examined before the first dose every day and 1 h, 2 h, 4 h, 24 h, 48 h, and 72 h after the last dose. If no irritation was observed at 72 h, the test may end. If mildly irritating, the observation period was extended to 7 days after the last administration; moderate irritation, the observation period was extended to 14 days after the last administration; intensity stimulation, the observation period was extended to 21 days after the last administration . Administration 3 times a day, at least 4 hours apart.
  • the rabbits were sacrificed by intraperitoneal injection of 20% urethane solution, and then the eyeballs were removed and fixed in the eyeball fixative for pathological material extraction, dehydration, embedding, staining and light microscopy.
  • the stimulation scores of the cornea, iris and conjunctiva of each animal at each observation time point were added to the total score, and the sum of the integrals of one group was divided by the number of animals to obtain the final score.
  • the degree of irritation was determined according to Table 17, and combined with the results of histopathological examination for comprehensive judgment.
  • the results of rabbit eye examination were as follows: The first group, the second group, the third group and the fourth group of rabbits had a small amount of secretions in the left and right eyes, the conjunctival hyperemia was bright red, and the iris was slightly hyperemia around the cornea. All eyes of the rabbits were normal at the end of 72 hours of administration, and no abnormal changes were observed. No serotonin was stained with yellow-green. The stimulating scores of the left and right eyes of each group were in the range of 0-3, and the eye irritation results were judged to be non-irritating.
  • Example 2 The pathological examination results showed that the rabbits of Example 2, Example 7, Example 15 and Comparative Example 3 were given to the rabbits of the genus Inoculum ophthalmic ointment, the oxytetracycline eye ointment, the bifidobacteria extract, and the white eye ointment.
  • Conjunctiva There is no degeneration and necrosis of epithelial cells in all parts, no expansion of interstitial blood vessels, congestion, and no inflammatory cell infiltration; cornea: corneal epithelial cells without degeneration, necrosis, stromal collagen fibers without swelling, rupture, no inflammatory cell infiltration Corneal endothelial cells have no necrosis and no proliferation; iris: iris ciliary body structure is clear, interstitial no hyperemia, no inflammatory cell infiltration; Hastelloy and lacrimal gland: glandular epithelial cells are normal, interstitial no congestion, edema and inflammation Cell infiltration.
  • Example 6 Example 7, Example 15 and Comparative Example 3 of B. obliquus ophthalmic ointment, oxytetracycline eye ointment, Inonotus obliquus extract 1 eye ointment and blank eye ointment on rabbit conjunctiva
  • the cornea, iris, Hastelloy and lacrimal gland were not irritating, indicating that the eye drops prepared by the two compounds of Inoculum and Trametic acid were safe.
  • the thermal stability of two compounds of Inoculum and Trametes was tested, and lanosterol was used as a reference.
  • the evaluation method was carried out by self-contrast method, and each compound was placed at 4 ° C and 80 ° C for the same time, and HPLC (test concentration: 0.3 mg/ml, methanol dissolution) was used to compare the inoculum, tausel and lanosterol.
  • HPLC test concentration: 0.3 mg/ml, methanol dissolution
  • the initial purity of each of the lanosterol, betulin, and ceric acid storage at 4 degrees was 96%, 94%, and 98%, respectively.
  • the content of lanosterol, betulin, and oleic acid was 73%, 88%, and 97% after being placed at 80 ° C for 1 day; lanosterol, betulin, and oleic acid were placed at 80 ° C for 2 days.
  • the content was changed to 48%, 68%, and 96%, respectively; the content of lanosterol, betulin, and oleic acid at 80 ° C for 3 days was 44%, 67%, and 96%, respectively.
  • the powder After the lanosterol was left at 80 ° C for 1 day, the powder became hard and formed into a large piece. The color was gradually yellowed. When placed for 1 week, one third of the lanosterol turned yellow; the inoculum and the oleic acid After standing at 80 ° C for 1 week, there was no change in appearance compared to 4 ° C. After lanosterol was placed at 80 °C for 2 weeks, all of them turned yellow, and some of them showed crystal transformation. The agglomeration was serious and the purity was only 32%. The inoculum was placed at 80 °C for 2 weeks and began to change compared with 4 °C. Yellow; thiocyanate was placed at 80 ° C for 2 weeks and placed at 4 ° C, there was no significant change in appearance, the purity remained above 93%.
  • Example 19 a series of chemical modification and derivative synthesis were carried out based on two compounds of Inoculum and Trametic acid, and some specific compounds obtained therein were identified and characterized.
  • Compound 1 is an inoculum.
  • Red aluminum solution (70% toluene solution, 0.1 mL) was added to tetrahydrofuran (10 mL), and compound 3 (200 mg) was added thereto, and the reaction was carried out at a temperature of -10 ° C for 5 hours. It was poured into dilute hydrochloric acid, extracted with ethyl acetate, dried over sodium sulfate,
  • Compound 1 is stearyl acid.
  • Red aluminum solution (70% toluene solution, 0.1 mL) was added to tetrahydrofuran (10 mL), and then compound 3' (200 mg) was added thereto, and the mixture was reacted at -20 ° C for 5 hours. It was poured into dilute hydrochloric acid, extracted with ethyl acetate, dried over sodium sulfate, and then evaporated
  • Example 19 a part of the specific compounds obtained in Example 19 and Example 20 were selected, and an intracellular mutated lens protein ( ⁇ B-crystallin R120G) aggregation inhibition experiment (experimental method such as Chinese patent CN201580057679) was used to evaluate the present invention.
  • the compound of the formula I and the compound of the formula II of the invention inhibit the aggregation activity of the lens protein mutant, thereby screening for a compound effective for treating a related disease such as a cataract.
  • the tumor cell line HeLa was selected as the host cell transfected with the mutant lens protein ( ⁇ B-crystallin R120G) expression plasmid.
  • the medium was DMEM containing 10% FBS, and the culture condition was 37 ° C, saturated humidity, 5% carbon dioxide cell incubator. Mycoplasma was negative.
  • the transfection process is to first inoculate a suitable amount of HeLa cells on the pre-coated slides. When the fully adherent cells grow to a cell density of about 75%, the expression vector is transfected with the liposome, and transfected for 4 hours. The liquid normal medium was further cultured in a cell culture incubator for 13 hours to express the foreign mutant lens protein.
  • the replaced new medium contained various compounds of the formula I and formula II of the present invention to be tested (all mother liquors were treated with DMSO as a solvent), wherein each compound had a final concentration of 10 ⁇ M and the negative control was 1.5% DMSO. Solvent, the positive control was lanosterol (10 ⁇ M).
  • the HeLa cells were treated with the medium containing the test compound, 1.5% DMSO solvent and lanosterol for 3.5 hours, respectively, and the normal fresh medium containing no DMSO, test compound or lanosterol was replaced for 6.5 hours, followed by Anti ⁇
  • the p62 antibody and DAPI were subjected to routine cellular immunofluorescence assay. The fluorescence field of each position was randomly selected from each slide, and the proportion of cells containing aggregated crystal proteins (crystal protein aggregates) was calculated to determine the mutation of each test compound.
  • the effect of intracellular aggregation of lens proteins was as shown in Table 19.
  • the experimental method was as follows: Inonotus montanic acid was used as a test compound, and its dissolution effect on wild-type ⁇ B crystal protein aggregate (WT ⁇ B) was measured, and lanosterol was used as a positive control.
  • the pharmaceutical composition dosage form used in the test group and the control group is a liposome, wherein the test formulation of the test group is 40 mg of DPPC, 10 mg of cholesterol and 10 mg of the test compound; and the formulation of the positive control group is 40 mg.
  • DPPC 10 mg of cholesterol and 10 mg of lanosterol
  • Formulation formulation of blank control group 40 mg of DPPC and 10 mg of cholesterol.
  • the liposome preparation was prepared as follows: 40 mg of DPPC, 10 mg of cholesterol and 10 mg of the corresponding compound were weighed and placed in a pear-shaped flask, and the blank liposome contained only 40 mg of DPPC and 10 mg of cholesterol. An appropriate amount of chloroform was added to sufficiently dissolve the mixture of DPPC, cholesterol and the corresponding compound, and an appropriate amount of glass beads was added thereto, followed by vacuum spinning at 50 °C. After the solution is evaporated to dryness, the mixture is further steamed for 10 minutes to obtain a relatively dry film, which is placed at room temperature overnight in a vacuum drying oven to sufficiently evaporate the organic solvent.
  • the obtained liposome solution was subjected to demulsification treatment with 4 to 5 volumes of a methanol solution to obtain a clear solution.
  • the concentration of the compound in the liposome was determined by HPLC method, and the chromatographic conditions were RP C18 (4.6 mm x 150 mm, 5 ⁇ m) column, column temperature 35 ° C, detection wavelength 210 nm, flow rate 1.0 ml / min, injection volume 25 ⁇ l.
  • methanol was used as the mobile phase
  • methanol:water 90:10 was used as the mobile phase.
  • the concentration of the corresponding compound in the liposome preparation was determined.
  • Protein aggregates were induced by incubating a solution containing 1.2 mg/ml of wild-type ⁇ B crystallin at 48 ° C for 48 hours, and adding 0.02% NaN 3 to prevent bacterial infection, thereby forming ⁇ B crystal protein aggregates ( WT ⁇ B). Then, an inoculum preparation or a lanosterol preparation (liposome form, the final concentration of both compounds was 125 ⁇ M) was added to the solution, respectively, and left to stand for 24 hours. The control group was treated with blank liposomes for the same treatment. After the treatment, ThT fluorescence was used to detect the effect of different compounds on ⁇ B crystal protein aggregates.
  • inosmolin can extremely efficiently re-dissolve ⁇ B crystal protein aggregates at a low or near-in vivo level of compound concentration (*: p ⁇ 0.05 compared to the blank), The effect is far superior to lanosterol.
  • the compounds of formula I of the present invention such as inoguin, have better prospects for cataract treatment/preventive drug development than lanosterol.

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Abstract

本发明涉及一种三萜类化合物的新用途。具体地，本发明提供了式I化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药的用途，它们被用于制备药物组合物或制剂，所述药物组合物或制剂用于预防和/或治疗晶状体病变引起的眼部疾病。本发明首次发现了式I化合物能够预防和/或治疗晶状体病变引起的眼部疾病，如白内障的预防和治疗，且安全性高，毒副作用小并具有显著的治疗效果和开发应用前景。

Description

含三萜类化合物的药物组合物及其用途 技术领域
本发明涉及医药领域，具体地，本发明涉及一种三萜类化合物或桦褐孔菌提取物的新用途。
背景技术
正常人眼的晶状体由众多排列有序的晶状体蛋白构成，一旦这些晶状体蛋白排列错误或者发生错误堆积，形成蛋白聚集体，便会影响晶状体正常的透明度和折光率。其中，白内障是一种主要的表现形式，也是世界范围内致盲最高的疾病。因此，晶状体疾病，尤其是白内障的治疗和预防，就甚为重要。遗憾的是，目前针对白内障临床上依然以手术和替换人工晶体为主要治疗手段。
如何简单利用药物，就可以预防、抑制病情发展甚至治疗白内障一直是眼科研究的热点。之前有研究组报道利用羊毛甾醇可以治疗动物的白内障，羊毛甾醇的结构式如下式所示：
Figure PCTCN2018125075-appb-000001
然而值得注意的是，缓释的羊毛甾醇药物只有结合高频次玻璃腔注射给药方式的同时，才对狗有白内障的治疗效果；并且，有不同研究者也分别指出对眼球或晶体直接给予羊毛甾醇无法恢复或改善灵长类动物的白内障晶体的透明度。
因此，本领域亟需开发一种能够有效治疗晶状体病变或与晶状体病变相关的眼部疾病的药物，该药物尤其需要在灵长类动物药效测试中得到令人满意的结果。
发明内容
本发明的目的就是提供一种能够有效治疗晶状体病变或与晶状体病变相关的眼部疾病的药物及其应用。
本发明的第一方面，提供了一种式I化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药、或其氘代化合物的用途，它们被用于制备药物组合物或制剂，所述药物组合物或制剂用于预防和/或治疗晶状体病变引起的眼部疾病；
Figure PCTCN2018125075-appb-000002
式中，
q为0、1或2；
R1a、R1b、R2a、R2b、R3a和R3b各自独立地选自：氢、取代或未取代的C1‐C20烷基、取代或未取代的C2‐C20烯基、取代或未取代的C2‐C20炔基、取代或未取代的C3‐C10环烷基、‐OH、取代或未取代的C1‐C10烷氧基、‐COOH、‐CHO、取代或未取代的C1‐C10酯基、‐SH、取代或未取代的C1‐C10烷硫基、‐A‐B，
或者R1a和R1b、R2a和R2b、和/或R3a和R3b构成＝O；
其中，A为无、二价连接基团；而B为H、‐OH、‐SH、C1‐C3烷氧基、C1‐C3烷硫基、‐CHO、‐COOH、C1‐C4酯基、C3‐C10环烷基、芳基、C3‐C10的5‐6元杂芳基、或苄基；
附加条件是R1a、R2a、R3a、R1b、R2b和R3b中至少一个为含O或S的基团；
Z选自下组：H、取代或未取代的C1‐C20烷基、取代或未取代的C2‐C20烯基、取代或未取代的C2‐C20炔基、取代或未取代的C3‐C10环烷基、取代或未取代的芳基、取代或未取代的5‐8元杂芳基、取代或未取代的‐(C1‐C6亚烷基)‐芳基、取代或未取代的‐(C1‐C6亚烷基)‐5‐8元杂芳基、‐OH、取代或未取代的C1‐C20烷氧基、‐SH、取代或未取代的C1‐C20烷硫基、取代或未取代的C1‐C10酯基、取代或未取代的C1‐C10酰基、取代或未取代的‐O‐芳基；
R4为氢、取代或未取代的C1‐C4烷基；
R7、R12和R15各自独立地选自：无、氢、取代或未取代的C1‐C4烷基；
R13a和R13b各自独立地选自：无、氢、取代或未取代的C1‐C8烷基、‐OH、取代或未取代的C1‐C8烷氧基、‐SH、取代或未取代的C1‐C8烷硫基、卤素、取代或未取代的C1‐C3酰基，或R13a和R13b构成＝O，且R13a和R13b中至多一个为无；
R11a和R11b各自独立地选自：无、氢、取代或未取代的C1‐C6烷基、‐OH，或R11a和R11b构成＝O，且R11a和R11b中至多一个为无；
R10a、R10b各自独立地选自：氢、‐OH、取代或未取代的C1‐C8烷氧基、‐SH、取代或未取代的C1‐C8烷硫基、‐OSO 3H、‐OCO‐取代或未取代的C1‐C7烷基、‐OPO 3H、‐COOH、‐(取 代或未取代的C1‐C7亚烷基)‐COOH、‐CHO、‐(取代或未取代的C1‐C7亚烷基)‐CHO，或R10a和R10b共同构成＝O；和
R5、R6、R8、R9a、R9b、R14、R16、R17a和R17b各自独立地选自：氢、OH、取代或未取代的C1‐C8烷基、取代或未取代的C1‐C8烷氧基、卤素。
在另一优选例中，所述的“取代”指有一个或多个氢被选自下组的基团所取代：卤素、‐OH、‐SH、‐COOH、‐(C1‐C7亚烷基)‐COOH、＝O、‐CHO、‐(C1‐C7亚烷基)‐CHO、C1‐C7烷基‐OCO‐、C1‐C3烷基、C3‐C6环烷基、氰基、硝基、氰酸酯基、异氰酸酯基、异硫氰酸酯基、磺酰胺基、肟基、NRaRb，其中，Ra和Rb各自独立地为H、C1‐C6烷基、C1‐C6卤代烷基、C3‐C6环烷基、或苄基。
在另一优选例中，所述的“取代”指有一个或多个氢被选自下组的基团所取代：卤素、‐OH、‐SH、‐COOH、‐(C1‐C7亚烷基)‐COOH、＝O、‐CHO、‐(C1‐C7亚烷基)‐CHO、C1‐C7烷基‐OCO‐、C1‐C3烷基、C3‐C6环烷基、NRaRb，其中，Ra和Rb各自独立地为H、C1‐C6烷基、C1‐C6卤代烷基、C3‐C6环烷基、或苄基。
在另一优选例中，在所述结构式中，所述的虚线
Figure PCTCN2018125075-appb-000003
所表示的双键的总数为0、1、2、3或4个。
在另一优选例中，所述的双键位于以下位点之间：a和b、b和c、c和d、e和f、c和f、f和g、g和h、h和a、b和i、i和j、a和k、和/或k和l。
在另一优选例中，所述的双键位于以下位点之间：a和b。
在另一优选例中，所述的双键位于以下位点之间：h和a、和/或b和i。
在另一优选例中，所述的结构式中，各C的价态符合化学稳定结构的要求(即C为四价)。
当R5、R6、R7、R8、R12、R14、R15和/或R16连接于含双键的C时，所述C的价位为4价。
在另一优选例中，所述的二价连接基团具有1‐10个选自下组的连接单元：‐CRaRb‐；‐C(OH)Ra‐、‐NRa‐、‐O‐、‐CO‐；其中，Ra和Rb各自独立地为H、C1‐C6烷基、C1‐C6卤代烷基、C3‐C6环烷基、或苄基。
在另一优选例中，所述的R1a、R1b、R2a、R2b、R3a和R3b各自独立地选自：氢、‐OH、‐CHO、‐COOH、‐SH、取代或未取代的C1‐C3烷基‐OCO‐、取代或未取代的C1‐C4烷基、‐A‐B、或R1a和R1b、R2a和R2b、和/或R3a和R3b构成＝O；
其中，A各自独立为取代或未取代的C1‐C4亚烷基，而B各自独立地为‐OH、‐SH、C1‐C3烷氧基、‐CHO、‐COOH、＝O。
在另一优选例中，所述的Z为(L1)m‐L2＝Y，m为0、1、2、3或4，各L1各自独立地为‐CH 2‐、‐CO‐、‐O‐、‐C(C1‐C3烷基)H‐、C(C1‐C3烷基)2‐、‐C(C1‐C3烷基)O‐、或‐NH‐，L2为‐CH＝、或‐C(C1‐C3烷基)＝；
Y为＝CH 2、＝CH‐取代或未取代的C1‐C7烷基、＝C‐(取代或未取代的C1‐C7烷基) 2、＝ CH‐取代或未取代的C2‐C7烯基、或＝CH‐取代或未取代的C2‐C7炔基。
在另一优选例中，对于Y而言，所述的取代表示一个或多个氢被选自下组的基团所取代：卤素、‐OH、‐SH、‐COOH、‐(C1‐C7亚烷基)‐COOH、＝O、‐CHO、‐(C1‐C7亚烷基)‐CHO、C1‐C7烷基‐OCO‐、C1‐C3烷基、C3‐C6环烷基、NRaRb，其中，Ra和Rb各自独立地为H、C1‐C6烷基、C1‐C6卤代烷基、C3‐C6环烷基、或苄基。
在另一优选例，Y为具有羟基或SH取代基的基团。
在另一优选例，所述的R4为氢、甲基。
在另一优选例中，R7、R12和R15各自独立选自：无、氢、甲基。
在另一优选例中，所述的R13a和R13b各自独立地选自：氢、甲基、‐OH、‐SH、＝O、卤素，且R13a和R13b中至多一个为无。
在另一优选例中，R13a和R13b中至少一个为‐OH、或‐SH。在另一优选例中，所述R10a、R10b各自独立地选自：氢、‐OH、‐SH、‐OSO 3H、‐OPO 3H、‐COOH、‐CHO；或R10a和R10b构成羰基。
在另一优选例中，所述R5、R6、R8、R9a、R9b、R14、R16、R17a和R17b各自独立选自氢、甲基。
在另一优选例中，所述R9a、R9b各自独立选自氢、甲基和卤素。
在另一优选例中，所述R9a为卤素(F、Cl、或Br)，而R9b选自氢或甲基。
在另一优选例中，R1a和R1b中至少一个为含O或S的基团。
在另一优选例中，R2a和R2b中至少一个为含O或S的基团。
在另一优选例中，R3a和R3b中至少一个为含O或S的基团。
在另一优选例中，所述的Z不是‐CH 2C(CH 3) 3；
在另一优选例中，Z为‐CH 2C(CH 3) 2‐OH。
在另一优选例中，q为1或2。
在另一优选例中，所述的式I化合物为式I‐1化合物：
Figure PCTCN2018125075-appb-000004
其中，q、R1a、R1b、R2a、R2b、R3a、R3b、R4、R7、R10a、R10b、R11a、R11b、R12、 R13a、R13b、R15和Z的定义如上。
在另一优选例中，R11a和/或R11b为甲基。
在另一优选例中，所述的式I化合物为式I‐2化合物：
Figure PCTCN2018125075-appb-000005
其中，q、R1a、R1b、R2a、R2b、R3a、R3b、R4、R7、R11a、R11b、R13a、R13b、R15和Z的定义如上。
在另一优选例中，在a和b之间为双键(＝)，而b和c，a和h，b和i，a和k以及k和i之间均为单键。
在另一优选例中，在b和i以及a和h之间均为双键(＝)，而a和b、b和c，a和k以及k和i之间均为单键。
在另一优选例中，所述的式I化合物选自下组化合物：
Figure PCTCN2018125075-appb-000006
Figure PCTCN2018125075-appb-000007
其中，R1a、R1b、R2a、R2b和Z的定义如上。
在另一优选例中，所述的式I化合物选自下组化合物：
Figure PCTCN2018125075-appb-000008
其中，R1a、R1b、R2a、R2b的定义如上。
在另一优选例中，所述的式I化合物为具有式I‐7所示的结构：
Figure PCTCN2018125075-appb-000009
其中，R1a、R1b、R2a、R2b的定义如上文所述。
在另一优选例中，所述的式I化合物为具有式I‐8所示的结构：
Figure PCTCN2018125075-appb-000010
其中，R1a、R1b、R2a、R2b的定义如上文所述。
在另一优选例中，所述式I所示的化合物选自下组(表A)：
表A
Figure PCTCN2018125075-appb-000011
Figure PCTCN2018125075-appb-000012
Figure PCTCN2018125075-appb-000013
Figure PCTCN2018125075-appb-000014
Figure PCTCN2018125075-appb-000015
Figure PCTCN2018125075-appb-000016
Figure PCTCN2018125075-appb-000017
Figure PCTCN2018125075-appb-000018
在另一优选例中，所述的式I化合物选自下组：
Figure PCTCN2018125075-appb-000019
在另一优选例中，所述式I所示的化合物选自下组：
Figure PCTCN2018125075-appb-000020
在另一优选例中，所述的晶状体病变引起的眼部疾病选自下组：白内障、老花眼、近视眼、皮质混浊、老视核硬化、糖尿病引发的眼部并发症。
在另一优选例中，所述的眼部疾病选自下组：先天性白内障、获得性白内障。
在另一优选例中，所述的眼部疾病选自下组：成熟期白内障、非成熟期白内障。
在另一优选例中，所述的白内障选自下组：外伤性白内障、代谢型白内障、老年型白内障、先天型白内障、自发型白内障、并发类白内障、或其组合。
在另一优选例中，所述的代谢型白内障包括糖尿病代谢型白内障。
在另一优选例中，所述的外伤性白内障包括手术相关的白内障。
在另一优选例中，所述的自发型白内障包括老龄自发性白内障。
在另一优选例中，所述药物组合物或制剂中包括：(a)作为第一活性成分的有效治疗量的式I所示的化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药；(b)药学上可接受的载体；
其中，第一活性成分的含量为0.001‐99wt％，较佳的0.01‐70wt％，更佳的为0.05‐40wt％的，按照组合物的总重量计。
在另一优选例中，所述药物组合物或制剂中包括：(a)作为第一活性成分的有效治疗量的式I所示的化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药、或其氘代化合物；(b)药学上可接受的载体；
其中，第一活性成分的含量为0.001‐99wt％，较佳的0.01‐70wt％，更佳的为0.05‐40wt％的，按照组合物的总重量计。
在另一优选例中，所述的第一活性成分的浓度优选为1-500mM或10～200mM，较佳地15～150mM，更佳地20～50mM；最佳地20～30mM。
在另一优选例中，所述的药用组合物或制剂的剂型选自：注射剂、外用制剂、口服制剂。
在另一优选例中，所述的药用组合物或制剂的剂型为眼用制剂，所述的眼用制剂为眼药水、乳剂、凝胶、眼用膏、缓释微球、眼内缓释植片、药用缓释药膜。
在另一优选例中，所述的药用组合物或制剂还包括：(c)第二活性成分，其中所述的第 二活性成分选自下组：羊毛脂类化合物、羊毛甾醇、所有锈革孔菌目或多孔菌目中包含菌类中含有的任一化合物(尤其是类固醇类化合物、萜类化合物)、唑类化合物、糖皮质激素类化合物、抗生素、或其组合。
在另一优选例中，所述的第二活性成分选自下组：羊毛脂类化合物、羊毛甾醇、所有锈革孔菌目或多孔菌目中包含菌类中含有的任一化合物(尤其是类固醇类化合物、萜类化合物)、唑类化合物、淀粉样蛋白调节剂、糖皮质激素类化合物、抗生素、或其组合。
在另一优选例中，所述的第二活性成分的含量为0.01-20wt％，较佳地为5-15wt％，按照组合物的总重量计。
在另一优选例中，所述的“第二活性成分”是羊毛脂类化合物或唑类化合物时，其浓度优选为10～200mM，较佳地15～150mM，更佳地20～50mM；最佳地20～30mM。
在另一优选例中，所述的羊毛脂类化合物选自：羊毛甾醇、二氢羊毛甾醇、25‐羟基胆固醇，或其组合。
在另一优选例中，所述的淀粉样蛋白调节剂选自：TPPB(CAS.497259‐23‐1)、迷迭香酸、强力霉素。
在另一优选例中，所述的糖皮质激素化合物选自下组：地塞米松、氢化可的松、或其组合。
在另一优选例中，所述的抗生素选自下组：妥布霉素、硫酸庆大霉素、金霉素、氯霉素、或其组合。
在另一优选例中，所述的唑类化合物选自益康唑、异康唑、联苯苄唑、克霉唑、阿立哌唑、酮康唑、氟康唑、苯基咪唑、咪康唑、环菌唑、三唑醇、戊唑醇、丙环唑、或其组合。
在另一优选例中，所述的唑类化合物选自益康唑、氟康唑、异康唑、戊唑醇、丙环唑、或其组合。
在另一优选例中，式I化合物与唑类化合物的质量比为50:1至1:50，较佳地10:1至1:10。
在另一优选例中，所述的式I化合物还包括式I化合物的氘代化合物。
在另一优选例中，所述的药物组合物或制剂还用于(b)抑制、逆转(溶解或解聚)晶状体蛋白聚集。
在另一优选例中，所述的药物组合物或制剂还用于(c)预防和/或治疗与晶状体蛋白聚集相关的疾病。
在另一优选例中，所述的晶状体蛋白包括αB晶状体蛋白。
在另一优选例中，所述与晶状体蛋白聚集相关的疾病选自下组：白内障、老花眼、近视眼、皮质混浊、老视核硬化、糖尿病引发的眼部并发症。
在本发明的第二方面，提供了一种体外非治疗性和/或非诊断性的改善或维持晶状体透明度的方法，它包括：将晶状体与式I化合物、或其光学异构体或其外消旋体、或其溶 剂化物、或其药学上可接受的盐、或其前药、或其氘代化合物接触，从而改善(如提高)或维持晶状体透明度，其中，所述的式I化合物如上所述。
在本发明的第三方面，提供了一种预防和/或治疗晶状体病变引起的眼部疾病的方法，包括步骤：给予所需对象式I所示的化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药、或其氘代化合物，其中所述式I化合物如上所定义。
在另一优选例中，所述对象为人和非人哺乳动物。代表性地，所述的非人哺乳动物包括(但并不限于)：宠物(如狗、猫)、家畜(如牛、羊、马、猪)、各种动物园动物(熊猫、大象)等。
在另一优选例中，所述的对象还包括除人和非人哺乳动物外的其他动物，如非哺乳动物。
在本发明的第四方面，提供了一种新颖的式II化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药、或其氘代化合物，
Figure PCTCN2018125075-appb-000021
式II中，
P为0、1或2；
R18a、R18b各自独立地为氢、取代或未取代的C1-C4烷基、或-COOH；
R19a、R19b各自独立地为氢、取代或未取代的C1-C4烷基、或卤素；
R20为取代或未取代的C1‐C6酰基、取代或未取代的C1-C6烷基、取代或未取代的糖基-O-(C1-C6烷基)-、取代或未取代的肟基、取代或未取代的C3-C6杂环烷基、或取代或未取代的C2-C6烯基；
R21a、R21b各自独立地为氢、羟基、巯基、或取代或未取代的C1-C4烷基；
R22a、R22b各自独立地为氢、或取代或未取代的C1-C4烷基；
R23a、R23b各自独立地为氢、羟基、巯基、糖基、或取代或未取代的C1-C烷氧基；
R24a、R24b各自独立地为氢、羟基、或卤素；
所述的“取代”指有一个或多个氢被选自下组的基团所取代：卤素、‐OH、-SH、‐COOH、＝O、‐CHO、C1‐C4烷基、C3‐C6环烷基、氨基、糖基。
在另一优选例中，所述的糖基为单糖(如戊糖或己糖)、或双糖。
在另一优选例中，在式II中，所述的虚线
Figure PCTCN2018125075-appb-000022
所表示的双键的总数为0、1或2个。
在另一优选例中，所述的双键位于m和n之间。
在另一优选例中，所述的双键位于m和l，和/或o和n之间。
在另一优选例中，R22a和/或R22b为甲基。
在另一优选例中，p为1。
在另一优选例中，p为2。
在另一优选例中，R23a为氢，而R23b为羟基、巯基、或糖基。
在另一优选例中，R20为糖基-O-(C1-C6烷基)-。
在另一优选例中，所述的糖基为戊糖基或己糖基，优选地选自下组：葡萄糖基、果糖基、甘露糖基、阿拉伯糖基、核糖基、或其组合。
在另一优选例中，所述的杂环烷基含有1‐3个选自下组N、O和S的杂原子。
在另一优选例中，所述的杂环烷基是含1个O的杂原子的C3‐C6杂环烷基。
在另一优选例中，所述的式II化合物具有选自下组的一种或多种特征：
R18a、R18b各自独立地为-COOH、或甲基；
R19a、R19b各自独立地为氢、或卤素；
R20为羟基丁酰基、羟丙基、肟基、甲氧基丙基、二甲基环氧乙基、HOOC-CH＝CH-、或六元单糖丙基；
R21a、R21b各自独立地为氢、羟基、或巯基；
R22a、R22b各自独立地甲基；
R23a、R23b各自独立地为氢、羟基、巯基或六元单糖基；和
R24a为氢，而R24b为氟。
在另一优选例中，同一碳原子上不含有2个或2个以上的羟基。
在另一优选例中，同一碳原子上不含有2个或2个以上的巯基。
在另一优选例中，所述的六元单糖为葡萄糖。
在另一优选例中，所述的式II化合物选自下组(表B)：
表B
Figure PCTCN2018125075-appb-000023
Figure PCTCN2018125075-appb-000024
Figure PCTCN2018125075-appb-000025
本发明的第五方面，提供了式II化合物或其药学上可接受的盐或其前药的用途，它被用于制备药物组合物或制剂，所述药物组合物或制剂用于：(a)预防和/或治疗晶状体病变引起的眼部疾病；(b)抑制和/或逆转(溶解或解聚)晶状体蛋白聚集；和/或(c)预防和/或治疗与晶状体蛋白聚集相关的疾病。
本发明的第六方面，提供一种桦褐孔菌提取物的用途，用于制备药物组合物或制剂，所述药物组合物或制剂用于：(a)预防和/或治疗晶状体病变引起的眼部疾病。
在另一优选例中，所述的提取物包括脂溶性提取物。
在另一优选例中，所述的提取物包括醇提取物。
在另一优选例中，所述的提取物包括非水性溶剂的提取物。
在另一优选例中，所述的提取物为非水溶性提取物或水难溶性提取物。
在另一优选例中，所述的提取物含有以下组分：萜类、甾类、或其组合。
在另一优选例中，所述的提取物包括萜类提取物。
在另一优选例中，所述的提取物包括三萜类化合物，较佳地四环三萜类提取物。
在另一优选例中，所述的提取物中含有选自下表C中一种或多种化合物：
表C
Figure PCTCN2018125075-appb-000026
Figure PCTCN2018125075-appb-000027
Figure PCTCN2018125075-appb-000028
更佳地，所述提取物中含有选自下组的化合物：
Figure PCTCN2018125075-appb-000029
在另一优选例中，所述提取物中含有选自下组的化合物：
Figure PCTCN2018125075-appb-000030
Figure PCTCN2018125075-appb-000031
或其组合。
在另一优选例中，所述的提取物中，所述的桦褐孔菌醇的重量百分比为0.01-99.99wt％，较佳地1-99wt％。
在另一优选例中，所述的提取物中，所述的栓菌酸的重量百分比为0.01-99.99wt％，较佳地1-99wt％。
在另一优选例中，在所述的提取物中，桦褐孔菌醇和栓菌酸占四环三萜类含量的5-100wt％，优选地10-100wt％，更优选地20-100wt％，更优选地30-100wt％，更优选地70-100wt％，最优选地80-100wt％。
在另一优选例中，所述的提取物不是纯羊毛甾醇。
在另一优选例中，所述的提取物中桦褐孔菌醇(Inotodiol)与羊毛甾醇的重量比≥2:1，较佳地≥5:1，更佳地≥10:1。
在另一优选例中，所述的提取物为桦褐孔菌的C1-C6醇的提取物。
在另一优选例中，所述的C1-C6醇包括甲醇、乙醇、丙醇或其组合。
在另一优选例中，所述的提取物在进行醇提取后，还进行进一步纯化。
在另一优选例中，所述的进一步纯化包括：用石油醚和/或酯类溶剂进行萃取。
在另一优选例中，所述桦褐孔菌提取物包括桦褐孔菌的子实体的脂溶性提取物或水溶性提取物。
在另一优选例中，所述的眼部疾病选自下组：白内障、老花眼、近视眼、皮质混浊、老视核硬化、糖尿病引发的眼部并发症。
在另一优选例中，所述的眼部疾病选自下组：先天性白内障、获得性白内障。
在另一优选例中，所述的眼部疾病选自下组：成熟期白内障、非成熟期白内障。
在另一优选例中，所述的白内障选自下组：外伤型白内障、代谢型白内障、并发型白内障、老年型白内障、自发型白内障，或其组合。
在另一优选例中，所述的外伤型白内障包括手术相关的白内障。
在另一优选例中，所述的自发型白内障包括老龄自发性白内障。
在另一优选例中，所述的代谢型白内障包括糖尿病代谢型白内障。
在另一优选例中，所述的桦褐孔菌提取物是通过以下方法制备的：
(1)用醇溶剂对桦褐孔菌子实体的粗粉进行回流提取，得醇提取物；
(2)将所述的醇提取物加水混悬后用石油醚萃取，获得萃取物，即为桦褐孔菌提取物；
和任选的步骤(3)将所述的桦褐孔菌提取物进行硅胶柱层析，收集洗脱液，分离得到纯化的桦褐孔菌提取物。
在另一优选例中，所述的桦褐孔菌提取物是通过以下方法制备的：
(1)用乙醇对桦褐孔菌子实体的粗粉进行回流提取，得乙醇提取物；
(2)将所述的乙醇提取物加水混悬后用石油醚萃取，分离溶液，得桦褐孔菌提取物。
在另一优选例中，所述的制备方法在步骤(2)后还包括步骤(3)将所述的桦褐孔菌提取物进行硅胶柱层析，收集洗脱液，分离得到纯化的桦褐孔菌提取物。
在另一优选例中，所述的步骤(1)中，桦褐孔菌子实体的粗粉与醇溶剂的重量比为1:30‐100，较佳地1：60‐90。
在另一优选例中，所述的醇溶剂包括乙醇。
在另一优选例中，所述的醇溶剂包括90‐100％(v/v)乙醇水溶液。
在另一优选例中，所述的子实体为干燥子实体。
在另一优选例中，所述药物组合物或制剂包括：(a)作为第一活性成分有效治疗量的桦褐孔菌提取物；(b)药学上可接受的载体。
其中，第一活性成分的含量为0.001-99wt％，较佳的0.01-70wt％，更佳的为0.05-40wt％的，按照组合物或制剂的总重量计。
在另一优选例中，所述的药用组合物或制剂的剂型选自：口服制剂、注射剂、外用制剂。
在另一优选例中，所述的药用组合物或制剂的剂型为眼用制剂，所述的眼用制剂为眼药水、乳剂、凝胶、眼用膏、缓释微球、眼内缓释植片、药用缓释药膜。
在另一优选例中，所述的药用组合物或制剂还包括：(c)第二活性成分，其中所述的第二活性成分选自下组：羊毛甾醇类化合物、唑类化合物、糖皮质激素类化合物、抗生素、或其组合。
在另一优选例中，所述的药用组合物或制剂还包括：(c)第二活性成分，其中所述的第二活性成分选自下组：羊毛甾醇类化合物、唑类化合物、糖皮质激素类化合物、淀粉样蛋白调节剂、抗生素、或其组合。
在另一优选例中，所述的第二活性成分的含量为0.01-20wt％，较佳地为5-15wt％，按照组合物的总重量计。
在另一优选例中，所述的“第二活性成分”是羊毛甾醇类化合物时，其浓度优选为10～200mM，较佳地15～150mM，更佳地20～50mM；最佳地20～30mM。
在另一优选例中，所述的羊毛甾醇类化合物选自：羊毛甾醇。
在另一优选例中，所述的糖皮质激素化合物选自下组：地塞米松、氢化可的松、或其组合。
在另一优选例中，所述的淀粉样蛋白调节剂选自：TPPB(CAS.497259‐23‐1)、迷迭香酸、强力霉素。
在另一优选例中，所述的抗生素选自下组：妥布霉素、硫酸庆大霉素、金霉素、氯霉素、或其组合。
在另一优选例中，所述的唑类化合物选自益康唑、异康唑、联苯苄唑、克霉唑、阿立哌唑、酮康唑、氟康唑、苯基咪唑、咪康唑、环菌唑、三唑醇、戊唑醇、丙环唑、或其组合。
在另一优选例中，桦褐孔菌提取物与唑类化合物的质量比为50:1至1:50，较佳地10:1至1:10。
在另一优选例中，所述的药物组合物或制剂还用于：(b)抑制、逆转晶状体蛋白聚集；和/或(c)预防和/或治疗与晶状体蛋白聚集相关的疾病。
在另一优选例中，所述的晶状体蛋白包括αB晶状体蛋白。
在另一优选例中，所述与晶状体蛋白聚集相关的疾病选自下组：白内障、老花眼、近视眼、皮质混浊、老视核硬化、糖尿病引发的眼部并发症。
本发明的第七方面，提供了一种体外非治疗性和/或非诊断性的改善或维持晶状体透明度的方法，包括步骤：将晶状体与桦褐孔菌提取物接触，从而改善(或提高)或维持晶状体透明度，其中，所述的桦褐孔菌提取物如本发明第四方面所述。
本发明的第八方面，提供了一种预防和/或治疗晶状体病变引起的眼部疾病的方法，包括步骤：给予所需对象桦褐孔菌提取物，其中，所述的桦褐孔菌提取物如本发明第六方 面所述。
在另一优选例中，所述对象为人和非人哺乳动物。代表性地，所述的非人哺乳动物包括(但并不限于)：宠物(如狗、猫)、家畜(如牛、羊、马、猪)、各种动物园动物(熊猫、大象)等。
在另一优选例中，所述的对象还包括除人和非人哺乳动物外的其他动物，如非哺乳动物。
在本发明的第九方面，提供了一种药物组合物，其包括：
(a)作为第一活性成分的有效治疗量的式I或式II所示的化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药、或其氘代化合物；
(b)药学上可接受的载体；和
(c)第二活性成分，其中所述的第二活性成分选自下组：唑类化合物、淀粉样蛋白调节剂、或其组合。
其中，第一活性成分的含量为0.001‐99wt％，较佳的0.01‐70wt％，更佳的为0.05‐40wt％的，按照组合物的总重量计。
在另一优选例中，所述的第一活性成分的浓度优选为1-500mM或10～200mM，较佳地15～150mM，更佳地20～50mM；最佳地20～30mM。
在另一优选例中，所述的第二活性成分的含量为0.01-20wt％，较佳地为5-15wt％，按照组合物的总重量计。
在另一优选例中，所述的“第二活性成分”是唑类化合物时，其浓度优选为10～200mM，较佳地15～150mM，更佳地20～50mM；最佳地20～30mM。
在另一优选例中，所述的唑类化合物选自益康唑、异康唑、联苯苄唑、克霉唑、阿立哌唑、酮康唑、氟康唑、苯基咪唑、咪康唑、环菌唑、三唑醇、戊唑醇、丙环唑、或其组合。
在另一优选例中，所述的唑类化合物选自益康唑、氟康唑、异康唑、戊唑醇、丙环唑、或其组合。
在另一优选例中，所述的淀粉样蛋白调节剂选自：TPPB(CAS.497259‐23‐1)、迷迭香酸、强力霉素、或其组合。
在另一优选例中，式I化合物(或式II化合物)与唑类化合物的质量比为50:1至1:50，较佳地10:1至1:10。
在另一优选例中，所述的式I化合物选自下组：桦褐孔菌醇、栓菌酸、或其组合。
在另一优选例中，所述的药用组合物或制剂的剂型选自：注射剂、外用制剂、口服制剂。
在另一优选例中，所述的药用组合物或制剂的剂型为眼用制剂，所述的眼用制剂为眼药水、乳剂、凝胶、眼用膏、缓释微球、眼内缓释植片、药用缓释药膜。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
附图说明
图1为本发明实施例1组选取的白内障造模成功后大鼠晶状体的上光源的形态。
图2为本发明实施例1组选取的白内障造模成功后大鼠晶状体的下光源的形态。
图3为本发明对比例1组选取的白内障造模成功后大鼠晶状体的上光源的形态。
图4为本发明对比例1组选取的白内障造模成功后大鼠晶状体的下光源的形态。
图5为本发明实施例1组选取的实验结束后大鼠晶状体的上光源的形态。
图6为本发明实施例1组选取的实验结束后大鼠晶状体的下光源的形态。
图7为本发明对比例1组选取的实验结束后大鼠晶状体的上光源的形态。
图8为本发明对比例1组选取的实验结束后大鼠晶状体的下光源的形态。
图9为本发明实施例2组选取的白内障造模成功后大鼠晶状体的上光源的形态。
图10为本发明实施例2组选取的白内障造模成功后大鼠晶状体的下光源的形态。
图11为本发明对比例2组选取的白内障造模成功后大鼠晶状体的上光源的形态。
图12为本发明对比例2组选取的白内障造模成功后大鼠晶状体的下光源的形态。
图13为本发明实施例2组选取的实验结束后大鼠晶状体的上光源的形态。
图14为本发明实施例2组选取的实验结束后大鼠晶状体的下光源的形态。
图15为本发明对比例2组选取的实验结束后大鼠晶状体的上光源的形态。
图16为本发明对比例2组选取的实验结束后大鼠晶状体的下光源的形态。
图17为本发明实施例8中桦褐孔菌醇眼药膏对食蟹猴先天型白内障(左眼)治疗前后的眼球的形态，其中17A为给药前，17B为给药后。
图18为本发明实施例8中桦褐孔菌醇眼药膏对食蟹猴先天型白内障(右眼)治疗前后的眼球的形态，其中18A为给药前，18B为给药后。
图19为本发明实施例9中桦褐孔菌醇眼药膏对食蟹猴(编号013321)自发型白内障(左眼)治疗前后的眼球的形态，其中19A为给药前，19B为给药后。
图20为本发明实施例9中桦褐孔菌醇眼药膏对食蟹猴(编号990447)自发型白内障(左眼)治疗前后的眼球的形态，其中20A为给药前，20B为给药后。
图21为本发明实施例9中羊毛甾醇眼药膏对食蟹猴(编号993747)自发型白内障(左眼)治疗前后的眼球的形态，其中21A为给药前，21B为给药后。
图22为本发明实施例10中桦褐孔菌醇眼药膏对大鼠外伤型白内障的(选取一只雌性大鼠的右眼作为拍照对比)治疗前后的眼球的形态，其中22A为给药前，22B为给药后。
图23为本发明实施例10中栓菌酸眼药膏对大鼠外伤型白内障的(选取一只雄性大鼠的右眼作为拍照对比)治疗前后的眼球的形态，其中23A为给药前，23B为给药后。
图24为本发明实施例10中羊毛甾醇眼药膏对大鼠外伤型白内障的(选取一只雌鼠的左眼作为拍照对比)治疗前后的眼球的形态，其中24A为给药前，24B为给药后。
图25为本发明实施例11中桦褐孔菌醇眼药膏对糖尿病大鼠代谢型白内障的(选取一只雄性大鼠的右眼作为拍照对比)治疗前后的眼球的形态，其中25A为给药前，25B为给药后。
图26为本发明实施例11中栓菌酸眼药膏对糖尿病大鼠代谢型白内障的(选取一只雄性大鼠的右眼作为拍照对比)治疗前后的眼球的形态，其中26A为给药前，26B为给药后。
图27为本发明实施例11中羊毛甾醇眼药膏对糖尿病大鼠代谢型白内障的(选取一只雄性大鼠的右眼作为拍照对比)治疗前后的眼球的形态，其中27A为给药前，27B为给药后。
图28为本发明实施例11中空白眼药膏对糖尿病大鼠代谢型白内障的(选取一只雄性大鼠的右眼作为拍照对比)治疗前后的眼球的形态，其中28A为给药前，28B为给药后。
图29为本发明实施例11中阳性对照组吡诺克辛钠滴眼液对糖尿病大鼠代谢型白内障的右眼治疗前后的眼球的形态，其中图29A为给药前，图29B为给药后。
图30为本发明实施例14制备的桦褐孔菌提取物2的的核磁共振(NMR)图。
图31为本发明实施例14制备的桦褐孔菌提取物2的的质谱(MS)图。
图32为本发明实施例14制备的桦褐孔菌提取物2的的高效液色谱(HPLC)图。
图33为本发明实施例22中桦褐孔菌醇显著降低晶状体蛋白聚集体ThT荧光信号检测结果。
具体实施方式
本发明人经过广泛而又深入的研究，首次意外地发现了一种式I所示化合物(包括纯净物、混合物或相应提取物)可以显著预防和/或治疗晶状体病变引起的眼部疾病。本发明实验结果表明式I所示化合物以简单的给药方式，快速达到治愈和预防晶状体病变引起的眼部疾病效果。在此基础上完成了本发明。
术语
如本文所用，“R1”、“R 1”和“R 1”的含义相同。其它类似定义的含义相同。
如本文所用，术语“C1‐C20烷基”或“C1‐C10烷基”或“C1‐C8烷基”指具有1‐20个或1‐10个或1‐8个碳原子的直链或支链烷基，例如甲基、乙基、丙基、异丙基、丁基、异丁基、仲丁基、叔丁基、戊基、己基、庚基、辛基，或类似基团。
如本文所用，术语“C1‐C8烷氧基”或“C1‐C4烷氧基”指具有1‐8个和1‐4个碳原子的直链或支链的烷氧基，例如甲氧基、乙氧基、丙氧基、异丙氧基、丁氧基、异丁氧基、仲丁氧基、叔丁氧基、戊氧基、己氧基、庚氧基、辛氧基，或类似基团。
如本文所用，术语“C1‐C8烷硫基””或“C1‐C4烷硫基”指具有1‐8个和1‐4个碳原子的直链或支链的烷硫基，例如甲硫基、乙硫基、丙硫基、异丙硫基、丁硫基、异丁硫基、仲丁 硫基、叔丁硫基、戊硫基、己硫基、庚硫基、辛硫基，或类似基团。
如本文所用，术语“C3‐C10环烷基”指具有3‐10个碳原子的环烷基，例如环丙基、环丁基、甲基环丁基、环戊基，或类似基团。
如本文所用，术语“C1‐C8亚烷基”或“C1‐C7亚烷基”或“C1‐C6亚烷基”指的是具有1‐8个和1‐7个和1‐6个碳原子的支链或直链的烷基分子的一个碳原子去掉2个氢原子形成的中有‐CH2‐的基团，例如亚甲基(‐CH2‐)，或类似基团。
如本文所用，术语“C2‐C10烯基”指具有1个或多个双键的2‐10个碳原子的直链或支链烯烃分子少掉一个和双键连接的氢原子形成的烃基，例如乙烯基(CH2＝CH‐)、(C(CH3) 2＝CH‐)，或类似基团。
如本文所用，术语“C2‐C8炔基”指具有1个或多个三键的2‐8个碳原子的直链或支链炔基分子少掉一个和三键连接的氢原子形成的烃基，例如乙炔基(CH三CH‐)、(H3C‐C三CH‐)，或类似基团。
如本文所用，术语“C1‐C6卤代烷基”指具有1‐6个碳原子的直链或支链烷基‐卤素结果的取代基，如‐CH 2Cl、‐CH 2CH 2Br、CH 2CHCH 2Cl，或类似基团。
如本文所用，术语“卤素”指F、Cl、Br和I。
如本文所用，术语“芳基”指C6‐C20的芳基，包括单环或双环或三环的芳基，例如苯基、萘基、蒽基。
如本文所用，术语“C6‐C10元芳基”指具有芳香结构的环状基团，例如苯基、萘基。
如本文所用，术语“5‐8元杂芳基”具有5～8元的单环或稠合多环且环系上具有N、O或S的杂芳基，例如吡咯基、吡啶基、呋喃基，或类似基团。
如本文所用，术语“氰酸酯基”结构式：-O-C≡N。
如本文所用，术语“异氰酸酯基”结构式：-N＝C＝O。
如本文所用，术语“异硫氰酸酯基”结构式：-N＝C＝S。
如本文所用，术语“磺酰胺基”结构式：
Figure PCTCN2018125075-appb-000032
或类似结构。
如本文所用，术语“肟基”结构式：
Figure PCTCN2018125075-appb-000033
其中，R 1和R 2分别表示连接的基团，且R 1和R 2可以相同或不同。
如本文所用，术语“糖基(glycosyl)”指通过从单糖(或双糖)的环状形式除去半缩醛羟基而形成的一价取代基。代表性的单糖包括戊糖和己糖。优选的糖基是单糖取代式I化合物上一个或多个OH，而形成的‐O‐糖基。如本文所用，术语“包含”、“包括”、“含有”可互换使用，不仅包括封闭式定义，还包括半封闭、和开放式的定义。换言之，所述术语包括了“由……构成”、“基本上由……构成”。
如本文所用，术语“氘(D)代”指化合物或基团中的一个或多个氢被氘所取代。氘代可以是一取代、二取代、多取代或全取代。在另一优选例中，氘在氘取代位置的氘同位素含 量是大于天然氘同位素含量(0.015％)，更佳地大于50％，更佳地大于75％，更佳地大于95％，更佳地大于97％，更佳地大于99％，更佳地大于99.5％。在另一优选例中，所述化合物中,D在氢原子所在位置的同位素含量≥95％，更佳地≥99％。
如本文所用，术语“老花眼”又称为“老视”指的是眼睛的晶状体失去其柔性的视觉状态，从而难以聚焦于近距离物体。典型地，所述的晶状体病变引起的眼部疾病不包括结膜炎、因病原体(如细菌、病毒等微生物)引起的眼睛感染或眼部炎症。
如本文所用，术语“预防”指在眼部疾病之前施用治疗有效量式I化合物(包括纯净物、混合物)和/或桦褐孔菌提取物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药或其氘代化合物，从而使得所述眼部疾病被预防，发生时间延迟、或仍然发生，但是比不施用式I化合物和/或桦褐孔菌提取物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药的程度减轻。如本文所用，术语“治疗”、指施用治疗有效量的式I化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药，从而减少、减轻或减缓眼部疾病、障碍或病症的进展或发展。在另一实施方式中，“治疗”指减轻、减缓眼部疾病、障碍或病症的进程，或改善眼部疾病、障碍或病症的一种或多种体征或症状。
本发明所述的“预防”和“治疗”包括延缓和终止疾病的进展，或消除晶状体病变引起的眼部疾病，并不需要100％抑制、消灭和逆转。在一些实施方案中，与不存在本发明所述式I所述化合物和/或桦褐孔菌提取物和其组合物或制剂时(例如，在未暴露于在本发明式I所述化合物和/或桦褐孔菌提取物和其组合物或制剂生物学匹配的对照受试者或标本中)观察到的水平相比，本发明所述式I所述化合物和/或桦褐孔菌提取物和其组合物或制剂将晶状体病变引起的眼部疾病(如白内障)减轻、预防、抑制和/或逆转了例如至少约1％、至少约10％、至少约30％、至少约50％、或至少约80％。
在另一优选例中，术语“白内障”是指在晶状体的表面和/或内部表现出引起浑浊(混浊)或不透明，或诱发晶状体肿胀的疾病或病症，包括先天性白内障以及获得性白内障。代表性的，所述白内障是包括(但不限于)：年龄相关性白内障、糖尿病性白内障、与手术相关的白内障、放射线照射引起的白内障、遗传性疾病引起的白内障、感染引起的白内障，或药物引起的白内障。
活性成分
本发明首次发现了式I化合物在晶状体病变相关眼病治疗方面的用途，所述式I化合物用于制备药物组合物或制剂，所述药物组合物或制剂用于预防和/或治疗晶状体病变引起的眼部疾病。
如本文所用，术语“本发明化合物”、“式I化合物”、可互换使用，指式I化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药。此外，该 术语还包括相应的氘代化合物。应理解，该术语还包括上述组分的混合物，在式I化合物中，如果存在手性碳原子，则手性碳原子可以为R构型，也可以为S构型，或二者的混合物。
式I化合物的结构如上述本发明的第一方面中所述。
在另一优选例中，在所述结构式中，所述的虚线
Figure PCTCN2018125075-appb-000034
所表示的双键的总数为0、1、2、3或4个。
在另一优选例中，所述的双键位于以下位点之间：a和b、b和c、c和d、e和f、c和f、f和g、g和h、h和a、b和i、i和j、a和k、和/或k和l。
在另一优选例中，所述的结构式中，各C的价态符合化学稳定结构的要求(即C为四价)。
在另一优选例中，所述的R1a、R1b、R2a、R2b、R3a和R3b各自独立地选自：氢、‐OH、‐CHO、‐COOH、‐SH、取代或未取代的C1‐C3烷基‐OCO‐、取代或未取代的C1‐C4烷基、‐A‐B、或R1a和R1b、R2a和R2b、和/或R3a和R3b构成＝O；
其中，A各自独立为取代或未取代的C1‐C4亚烷基，而B各自独立地为‐OH、‐SH、C1‐C3烷氧基、‐CHO、‐COOH、＝O。
在另一优选例，Y为具有羟基或SH取代基的基团。
在另一优选例，所述的R4为氢、甲基。
在另一优选例中，R7、R12和R15各自独立选自：无、氢、甲基。
在另一优选例中，所述的R13a和R13b各自独立地选自：氢、甲基、‐OH、‐SH、＝O、卤素、且R13a和R13b中至多一个为无。
在另一优选例中，所述R10a、R10b各自独立地选自：氢、‐OH、‐SH、‐0SO 3H、‐OPO 3H、‐COOH、‐CHO，或R10a和R10b构成羰基。
在另一优选例中，所述R5、R6、R8、R9a、R9b、R14、R16、R17a和R17b各自独立选自氢、甲基。
在另一优选例中，所述的Z不是‐CH 2C(CH 3) 3。
在另一优选例中，所述的Z是‐CH 2C(CH 3) 2‐OH。
在另一优选例中，所述的式I化合物为式I‐1、I‐2、I‐3、I‐4、I‐5、I‐6、I‐7和I‐8中任一化合物。
在另一优选例中，所述式I所示的化合物选自下组：
Figure PCTCN2018125075-appb-000035
在本发明中，术语“药学上可接受的盐”指本发明化合物与酸或碱所形成的适合用作药物的盐。药学上可接受的盐包括无机盐和有机盐。一类优选的盐是本发明化合物与酸形成的盐。适合形成盐的酸包括但并不限于：盐酸、氢溴酸、氢氟酸、硫酸、硝酸、磷酸等无机酸，甲酸、乙酸、丙酸、草酸、丙二酸、琥珀酸、富马酸、马来酸、乳酸、苹果酸、酒石酸、柠檬酸、苦味酸、甲磺酸、苯甲磺酸，苯磺酸等有机酸；以及天冬氨酸、谷氨酸等酸性氨基酸。一类优选的盐是本发明化合物与碱形成的盐。适合形成盐的酸包括但并不限于：氢氧化钠、氢氧化钾、碳酸钠、碳酸氢钠、磷酸钠等无机碱，氨水、三乙胺、二乙胺等有机碱。
本发明中，术语“前药”也称为前体药物、药物前体、前驱药物等，是指经过生物体内转化后才具有药理作用的化合物。前体药物本身没有生物活性或活性很低，经过体内代谢后变为有活性的物质，这一过程的目的在于增加药物的生物利用度，加强靶向性，降低药物的毒性和副作用。如本发明中，式A所示化合物的前药可代谢成式I化合物(代表性的如桦褐孔菌醇)。在本发明中，一种优选地的前药为本发明式I化合物与含有羟基的化合物或含有羧基的化合物经酯化反应形成的酯。代表性地，含有羟基的化合物包括C1-C6低级醇(如乙醇、丙醇等)、低级糖(如葡萄糖、乳糖等)。代表性地，含有羧基的化合物包括C1-C6低级酸。典型地，C1-C6低级酸包括有机酸或无机酸，例如，C1-C6低级酸为盐酸、硫酸、乙酸、丙酸、草酸、富马酸、马来酸、苹果酸、酒石酸等。另一类优选的前药是糖基化的化合物，即式I化合物中的一个或多个OH基团(例如式1中的R10a、R10b、Z或其他位置)被糖基所取代，而形成的O‐糖基化的产物。
本发明的式I化合物可采用现有技术中本领域技术人员熟知的方法进行制备，对各个步骤的反应参数没有特别限制。此外，本发明的化合物也可通过市场购买获得。典型的，本发明所述式I化合物从桦褐孔菌中提取、分离和纯化制备所得。
本发明中，发明人对不同的式I化合物进行药代动力学研究，结果发现，式I化合物结构中，R1a、R2a、R3a、R1b、R2b和R3b中至少一个为含O或S的基团(如‐OH或‐SH)时，能够明显延长式I化合物在体内的半衰期，尤其是可以提高在眼部的聚集浓度，延长房水内的滞留时间，提高对眼部疾病的治疗效果。
本发明所述化合物的用途
本发明上述式I化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药、或其氘代化合物用于制备药物组合物或制剂，所述药物组合物或制剂用于：(a)预防和/或治疗晶状体病变引起的眼部疾病。
在另一优选例中，所述的药物组合物或制剂还用于：(b)抑制、逆转(溶剂或解聚)晶状体蛋白聚集；和/或(c)预防和/或治疗与晶状体蛋白聚集相关的疾病。
在另一优选例中，所述的眼部疾病包括(但并不限于)：白内障、老花眼、近视眼、皮质混浊、老视核硬化、糖尿病引发的眼部并发症。
桦褐孔菌提取物及其制备方法
本发明首次发现了桦褐孔菌提取物在晶状体病变相关眼病治疗方面的用途，用于制备药物组合物或制剂，所述药物组合物或制剂用于预防和/或治疗晶状体病变引起的眼部疾病。
如本文所用，术语“提取物”或“有效部位”包括水溶性的和/或脂溶性的提取物。该术语还包括醇提物或水提物，尤其是非水溶性、或水难溶性的提取物或脂溶性的提取物。此外，该术语还包括有效部位群，即含有脂溶性有效部位和水溶性有效部位的萃取物或其混合物。
经分析，本发明有效部位所含的化学成分至少包括选自下组的物质：萜类、甾体或其组合。
可用于制备本发明的桦褐孔菌提取物的方法没有特别限制。可以用常规方法，以桦褐孔菌为原料，获得水溶性的和/或脂溶性的提取物。
在本发明的一个优选例中，有效部位的制备通过溶剂提取法、萃取法、和/或色谱法进行。
在本发明中，对于溶剂提取法，其所用的溶剂没有特别限制，代表性的例子包括(但并不限于)：水、乙醇、甲醇、丙酮、乙酸乙酯中的一种或几种溶剂的混合溶剂。提取次数可以是一次或多次。
在本发明中，对于溶剂萃取法，其所用的溶剂没有特别限制，代表性的例子包括(但并不限于)：正丁醇、乙酸乙酯、二氯甲烷、氯仿、环己烷、石油醚中的一种或几种溶剂的混合溶剂。萃取次数可以是一次或多次；
在本发明中，对于柱色谱法，其柱色谱没有特别限制，代表性的例子包括(但并不限于)：活性碳、硅胶、反相硅胶、大孔树脂、葡聚糖凝胶中的一种或几种的组合。
在一个优选例中，所述的提取物包括萜类组分、例如三萜类组分。优选地，所述的提取物包括四环三萜类组分。
在一个优选例中，所述的提取物包括对桦褐孔菌使用不同提取方法获得的两种或两种以上的提取物的重新组合后产生的混合物。
在另一优选例中个，所述提取物中含有选自下组的化合物：
Figure PCTCN2018125075-appb-000036
在另一优选例中，所述的提取物中，所述的桦褐孔菌醇的重量百分比为0.01-99.99wt％。在另一优选例中，所述的提取物中，所述的栓菌酸的重量百分比为0.01-99.99wt％。
在本发明的一个实施例中，提供了一种桦褐孔菌提取物的制备方法，包括步骤：
(1)用乙醇对桦褐孔菌干燥子实体的粗粉进行回流提取，得乙醇提取物；
(2)将所述的乙醇提取物加水混悬后用石油醚萃取，分离溶液，得所述的桦褐孔菌提取物。
在另一优选例中，所述的制备方法在步骤(2)后还包括步骤(3)将所述的桦褐孔菌提取物进行硅胶柱层析，收集洗脱液，分离得到纯化的桦褐孔菌提取物。
在另一优选例中，提供了一种从桦褐孔菌中提取桦褐孔菌醇的方法，包括步骤：
首先，将桦褐孔菌干燥子实体的粗粉用乙醇回流提取，提取液减压蒸馏除去溶剂，得乙醇提取物。再将乙醇提取物加水混悬，依次用石油醚和乙酸乙酯萃取，将提取液分别减压蒸馏除去溶剂，得石油醚提取和乙酸乙酯提取物。将石油醚提取物进行硅胶柱层析，用石油醚‐乙酸乙酯梯度洗脱(如15:1‐2:1)，每次收集洗脱液根据薄层色谱检测结果，合并相同流分，减压蒸馏除去溶剂后对单一成分进行重结晶。在石油醚‐乙酸乙酯(如约5:1)时洗脱得到桦褐孔菌醇；或将乙酸乙酯提取物进行硅胶柱层析，先用氯仿‐甲醇梯度洗脱(如100:1‐10:1)，在氯仿‐甲醇(如70:1)时洗脱得到混合物Ⅰ，再用石油醚‐乙酸乙酯梯度洗脱(如30:1‐1:1)，在石油醚‐乙酸乙酯(如8:1)时洗脱得到桦褐孔菌醇。
在另一优选例中，还提供了一种从桦褐孔菌中提取栓菌酸的方法，包括步骤：
首先，将桦褐孔菌干燥子实体的粗粉用乙醇回流提取，提取液减压蒸馏除去溶剂，得乙醇提取物。再将乙醇提取物加水混悬，依次用石油醚和乙酸乙酯萃取，将提取液分别减压蒸馏除去溶剂，得石油醚提取和乙酸乙酯提取物。将乙酸乙酯提取物进行硅胶柱层析，先用氯仿‐甲醇梯度洗脱(如100:1‐10:1),在氯仿‐甲醇(如50:1)时洗脱得到混合物Ⅱ。再将混合物Ⅱ进行硅胶柱层析，用石油醚‐乙酸乙酯梯度洗脱(如20:1‐1:2)，在石油醚‐乙酸乙酯(如3:1)时洗脱得到混合物Ⅲ。将混合物Ⅲ进行硅胶柱层析，用石油醚‐乙酸乙酯梯度洗脱(如5:1‐1:2)，在石油醚‐乙酸乙酯(如2:1)时洗脱得到栓菌酸。
桦褐孔菌提取物用途
本发明首次提供了所述的桦褐孔菌提取物用于制备药物组合物或制剂，所述药物组合物或制剂用于预防和/或治疗晶状体病变引起的眼部疾病。
在另一优选例中，所述的药物组合物或制剂还用于：(b)抑制、逆转晶状体蛋白聚集；和/或(c)预防和/或治疗与晶状体蛋白聚集相关的疾病。
在另一优选例中，所述的眼部疾病(但并不限于)：白内障、老花眼、近视眼、皮质混浊、老视核硬化、糖尿病引发的眼部并发症。
药物组合物或制剂和施用方法
由于本发明化合物和桦褐孔菌提取物具有优异的预防和/或治疗晶状体病变引起的眼部疾病，因此含有本发明化合物的活性成分和/或桦褐孔菌提取物的药物组合物或制剂可用于：(a)预防和/或治疗晶状体病变引起的眼部疾病；(b)抑制、逆转(溶解或解聚)晶状体蛋白聚集；和/或(c)预防和/或治疗与晶状体蛋白聚集相关的疾病。所述药物组合物或制剂中包括：本发明式I化合物和/或桦褐孔菌提取物；和药学上可接受的载体。
其中，第一活性成分的含量为0.001‐99wt％，较佳的0.01‐70wt％，更佳的为0.05‐40wt％的，按照组合物的总重量计。
在另一优选例中，所述的药用组合物或制剂还可以包括：(c)第二活性成分，其中所述的第二活性成分选自下组：羊毛脂类化合物、羊毛甾醇、所有锈革孔菌目或多孔菌目中包含菌类中含有的任一化合物、唑类化合物、淀粉样蛋白调节剂、糖皮质激素类化合物、抗生素、或其组合。
在另一优选例中，所述的第二活性成分的含量为0.01-20wt％，较佳地为5-15wt％，按照组合物的总重量计。
在另一优选例中，所述的“第二活性成分”是羊毛脂类化合物时，其浓度优选为10～200mM，较佳地15～150mM，更佳地20～50mM；最佳地20～30mM。
在另一优选例中，所述的羊毛脂类化合物选自：羊毛甾醇。
在另一优选例中，所述的糖皮质激素化合物选自下组：地塞米松、氢化可的松、或其组合。
在另一优选例中，所述的抗生素选自下组：妥布霉素、硫酸庆大霉素、金霉素、氯霉素、或其组合。
在另一优选例中，所述的唑类化合物选自益康唑、异康唑、联苯苄唑、克霉唑、阿立哌唑、酮康唑、氟康唑、苯基咪唑、咪康唑、环菌唑、三唑醇、戊唑醇、丙环唑、或其组合。
如本文所用，术语“淀粉样蛋白调节剂”，又称类淀粉蛋白调节剂、淀粉样蛋白抑制剂、类淀粉蛋白抑制剂，具体是指可以调控或抑制蛋白堆积、粘附、沉淀，影响蛋白形成 原纤维状蛋白聚集体的一类分子或化合物。
如本文所用，术语“有效治疗量”，是指对人和/或动物产生功能或活性的且可被人和/或动物所接受的量。本领域的普通技术人员应该理解，所述的“有效量”或“有效剂量”可随着药物组合物的形式、给药途径、所用药物的辅料、疾病的严重程度以及与其他药物联合用药等情况的不同而有所不同。
术语“药学上可接受的载体”指的是：一种或多种相容性固体、半固体、液体或凝胶填料，它们适合于人体或动物使用，而且必须有足够的纯度和足够低的毒性。“相容性”是指药物组合物中的各组分和药物的活性成分以及它们之间相互掺和，而不明显降低药效。
应理解，在本发明中，所述的载体没有特别的限制，可选用本领域常用材料，或用常规方法制得，或从市场购买得到。
药学可接受的载体部分例子有纤维素及其衍生物(如甲基纤维素、乙基纤维素、羟丙甲基纤维素、羧甲基纤维素钠等)、明胶、滑石粉、固体润滑剂(如硬脂酸、硬脂酸镁)、硫酸钙、植物油(如豆油、芝麻油、花生油、橄榄油等)、多元醇(如丙二醇、甘油、甘露醇、山梨醇等)、乳化剂(如吐温)、润湿剂(如十二烷基硫酸钠)、缓冲剂、螯合剂、增稠剂、pH调节剂、透皮促进剂、着色剂、调味剂、稳定剂、抗氧化剂、防腐剂、抑菌剂、无热原水、脂质体等。
除了活性药物成分外，液体剂型可包含本领域中常规采用的惰性稀释剂，如水或其它溶剂，增溶剂和乳化剂，例如，乙醇、异丙醇、碳酸乙酯、乙酸乙酯、丙二醇、1,3‐丁二醇、二甲基甲酰胺以及油，特别是棉籽油、花生油、玉米胚油、橄榄油、蓖麻油和芝麻油或这些物质的混合物等。
除了这些惰性稀释剂外，组合物也可包含助剂，如润湿剂、乳化剂和悬浮剂等
在本发明中，药物组合物的剂型包括(但不限于)口服制剂、注射剂、外用制剂。代表性的包括(但不限于)：片剂、注射剂、输液剂、膏剂、凝胶剂、溶液剂、微球、膜剂。
一种优选的的剂型为眼用制剂。代表性的，所述的眼用制剂为眼药水、乳剂、凝胶、眼用膏、缓释微球、眼内缓释植片、药用缓释药膜。
所述眼用制剂包含药学上可接受的药物载体，代表性的来自包括但不限于：溶剂或稀释剂、表面活性剂、增稠剂，渗透压调节剂、pH调节剂、抑菌剂、螯合剂。
溶剂
在眼用制剂中(如滴眼剂、眼药水)使用是溶剂或稀释剂包括水溶剂或非水溶剂。所述的水溶剂包括蒸馏水、生理盐水、注射用水等；所用的非水溶剂包括乙醇、丙二醇、甘油、植物油(如橄榄油、蓖麻油、玉米油、注射用大豆油)等。
表面活性剂
在眼用制剂中，表面活性剂选自阴离子表面活性剂、阳离子表面活性剂、非离子表面活性剂、离液型(chaotropic)表面活性剂或其组合。其中非离子表面活性剂选自：吐温、司盘、脂肪酸甘油酯、聚氧乙烯类，聚氧乙烯-聚氧丙烯共聚物，或其组合。一般表面活性 剂的用量(或含量)为0-2wt％，更佳地0.1-1wt％。
增稠剂
在眼用制剂中，增稠剂可用于提高物系粘度，使物系保持均匀的稳定的悬浮状态或乳浊状态，通过添加适量增稠剂，增加药物在眼部的滞留时间，从而增加眼部对于有效小成分的吸收。
眼用制剂中，增稠剂优选为壳聚糖、羟丙基甲基纤维素(HPMC)，甲基纤维素(MC)，和聚维酮(PVP)，明胶、羧甲基纤维素钠(CMC-Na)等。一般，增稠剂的用量(或含量)为0～6wt％，较佳地为0.1-5wt％。
渗透压调节剂
在眼部制剂添加适当的渗透压调节剂，使眼用制剂的渗透压人体眼部环境的渗透压相似，从而减少对眼部的刺激。代表性，常用的渗透压调节剂包括(但不局限于)乙酸、乙酸钠、碳酸氢钠。一般的，渗透压调节剂的用量(或含量)使眼用制剂保持在一个等渗或等张环境中。
pH调节剂
在眼部制剂添加适当的pH调节剂，使眼用制剂的pH值保持在一个适当范围，与人体眼部环境的pH相似，从而减少对眼部的刺激。代表性，常用的pH调节剂包括(但不局限于)氯化钠、氯化钾、葡萄糖。一般的，pH调节剂用量(或含量)使眼用制剂的pH值保持在5-9.
抑菌剂
在眼用制剂中，抑菌剂能够杀死或抑制霜剂中细菌的生长，阻止细菌滋生过多，危害人体健康。在本发明中，抑菌剂并没有特别的限制，可以是尼泊金醇、尼泊金酯中的一种或多种组合。代表性的，本发明中的抑菌剂选自下组：尼泊金甲酯、尼泊金乙酯、尼泊金丙酯，或其组合。
螯合剂
在眼部制剂中，适当加入一定量的螯合剂，如EDTA，可增加制剂的稳定性。通常，螯合剂的浓度范围为0～0.05wt％。本发明所述组合物或制剂施用方式没有特别限制，代表性的施用方式包括(但并不限于)：局部给药、口服、注射等方式。
一种优选的施用方式是将所述组合物或制剂进行眼部局部给药，包括(但不限于)结膜、眼球后、眼周、视网膜、脉络膜上或眼内给药等，代表性的如滴加眼药水、眼内注射、眼部黏膜注射、眼部黏膜涂膜等方式。
药物制剂应与给药方式相匹配。本发明药剂还可与其他协同治疗剂一起使用(包括之前、之中或之后使用)。使用药物组合物或制剂时，是将安全有效量的药物施用于所需对象(如人或非人哺乳动物)，所述安全有效量通常至少约10微克/千克体重，而且在大多数情况下不超过约8毫克/千克体重，较佳地该剂量是约10微克/千克体重-约1毫克/千克体重。当然，具体剂量还应考虑给药途径、病人健康状况等因素，这些都是熟练医师技能范围之内的。
体外非治疗性和/或非治疗性的改善晶状体透明度的方法
本发明提供了一种体外非治疗性的改善或维持晶状体透明度的方法，包括将晶状体与式I化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药、或其氘代化合物和/或桦褐孔菌提取物接触，从而抑制晶状体病变。式I化合物如本发明所述。
在体外非治疗性的改善晶状体透明度的方法中，晶状体病变包括混浊或氧化等，抑制晶状体病变包括改善或维持晶状体透明度。
预防和/或治疗眼部疾病的方法
本发明还提供了一种预防和/或治疗眼部疾病的方法，该方法包括步骤，给予所需对象施用本发明所述式I化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药、或其氘代化合物和/或桦褐孔菌提取物。
在另一优选例中，所述对象为人和非人哺乳动物。代表性地，所述的非人哺乳动物包括(但并不限于)：宠物(如狗、猫)、家畜(如牛、羊、马、猪)、各种动物园动物(熊猫、大象、虎)等。
在另一优选例中，所述的对象还包括除人和非人哺乳动物外的其他动物，如非哺乳动物。
本发明的主要优点包括：
(a)本发明首次发现了式I化合物和桦褐孔菌提取物能够预防和/或治疗眼部疾病，如白内障的预防和治疗，并具有显著的治疗效果。
(b)本发明的式I化合物和桦褐孔菌提取物具有优良的安全性，毒副作用很小或几乎无毒副作用。
(c)本发明式I化合物和桦褐孔菌提取物体内环境代谢慢，生物利用度高，适合口服、局部等多种方式给药。
(d)本发明式I化合物和桦褐孔菌提取物对于对眼部疾病的治疗具有良好的开发应用前景。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数按重量计算。在本发明实施例中使用的实验动物，均通过动物饲养基地所在地的伦理委员会审核、批准。
实施例1
桦褐孔菌醇眼药水配方：
表1 实施例1中桦褐孔菌醇眼药水配方
组分 含量
羟丙基-β-环糊精 4g
Tween-80 10mg
桦褐孔菌醇 0.125g
EDTA-2Na 10mg
PBS溶液 定容至10mL
制备方法
将10mg Tween-80和0.125g桦褐孔菌醇依次加入到2mL的PBS溶液中，超声至混合均匀，制得组分A。再将4g羟丙基‐β‐环糊精加入到5mL PBS溶液中，制得组分B。将组分B分四次加入组分A，超声至混合均匀，再加入10mg EDTA-2Na，加PBS溶液定容至10mL。
对比例1
羊毛甾醇眼药水配方：
表C1 对比例C1中羊毛甾醇眼药水配方
组分 含量
羟丙基-β-环糊精 4g
Tween-80 10mg
羊毛甾醇 0.125g
EDTA-2Na 10mg
PBS溶液 定容至10mL
制备方法
将10mg Tween-80和0.125g羊毛甾醇依次加入到2mL的PBS溶液中，超声至混合均匀，制得组分A。再将4g羟丙基‐β‐环糊精加入到5mL PBS溶液中，制得组分B。将组分B分四次加入组分A，超声至混合均匀，再加入10mg EDTA-2Na，加PBS溶液定容至10mL。
实施例2
栓菌酸眼药水配方：
表2 栓菌酸眼药水配方
组分 含量
羟丙基-β-环糊精 4g
Tween-80 10mg
栓菌酸 0.125g
EDTA-2Na 10mg
PBS溶液 定容至10mL
制备方法
将10mg Tween‐80和0.125g栓菌酸依次加入到2mL的PBS溶液中，超声至混合均匀，制得组分A。再将4gβ‐环糊精加入到5mL PBS溶液中，制得组分B。将组分B分四次加入组分A，超声至混合均匀，再加入10mg EDTA‐2Na，加PBS溶液定容至10mL。
对比例2
羊毛甾醇眼药水配方：
表C2 对比例C2中眼药水配方
组分 含量
羟丙基-β-环糊精 4g
Tween-80 10mg
羊毛甾醇 0.125g
EDTA-2Na 10mg
PBS溶液 定容至10mL
制备方法
将10mg Tween‐80和0.125g羊毛甾醇依次加入到2mL的PBS溶液中，超声至混合均匀，制得组分A。再将4gβ‐环糊精加入到5mL PBS溶液中，制得组分B。将组分B分四次加入组分A，超声至混合均匀，再加入10mg EDTA‐2Na，加PBS溶液定容至10mL。
实施例3核型白内障治疗效果考察
在本实施例3中，比较实施例1和对比例1的两种活性成分对晶状体病变(白内障)的治疗效果。
3.1白内障造模
大鼠白内障模型建立。选出生后10-13天的Wistar大鼠，雌雄不限，用亚硒酸钠造模，每只大鼠按照20μmol/kg体重，于背颈部皮下注射。亚硒酸钠用医用生理盐水配制，配好后的溶液用0.22μm的膜过滤除菌后常温避光保存。大鼠睁眼后即可观察到眼部有明显核型白内障。
3.2白内障治疗效果考察
将8只白内障大鼠模型，按照每只大鼠的左右眼平均分成2组，每组8个对应的眼球晶状体，分别用实施例1中配方(实施例1组)和对比例1中配方(对比例1组)浸泡处理，实验开始前，分别观察两个组大鼠眼球晶状体的形态，并拍照，其中实施例1组中白内障造模成功后选取的一个大鼠眼球晶状体的上光源和下光源的形态分别如图1和图2所示；对 比例1组中白内障造模成功后选取的一个大鼠眼球晶状体的上光源和下光源的形态分别如图3和图4所示。所有大鼠的晶状体取出后拍照评分均为5分
实施例1组和对比例1组中的大鼠左右眼晶状体分别浸泡在桦褐孔菌醇眼药水和相同浓度结构类似物羊毛甾醇眼药水中，两周后观察效果并评分，其中，实施例1组实验结束后选取的一个大鼠眼球晶状体的上光源和下光源的形态分别如图5和图6所示；对比例1组实验结束后选取的一个大鼠眼球晶状体的上光源和下光源的形态分别如图7和图8所示；评分标准是：
5分-白内障核饱满、边缘光滑、完全不透光；
4分-白内障核中心饱满、边缘出现弥散、完全不透光；
3分-白内障核体积变小但不明显、不透光区密度下降；
2分-白内障核显著缩小；
1分-白内障核消失，皮质透明，晶体恢复正常。
结果如下：
表3 实施例1和对比例1对白内障的治疗效果评分
组别 大鼠A 大鼠B 大鼠C 大鼠D 大鼠E 大鼠F 大鼠G 大鼠H
实施例1 2 2 3 2 3 3 3 2
对比例1 5 4 5 3 5 4 5 4
从表3中可看出，相对于结构类似物羊毛甾醇，以桦褐孔菌醇为代表的本发明式I化合物(在特定位置具有O或S基团的化合物)，即使以简单的给药方式，也具有优异的治疗白内障的效果，可以快速缓解、治愈和预防动物白内障。
实施例4
在本实施例4中，比较实施例2和对比例2的两种活性成分对晶状体病变(白内障)的治疗效果。
4.1白内障造模
大鼠白内障模型建立。选出生后10‐13天的Wistar大鼠，雌雄不限，用亚硒酸钠造模，每只大鼠按照20μmol/kg体重，于背颈部皮下注射。亚硒酸钠用医用生理盐水配制，配好后的溶液用0.22μm的膜过滤除菌后常温避光保存。大鼠睁眼后即可观察到眼部有明显核型白内障。
4.2白内障治疗效果考察
将8只白内障大鼠模型，按照每只大鼠的左右眼平均分成2组，每组8个对应的眼球晶状体，分别用实施例2中配方(实施例2组)和对比例2中配方(对比例2组)处理，实验开 始前，分别观察两个组大鼠眼球晶状体的形态，并拍照，其中实施例2组中白内障造模成功后选取的一个大鼠眼球晶状体的上光源和下光源的形态分别如图9和图10所示；对比例2组中白内障造模成功后选取的一个大鼠眼球晶状体的上光源和下光源的形态分别如图11和图12所示。所有大鼠的晶状体取出后拍照评分均为5分。
实施例2组和对比例2组中的每只大鼠左右眼晶状体分别浸泡在栓菌酸眼药水和相同浓度结构类似物羊毛甾醇眼药水中，第9天观察效果并评分(表4)。晶状体连续浸泡三周后，实施例2组实验结束后选取的一个大鼠眼球晶状体的上光源和下光源的形态分别如图13和图14所示；对比例2组实验结束后选取的一个大鼠眼球晶状体的上光源和下光源的形态分别如图15和图16所示；
评分标准同实施例3。
结果如下：
表4 实施例2和对比例2对白内障的治疗效果评分
组别 大鼠A 大鼠B 大鼠C 大鼠D 大鼠E 大鼠F 大鼠G 大鼠H
实施例2 3 3 3 2 4 2 3 2
对照例2 5 5 5 5 5 4 5 5
从表4和图9-16中可看出，相对于羊毛甾醇，以栓菌酸为代表的本发明式I化合物(在特定位置具有O或S基团的化合物)，即使以简单的给药方式，也具有优异的治疗白内障的效果(经过栓菌酸眼药水处理后的核性白内障晶状体，其中心位置的混浊面积明显缩小)，可以快速缓解、治愈和预防动物白内障。
实施例5药代动力学考察
实验方法及结果
取10周龄左右SD大鼠，雌雄不限，经腹腔注射戊巴比妥钠麻醉并扩瞳后，行眼部玻璃腔注射术，所有动物双眼均注射相同种类药物，每个玻璃腔内药物注射量为5微升，每种药物配方(实施例1，实施例2和对比例1)受试动物数量为4只，术后10天检测房水中药物代谢情况。
经与刚注射后药物浓度初始值比较，羊毛甾醇浓度下降了82％±14.7％，而桦褐孔菌醇浓度下降了11％±2.3％，栓菌酸浓度只下降了8％±1.5％。
上述结果提示，对于晶状体病变相关的眼部疾病，桦褐孔菌醇和栓菌酸均具有更为优异的药代动力学性能，可以长时间在房水中保持疾病治疗或预防要求的浓度。
实施例6
桦褐孔菌醇眼药膏配方及其制备方法：
表5 桦褐孔菌醇眼药膏配方
组分 含量
桦褐孔菌醇 1.25g
白凡士林 75g
液体石蜡 12.5g
制备方法
称取桦褐孔菌醇1.25g，白凡士林75g，液体石蜡12.5g，置于容器中，70℃水浴中超声处理，至所有化合物均匀分散。快速转移至洁净、无菌的眼药管中，冰浴急冷至药膏凝固，4℃保存。
实施例7
栓菌酸眼药膏配方及其制备方法：
表6 栓菌酸眼药膏配方
组分 含量
栓菌酸 1.25g
白凡士林 75g
液体石蜡 12.5g
制备方法
称取栓菌酸1.25g，白凡士林75g，液体石蜡12.5g，置于容器中，70℃水浴中超声处理，至化合物均匀分散。快速转移至洁净、无菌的眼药管中，冰浴急冷至药膏凝固，4℃保存。
对比例3
空白眼药膏配方及其制备方法：
表C3 空白眼药膏配方
组分 含量
白凡士林 75g
液体石蜡 12.5g
制备方法
称取白凡士林75g，液体石蜡12.5g，置于容器中，70℃水浴中超声处理，至所有物质均匀分散。快速转移至洁净、无菌的眼药管中，冰浴急冷至药膏凝固，4℃保存。
对比例4
羊毛甾醇眼药膏配方及其制备方法：
表C4 羊毛甾醇眼药膏配方
组分 含量
羊毛甾醇 1.25g
白凡士林 75g
液体石蜡 12.5g
制备方法
称取羊毛甾醇1.25g，白凡士林75g，液体石蜡12.5g，置于容器中，70℃水浴中超声处理，至所有物质均匀分散。快速转移至洁净、无菌的眼药管中，冰浴急冷至药膏凝固，4℃保存。
对比例5:
低剂量羊毛甾醇眼药膏配方及其制备方法：
表C5 低剂量羊毛甾醇眼药膏配方
组分 含量
羊毛甾醇 0.45g
白凡士林 75g
液体石蜡 12.5g
制备方法:
称取羊毛甾醇0.45g，白凡士林75g，液体石蜡12.5g，置于容器中，70℃水浴中超声处理，至所有化合物均匀分散。快速转移至洁净、无菌的眼药管中，冰浴急冷至药膏凝固，4℃保存。
实施例8
在本实施例8中，以灵长类动物食蟹猴为动物模型，考察实施例6中的桦褐孔菌醇眼药膏对先天型白内障的治疗效果。
先天型白内障动物的给药方式和治疗效果考察
选择自然发生的先天型白内障食蟹猴，麻醉、扩瞳后裂隙灯检查双眼并拍照，排除眼部无其它异常或炎症反应。每日早9:00、中午13:00和傍晚17:00三次双眼同时上药。给药方式为实施例6中的桦褐孔菌醇眼药膏直接涂于食蟹猴左右两只眼球角膜表面，给药时猴子面部朝上，并在给药后保定动物3-5分钟。左右双眼连续给药95天后，再次麻醉、扩瞳、裂隙灯检查并拍照。桦褐孔菌醇眼药膏给药前后食蟹猴的左、右眼的形态对比分别如图17和图18所示，从图17和图18可以看出：经桦褐孔菌醇眼药膏治疗后，用近似相同的裂隙灯检查条件拍射对比图片，受试的食蟹猴的左、右眼晶状体的白内障混浊程度明显减轻,部分给药前混浊区域在桦褐孔菌醇处理后混浊完全消失，混浊区域持续减小，整个晶体没有新的混浊区域出现，原先晶体中心极性位置先天型白内障区域透光度均出现明显的 提高，左、右眼的晶状体明显的向透明(正常)状态恢复，这一现象首次证实化合物桦褐孔菌醇能够逆转灵长类动物先天型白内障的混浊程度，并且，整个治疗过程的给药方式是无创伤性的。上述结果表明桦褐孔菌醇对灵长类动物的先天型白内障具有明显的治疗效果。
实施例9
在本实施例9中，以老年灵长类动物食蟹猴为动物模型，考察实施例6中的桦褐孔菌醇眼药膏对自发型白内障的治疗效果，同时以对比例4作为对照组。所有实验中使用的猴子都为雄性。
自发型白内障治疗效果考察
对一只17岁大小的食蟹猴(编号013321)麻醉、扩瞳后进行眼部裂隙灯检查并拍照，排除无其它眼部疾病后，确定其右眼晶状体正常无白内障，左眼为中央下瞳孔区前囊下皮质混浊，属于自发型白内障。对一只19岁大小的猴(编号990447)扩瞳后进行眼部裂隙灯检查并拍照，排除无其它眼部疾病后，确定其左、右眼均为后囊下皮质混浊，属于自发型白内障。两只猴子的血糖值均在正常范围内，无糖尿病病史和眼部外伤病史。每日早9:00、中午13:00和傍晚17:00三次上药。给药方式为实施例6中的桦褐孔菌醇眼药膏分别、直接涂于编号013321和990447的猴子左侧患病眼球角膜表面，上药时猴子面部朝上，并在上药后保定动物3-5分钟。两只猴子的左侧患眼连续给药42天后，再次按照相同方法检查它们的左眼的眼球并拍照。实验组进行的同时，取一只19岁大小患自发型白内障的猴(编号993747)作为对照组，用对比例4中的羊毛甾醇眼药膏做相同给药处理和检查、拍照。桦褐孔菌醇眼药膏给药前后的眼部对比结果见图19和图20，从图19和图20中可以看出，桦褐孔菌醇眼药膏治疗后，老龄猴子患眼的晶状体混浊程度明显降低，原先白内障混浊严重的区域透光度得到明显提升，甚至多个部位已经完全恢复透明，整个晶体白内障区域的混浊面积也出现明显减小，给药处理的晶状体白内障等级出现向透明(正常)水平的逆转。对照组羊毛甾醇的治疗效果见图21(同为给药对象的左眼)，从给药前后的对比图片可以明显观察到，羊毛甾醇没有减缓老年灵长类动物自发型白内障的病情进展，和用药前相比，经过羊毛甾醇处理后晶体内部的混浊面积增加了近50％，晶体整体的透光性进一步下降。上述结果表明，桦褐孔菌醇对老年灵长类动物自发型白内障有明显、独特的治疗效果。
实施例10
在本实施例10中，以大鼠为动物模型，考察实施例6、实施例7中的桦褐孔菌醇眼药膏和栓菌酸眼药膏对外伤型白内障的治疗效果，同时以对比例3中的空白眼药膏作为空白对照，以对比例4的羊毛甾醇眼药膏作为阳性对照组。
外伤型白内障治疗效果考察
将16只周龄为4‐6周的雌雄各半的SD大鼠平均分成四组，每组含雌雄SD大鼠各2只(每组大鼠的编号分别是A、B、C和D，其中A和B为雄鼠，C和D为雌鼠)。用戊巴比妥钠麻醉每组SD大鼠后，扩瞳并用注射针头刺穿角膜及晶状体囊，在显微镜观察下针头反复在晶体皮质内滑动，直至晶体内出现明显混浊。造模手术结束2周后重新麻醉大鼠并扩瞳、拍照、评估大鼠外伤型白内障及其程度。晶状体皮质用裂隙灯观察，打分评估参考Lens opacities classification system II(LOCS II)标准，由于是外伤引发的晶体局部白内障，首次打分时忽略混浊面积等因素只考察皮质混浊深浅程度，后期通过皮质混浊深浅程度的变化情况来评估药效。首次拍照隔天开始给药，四组SD大鼠分别给予实施例6(实验组1)、实施例7(实验组2)、对比例3(溶剂对照组3)和对比例4中的桦褐孔菌醇眼药膏、栓菌酸眼药膏、空白眼药膏和羊毛甾醇眼药膏，工作日连续给药，非工作日不给药，工作日给药频率每天一次，每个眼上药半个绿豆大小药物剂量，直接涂于大鼠角膜之上。三个月后再次麻醉大鼠并扩瞳、拍照(除溶剂对照组3外)、评估大鼠外伤型白内障及其程度，实施例6、实施例7、对比例3和对比例4中的桦褐孔菌醇眼药膏、栓菌酸眼药膏、空白眼药膏和羊毛甾醇眼药膏对外伤型白内障治疗效果考察结果如表7‐10和图22‐24所示。
表7 实施例6中桦褐孔菌醇眼药膏对大鼠外伤型白内障的治疗效果
Figure PCTCN2018125075-appb-000037
表8 实施例7中栓菌酸眼药膏对大鼠外伤型白内障的治疗效果
Figure PCTCN2018125075-appb-000038
表9 对比例3中空白眼药膏对大鼠外伤型白内障的治疗效果
Figure PCTCN2018125075-appb-000039
表10 对比例4中羊毛甾醇眼药膏对大鼠外伤型白内障的治疗效果
羊毛 A鼠左 A鼠右 B鼠左 B鼠右 C鼠左 C鼠右 D鼠左 D鼠右
甾醇 眼 眼 眼 眼 眼 眼 眼 眼
给药前 CIV CIII Ctr Ctr CII CI CV CIII
给药后 CIV CIII C0 C0 CI Ctr CV CIII
从图22中可以明显观察到，外伤型白内障患眼的皮质混浊区域在桦褐孔菌醇眼药膏给药治疗后完全恢复透明。
从图23中可以明显观察到，外伤型白内障患眼的皮质混浊区域在栓菌酸眼药膏给药处理后几乎恢复透明。
从图24和表10可以得出结论，在治疗外伤型白内障方面，羊毛甾醇对混浊程度轻微的晶体有恢复透明的效果，但对于严重的白内障没有观察到病情明显被逆转的药效。
综上，从表7‐10和图22‐24可以明显看出，相对于羊毛甾醇，桦褐孔菌醇和栓菌酸均对外伤型白内障具有更加优异的治疗效果，尤其针对白内障程度严重的患眼。
实施例11
在本实施例11中，以大鼠为动物模型，考察实施例6、实施例7中的桦褐孔菌醇眼药膏和栓菌酸眼药膏对糖尿病动物模型中代谢型白内障的治疗效果，同时以对比例3中的空白眼药膏作为空白对照，以对比例4中羊毛甾醇眼药膏作为阳性对照。由于商品吡诺克辛钠滴眼液(白内停)是治疗轻度糖尿病性白内障或并发性白内障的眼科用药，故亦作为阳性对照。
糖尿病代谢型白内障治疗效果考察
将12只周龄为8周的SD大鼠分成四组，每组含SD大鼠各3只(每组大鼠的编号分别为A、B和C)。同时增加一只8周的雄性SD大鼠，作为糖尿病模型造模成功后吡诺克辛钠滴眼液组治疗对象。所有糖尿病大鼠模型建立采用大鼠腹腔注射链脲佐菌素(STZ)方法。最后一次注射2周后采尾部静脉血，确定所有大鼠血糖值都在25.4mmol/L以上，显著高于正常大鼠血糖值(6.6‐9.7mmol/L)。模型诱导2月后用戊巴比妥钠麻醉所有13只大鼠，扩瞳并拍照评估大鼠右眼白内障及其程度。出现白内障动物的右眼晶状体皮质用裂隙灯观察，打分评估采用Lens opacities classification system II(LOCS II)标准(吡诺克辛钠滴眼液组不进行打分评估)。首次拍照隔天开始右眼给药，四组SD大鼠分别给予实施例6(实验组1)、实施例7(实验组2)、对比例3(溶剂对照组3)和对比例4(阳性对照组)中的桦褐孔菌醇眼药膏、栓菌酸眼药膏、空白眼药膏和羊毛甾醇眼药膏，工作日连续给药，非工作日不给药，工作日给药频率每天一次，上药剂量为半个绿豆大小，直接涂于大鼠右眼角膜之上，左眼不处理；另外一只吡诺克辛钠滴眼液组大鼠右眼按照商品说明书给药方式和剂量给药(非工作日不给药)，左眼不给药。所有动物模型处理2个月后再次麻醉，大鼠取血测定血糖值(均在18.6mmol/L以上)并拍照评估大鼠右眼晶状体及其皮质的白内障程度，实施例6、实施例7、对比例3和对比例4中的桦 褐孔菌醇眼药膏、栓菌酸眼药膏、空白眼药膏和羊毛甾醇眼药膏对糖尿病代谢型白内障治疗效果考察结果如表11‐14和图25‐28所示，吡诺克辛钠滴眼液组大鼠右眼治疗结果见图29。
表11 实施例6中桦褐孔菌醇眼药膏对大鼠糖尿病代谢型白内障的治疗效果
实验组1 A鼠 B鼠 C鼠
给药前 CII CII Ctr
给药后 Ctr Ctr Ctr
表12 实施例7中栓菌酸眼药膏对糖尿病代谢型白内障的治疗效果
实验组2 A鼠 B鼠 C鼠
给药前 CI Ctr Ctr
给药后 Ctr CI Ctr
表13 对比例3中空白眼药膏对大鼠糖尿病代谢型白内障的治疗效果
溶剂对照组3 A鼠 B鼠 C鼠
给药前 CI Ctr Ctr
给药后 CIII CII CII
表14 对比例4中羊毛甾醇眼药膏对大鼠糖尿病代谢型白内障的治疗效果
阳性对照组 A鼠 B鼠 C鼠
给药前 CII CI Ctr
给药后 CIII CI CIII
从图25中可以明显观察到，糖尿病代谢型白内障患眼的皮质混浊区域在桦褐孔菌醇眼药膏给药处理后混浊面积大范围减少，混浊程度也变为很浅的水平，整个晶状体几乎恢复透明状态。
从图26中可以明显观察到，糖尿病代谢型白内障患眼的皮质混浊区域在栓菌酸眼药膏给药处理后混浊面积大范围减少，混浊程度也变为极浅水平，整个晶状体几乎恢复透明状态。
从图27中可以明显观察到，大鼠晶状体内的混浊面积在羊毛甾醇给药处理后增加，原先没有白内障的区域也出现了混浊，晶体整体的透光性继续下降，病情恶化。羊毛甾醇针对糖尿病引发的代谢型白内障没有逆转恢复晶体透明的药效。
从图28中可以观察到空白眼药膏对照组的糖尿病大鼠模型白内障程度随着时间的增长而病情显著恶化，混浊范围增加，混浊程度加重。
从图29中可以观察到吡诺克辛钠滴眼液对照组的糖尿病大鼠模型白内障随着时间的增长而病情进一步恶化，晶体整体混浊范围增加，混浊程度加重，但病情进展速度好于空白眼药膏对照组。
从表11‐14和图25‐29可以明显看出，相对于羊毛甾醇和商品化的吡诺克辛钠滴眼液，桦褐孔菌醇和栓菌酸均对糖尿病代谢型白内障不仅具有明显的抑制病情发展的效果，还具有恢复白内障晶状体透明的药效(逆转效果)。
实施例12
在本实施例12中，以犬为动物模型，考察实施例6中的桦褐孔菌醇对老年型白内障的治疗效果。
老年型白内障治疗效果考察
对一只13岁大小的狗(金毛)扩瞳后进行眼部裂隙灯检查，确定受试动物无其它眼部疾病和确定其血糖水平正常后，按照Lens opacities classification system II(LOCS II)标准，确定其左、右眼为老年型白内障，皮质白内障评级打分分别是CIV级、CIII级。每天早、晚双眼同时各给予实施例6中的桦褐孔菌醇眼药膏1次，一天2次，每次2个黄豆大小给药剂量，药膏直接涂于角膜上，上药后保定动物头部5分钟以增加药物利用率，不间隔连续给药90天后，再次按照相同方法检查晶状体并评估皮质白内障程度，发现其左、右眼白内障分别降为CI级和CI级，结果证明，桦褐孔菌醇对老年型白内障有明显的治疗效果。
实施例13
桦褐孔菌提取物1的制备
桦褐孔菌提取物1：桦褐孔菌干燥子实体的粗粉1千克，按料液比1：75(g:ml)用95％乙醇回流提取，提取液减压蒸馏除去溶剂，得40克乙醇提取物。将全部乙醇提取物加水混悬，用石油醚萃取，将提取液分别减压蒸馏除去溶剂，得1.25克桦褐孔菌提取物1。
实施例14
桦褐孔菌提取物2制备
桦褐孔菌提取物2：将实施例13制备的桦褐孔菌提取物1进行硅胶柱层析，用石油醚‐乙酸乙酯梯度洗脱(15:1‐2:1)，每次收集洗脱液50mL并根据薄层色谱检测结果，合并相同流分，减压蒸馏除去溶剂后对单一成分进行重结晶，在石油醚‐乙酸乙酯5:1时洗脱得到桦褐孔菌提取物2。
桦褐孔菌提取物2的核磁共振(NMR)图、质谱(MS)图和高效液色谱(HPLC)图如图30‐32所示，经核磁、质谱和高效液相色谱分析可得：桦褐孔菌提取物2为单一桦褐孔菌醇(Inotodiol)成分，且纯度为98.36％，桦褐孔菌醇的结构式如下所示：
Figure PCTCN2018125075-appb-000040
实施例15
提取物眼用制剂的制备
在本实施例中，采用实施例13中提取物1作为活性成分，按常规方法制备眼药膏。配方如下：
桦褐孔菌提取物1眼药膏：
提取物1     1.25g
白凡士林    75g
液体石蜡    12.5g
制备方法:
称取提取物1，质量为1.25g，白凡士林75g，液体石蜡12.5g，置于容器中，70℃水浴中超声处理，至所有化合物均匀分散。快速转移至洁净、无菌的眼药管中，冰浴急冷至药膏凝固，4℃保存。
实施例16
桦褐孔菌提取物对获得型白内障(非先天型白内障)的治疗效果
在本实施例中，测试实施例13、实施例14中涉及的提取物所制备的眼用制剂的药学活性。
根据已发表的文献数据，桦褐孔菌的提取物1中存在少量羊毛甾醇，其质量分数约占16％。因为羊毛甾醇治疗白内障的效果和其给药剂量大小呈正相关的关系，所以为了排除少量羊毛甾醇对桦褐孔菌提取物治疗白内障效果的干扰，实验设计上除了论证桦褐孔菌提取物1眼药膏药效外(实施例15)，增加一组低剂量的羊毛甾醇眼药膏对照组(对比例5)，论证剂量大于2倍于桦褐孔菌提取物1其含量的羊毛甾醇能否对动物白内障有明显治疗效果。同时，实验设计羊毛甾醇(对比例4)和桦褐孔菌醇(实施例6)作为对照组和实验组。
选取金毛、泰迪品种后天自然出现眼部白内障的狗8只，排除没有其它眼部疾病后，扩瞳、裂隙灯观察动物左右眼晶状体并评分。之后随机分组，每2只狗对应一种药剂配方，分别是对比例4、实施例15、实施例6和对比例5，双眼同时用药，每天早、晚各施药一次，所有动物用药剂量相同。施药过程保证药膏完全分布于角膜。连续给药一个月后，所 有动物再用裂隙灯观察晶体并评价打分，结果如表15所示。所有受试动物给药过程均未出现眼部不适症状。
表15 不同组体内治疗获得性白内障药效评价
Figure PCTCN2018125075-appb-000041
其中评分标准：
分级 裂隙灯检查
I 小于1/5环面,小水泡、细毛刷样
II 小于1/3环面，簇样、片样、楔样
III 小于1/2环面，环状、楔样、大片样
IV 大于等于1/2环面，块样弥漫、放射样
V 核性混浊，小于等于1/2环面，块样弥漫、放射样
VI 全部混浊
VII 晶体萎缩、核下沉
从表15中可以看出，除羊毛甾醇外，桦褐孔菌提取物中含有大量对动物安全的可以 用来治疗动物白内障的成分，这些成分可以用来制备治疗、预防、中止病情发展和/或延缓病情发展的针对白内障的药物。
实施例17
在本实施例17中，以兔为动物模型，考察实施例6、实施例7、实施例15中的桦褐孔菌醇、栓菌酸和桦褐孔菌提取物1的眼部药物安全性，同时以对比例3中的空白眼药膏作为空白对照。
眼刺激性实验方法：
将16只体重范围2.3‐3.0kg的雌雄各半的兔分成四组，每组含雌雄兔各2只，采用自身对照，第一组左眼给予实施例6中的桦褐孔菌醇眼药膏，右眼给0.9％氯化钠注射液，第二组左眼给予实施例7中的栓菌酸眼药膏，右眼给0.9％氯化钠注射液，第三组左眼给予实施例15中的桦褐孔菌提取物1眼药膏，右眼给0.9％氯化钠注射液，第四组左眼给予对比例3中的空白眼药膏，右眼给0.9％氯化钠注射液，眼药膏给药体积为约黄豆大小眼药膏/眼，0.9％氯化钠注射液给药体积为0.1ml/眼。
试验前对每只兔的双眼进行荧光素钠检查，有眼刺激症状、角膜缺陷和结膜损伤的动物不能用于试验，检查合格的兔用于试验。每天第一次给药前以及最后一次给药后1h、2h、4h、24h、48h、72h对眼部进行检查，如果在72h未见任何刺激症状，试验则可结束。如果有轻度刺激性，观察期延长至最后一次给药后7d；有中度刺激性，观察期延长至最后一次给药后14d；有强度刺激性，观察期延长至最后一次给药后21d。每天给药3次，间隔至少4小时。
试验结束，用20％乌来糖溶液腹腔注射麻醉后颈动脉放血处死兔子，然后取出眼球固定于眼球固定液中，用于病理取材、脱水、包埋、染色和光镜检查。
其中，给药过程观察的眼刺激反应评分标准如表16所示：
表16眼刺激反应评分标准
Figure PCTCN2018125075-appb-000042
Figure PCTCN2018125075-appb-000043
按表16的要求，将每个观察时间点每个动物的眼角膜、虹膜和结膜的刺激反应分值相加得总积分，将一组的积分总和除以动物数，即得最后分值。按表17判定其刺激程度，并结合组织病理学检查结果作综合判定。
表17 眼刺激性评价
Figure PCTCN2018125075-appb-000044
刺激性实验结果：
连续七天给药，兔眼部检查结果如下：第一组、第二组、第三组和第四组部分兔左右眼见有少量分泌物，结膜充血呈鲜红色，虹膜可见角膜周围有轻度充血，末次给药72h观察时所有兔眼均正常，未见有异常变化，荧光素钠检查均未见角膜被染成黄绿色。各组兔左、右眼刺激性分值均在0‐3范围内，眼刺激性结果判定为无刺激性。
病理检查结果显示，给予实施例6、实施例7、实施例15和对比例3的桦褐孔菌醇眼药 膏、栓菌酸眼药膏、桦褐孔菌提取物眼药膏和空白眼药膏后兔眼结膜：各部位上皮细胞无变性、坏死，间质血管无扩张、充血，无炎细胞浸润等病变；角膜：角膜上皮细胞无变性、坏死，基质层胶原纤维无肿胀、断裂，无炎细胞浸润，角膜内皮细胞无坏死、无增生；虹膜：虹膜睫状体组织结构清晰，间质无充血，无炎细胞浸润；哈氏腺和泪腺：腺体上皮细胞正常，间质无充血、水肿和炎细胞浸润。
综上，实施例6、实施例7、实施例15和对比例3的桦褐孔菌醇眼药膏、栓菌酸眼药膏、桦褐孔菌提取物1眼药膏和空白眼药膏对兔眼结膜、角膜、虹膜、哈氏腺和泪腺等均无刺激性，表明桦褐孔菌醇和栓菌酸两种化合物制备而来的眼药安全。
实施例18
测试桦褐孔菌醇和栓菌酸两个化合物的热稳定性，同时以羊毛甾醇作为参照品。评估方法采取自身对比法，将每种化合物在4摄氏度和80摄氏度分别放置相同的时间，利用HPLC(测试浓度：0.3mg/ml，甲醇溶解)比较桦褐孔菌醇、栓菌酸和羊毛甾醇在80摄氏度放置不同实验时间后自身纯度变化百分比，结合各化合物外观属性直接观察结果，判定三种结构相似化合物的热稳定性和成药性。
羊毛甾醇、桦褐孔菌醇、栓菌酸4度存放时各自实验起始纯度分别为96％、94％、98％。羊毛甾醇、桦褐孔菌醇、栓菌酸在80℃放置1天后，含量分别变为73％、88％、97％；羊毛甾醇、桦褐孔菌醇、栓菌酸在80℃放置2天后，含量分别变为48％、68％、96％；羊毛甾醇、桦褐孔菌醇、栓菌酸在80℃放置3天后，含量分别变为44％、67％、96％。
羊毛甾醇在80℃放置1天后，粉末变硬，结成一大块，继续放置，颜色逐渐变黄，放置1周时，羊毛甾醇有三分之一变黄；桦褐孔菌醇和栓菌酸于80℃分别放置1周后，与4℃相比，外观上没有任何变化。羊毛甾醇在80℃放置2周后全部变黄，且有一部分呈现变晶，结块严重，纯度只有32％；桦褐孔菌醇在80℃放置2周与放置4℃相比，开始出现变黄；栓菌酸在80℃放置2周与放置4℃相比，外观上没有显著变化，纯度依然维持在93％以上。
上述结果证明，以桦褐孔菌醇和栓菌酸为代表的式I化合物较羊毛甾醇更加稳定，成药性更好。
实施例19
在本实施例19中，以桦褐孔菌醇和栓菌酸两个化合物为基础，进行了一系列的化学修饰和衍生物合成，并对其中获得的一些具体化合物进行了结构鉴定和表征。
以桦褐孔菌醇为基础进行的一系列的化学修饰和衍生物合成方法如下所示：
Figure PCTCN2018125075-appb-000045
Figure PCTCN2018125075-appb-000046
其中以桦褐孔菌醇为基础进行的一系列的化学修饰和衍生物合成的部分具体过程如下所示：
化合物1
化合物1为桦褐孔菌醇。
化合物2和化合物3的合成
化合物1桦褐孔菌醇(200mg)溶于二氯甲烷(4mL)，加入醋酸酐(180mg)，吡啶(1mL),室 温搅拌反应6h，TLC显示原料消失，浓缩除去溶剂，得化合物2(240mg)。
化合物2(200mg)溶于二氯甲烷(4mL)，冰浴下m‐CPBA(间氯过氧苯甲酸)(85％，85mg)和碳酸氢钠(42mg)间隔2h分三批加入至上述溶液中，碳酸氢钠水溶液洗涤一次，浓缩后柱层析得化合物3(154mg)。
化合物4的合成
高碘酸钠(87mg)加入到10mL乙醚中，搅拌下加入化合物3(200mg)，室温反应30min。水洗，分层，有机相浓缩后柱层析得化合物4(140mg)。
化合物5的合成
化合物4(200mg)溶于10mL乙醇中，加入盐酸羟胺(31mg)，碳酸钾(165mg)，50℃搅拌反应6h，降温后过滤浓缩，柱层析得化合物5(147mg)。
1H NMR(CD 3Cl，400MHz):δ(ppm)3.70(1H，m)，3.24(1H，dd，J＝11.6Hz，J＝4.4Hz)，0.73，0.82，0.88，0.99，1.01(各3H，s)，0.97(3H，d，J＝6.8)； 13C NMR(CDCl 3，100MHz):δ(ppm)153.2，134.5，134.2，79.0，73.4，50.4，49.4，47.2，44.8，41.6，38.9，37.0，35.6，30.9，29.1，28.0，27.8，27.2，26.5，26.0，24.3，21.0，19.2，18.2，18.0，15.7，15.4，12.6。
化合物6的合成
红铝溶液(70％甲苯溶液，0.1mL)加入四氢呋喃(10mL)中，加入化合物3(200mg)，控温‐10℃反应5h。倒入稀盐酸中，乙酸乙酯萃取，硫酸钠干燥，浓缩柱层析后得到化合物6(120mg)。
化合物7的合成
化合物6(200mg)溶于10mL四氢呋喃中，加入钠氢(16mg)，碘甲烷(58mg)室温搅拌反应12h。反应完后浓缩除去溶剂，加入二氯甲烷溶解，水洗，浓缩后柱层析得化合物7(85mg)。
化合物9的合成
化合物7(300mg)溶于甲醇(6mL)中，加入碳酸钾(150mg)，加热至50℃反应6h，过滤后浓缩，再加入二氯甲烷(6mL)，吡啶(1mL)，TBDMSCl(80mg)，室温反应至底物消失，再加入碘甲烷(80mg)，室温反应12h，水洗，除去溶剂，加入四氢呋喃(6mL)，四丁基氟化铵(280mg)，室温反应5h，浓缩除去溶剂，二氯甲烷溶解，水洗浓缩后柱层析得化合物9(90mg)。
1H NMR(CD 3Cl，400MHz):δ(ppm)3.37(3H，s)，3.16(3H，s)0.68(3H，s)； 13C NMR(CDCl 3，100MHz):δ(ppm)134.7，134.5，88.8，73.4，73.3，57.7，51.4，50.7，49.4，47.2，44.8，41.6，39.0，37.2，37.0，36.6，35.7，30.9，29.5，28.0，27.8，26.8，26.5，24.4，22.8， 21.2，19.2，18.2，18.0，16.3，12.6。
化合物10的合成
化合物2(200mg)溶于乙醇(4mL)，加入氢氧化钠(15mg)，室温反应6h，浓缩除去溶剂，柱层析后溶于二氯甲烷(4mL)，冰浴下m‐CPBA(间氯过氧苯甲酸)(85％，50mg)和碳酸氢钠(20mg)间隔2h分三批加入至上述溶液中，碳酸氢钠水溶液洗涤一次，浓缩后柱层析得化合物10(100mg)。
1H NMR(CD 3Cl，400MHz):δ(ppm)4.50(1H，m)，3.74(1H，m)3.24(1H，dd)2.68(1H，t，J＝6.4Hz)，0.69(3H，s)； 13C NMR(CDCl 3，100MHz):δ(ppm)171.2，134.5，134.2，81.0，72.2，63.5，58.4，52.0，50.4，49.9，44.8，41.6，37.9，37.0，35.5，30.9，29.1，26.0，27.8，25.8，25.1，25.0，24.3，23.0，21.0，19.2，18.9，18.8，18.7，18.4，12.4。
化合物12的合成
化合物3(200mg)溶于异丙醇(5mL)中，加入水(2mL)和次磷酸(0.5mL，50％水溶液)，回流反应3h，冷却后加入20mL水，过滤烘干得白色固体化合物12。
化合物13的合成
化合物1(200mg)溶于四氢呋喃水混合物(1:1，10mL)中，加入NBS(80mg)，室温反应2h，浓缩除去四氢呋喃，加入二氯甲烷萃取水相，浓缩除去溶剂后柱层析得化合物13(190mg)。
1H NMR(CD3Cl，400MHz):δ(ppm)4.0‐3.9(m，1H)，3.2(1H，m)，0.69(3H，s)；13C NMR(CDCl3，100MHz):δ(ppm)134.5，134.2，79，74，70.7，63.8，50.4，49.4，47.2，44.8，41.8，38.9，37，35.6，35.2，30.9，29.1，29.1，27.8，27.8，26.5，26，26，24.3，21，19.2，15.4，15.4，12.7，12.6。
化合物15的合成
化合物6(200mg)溶于甲醇(4mL)，加入碳酸钾(100mg)，加热至50℃反应6h。过滤后浓缩柱层析得化合物15(145mg)。
1H NMR(CD 3Cl，400MHz):δ(ppm)3.7(1H，m)，3.25(1H，m)0.69(3H，s)； 13C NMR(CDCl 3，100MHz):δ(ppm)134.5，134.2，79.0，73.4，70.3，50.4，49.4，47.2，44.8，41.6，40.0，38.9，37.0，35.6，30.9，29.5，29.1，28.0，27.8，26.8，26.5，26.0，24.3，21.0，19.2，18.2，18.0，15.7，15.4，12.6。
化合物17的合成
化合物12(200mg)溶于乙醇中(10mL)，加入碳酸钾(100mg)，室温反应16h，过滤后浓缩柱层析得化合物17(130mg)。
1H NMR(CD 3Cl，400MHz):δ(ppm)4.5‐4.4(m，1H)，3.35(1H，m)，0.68(3H，s)； 13C NMR(CDCl 3，100MHz):δ(ppm)134.5，134.2，79，75.4，74.2，68.3，50.4，49.4，47.2，44.8，42，38.9，37，35.6，34.3，30.9，29.1，29.1，27.8，27.8，26.5，25.7，25.7，24.3，21，19.2，15.4，15.4，12.7，12.6。
化合物19的合成
化合物15(200mg)溶于二氯甲烷(6mL)中，加入吡啶(1mL)，TBDMSCl(65mg)，室温反应至底物消失，水洗后加入戴斯‐马丁氧化剂(170mg)，室温反应6h，碳酸氢钠水溶液洗涤，浓缩除去溶剂，加入四氢呋喃(6mL)，四丁基氟化铵(170mg)，室温反应5h，浓缩后加入二氯甲烷，水洗后浓缩柱层析得化合物19(93mg)。
化合物20的合成
化合物12(200mg)溶于二氯甲烷(5mL)中，加入戴斯‐马丁氧化剂(170mg)，室温反应6h，碳酸氢钠水溶液洗涤，浓缩除去溶剂，加入乙醇10mL，碳酸钾(100mg)，室温反应16h，过滤后浓缩柱层析得化合物20(102mg)。 1H NMR(CD 3Cl，400MHz):δ(ppm)3.86(1H，m)，3.23(1H，d)，2.62‐2.51(2H，m)，0.69(3H，s)。
化合物21的合成
化合物15(400mg)溶于二氯甲烷(20mL)，冰浴下m‐CPBA(间氯过氧苯甲酸)(85％，180mg)和碳酸氢钠(60mg)间隔2h分三批加入至上述溶液中后室温反应过夜，碳酸氢钠水溶液洗涤一次，浓缩除去溶剂加入四氢呋喃(10mL)，加入100uL 40％的氢氟酸溶液，室温反应4天，加入碳酸氢钠水溶液，二氯甲烷萃取，有机相浓缩后柱层析得化合物21(260mg)。
1H NMR(CD 3Cl，400MHz):δ(ppm)5.49(1H，m)，5.33(1H，m)，3.24(1H，m)，0.58(3H，s)； 13C NMR(CDCl 3，100MHz):δ(ppm)146，142.8，120.3，116.5，79.1，73.4，70.3，51.2，50.5，49.2，44.5，41.6，41.6，38.8，38，37.5，36.8，31.6，29.5，29.1，27.9，26，25.7，25.7，23.1，23.1，22.9，21.3，15.9，12.6。
化合物22的合成
化合物19(200mg)溶于二氯甲烷(6mL)，加入三乙胺(1mL)，TBSOTf(340mg)室温反应2h，水洗后浓缩除去溶剂柱层析得化合物22(302mg)。
化合物23和化合物24的合成
化合物22(200mg)溶于DMF(6mL)，加入1‐氯甲基‐4‐氟‐1,4‐重氮化二环2.2.2辛烷双(四氟硼酸)盐(selectfluor，88mg)，室温反应1h，加入二氯甲烷，碳酸氢钠水溶液洗涤后减压浓缩除去溶剂，加入乙醇(6mL)，硼氢化钠(10mg)，室温反应1h，加入稀盐酸至上述体系，浓缩除去溶剂，乙酸乙酯萃取浓缩后柱层析得白色固体，加入四氢呋喃(6mL)，四丁基氟化铵(130mg)，室温反应5h，浓缩后加入二氯甲烷，水洗浓缩后通过制备液相分离化合物23(43)和化合物24(8mg)。
化合物23： 1H NMR(CD 3Cl，400MHz):δ(ppm)5.01‐4.86(1H，m)，3.68(1H，d)，0.67(3H，s)； 13C NMR(CDCl 3，100MHz):δ(ppm)134.7，133.9，91.4，76.7，73.0，71.1，50.5，49.9，47.2，46.3，42.5，41.6，35.8，35.6，31.0，30.9，29.7，29.5，29.4，28.4，28.2，26.2，25.4，24.3，21.3，21.2，20.3，18.8，15.4，12.6。
化合物24： 1H NMR(CD 3Cl，400MHz):δ(ppm)4.64‐4.50(1H，m)，3.28(1H，dd)，0.68(3H，s)； 13C NMR(CDCl 3，100MHz):δ(ppm)134.9，133.5，93.4，81.1，73.0，71.1，50.4，49.8，47.2，46.3，42.5，41.6，36.7，36.5，30.8，30.7，29.7，29.3，29.2，28.4，28.2，26.2，25.4，24.3，21.3，21.1，18.7，18.0，15.4，12.6。
化合物25的合成
化合物22(400mg)溶于二氯甲烷(10mL)，冰浴下m‐CPBA(间氯过氧苯甲酸)(85％，100mg)和碳酸氢钠(40mg)间隔2h分三批加入至上述溶液中，碳酸氢钠水溶液洗涤一次，浓缩后加入四氢呋喃(10mL)，四丁基氟化铵(400mg)，室温反应5h，浓缩后加入二氯甲烷，水洗浓缩柱层析得化合物25(144mg)。
1H NMR(CD 3Cl，400MHz):δ(ppm)3.65(1H，m)，2.93(1H，d)，0.64(3H，s)； 13C NMR(CDCl 3，100MHz):δ(ppm)135.48，135.14，83.97，73.4，70.3，69.6，51.49，50.61，47.2，45.38，44.98，41.6，41.6，40.02，38.91，31.88，31.57，29.5，29.1，28.95，28.91，27.18，26，24.55，21.97，21.84，19.12，19.09，16.18，12.6。
化合物27的合成
化合物4(200mg)溶于DMF(5mL)中，加入膦酰基丙酸三甲酯(78mg)，碳酸钾(60mg)，室温反应6h，倒入水中，二氯甲烷萃取，浓缩除去溶剂后，加入四氢呋喃(5mL)，氢氧化锂水溶液(10％，0.3mL)室温反应6h。浓缩除去四氢呋喃，加入水(5mL)，稀盐酸调pH至2.5，过滤，烘干得化合物27(132mg)。
1H NMR(CD 3Cl，400MHz):δ(ppm)6.8(1H，m)，3.77(1H，m)3.16(1H，m)0.66(3H，s)； 13C NMR(CDCl 3，100MHz):δ(ppm)170.5，140.1，134.5，134.2，128.8，78.6，72.3，50.4，49.8，46.6，44.3，41.1，38.7，36.9，35.6，34.8，31，30.7，27.7，27.5，27.3，26.4，24，20.9，18.9，18.2，15.5，15.2，12.2，11.5。
化合物32的合成
Wittig试剂的制备：2‐溴正丙醇(100mg)溶于二氯甲烷(5mL)，加入三苯基膦(189mg)，回流反应16h，过滤后得wittig试剂。
DMF(5mL)加入上述wittig试剂(160mg)，钠氢(16mg)，室温搅拌2h，再加入化合物4(200mg)于室温反应6h，倒入水中，过滤，滤饼四氢呋喃(5mL)溶解，氢氧化锂水溶液(10％，0.2mL)室温反应6h。浓缩除去四氢呋喃，加入水(5mL)，二氯甲烷萃取，浓缩后柱层析得化合物32(120mg)。 1H NMR(CD 3Cl，400MHz):
δ(ppm)5.85(1H，m)4.04(2H，s)3.7(1H，m)，3.25(1H，dd)0.63(3H，s)； 13C NMR(CDCl 3，100MHz):δ(ppm)137.6，134.5，134.2，121.4，79，73.4，68.7，50.4，49.4，47.2，44.8，41.6，38.9，37，35.6，33.9，30.9，30.9，29.1，28，27.8，26.5，24.3，21，19.2，18.2，15.7，15.4，12.9，11.8。
化合物34的合成
化合物4(200mg)溶于DMF(5mL)中，加入膦酰基乙酸三甲酯(78mg)，碳酸钾(60mg)，室温反应6h，倒入水中，二氯甲烷萃取，浓缩除去溶剂后，加入四氢呋喃(5mL)，氢氧化锂水溶液(10％，0.3mL)室温反应6h。浓缩除去四氢呋喃，加入水(5mL)，稀盐酸调pH至2.5，过滤，烘干得化合物34(138mg)。
1H NMR(CD 3Cl，400MHz):δ(ppm)7.02(1H，dt)，6.4‐6.7(1H，m)，3.8(1H，m)3.18(1H，m)0.67(3H，s)； 13C NMR(CDCl 3，100MHz):δ(ppm)170.5，145.1，134.5，134.2，121.8，78.6，72.3，50.4，49.8，46.6，44.3，41.1，38.7，36.9，35.6，31.8，31，30.7，27.7，27.5，27.3，26.4，24，20.9，18.9，18.2，15.5，15.2，11.5。
化合物42的合成
化合物1(600mg)溶于二氯甲烷(12mL)中，加入吡啶(1mL)，TBDMSCl(203mg)，室温反应6h，水洗后硫酸钠干燥，加入戴斯‐马丁氧化剂(170mg)，室温反应6h，碳酸氢钠水溶液洗涤，硫酸钠干燥后加入三乙胺(1mL)，TBSOTf(358mg)，室温反应2h，水洗后浓缩除去溶剂，加入DMF(20mL)，1‐氯甲基‐4‐氟‐1,4‐重氮化二环2.2.2辛烷双(四氟硼酸)盐(selectfluor，336mg)，室温反应1h，加入二氯甲烷，碳酸氢钠水溶液洗涤后减压浓缩除去溶剂，加入乙醇(20mL)，硼氢化钠(30mg)，室温反应1h，稀盐酸调pH至4.5，浓缩除去溶剂，乙酸乙酯萃取，硫酸钠干燥，浓缩后加入二氯甲烷(20mL)，三乙胺(1mL)，TBSOTf(358mg)，室温反应2h，水洗后浓缩除去溶剂并柱层析得化合物39(116mg)。化合物39(100mg)溶于二氯甲烷(4mL)，冰浴下m‐CPBA(间氯过氧苯甲酸)(85％，30mg)和碳酸氢钠(20mg)间隔2h分三批加入至上述溶液中，碳酸氢钠水溶液洗涤一次，浓缩除去溶剂后加到高碘酸钠(31mg)的乙醚(5mL)溶液中，室温反应30min。水洗，分层，有机相浓缩后加入DMF(5mL)，膦酰基乙酸 三甲酯(26mg)，碳酸钾(20mg)，加热至50℃反应2h。倒入水中，乙酸乙酯萃取，浓缩后加入四氢呋喃(6mL)，氢氧化锂水溶液(10％，0.1mL)室温反应16h，加入水后，调pH至2.5，浓缩除去四氢呋喃，乙酸乙酯萃取，浓缩后制备液相分离得化合物42(35mg)。
1H NMR(CD 3Cl，400MHz):δ(ppm)7.02(1H，dt)，6.4‐6.7(1H，m)，5.01‐4.86(1H，m)，3.8(1H，m)，3.7(1H，d)，0.67(3H，s)； 13C NMR(CDCl 3，100MHz):δ(ppm)170.5，145.1，134.5，134.2，121.8，91.4，76.7，72.3，50.4，49.8，46.6，44.3，41.1，38.7，36.9，35.6，31.8，31，30.7，27.7，27.5，26.4，24，20.9，18.9，18.2，15.5，15.2，11.5。
以栓菌酸为基础进行的一系列的化学修饰和衍生物合成方法如下所示：
Figure PCTCN2018125075-appb-000047
其中以桦栓菌酸为基础进行的一系列的化学修饰和衍生物合成的部分具体过程如下所示：
化合物1'
化合物1'为栓菌酸。
化合物3'的合成
化合物1'栓菌酸(200mg，提取于桦褐孔菌)溶于甲醇(4mL)，室温下加入氯化亚砜(52mg)，室温搅拌反应1h，浓缩除去溶剂，加入二氯甲烷(4mL)，醋酸酐(45mg)，室温反应6小时后，冰浴下m‐CPBA(间氯过氧苯甲酸)(85％，75mg)和碳酸氢钠(80mg)间隔2小时分三批加入至上述溶液中，碳酸氢钠水溶液洗涤一次，浓缩后柱层析得化合物3'(162mg)。
化合物6'的合成
红铝溶液(70％甲苯溶液，0.1mL)加入四氢呋喃(10mL)中，再加入化合物3'(200mg)，控温‐20℃反应5小时。倒入稀盐酸中，乙酸乙酯萃取，硫酸钠干燥，浓缩柱层析后得到化合物6'(106mg)。
化合物15'的合成
化合物6'(200mg)溶于四氢呋喃中(4mL)，加入氢氧化锂水溶液(10％，0.2mL)，室温反应6小时。浓缩除四氢呋喃后调pH至2.5，乙酸乙酯萃取，浓缩后柱层析得化合物15'(140mg)。
1H NMR(DMSO，400MHz):δ(ppm)3.04(1H，m)0.68(3H，s)； 13C NMR(DMSO，100MHz):δ(ppm)177.5，134.8，133.9，77.2，71.4，50.5，49.5，47.9，47，44.7，44.3，39，37，35.7，32.8，30.7，29.5，29.4，28.8，28.6，28，27.8，26.5，24.5，21.1，20.6，19.4，18.4，16.3，16.1。
化合物12'的合成
化合物3'(200mg)溶于异丙醇(5mL)中，加入水(2mL)和次磷酸(0.5mL，50％水溶液)，回流反应3小时，冷却后加入20mL水，过滤烘干得白色固体化合物12'(180mg)。
化合物20'的合成
化合物12'(200mg)溶于二氯甲烷(5mL)中，加入戴斯马丁氧化剂(200mg)，室温搅拌反应6小时。碳酸氢钠洗涤并浓缩后加入乙醇10mL，碳酸钾(100mg)，室温反应16小时，加入水，调pH 2.5后乙酸乙酯萃取，浓缩柱层析得化合物20'(102mg)。
1H NMR(DMSO，400MHz):δ(ppm)3.1(1H，dd)，2.17(1H，m)0.72(3H，s)； 13C NMR(DMSO，100MHz):δ(ppm)215.4，177.3，134.8，133.9，79，77.2，50.5，49.5，47.9，47.1，44.3，39，37，35.7，32.8，31，30.7，28.8，28.7，28.6，26.7，26.67，26.5，24.8，21.3，21.1，19.4，18.4，16.3，16.2。
化合物21'的合成
化合物6'(400mg)溶于二氯甲烷(20mL)，冰浴下m‐CPBA(间氯过氧苯甲酸)(85％，129mg)和碳酸氢钠(50mg)间隔2小时分三批加入至上述溶液中后室温反应过夜，碳酸氢钠水溶液洗涤一次，浓缩除去溶剂加入四氢呋喃(10mL)，加入100uL 40％的氢氟酸溶液，室温反应4天，加入碳酸氢钠水溶液，二氯甲烷萃取，有机相浓缩后加入四氢呋喃(10mL)，氢氧化锂水溶液(10％，0.4mL)，室温反应16小时，浓缩除去溶剂，加入水后调pH至2.5，乙酸乙酯萃取，硫酸钠干燥，浓缩后柱层析得化合物21'(106mg)。 1H NMR(DMSO，400MHz):δ(ppm)5.62(1H，d)，5.36(1H，d)，3.48(1H，m)，1.01(3H，s)； 13C NMR(DMSO，100MHz):δ(ppm)177.5，146，142.8，121，116.5，79.1，71.4，50.5，49.2，48.1，47，44.7，44.3，38.8，37.5，36.4，36，31.6，29.5，29.4，28.8，28.1，27.8，27.3，25.9，23.3，22.9，20.6，16.6，16.3。
实施例20
在本实施例20中，以干燥的桦褐孔菌为原料，粉碎后进行了一系列的提纯、萃取、硅胶柱层析等化学成分的分离操作，获得10个具体化合物。
表18 桦褐孔菌提取物化学结构鉴定和提取方法
Figure PCTCN2018125075-appb-000048
Figure PCTCN2018125075-appb-000049
Figure PCTCN2018125075-appb-000050
Figure PCTCN2018125075-appb-000051
Figure PCTCN2018125075-appb-000052
实施例21
在本实施例中，选择部分实施例19和实施例20中得到的具体化合物，进行胞内突变的晶状体蛋白(αB‐crystallin R120G)聚集抑制实验(实验方法如中国专利CN201580057679所述)，评估本发明式I化合物和本发明式II化合物对晶状体蛋白突变体聚集抑制活性，从而筛选有效治疗相关疾病(如白内障)的化合物。
实验方法
选择肿瘤细胞系HeLa做为突变晶状体蛋白(αB‐crystallin R120G)表达质粒转染的宿主细胞，培养基为含10％FBS的DMEM，培养条件是37摄氏度、饱和湿度、5％二氧化碳细胞培养箱。支原体检测为阴性。转染过程是先在预包被的玻片上接种合适数量的HeLa细胞，在完全贴壁的细胞生长到细胞密度约为75％时候，用脂质体转染表达载体，转染4小时后换液正常培养基，继续在细胞培养箱中培养13小时以使外源突变晶状体蛋白表达。之后换液，更换的新培养基中分别含有各种待测的本发明式I、式II化合物(所有母液用DMSO作为溶剂)，其中每种化合物的终浓度为10μM，阴性对照为1.5％DMSO溶剂，阳性对照为羊毛甾醇(10μM)。分别用含有待测化合物、1.5％DMSO溶剂和羊毛甾醇的培养基处理上述HeLa细胞3.5小时后，更换不含DMSO、待测化合物或羊毛甾醇的正常新鲜培养基继续培养6.5小时，后续利用Anti‐p62抗体和DAPI进行常规细胞免疫荧光检测，每张玻片随机选取3个位置的视野范围，计算含有聚集晶体蛋白(晶体蛋白聚集体)的细胞的比例，最终确定 每种待测化合物对突变的晶状体蛋白的细胞内聚集影响，结果如表19所示。
表19 不同结构本发明式I、式II化合物和桦褐孔菌提取物对晶状体蛋白突变体(αB‐crystallin R120G)在细胞内聚集的影响
化合物编号 抑制效果 化合物编号 抑制效果 化合物编号 抑制效果
化合物9 ++ 化合物10 + 化合物5 +
化合物17 ++ 化合物13 ++ 化合物15 +
化合物23 + 化合物21 + 化合物20 ++
化合物24 + 化合物25 + 化合物27 +
化合物32 + 化合物34 + 化合物42 +
化合物15' + 化合物20' ++ 化合物21' ++
化合物2″ ++ 化合物3″ ++ 化合物4″ +
化合物5″ ++ 化合物6″ ++ 化合物7″ ++
化合物8″ ++ 化合物17″ ++ 栓菌酸 +
桦褐孔菌醇 + 阴性对照 ++++ 羊毛甾醇 +++
注：表中抑制效果使用符号与含有聚集蛋白细胞百分比对应关系是：
“+”代表视野中有≤30％的细胞含有蛋白聚集体；
“++”代表视野中有＞30％且≤45％的细胞含有蛋白聚集体；
“+++”代表视野中有＞45％且≤60％的细胞含有蛋白聚集体；
“++++”代表视野中有＞60％的细胞含有蛋白聚集体。
从表19中可以看出除了桦褐孔菌醇、栓菌酸外，其它式I、式II所示化合物和桦褐孔菌提取物也有优于羊毛甾醇的降低胞内晶状体蛋白聚集的作用。
实施例22
低浓度的式I化合物可有效溶解晶状体蛋白聚集体
在本实施例中，研究较低浓度式I化合物对于较高浓度蛋白聚集体是否有显著的解聚作用。
实验方法如下：桦褐孔菌醇为测试化合物，测定其对野生型的αB晶状体蛋白聚集体(WTαB)的溶解作用，同时以羊毛甾醇作为阳性对照。
在本实施例中，测试组和对照组采用的药物组合物剂型为脂质体，其中，测试组的制剂配方为40mg的DPPC，10mg的胆固醇和10mg测试化合物；阳性对照组的制剂配方为40mg的DPPC，10mg的胆固醇和10mg羊毛甾醇；空白对照组的制剂配方：40mg的DPPC和10mg的胆固醇。
脂质体制剂的制法如下：分别称取40mg的DPPC，10mg的胆固醇和10mg的相应化合物， 置于梨形瓶中，空白脂质体则仅含40mg的DPPC和10mg的胆固醇。加入适量的氯仿，使得DPPC，胆固醇和相应的化合物组成的混合物充分溶解，加入适量的玻璃珠，在50℃条件下真空旋蒸。待溶液蒸干后，再继续旋蒸10min，得到较为干燥的薄膜，真空干燥箱中常温放置过夜，使有机溶剂充分挥发。次日，加入经过预热的PBS溶液10ml，在60℃水浴中，对脂质体薄膜进行旋转水化，薄膜完全脱离器皿后，再继续旋转1小时。将水化液置于冰浴中，超声处理40min。4℃条件下，12000rpm，离心10min，取上清，制得相应的化合物的脂质体溶液。
脂质体制剂中化合物浓度的分析：对于制得的脂质体溶液，用4～5倍体积的甲醇溶液进行破乳处理，得到的澄清溶液。脂质体中化合物浓度的确定采用HPLC方法测定，色谱条件为，RP C18(4.6mm x 150mm，5μm)色谱柱，柱温35℃，检测波长210nm，流速1.0ml/min，进样量25μl。在对羊毛甾醇脂质体样品及其标品进行分析时，使用甲醇作为流动相，对桦褐孔菌醇及其标品进行分析时，使用甲醇：水＝90:10，作为流动相。经过比对分析，确定脂质体制剂中的相应化合物浓度。
蛋白聚集体的诱导方式为，将含1.2mg/ml的野生型αB晶状体蛋白的溶液，在37℃孵育48小时，体系中加入0.02％的NaN 3防止染菌，从而形成αB晶状体蛋白聚集体(WTαB)。然后，向所述溶液中分别加入桦褐孔菌醇制剂或羊毛甾醇制剂(脂质体形式，两种化合物的终浓度都为125μM)，继续放置24小时。对照组使用空白脂质体进行相同处理。处理结束后，用ThT荧光检测不同化合物对αB晶状体蛋白聚集体的影响。
结果如图33所示，桦褐孔菌醇能够显著地降低ThT荧光信号，羊毛甾醇却使ThT荧光信号增加。这一结果提示，在低浓度或更接近体内水平的化合物浓度范围内，桦褐孔菌醇能够极其有效地使αB晶状体蛋白聚集体重新溶解(*:与空白对照相比，p＜0.05)，效果远远优于羊毛甾醇。这说明，相较于羊毛甾醇，本发明的式I化合物(如桦褐孔菌醇)具有更好的白内障治疗/预防药物开发前景。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。

Claims (37)

  1. 一种式I化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药的用途，其特征在于，用于制备药物组合物或制剂，所述药物组合物或制剂用于：(a)预防和/或治疗晶状体病变引起的眼部疾病；
    Figure PCTCN2018125075-appb-100001
    式中，
    q为0、1或2；
    R1a、R1b、R2a、R2b、R3a和R3b各自独立地选自：氢、取代或未取代的C1‐C20烷基、取代或未取代的C2‐C20烯基、取代或未取代的C2‐C20炔基、取代或未取代的C3‐C10环烷基、‐OH、取代或未取代的C1‐C10烷氧基、‐COOH、‐CHO、取代或未取代的C1‐C10酯基、‐SH、取代或未取代的C1‐C10烷硫基、‐A‐B，
    或者R1a和R1b、R2a和R2b、和/或R3a和R3b构成＝O；
    其中，A为无、二价连接基团；而B为H、‐OH、‐SH、C1‐C3烷氧基、C1‐C3烷硫基、‐CHO、‐COOH、C1‐C4酯基、C3‐C10环烷基、芳基、C3‐C10的5‐6元杂芳基、或苄基；
    附加条件是R1a、R2a、R3a、R1b、R2b和R3b中至少一个为含O或S的基团；
    Z选自下组：H、取代或未取代的C1‐C20烷基、取代或未取代的C2‐C20烯基、取代或未取代的C2‐C20炔基、取代或未取代的C3‐C10环烷基、取代或未取代的芳基、取代或未取代的5‐8元杂芳基、取代或未取代的‐(C1‐C6亚烷基)‐芳基、取代或未取代的‐(C1‐C6亚烷基)‐5‐8元杂芳基、‐OH、取代或未取代的C1‐C20烷氧基、‐SH、取代或未取代的C1‐C20烷硫基、取代或未取代的C1‐C10酯基、取代或未取代的C1‐C10酰基、取代或未取代的‐O‐芳基；
    R4为氢、取代或未取代的C1‐C4烷基；
    R7、R12和R15各自独立地选自：无、氢、取代或未取代的C1‐C4烷基；
    R13a和R13b各自独立地选自：无、氢、取代或未取代的C1‐C8烷基、‐OH、取代或 未取代的C1‐C8烷氧基、‐SH、取代或未取代的C1‐C8烷硫基、卤素、取代或未取代的C1‐C3酰基，或R13a和R13b构成＝O，且R13a和R13b中至多一个为无；
    R11a和R11b各自独立地选自：无、氢、取代或未取代的C1‐C6烷基、‐OH，或R11a和R11b构成＝O，且R11a和R11b中至多一个为无；
    R10a、R10b各自独立地选自：氢、‐OH、取代或未取代的C1‐C8烷氧基、‐SH、取代或未取代的C1‐C8烷硫基、‐OSO 3H、‐OCO‐取代或未取代的C1‐C7烷基、‐OPO 3H、‐COOH、‐(取代或未取代的C1‐C7亚烷基)‐COOH、‐CHO、‐(取代或未取代的C1‐C7亚烷基)‐CHO，或R10a和R10b共同构成＝O；和
    R5、R6、R8、R9a、R9b、R14、R16、R17a和R17b各自独立地选自：氢、OH、取代或未取代的C1‐C8烷基、取代或未取代的C1‐C8烷氧基、卤素；
    所述的“取代”指有一个或多个氢被选自下组的基团所取代：卤素、‐OH、‐SH、‐COOH、‐(C1‐C7亚烷基)‐COOH、＝O、‐CHO、‐(C1‐C7亚烷基)‐CHO、C1‐C7烷基‐OCO‐、C1‐C3烷基、C3‐C6环烷基、NRaRb，其中，Ra和Rb各自独立地为H、C1‐C6烷基、C1‐C6卤代烷基、C3‐C6环烷基、或苄基。
  2. 如权利要求1所述的用途，其特征在于，所述的二价连接基团具有1‐10个选自下组的连接单元：‐CRaRb‐；‐C(OH)Ra‐、‐NRa‐、‐O‐、‐CO‐；其中，Ra和Rb各自独立地为H、C1‐C6烷基、C1‐C6卤代烷基、C3‐C6环烷基、或苄基。
  3. 如权利要求1所述的用途，其特征在于，所述的R1a、R1b、R2a、R2b、R3a和R3b各自独立地选自：氢、‐OH、‐CHO、‐COOH、‐SH、取代或未取代的C1‐C3烷基‐OCO‐、取代或未取代的C1‐C4烷基、‐A‐B、或R1a和R1b、R2a和R2b、和/或R3a和R3b构成＝O；
    其中，A各自独立为取代或未取代的C1‐C4亚烷基，而B各自独立地为‐OH、‐SH、C1‐C3烷氧基、‐CHO、‐COOH、＝O。
  4. 如权利要求1所述的用途，其特征在于，所述的Z为(L1)m‐L2＝Y，m为0、1、2、3或4，各L1各自独立地为‐CH 2‐、‐CO‐、‐O‐、‐C(C1‐C3烷基)H‐、C(C1‐C3烷基)2‐、‐C(C1‐C3烷基)O‐、或‐NH‐，L2为‐CH＝、或‐C(C1‐C3烷基)＝；
    Y为＝CH 2、＝CH‐取代或未取代的C1‐C7烷基、＝C‐(取代或未取代的C1‐C7烷基) 2、＝CH‐取代或未取代的C2‐C7烯基、或＝CH‐取代或未取代的C2‐C7炔基。
  5. 如权利要求1所述的用途，其特征在于，所述R5、R6、R8、R9a、R9b、R14、R16、R17a和R17b各自独立选自氢、甲基。
  6. 如权利要求1所述的用途，其特征在于，R1a和R1b中至少一个为含O或S的基团；和/或
    R2a和R2b中至少一个为含O或S的基团；和/或
    R3a和R3b中至少一个为含O或S的基团。
  7. 如权利要求1所述的用途，其特征在于，所述的式I化合物为式I‐1化合物：
    Figure PCTCN2018125075-appb-100002
    其中，q、R1a、R1b、R2a、R2b、R3a、R3b、R4、R7、R10a、R10b、R11a、R11b、R12、R13a、R13b、R15和Z的定义如权利要求1所述；
    或者，所述的式I化合物为式I‐2化合物：
    Figure PCTCN2018125075-appb-100003
    其中，q、R1a、R1b、R2a、R2b、R3a、R3b、R4、R7、R11a、R11b、R13a、R13b、R15和Z的定义如权利要求1所述；
    或所述的式I化合物选自下组化合物：
    Figure PCTCN2018125075-appb-100004
    Figure PCTCN2018125075-appb-100005
    其中，R1a、R1b、R2a、R2b和Z的定义如权利要求1所述；
    或者，所述的式I化合物选自下组化合物：
    Figure PCTCN2018125075-appb-100006
    其中，R1a、R1b、R2a、R2b的定义如权利要求1所述；
    或者，所述的式I化合物为具有式I‐7所示的结构：
    Figure PCTCN2018125075-appb-100007
    其中，R1a、R1b、R2a、R2b的定义如上文所述；
    或者，所述的式I化合物为具有式I‐8所示的结构：
    Figure PCTCN2018125075-appb-100008
    其中，R1a、R1b、R2a、R2b的定义如上文所述。
  8. 如权利要求1所述的用途，其特征在于，Z为‐CH 2C(CH 3) 2‐OH。
  9. 如权利要求1所述的用途，其特征在于，所述式I所示的化合物选自下组(表A)：
    表A
    Figure PCTCN2018125075-appb-100009
    Figure PCTCN2018125075-appb-100010
    Figure PCTCN2018125075-appb-100011
    Figure PCTCN2018125075-appb-100012
    Figure PCTCN2018125075-appb-100013
    Figure PCTCN2018125075-appb-100014
    Figure PCTCN2018125075-appb-100015
    Figure PCTCN2018125075-appb-100016
  10. 如权利要求1所述的用途，其特征在于，所述的化合物选自下组：
    Figure PCTCN2018125075-appb-100017
  11. 如权利要求1所述的用途，其特征在于，所述式I所示的化合物选自下组：
    Figure PCTCN2018125075-appb-100018
  12. 如权利要求1所述的用途，其特征在于，所述的晶状体病变引起的眼部疾病选自下组：白内障、老花眼、近视眼、皮质混浊、老视核硬化、糖尿病引发的眼部并发症。
  13. 如权利要求12所述的用途，其特征在于，所述的白内障选自下组：外伤性白内障、代谢型白内障、老年型白内障、先天型白内障、自发型白内障、并发类白内障，或其组合。
  14. 如权利要求1所述的用途，其特征在于，所述的药物组合物或制剂还用于：(b)抑制和/或逆转(溶解或解聚)晶状体蛋白聚集；和/或(c)预防和/或治疗与晶状体蛋白聚集相关的疾病。
  15. 如权利要求14所述的用途，其特征在于，所述的晶状体蛋白包括αB晶状体蛋白。
  16. 如权利要求1所述的用途，其特征在于，所述的式I化合物还包括式I化合物的氘代化合物。
  17. 如权利要求1所述的用途，其特征在于，所述药物组合物或制剂中包括：(a)作为第一活性成分的有效治疗量的式I化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药；(b)药学上可接受的载体。
  18. 如权利要求1所述的用途，其特征在于，所述的药用组合物或制剂的剂型选自：注射剂、外用制剂、口服制剂。
  19. 如权利要求1所述的用途，其特征在于，所述的药用组合物或制剂的剂型为眼用制剂，所述的眼用制剂为眼药水、乳剂、凝胶、眼用膏、缓释微球、眼内缓释植片、药用缓释药膜。
  20. 一种体外非治疗性和非诊断性的改善或维持晶状体透明度的方法，其特征在于，将晶状体与式I化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药接触，从而改善或维持晶状体透明度，其中，所述的式I化合物如权利要求1所述。
  21. 一种预防和/或治疗晶状体病变引起的眼部疾病的方法，其特征在于，给予所需对象式I所示的化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药、或其氘代化合物，其中所述式I化合物如权利要求1所述。
  22. 一种式II化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药、或其氘代化合物，
    Figure PCTCN2018125075-appb-100019
    式II中，
    P为0、1或2；
    R18a、R18b各自独立地为氢、取代或未取代的C1-C4烷基、或-COOH；
    R19a、R19b各自独立地为氢、取代或未取代的C1-C4烷基、或卤素；
    R20为取代或未取代的C1‐C6酰基、取代或未取代的C1-C6烷基、取代或未取代的糖基-O-(C1-C6烷基)-、取代或未取代的肟基、取代或未取代的C3-C6杂环烷基、或取代或未取代的C2-C6烯基；
    R21a、R21b各自独立地为氢、羟基、巯基、或取代或未取代的C1-C4烷基；
    R22a、R22b各自独立地为氢、或取代或未取代的C1-C4烷基；
    R23a、R23b各自独立地为氢、羟基、巯基、糖基、或取代或未取代的C1-C烷氧基；
    R24a、R24b各自独立地为氢、羟基、或卤素；
    所述的“取代”指有一个或多个氢被选自下组的基团所取代：卤素、‐OH、-SH、‐COOH、＝O、‐CHO、C1‐C4烷基、C3‐C6环烷基、氨基、糖基。
  23. 如权利要求22所述的式II化合物，其特征在于，所述的式II化合物具有选自下组的一种或多种特征：
    R18a、R18b各自独立地为-COOH、或甲基；
    R19a、R19b各自独立地为氢、或卤素；
    R20为羟基丁酰基、羟丙基、肟基、甲氧基丙基、二甲基环氧乙基、HOOC-CH＝CH-、或六元单糖丙基；
    R21a、R21b各自独立地为氢、羟基、或巯基；
    R22a、R22b各自独立地甲基；
    R23a、R23b各自独立地为氢、羟基、巯基或六元单糖基；和
    R24a为氢，而R24b为氟。
  24. 如权利要求22所述的式II化合物，其特征在于，所述的式II化合物选自下组：
    表B
    Figure PCTCN2018125075-appb-100020
    Figure PCTCN2018125075-appb-100021
  25. 一种桦褐孔菌提取物的用途，其特征在于，用于制备药物组合物或制剂，所述药物组合物或制剂用于：(a)预防和/或治疗晶状体病变引起的眼部疾病。
  26. 如权利要求25所述的用途，其特征在于，所述的提取物包括脂溶性提取物。
  27. 如权利要求25所述的用途，其特征在于，所述的提取物包括萜类提取物。
  28. 如权利要求25所述的用途，其特征在于，所述的提取物中含有选自下表C中一种或多种化合物：
    表C
    Figure PCTCN2018125075-appb-100022
    Figure PCTCN2018125075-appb-100023
    Figure PCTCN2018125075-appb-100024
  29. 如权利要求25所述的用途，其特征在于，所述提取物中含有选自下组的化合物：
    Figure PCTCN2018125075-appb-100025
  30. 如权利要求25所述的用途，其特征在于，所述提取物中含有选自下组的化合物：
    Figure PCTCN2018125075-appb-100026
    Figure PCTCN2018125075-appb-100027
    或其组合。
  31. 如权利要求25所述的用途，其特征在于，所述的眼部疾病选自下组：白内障、老花眼、近视眼、皮质混浊、老视核硬化、糖尿病引发的眼部并发症。
  32. 如权利要求25所述的用途，其特征在于，所述的桦褐孔菌提取物是通过以下方法制备的：
    (1)用醇溶剂对桦褐孔菌子实体的粗粉进行回流提取，得醇提取物；
    (2)将所述的醇提取物加水混悬后用石油醚萃取，获得萃取物，即为桦褐孔菌提取物；
    和任选的步骤(3)将所述的桦褐孔菌提取物进行硅胶柱层析，收集洗脱液，分离得到纯化的桦褐孔菌提取物。
  33. 如权利要求25的用途，其特征在于，所述的药物组合物或制剂还用于：(b)抑制、逆转(溶解或解聚)晶状体蛋白聚集；和/或(c)预防和/或治疗与晶状体蛋白聚集相关的疾病。
  34. 如权利要求33所述的用途，其特征在于，所述的晶状体蛋白包括αB晶状体蛋白。
  35. 如权利要求25所述的用途，其特征在于，所述的药物组合物或制剂的剂型为眼用制剂，所述的眼用制剂为眼药水、乳剂、凝胶、眼用膏、缓释微球、眼内缓释植片、药物缓释药膜。
  36. 一种体外非治疗性和非诊断性的改善或维持晶状体透明度的方法，其特征在于，将晶状体与桦褐孔菌提取物接触，从而改善或维持晶状体透明度。
  37. 一种药物组合物，其特征在于，包括：
    (a)作为第一活性成分的有效治疗量的式I或式II所示的化合物、或其光学异构体或其外消旋体、或其溶剂化物、或其药学上可接受的盐、或其前药、或其氘代化合物；
    (b)药学上可接受的载体；和
    (c)第二活性成分，其中所述的第二活性成分选自下组：唑类化合物、淀粉样蛋白调节剂、或其组合。
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