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WO2019001171A1 - Phosphorus-containing compound and preparation and use thereof - Google Patents

Phosphorus-containing compound and preparation and use thereof Download PDF

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Publication number
WO2019001171A1
WO2019001171A1 PCT/CN2018/087629 CN2018087629W WO2019001171A1 WO 2019001171 A1 WO2019001171 A1 WO 2019001171A1 CN 2018087629 W CN2018087629 W CN 2018087629W WO 2019001171 A1 WO2019001171 A1 WO 2019001171A1
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WO
WIPO (PCT)
Prior art keywords
group
compound
alkyl
phosphorus
amino
Prior art date
Application number
PCT/CN2018/087629
Other languages
French (fr)
Chinese (zh)
Inventor
沈旺
丁悦
姜浩
陈福利
汪江峰(
吴兴龙
李寸飞
杨立国
胡彪
姜启阳
安治星
党奎峰
Original Assignee
维眸生物科技上海有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from CN201810291023.0A external-priority patent/CN109134533B/en
Application filed by 维眸生物科技上海有限公司 filed Critical 维眸生物科技上海有限公司
Priority to JP2019539245A priority Critical patent/JP6949967B2/en
Priority to EP18823397.7A priority patent/EP3647315A4/en
Priority to CA3092885A priority patent/CA3092885C/en
Priority to AU2018291666A priority patent/AU2018291666B2/en
Priority to US16/606,774 priority patent/US11325929B2/en
Priority to KR1020197028357A priority patent/KR102208639B1/en
Publication of WO2019001171A1 publication Critical patent/WO2019001171A1/en

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/66—Phosphorus compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/66—Phosphorus compounds
    • A61K31/662—Phosphorus acids or esters thereof having P—C bonds, e.g. foscarnet, trichlorfon
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/28—Phosphorus compounds with one or more P—C bonds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/28—Phosphorus compounds with one or more P—C bonds
    • C07F9/30—Phosphinic acids [R2P(=O)(OH)]; Thiophosphinic acids ; [R2P(=X1)(X2H) (X1, X2 are each independently O, S or Se)]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/28—Phosphorus compounds with one or more P—C bonds
    • C07F9/30—Phosphinic acids [R2P(=O)(OH)]; Thiophosphinic acids ; [R2P(=X1)(X2H) (X1, X2 are each independently O, S or Se)]
    • C07F9/32—Esters thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/28—Phosphorus compounds with one or more P—C bonds
    • C07F9/50—Organo-phosphines

Definitions

  • the present invention relates to the field of medical chemistry, and in particular to a phosphorus-containing compound, a preparation method thereof, and an application for treating dry eye.
  • Tears provide long-lasting moisturization and lubrication to the eyes, which is the key to maintaining vision and eye comfort. Tears are composed of water, oil, mucus, antibodies, and some specific proteins that are used to fight infection. These components are secreted by specific glands located around the eye. When there is an imbalance in the tear system, people will feel dry eyes.
  • Dry eye syndrome is a common ocular surface inflammatory disease. People with dry eye may experience eye pain, photosensitivity, itching, redness and blurred vision. Dry eye syndrome has a variety of predisposing factors, including age, gender, environment, drugs, surgery, and systemic diseases such as autoimmune diseases, diabetes, thyroid disease, lymphoma and the like. Dry eye disease without proper diagnosis and treatment may lead to further complications such as infection, keratinization of the ocular surface, corneal ulceration and conjunctival squamous.
  • dry eye syndrome is a very serious disease that afflicts 5-10% of the population, especially those who work long hours before and after the middle age. More than 30% of today's ophthalmologist clinics are dry eye patients. Despite the large number of patients with dry eye syndrome, there are no drugs approved for the treatment of dry eye syndrome. The patient can only have temporary relief from artificial tears. Therefore, there is an urgent need to treat drugs for dry eye syndrome.
  • Dry eye syndrome is a continuous pathological process in which the condition progresses from light to heavy, and there is no obvious boundary between light, medium and severe.
  • the invention is a novel immune cell migration inhibitor. It has good hydrophilicity and can be developed into eye drops. It has a strong inhibitory effect on immune cell migration and can alleviate the symptoms of most dry eye patients.
  • R 1 is selected from the group consisting of alkyl, aryl, benzyl and derivatives thereof;
  • the alkyl group in R 1 is selected from the group consisting of a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, a pentyl group, a cyclopentyl group, a hexyl group, a cyclohexyl group and the like having 1 to 12 carbon atoms. , branched or cyclic alkyl;
  • the aryl group is selected from, for example, a phenyl group, a naphthyl group, a quinolyl group, or an alkyl group/phenyl group/halogen/nitro/amino/sulfonyl group having any one or more carbons of 1 to 3 carbon atoms.
  • the benzyl group is selected from, for example, an alkyl group having at least one or several carbons having a carbon number of 1-3, a phenyl group/halogen/nitro group/amino group/sulfonyl group and an alkyl group having 1 to 3 carbon atoms.
  • R 2 is selected from the group consisting of a hydroxyl group, an alkyl group, a hydrogen, an alkoxy group, an amine group, and an alkylamine group;
  • the alkyl group in R 2 is selected from, for example, methyl group, ethyl group, propyl group, isopropyl group, cyclopropyl group, butyl group, isobutyl group, pentyl group, cyclopentyl group, hexyl group, cyclohexyl group and the like, and the number of carbon atoms is 1- a branched, branched or cyclic alkyl group of 12;
  • the alkoxy group is selected from the group consisting of a branched, branched or cyclic alkoxy group having a carbon number of from 1 to 12 such as a methoxy group or an ethoxy group;
  • n is selected from 0 or 1;
  • X is selected from the group consisting of carbon, oxygen, and nitrogen;
  • X when it is carbon, it may be (-CH 2 -), or (-C(R1R2)-, wherein R1, R2 may be the same or different any substituent group, such as: hydrogen An alkyl group such as a methyl group or an ethyl group; an aromatic group such as a phenyl group or a benzyl group; a hydroxyl group, an alkoxy group, a halogen, etc.;
  • X when it is nitrogen, it may be (-NH-), or (-N(R N )-, wherein R N may be any substituent such as methyl, ethyl or the like.
  • An aromatic group such as a phenyl group or a benzyl group;
  • Z is selected from the group consisting of carbon, oxygen, sulfur, and nitrogen;
  • Z when it is nitrogen, it may be (-NH-), or (-N(R N )-, wherein R N may be any substituent such as methyl, ethyl or the like.
  • An aromatic group such as a phenyl group or a benzyl group;
  • R 3 is any optionally substituted hydrogen, alkyl, alkoxy, halogen, amino, cyano, hydroxy, nitro, aryl;
  • Y is selected from the group consisting of carbon, oxygen, and nitrogen;
  • Y when it is carbon, it may be (-CH 2 -), or (-C(R1R2)-, wherein R1, R2 may be the same or different any substituent group, such as: hydrogen , an alkyl group such as a methyl group or an ethyl group, an aromatic group such as a phenyl group or a benzyl group, a halogen or the like;
  • Y when it is nitrogen, it may be (-NH-), or (-N(R N )-, wherein R N may be any substituent such as methyl, ethyl or the like. a group, an aromatic group such as a phenyl group or a benzyl group, etc.);
  • R 4 is selected from the group consisting of alkyl, alkoxy, aryl, benzyl and derivatives thereof;
  • R 5 is selected from the group consisting of hydrogen, alkyl, aryl, benzyl and derivatives thereof;
  • the substituent groups represented by G1 and G2 are disposed on the benzene ring in a meta, para or ortho position.
  • the present invention provides a phosphorus-containing compound which is further characterized in that the above aryl group is selected from the group consisting of a phenyl group and a derivative thereof, a naphthyl group and a derivative thereof, an N or O heterocyclic phenyl group, and a derivative, an N or O heterocyclonaphthyl group and a derivative thereof;
  • the above derivative means a hydrogen, an alkyl group, an alkoxy group, a halogen group, an amino group, a cyano group, a hydroxyl group, a nitro group, an aryl group, an alkylsulfonyl group or a phenylsulfonyl group which is optionally substituted on the aromatic ring.
  • the present invention provides a phosphorus-containing compound which is further characterized in that X is selected from the group consisting of ammonia (-NH-) and amine (-N(R3)-);
  • Y is selected from (-NH-), amine (-N(R N )-), ammonium (-N + (R4R5)-, wherein R N may be any substituent group such as methyl, ethyl, etc.
  • R N may be any substituent group such as methyl, ethyl, etc.
  • R4 and R5 may be the same or different substituent groups, such as an alkyl group such as a methyl group or an ethyl group, or an aromatic group such as a phenyl group or a benzyl group.
  • the anion coordinated to N + may be selected from halogens).
  • the phosphorus-containing compound provided by the present invention further has the following characteristics:
  • R 4 is selected from the group consisting of the following structures:
  • n is selected from an integer from 0 to 5;
  • the above A is selected from the group consisting of sulfur, carbon, nitrogen, and oxygen;
  • R 42 is selected from the group consisting of aryl, alkyl, alkylamino, alkylsulfonylamino, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl;
  • aryl group is selected from the group consisting of a 6-12 membered aromatic group and a derivative thereof, a 5-12 membered aromatic ring, and any or any of several carbon atoms is a heteroaryl group substituted with oxygen, nitrogen or sulfur;
  • the above derivative means a hydrogen, an alkyl group, an alkoxy group, a halogen group, an amino group, a cyano group, a hydroxyl group, a nitro group, a sulfonyl group, an alkylsulfonyl group or a phenylsulfonyl group which is optionally substituted on the aromatic ring.
  • the above heteroaryl group may further have a structure of -NR 422 ;
  • R 422 is a sulfonyl group, an alkylsulfonyl group, an alkyl group, or a hydroxyl group;
  • the above cycloalkyl group is a 3-12 membered cycloalkyl group
  • the above substituted cycloalkyl group is a sulfonyl group, an alkylsulfonyl group, an alkyl group, an alkoxy group, a hydroxyl group, an amino group, a nitro group;
  • heterocycloalkyl group is a 3-12 membered heterocycloalkyl group, and any or any of several carbon atoms is replaced by oxygen, nitrogen or sulfur;
  • the above substituted heterocycloalkyl group is a quaternary ring of any or any of a plurality of sulfonyl groups, alkylsulfonyl groups, alkyl groups, alkoxy groups, hydroxyl groups, amino groups, nitro groups, carbonyl-substituted aza, oxa or thia groups. a five-, six- or seven-membered cycloalkyl group;
  • the above substituted heterocycloalkyl group may further have a structure of -NR 422 ;
  • R 422 is a sulfonyl group, an alkylsulfonyl group, an alkyl group, or a hydroxyl group;
  • R 42 may also be selected from the group consisting of the following structures:
  • R 43 and R 44 are the same or different alkyl group, a hydroxyl group or a hydroxyl group substituted with an alkyl group having not more than 5 carbon atoms;
  • G3 is a ring of 3-12 yuan
  • R 45 above is an alkyl group, a hydroxyl group, an alkoxy group or an amino group substituted on the G 3 ring.
  • the phosphorus-containing compound provided by the present invention further has the following characteristics: that is, a compound represented by the following structure:
  • C 1 and C 2 are each a carbon atom, and the carbon bond therebetween is a single bond, a double bond or a triple bond.
  • the phosphorus-containing compound provided by the present invention further has the following characteristics: that is, a compound represented by the following structure:
  • R 11 is selected from any one or more substituted hydrogen, alkyl, alkoxy, halogen, amino, cyano, hydroxy, nitro;
  • R 2 is selected from the group consisting of hydroxyl, alkyl, alkoxy, halogen
  • R 3 is any optionally substituted hydrogen, alkyl, alkoxy, halogen, amino, cyano, hydroxy, nitro, aryl;
  • Y 1 is selected from the group consisting of hydrogen, alkyl, and aryl;
  • R 41 is selected from any one or more substituted hydrogen, alkyl, alkoxy, alkylsulfonyl, arylsulfonyl, halogen, amino, cyano, hydroxy, nitro;
  • R 5 is selected from the group consisting of hydrogen, alkyl, aryl, benzyl and derivatives thereof.
  • the phosphorus-containing compound provided by the present invention further has the following characteristics: that is, a compound represented by the following structure:
  • X 1 , X 2 , X 3 and X 4 are selected from the group consisting of hydrogen, alkyl, halogen and hydroxyl.
  • the phosphorus-containing compound provided by the present invention has the feature that the phosphorus-containing compound is selected from the following compounds:
  • the present invention also provides a method for preparing the above phosphorus-containing compound, which is characterized in that:
  • the above compound C is a compound represented by the following structure:
  • the above compound B is a compound represented by the following structure:
  • L 1 L 1 'and L 2 and L 2' respectively, a pair of reactive groups can react with each other, in the course of the reaction, by L 1 and L 1 'reaction, L 2 and L 2' is reacted with Obtaining the target product;
  • R 1 is selected from the group consisting of alkyl, aryl, benzyl and derivatives thereof;
  • R 2 is selected from the group consisting of hydroxyl, alkyl, alkoxy, halogen
  • n is selected from a natural number of 1-3;
  • R 3 is any optionally substituted hydrogen, alkyl, alkoxy, halogen, amino, cyano, hydroxy, nitro, aryl;
  • R 4 is selected from the group consisting of alkyl, alkoxy, aryl, benzyl and derivatives thereof;
  • R 5 is selected from the group consisting of hydrogen, alkyl, aryl, benzyl and derivatives thereof.
  • the method for preparing a phosphorus-containing compound provided by the present invention further has the characteristics that the above L 1 and L 1 ' and between L 2 and L 2 ' can occur, and the substitution reaction, the addition reaction, and the elimination reaction are carried out. Or the displacement reaction, thereby forming a linkage between L 1 and L 1 'and L 2 and L 2 '.
  • the method for preparing a phosphorus-containing compound provided by the present invention further has the feature that the above L 1 is selected from the group consisting of halogen, amino, cyano, thio, hydroxy, alkoxy;
  • the above L 1 ' is selected from the group consisting of halogen, alkynyl, carboxy, amino, cyano, ester, alkoxy, sulfonylamino, alkoxysulfonyl;
  • L 2 is selected from the group consisting of halogen, carboxyl, amino, cyano, ester, alkoxy, sulfonylamino, alkoxysulfonyl;
  • the above L 2 ' is selected from the group consisting of halogen, amino, thio, hydroxy and alkoxy.
  • the method for preparing a phosphorus-containing compound provided by the present invention has the characteristic that the molar ratio of the above compound A to the compound C is 1:0.1-10;
  • the molar ratio of the above compound C to the compound B is 1:0.1-10.
  • the method for preparing a phosphorus-containing compound provided by the present invention further has the following characteristics: that is, the specific process steps are as follows:
  • Step 1 adding a halogenating reagent to the derivative of the phosphoric acid diester, reacting at a temperature of 50-100 ° C for 1-5 hours, and directly spinning to obtain a substrate 1;
  • this step it is intended to prepare a derivative of a phosphoric acid diester into a substrate having an active reactive group, and if the compound A having a reactive group L 1 is directly selected, the first step can be omitted.
  • the derivative of the phosphodiester is a compound represented by the following structure:
  • R 111 , R 112 , and R 113 are selected from an aryl group (e.g., an aromatic group such as a phenyl group, a naphthyl group, or a quinolyl group), an alkyl group (e.g., a methyl group, an ethyl group, a propyl group, an isopropyl group, or a cyclohexyl group). , an alkyl group such as a cyclopentyl group);
  • an aryl group e.g., an aromatic group such as a phenyl group, a naphthyl group, or a quinolyl group
  • an alkyl group e.g., a methyl group, an ethyl group, a propyl group, an isopropyl group, or a cyclohexyl group.
  • an alkyl group such as a cyclopentyl group
  • the halogenating reagent generally uses a reagent for providing halogen such as chlorosulfoxide, phosgene or bromine;
  • the reaction is preferably carried out under the protection of a shielding gas such as nitrogen, argon or helium.
  • a shielding gas such as nitrogen, argon or helium.
  • the amount of the halogenating agent added is 0.5 to 4 ml per 100 mg of the derivative of the phosphodiester.
  • Step 2 in the derivative of methyl ethynylbenzoate at a temperature below 0 ° C, the Grignard reagent and the substrate 1 are sequentially added, the reaction is carried out for 0.1-2 hours, the reaction is quenched with an acid solution, and the organic phase is extracted. Dry to obtain intermediate product 1;
  • the derivative of methyl ethynylbenzoate is a compound represented by the following structure:
  • R 311 is selected from any or a plurality of substituted halogen, nitro, aryl (eg, aryl, naphthyl, quinolyl, etc.), alkyl (eg, methyl, ethyl, propyl)
  • alkyl eg, methyl, ethyl, propyl
  • An alkyl group such as an isopropyl group, a cyclohexyl group or a cyclopentyl group;
  • the ethynyl and methyl formate groups may be para, ortho or meta;
  • the mass ratio of the derivative of methyl ethynylbenzoate to Grignard reagent and substrate 1 is 1:0.01-10:1:-10;
  • the reaction is preferably carried out under the protection of a protective gas such as nitrogen, argon or helium;
  • the reaction is preferably carried out in an ether solvent
  • the acid used for the quenching reaction is preferably a mineral acid, the concentration of the acid is preferably from 0.5 to 1.5 mol/L, and the amount of the acid is preferably from 0.01 to 10 times the total amount of the reactant;
  • the reagent for extraction is preferably an ester solvent.
  • Step 3 the intermediate product 1 and the de-esterification reagent, reacted at a temperature of 100-150 degrees Celsius for 2-5 hours, adding an acid solution, extracting the organic phase to spin dry, to obtain an intermediate product 2;
  • the mass ratio of the intermediate product 1 and the deesterification reagent is 1:0.5-3;
  • the reaction is preferably carried out under the protection of a protective gas such as nitrogen, argon or helium;
  • the acid used for the quenching reaction is preferably a mineral acid, the concentration of the acid is preferably from 0.5 to 1.5 mol/L, and the amount of the acid is preferably from 0.01 to 10 times the total amount of the reactant;
  • the reagent for extraction is preferably an ester solvent.
  • Step 4 in the intermediate product 2, sequentially add L 2 as the amino group of the compound C, a basic catalyst, at a temperature of 20-50 ° C, the reaction is 1-10 hours, quench the reaction with an acid solution, extract the organic phase spin dry A phosphorus-containing compound containing an alkynyl group is obtained.
  • the molar ratio of the intermediate product 2, the compound C and the basic catalyst is 1:1-5:5-20;
  • the acid used for the quenching reaction is preferably a mineral acid, the concentration of the acid is preferably from 0.5 to 1.5 mol/L, and the amount of the acid is preferably from 0.01 to 10 times the total amount of the reactant;
  • the reagent for extraction is preferably an ester solvent.
  • the method for preparing a phosphorus-containing compound provided by the present invention has the feature that the alkynyl-containing phosphorus-containing compound undergoes a reduction reaction, an esterification reaction, an amidation reaction, a substitution reaction, and an addition reaction. After one or more of the reactions, the corresponding phosphorus-containing product can be obtained.
  • the present invention provides the use of the above phosphorus-containing compound, which is characterized in that it can be used as an immune cell migration inhibitor.
  • the present invention provides the use of the above phosphorus-containing compound, characterized in that the eye drops containing the above phosphorus-containing compound are useful for alleviating and treating dry eye.
  • the preparation method of the ophthalmic preparation can be any conventional preparation method.
  • the above compound is added to 10-200 times by weight of sterile physiological saline, 0.01-1 times of alkali solution is added, and stirred to obtain a transparent solution; and the buffer solution is added to the solution obtained above until the pH of the solution is 6.5. Between -7.5; add sterile physiological saline to the obtained aqueous solution until the total volume reaches 1.5-20 times the original volume. The above solution was again purged with nitrogen, bubbling for 0.5-10 hours, and the resulting solution was sealed and stored at 5 ° C in the dark. Dispense into a disposable eye drop bag for use. Wherein the sodium hydroxide and a saturated aqueous solution of NaH 2 PO 4 may be replaced by other buffer solutions.
  • a new class of phosphorus-containing compounds is synthesized, which is a novel immune cell migration inhibitor. It has good hydrophilicity, is easy to develop into eye drops, has a strong inhibitory effect on immune cell migration, and may alleviate the symptoms of most dry eye patients.
  • Step B Methyl 2,6-dichloro-4-((phenyl(methoxy)phosphoryl)ethynyl)benzoate (Compound 1.2)
  • Step C 2,6-Dichloro-4-((hydroxy(phenyl)phosphoryl)ethynyl)benzoic acid (Compound 1.3)
  • Step D (2s)-2-(2,6-dichloro-4-((hydroxy(phenyl)phosphoryl)ethynyl) Benzyl)-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester (Compound 1.4)
  • Step E (2s)-2-(2,6-Dichloro-4-(2-(hydroxy(phenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl) Phenyl)propionic acid (compound 1)
  • Step A (m-methoxyphenyl)ethoxyphosphoryl chloride (Compound 2.1)
  • Step B Methyl 2,6-dichloro-4-((m-methoxyphenyl)(ethoxy)phosphoryl)ethynyl)benzoate (Compound 2.2)
  • Step C 2,6-Dichloro-4-((hydroxy(m-methoxyphenyl)phosphoryl)ethynyl)benzoic acid (Compound 2.3)
  • Step D (2s)-2-(2,6-Dichloro-4-((hydroxy(m-methoxyphenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl) Phenyl) benzyl propionate (compound 2.4)
  • Step E (2s)-2-(2,6-Dichloro-4-((hydroxy(3-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl) Phenyl) propionic acid (compound 2)
  • Step A (2s)-2-(2,6-Dichloro-4-((methoxy(phenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl) Phenyl) benzyl propionate (Compound 4.1)
  • Step B (2s)-2-(2,6-Dichloro-4-(2-(methoxy(phenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonate) Acyl)phenyl)propionic acid (compound 4)
  • Step A 2,6-Dichloro-4-(((hydroxyphenyl)(ethoxy)phosphoryl)ethynyl)benzoic acid (Compound 6.1)
  • Compound 2.2 was dissolved in DCM, and 1 mol/L of boron tribromide (10 eq) was added at a low temperature, and stirred at 0 °C for 30 minutes, then quenched at -40 °C, extracted with EA three times, and combined with organic phase rotation. Just do it.
  • Step B (2s)-2-(2,6-Dichloro-4-((ethoxy(m-hydroxyphenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonate) Benzyl)phenyl)propionic acid benzyl ester (Compound 6.2)
  • Step C (2s)-2-(2,6-Dichloro-4-(2-(ethoxy(m-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3-( Methanesulfonyl)phenyl)propionic acid (compound 6)
  • Step A (2s)-2-(2,6-Dichloro-4-((methoxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-( Methyl sulfonyl)phenyl)propanoate (Compound 7.1)
  • Step B (2s)-2-(2,6-Dichloro-4-((methoxy(3-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonate) Acyl)phenyl)propionic acid (compound 7)
  • Step B Methyl 2,6-dichloro-4-((phenyl(methyl)phosphoryl)ethynyl)benzoate (Compound 9.2)
  • Step D (2s)-2-(2,6-Dichloro-4-((methyl(phenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl)benzene Base) benzyl propionate (compound 9.4)
  • Step E (2s)-2-(2,6-Dichloro-4-((methyl(phenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl)benzene Propionate (compound 9)
  • Step C Methyl 2,6-dichloro-4-((m-methoxyphenyl)(methyl)phosphoryl)ethynyl)benzoate (Compound 11.3)
  • Step D 2,6-Dichloro-4-((methyl(m-hydroxyphenyl)phosphoryl)ethynyl)benzoic acid (Compound 11.4)
  • Step E (2s)-2-(2,6-Dichloro-4-((methyl(3-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl) Phenyl) benzyl propionate (compound 11.5)
  • Step F (2s)-2-(2,6-Dichloro-4-((methyl(3-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl) Phenyl) propionic acid (compound 11)
  • Example 15 was prepared by the same procedure as in Example 11 except that "dimethoxymethoxyphosphate” was replaced with “p-methoxyphosphoric acid diethyl ester”.
  • Example 16 was prepared by the same procedure as in Example 13 except that "Compound 7.1” was replaced with “Compound 1.4".
  • Step A 3-(2-Thienylamino)-N-[(1,1-dimethylethoxy)carbonyl]-L-alanine methyl ester (Compound 17.1)
  • Step B (S)-2-Amino-3-(2-thienamido)propionic acid methyl ester (Compound 17.2)
  • Step C Methyl 2,6-dichloro-4-((m-methoxyphenyl)(ethoxy)phosphoryl)ethyl)benzoate (Compound 17.3)
  • Step D 2,6-Dichloro-4-(((hydroxyphenyl)(ethoxy)phosphoryl)ethyl)benzoate (Compound 17.4)
  • Step E (2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzoylamino)-3-(2-thiophene) Amido) methyl propionate (compound 17.5)
  • Step F (2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzoylamino)-3-(2-thiophene) Amido)propionic acid (compound 17)
  • Step A (S)-2-((tert-Butyloxycarbonyl)amino)-3-(1-methyl-1,2,5,6-tetrahydropyridin-3-yl)propanoic acid methyl ester) ( Compound 18.1)
  • Step B (S)-2-((tert-Butyloxycarbonyl)amino)-3-(1-(methanesulfonamide)-1,2,5,6-tetrahydropyridin-3-yl)propanoic acid Ester (Compound 18.2)
  • Step C (S)-2-Amino-3-(1-(methylsulfonamide)-1,2,5,6-tetrahydropyridin-3-yl)propanoic acid methyl ester (Compound 18.3)
  • Step D (2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzoylamino)-3-(1-( Methanesulfonamide)-1,2,5,6-tetrahydropyridin-3-yl)propionic acid (Compound 18)
  • Step A 1-Cyano-9,9-dimethyl-2-(methylthio)-7-carbonyl-8-oxy-1,3,6-triazin-1-ene-5-carboxylate Methyl ester (compound 19.1)
  • Step B 1-cyano-2-((R)-3-hydroxytetrahydropyrrol-1-yl)-9,9-dimethyl-7-carbonyl-8-oxy-1,3,6- Methyl triazain-1-ene-5-carboxylate.
  • the obtained compound 19.1 was dissolved in acetonitrile (20 ml), and (R)-pyrrolidin-3-ol (20 mmol) and silver nitrate (20 mmol) were added. The reaction was refluxed for 8 hours, filtered over silica gel and dried. The crude product was separated on a silica gel column eluting with 100/5/1 DCM/MeOH/EtOAc.
  • Step C (2s)-2-Amino-3-((R)-N'-cyano-3-hydroxytetrahydropyrrole-1-carbenyl)benzoylamino)propanoic acid (Compound 19.3)
  • Step A Ethyl (3-methoxyphenyl) phosphate (compound 20.1)
  • Step C (2s)-2-(2,6-Dichloro-4-(ethyl(3-hydroxyphenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl) Phenyl) propionic acid (compound 20)
  • Example 21 The exact same procedure as in Preparation Example 3 was used to prepare Example 21, wherein Compound 20 was substituted for Compound 2.
  • Example 22 The exact same procedure as in Preparation Example 21 was used to prepare Example 22, wherein the cyclopropyl Grignard reagent was substituted for the ethyl Grignard reagent.
  • Example 23 The exact same procedure as in Preparation Example 21 was used to prepare Example 23, wherein the butyl Grignard reagent was substituted for the ethyl Grignard reagent.
  • Step A Diethyl benzofuran-6-ylphosphoric acid (Compound 24.1)
  • Step B Methyl (benzofuran-6-yl)phosphoric acid ethyl ester (Compound 24.2)
  • the compound 24.2 was dissolved in thionyl chloride (5 ml), and the resulting solution was refluxed for 5 hours, and the reaction solution was dried to give the desired product, which was used directly in the next step.
  • Step D (2s)-2-(2,6-Dichloro-4-((methyl(benzofuran-6-yl)phosphoryl)ethynyl)benzylamino)-3-(3-(A) Sulfonyl)phenyl)propionic acid (compound 24)
  • Example 2 The exact same procedure as in Preparation Example 3 was used to prepare Example 2, in which Compound 24 was substituted for Compound 2.
  • Step A Methyl 2,6-dichloro-(4-hydroxymethyl)benzoate (Compound 28.1) Dimethyl 2,6-dichloroterephthalate (2.63 g, 10 mmol) dissolved in THF (50 mL) The lithium borohydride (12 mm ml) was slowly added, and the mixture was stirred for 1 hour, then acetone (1 ml) and EtOAc (100 ml). The resulting solution was washed twice with water, dried over anhydrous Na.sub.2SO4, filtered and evaporated.
  • Step B 2,6-Dichloro-(4-hydroxymethyl)benzoic acid (Compound 28.2)
  • the compound 28.1 was dissolved in pyridine (20 ml), lithium iodide (15 mmml) was added, and the reaction was stirred at reflux for 5 hours, then dried, and the crude product was purified using silica gel column, mobile phase 95/5/0.5 (v/v/v) DCM/MeOH/AcOH.
  • Step C (S)-2-(2,6-Dichloro-4-(hydroxymethyl)benzamide)-3-(3-(3-(3-(methylsulfonyl)phenyl)propyl Methyl ester (compound 28.3)
  • Step D (S)-2-(2,6-dichloro-4-(hydroxymethyl)benzamide)-3-(3-(3-(3-(methylsulfonyl)phenyl)propanoic acid (Compound 28.4)
  • Step E (2S)-2-(2,6-Dichloro-4-(((methyl(phenyl)phosphoryl)oxy)methyl)benzoylamino)-3-(3-(A) Sulfonyl)phenyl)propionic acid (Compound 28).
  • Step A (Chloromethyl)(3-methoxyphenyl)(methyl)phosphorus oxide (Compound 29.1)
  • Step B Methyl 2,6-dichloro-4-(((3-methoxyphenyl)(methyl)phosphoryl)methyl)amino)benzoate (Compound 29.2)
  • Step C 2,6-Dichloro-4-(((3-methoxyphenyl)(methyl)phosphoryl)methyl)amino)benzoic acid (Compound 29.3)
  • Step D (2S)-2-(2,6-Dichloro-4-(((3-hydroxyphenyl)(methyl)phosphoryl)methyl)amino)benzamide)-3-( Methyl 3-(methylsulfonyl)phenyl)propanoate (Compound 29.4)
  • Step E (2S)-2-(2,6-Dichloro-4-((3-hydroxyphenyl)(methyl)phosphoryl)methyl)amino)benzamide)-3-(3) -(Methanesulfonyl)phenyl)propionic acid (Compound 29)
  • Step A (E)-2,6-Dichloro-4-(2-(diethoxyphosphono)vinyl)benzoic acid methyl ester (Compound 32.1)
  • Step B (E)-2,6-Dichloro-4-(2-(chloroethoxyphosphonyl)vinyl)benzoic acid methyl ester (Compound 32.2)
  • Step D (E)-4-(2-(3-Hydroxyphenyl)ethoxyphosphonovinyl)-2,6-dichlorobenzoic acid (Compound 32.4)
  • Step E (S,E)-2-(4-(2-(3-hydroxyphenyl)ethoxyphosphonylvinyl)-2,6-dichlorobenzamide)-3-(3-(methylsulfonate) Benzyl)phenyl)propionic acid benzyl ester (compound 32.5)
  • Step F (S,E)-2-(4-(2-(3-hydroxyphenyl)ethoxyphosphonylvinyl)-2,6-dichlorobenzamide)-3-(3-(methylsulfonate) Acyl)phenyl)propionic acid (compound 32)
  • Example 16 was converted to Example 33 using the exact same procedure as in Example 14.
  • Example 34 was prepared by substituting 2-thiophenecarboxylic acid in Example 17 to 3-hydroxybenzoic acid.
  • Example 35 was prepared by substituting the 2-thiophenecarboxylic acid of Example 17 with 3,5-dihydroxybenzoic acid.
  • Example 43 was prepared by substituting 2-thiophenecarboxylic acid in Example 17 with (s)-3-hydroxypyrrolidine-1-carbonyl chloride.
  • Example 45 was prepared by substituting (R)-3-pyrrolidinol in Example 19 with (trans)-3,4-pyrrolidine diol.
  • Example 46 was prepared by replacing (R)-3-pyrrolidinol in Example 19 with diethanolamine.
  • Step A Methyl 2,6-dichloro-4-((o-methoxyphenyl)(ethoxy)phosphoryl)ethynyl)benzoate (compound 47.2)
  • Step B 2,6-Dichloro-4-((hydroxy(o-methoxyphenyl)phosphoryl)ethynyl)benzoic acid (Compound 47.3)
  • Step C (2s)-2-(2,6-Dichloro-4-((hydroxy(o-methoxyphenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl) Phenyl) benzyl propionate (compound 47.4)
  • Step D (2s)-2-(2,6-Dichloro-4-((methoxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-( Methyl sulfonyl)phenyl)propanoate (Compound 47.5)
  • Step E (2s)-2-(2,6-Dichloro-4-((methoxy(2-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonate) Acyl)phenyl)propionic acid (compound 47)
  • Example 48 was prepared by the same procedure as in Example 11 except that "dimethoxym-methoxyphosphate” was replaced with "4-methoxy-3-chlorophosphate diethyl ester".
  • Step A 2,6-Dichloro-4-(((benzofuran-6-yl)(ethoxy)phosphoryl)ethynyl)benzoic acid methyl ester (Compound 49.1)
  • Step B 2,6-Dichloro-4-((hydroxy(benzofuran-6-yl)phosphoryl)ethynyl)benzoic acid (Compound 49.2)
  • Step C (2s)-2-(2,6-Dichloro-4-((hydroxy(benzofuran-6-yl)phosphoryl)ethynyl)benzamide)-3-(3-(A) Benzyl sulfonyl)phenyl)propionate (Compound 49.3)
  • Step D (2s)-2-(2,6-Dichloro-4-((hydroxy(benzofuran-6-yl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonate) Acyl)phenyl)propionic acid (compound 49)
  • Step A Methyl 2,6-dichloro-4-(1-hydroxy 2-(methoxy(3-methoxyphenyl)phosphoryl)ethyl)benzoate (Compound 50.1)
  • Step B 2,6-Dichloro-4-(1-hydroxy 2-(hydroxy(3-hydroxyphenyl)phosphoryl)ethyl)benzoic acid (Compound 50.2)
  • Step C (2s)-2-(2,6-Dichloro-4-(1-hydroxy2-(hydroxy(3-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3) -(Methanesulfonyl)phenyl)propanoic acid benzyl ester (Compound 50.3)
  • Step D (2s)-2-(2,6-Dichloro-4-(1-hydroxy2-(hydroxy(3-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3) -(Methanesulfonyl)phenyl)propionic acid (Compound 50)
  • the compound 32 was dissolved in 5 ml of tetrahydrofuran, and 1 ml of a LiOH aqueous solution was added thereto, and the mixture was stirred at room temperature for 5 hr.
  • Step A (E)-4-(2-(bis(3-methoxyphenyl)phosphono)vinyl)-2,6-dichlorobenzoic acid methyl ester (compound 52.1)
  • Step B (E)-4-(2-(bis(3-hydroxyphenyl)phosphono)vinyl)-2,6-dichlorobenzoic acid (Compound 52.2)
  • Step C (S,E)-2-(4-(2-(bis(3-hydroxyphenyl)phosphono)vinyl)-2,6-dichlorobenzamide)-3-(3-(A Benzyl sulfonyl)phenyl)propionate (compound 52.3)
  • Step D (S,E)-2-(4-(2-(bis(3-hydroxyphenyl)phosphono)vinyl)-2,6-dichlorobenzamide)-3-(3-(A Sulfonyl)phenyl)propionic acid (compound 52)
  • Example 54 The exact same procedure as in Preparation Example 32 was used to prepare Example 54, wherein p-methoxymagnesium bromide was substituted for m-methoxymagnesium bromide.
  • Step A dimethyl (benzofuran-3-yl) phosphine oxide (compound 55.1)
  • Step B (2s)-2-(2,6-Dichloro-4-(1-hydroxy2-(methyl(benzofuran-3-yl)phosphoryl)ethyl)benzamide)-3 -(3-(methylsulfonyl)phenyl)propionic acid (Compound 55)
  • Example 56 The exact same procedure as in Preparation Example 21 was used to prepare Example 56, wherein the isopropyl Grignard reagent was substituted for the ethyl Grignard reagent.
  • Step A (E)-4-(2-(3-Methoxyphenyl)chlorophosphonovinyl)methyl-2,6-dichlorobenzoate (Compound 57.1)
  • Step B (E)-4-(2-(3-Methoxyphenyl)cyclopropylphosphonovinyl)methyl-2,6-dichlorobenzoate (Compound 57.2)
  • Step C (E)-4-(2-((3-Hydroxyphenyl)cyclopropylphosphono)vinyl)-2,6-dichlorobenzoic acid (Compound 57.3)
  • Step D (S,E)-2-(4-(2-(3-hydroxyphenyl)cyclopropylphosphono)vinyl)-2,6-dichlorobenzamide)-3-(3-( Methyl sulfonyl)phenyl)propanoate (compound 57.4)
  • Step E (S,E)-2-(4-(2-(3-hydroxyphenyl)cyclopropylphosphono)vinyl)-2,6-dichlorobenzamide)-3-(3-( Methanesulfonyl)phenyl)propionic acid (compound 57)
  • Example 58 The exact same procedure as in Preparation Example 57 was used to prepare Example 58 wherein the methyl Grignard reagent was substituted for the cyclopropyl Grignard reagent.
  • T-cell adhesion assay using human T lymphocyte strain Jurkat (ATCC TIB-152): goat anti-Human IgG (Fc specific) (Sigma I8885) was diluted to 10 ⁇ g/ml in PBS at 100 °L per well at 4 °C The cells were incubated for 12 hours in a /96 well plate. The liquid in the well plate was poured out, blocked with 200 uL of 1% BSA at 37 ° C for 90 minutes, and washed three times with PBS. 50 uL of 1 ug/mL ICAM-1 (containing 0.1% BSA, 0.01% Tween 20) was added to each well and incubated at 37 ° C for 3 hours. The plate was washed 3 times with assay buffer (20 mM HEPES pH 7.6, 140 mM NaCl, 1 mM MgCl 2 , 1 mM MnCl 2 , 0.2% glucose).
  • assay buffer 20 mM HEPES pH 7.6, 140 mM NaC
  • the Jurkat cytometer was centrifuged at 100-G, and the cells were resuspended in an assay buffer (20 mM HEPES pH 7.6, 140 mM NaCl, 1 mM MgCl 2 , 1 mM MnCl 2 , 0.2% glucose) at 37 ° C.
  • an assay buffer (20 mM HEPES pH 7.6, 140 mM NaCl, 1 mM MgCl 2 , 1 mM MnCl 2 , 0.2% glucose
  • the inhibitor was diluted to a final concentration of 2X in assay buffer, and 50 uL of the compound solution and 60 uL of Jurkat cells were mixed at room temperature, and incubated at 37 ° C for 30 minutes. 100 ⁇ L/well of cells and inhibitor were added to the plate and incubated for 1 hour at room temperature. The total fluorescence was measured by a fluorometer: ex: 485; em: 530; cutoff: 530 to measure the total fluorescence. The plate was washed once with the assay buffer and the fluorescence was measured with a fluorometer: ex: 485; em: 530; cutoff:. Results obtained are plotted inhibition - concentration diagram, and the EC 50 calculated by standard methods. Table 1 shows the partial EC 50 values measured by this method.
  • Example EC50(nM) Example EC50(nM) 1 30 twenty one 7.3 2 9.4 twenty two twenty two 3 8.5 twenty three 61 4 11 twenty four 10.2 5 twenty two 25 63 6 17 26 twenty three 7 7.2 27 NA * 8 6.2 28 NA * 9 11.8 29 NA * 10 78 30 7.2 11 13.5 31 7.1 12 29 32 3.7 13 29 33 10.8 14 8.5 34 12.5 15 1.8 35 3.8 16 7.2 36 30 17 15 37 >1000 18 NA * 38 twenty four 19 230 39 NA * 20 NA * 40 NA *
  • Example EC50(nM) Example EC50(nM) 41 NA 51 4.2 42 150 52 74 43 69 53 twenty two 44 NA * 54 9.6 45 >1000 55 19 46 >1000 56 63 47 340 57 5.3 48 6.8 58 1.9 49 10.8 50 16
  • Example 11 The compound obtained in Example 11 was added to 90 mL of sterile physiological saline, and 0.7 g of NaOH was added thereto, followed by stirring to obtain a transparent solution; and a saturated aqueous solution of NaH 2 PO 4 was added to the solution obtained above until the pH of the solution was 6.75. Between -7.25. Sterile physiological saline was added to the obtained aqueous solution until the total volume reached 100.0 mL. The above solution was purged with nitrogen, and bubbled for 1 hour, and the resulting solution was sealed and stored at 5 ° C in the dark. Dispense into disposable eye drops bags, each containing 60 mL of formulation solution. The method and specific ratio of the formulation can also be adjusted as needed, depending on the nature of the particular compound and the requirements of the application.

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Abstract

Provided is a phosphorus-containing compound characterized by being a compound represented by the following structure: the compound is a novel immunocyte migration inhibitor. The compound has good hydrophilicity and can be developed into eye drops. The compound has a strong inhibitory ability to immunocyte migration and can relieve the symptoms of most dry-eye patients.

Description

一种含磷化合物及其制备和应用Phosphorus-containing compound and preparation and application thereof 技术领域Technical field
本发明涉及医药化学领域，具体地，涉及一种含磷化合物及其制备方法和在治疗干眼症上的应用。The present invention relates to the field of medical chemistry, and in particular to a phosphorus-containing compound, a preparation method thereof, and an application for treating dry eye.
背景技术Background technique
眼泪给眼睛持久的润湿和润滑，是维持视力和眼睛舒适度的关键。眼泪由水、油脂、粘液、抗体和一些特定蛋白质(用于抵抗感染)的组成。这些组分由位于眼周围的特殊的腺体分泌。当该眼泪体系中存在不平衡时，人会感觉眼干。Tears provide long-lasting moisturization and lubrication to the eyes, which is the key to maintaining vision and eye comfort. Tears are composed of water, oil, mucus, antibodies, and some specific proteins that are used to fight infection. These components are secreted by specific glands located around the eye. When there is an imbalance in the tear system, people will feel dry eyes.
干眼综合征为常见的眼表面炎性疾病。患有干眼病的人可能感觉到眼睛疼痛、光敏性、发痒、发红和视力模糊。干眼综合征有多种诱发因素，包括年龄、性别、环境、药物、手术，和全身疾病如自身免疫系统疾病、糖尿病、甲状腺疾病、淋巴瘤等。干眼病如得不到恰当的诊断和治疗可能会导致进一步的并发症如感染、眼表面角质化、角膜溃疡和结膜鳞状化等。Dry eye syndrome is a common ocular surface inflammatory disease. People with dry eye may experience eye pain, photosensitivity, itching, redness and blurred vision. Dry eye syndrome has a variety of predisposing factors, including age, gender, environment, drugs, surgery, and systemic diseases such as autoimmune diseases, diabetes, thyroid disease, lymphoma and the like. Dry eye disease without proper diagnosis and treatment may lead to further complications such as infection, keratinization of the ocular surface, corneal ulceration and conjunctival squamous.
故而，干眼症是困扰5-10％人口的很大的疾病，特别是长期工作于计算机前，及中年以后的人群。现今眼科医生门诊超过30％是干眼症病人。中国尽管干眼症患者众多，但迄今没有被批准用于治疗干眼症的药物。病人只能已人造眼泪得到暂时的缓解。因此，急需治疗干眼症的药物。Therefore, dry eye syndrome is a very serious disease that afflicts 5-10% of the population, especially those who work long hours before and after the middle age. More than 30% of today's ophthalmologist clinics are dry eye patients. Despite the large number of patients with dry eye syndrome, there are no drugs approved for the treatment of dry eye syndrome. The patient can only have temporary relief from artificial tears. Therefore, there is an urgent need to treat drugs for dry eye syndrome.
干眼症的发病率和年龄与呈正比，50岁以上人群约20％有不同程度的干眼症；性别也影响干眼症，女性，特别是高龄女性患干眼症远高于男性，这可能和性荷尔蒙的分泌有关；白领工作人员长时间在空调环境下，另外长时间使用屏幕也造成该人群干眼症发病率高。干眼症是一连续的病理过程，病情由轻到 重连续发展，且轻、中、重度之间无明显的分界线。尽管干眼症病因复杂，研究发现各种原因引发的干眼症的病理类似：免疫细胞侵润眼睛表面组织引发慢性炎症，造成眼表伤害。目前，欧美市场有两个药物被批准：(1)环孢素A悬浮液。该药是很强大免疫系统抑制药，因此有可能造成对免疫系统的伤害。同时，因为其是悬浮液，存在长期存放稳定性，以及使用时对病人眼睛刺激等问题；(2)Lifitigrast,该药2016年12月或得美国FDA批准，是免疫细胞迁徙抑制剂，通过阻止免疫细胞进入炎症部位达到治疗效果；但该药亲脂性很高，而且在临床上对>50％的病人没有效果。The incidence and age of dry eye syndrome are directly proportional to the fact that about 20% of people over 50 years old have different degrees of dry eye syndrome; gender also affects dry eye syndrome, and women, especially older women, suffer from dry eye syndrome much higher than men. May be related to the secretion of sex hormones; white-collar workers for a long time in the air-conditioned environment, and the long-term use of the screen also caused a high incidence of dry eye syndrome in this population. Dry eye syndrome is a continuous pathological process in which the condition progresses from light to heavy, and there is no obvious boundary between light, medium and severe. Despite the complex etiology of dry eye, the study found that the pathology of dry eye caused by various causes is similar: immune cells invade the surface tissue of the eye to cause chronic inflammation, causing ocular surface damage. Currently, two drugs are approved in the European and American markets: (1) cyclosporin A suspension. This medicine is a very powerful immune system inhibitor, so it may cause damage to the immune system. At the same time, because it is a suspension, there are long-term storage stability, and the eye irritation of the patient during use; (2) Lifitigrast, the drug was approved by the US FDA in December 2016, is an immune cell migration inhibitor, by blocking The immune cells enter the site of inflammation to achieve therapeutic effects; however, the drug is highly lipophilic and has no clinical effect on >50% of patients.
发明内容Summary of the invention
本发明为新型的免疫细胞迁徙抑制剂。有很好的亲水性，可以开发成眼药水。对免疫细胞迁徙有很强的抑制能力，可以缓解大多数干眼症病人的症状。The invention is a novel immune cell migration inhibitor. It has good hydrophilicity and can be developed into eye drops. It has a strong inhibitory effect on immune cell migration and can alleviate the symptoms of most dry eye patients.
本发明提供的一种含磷化合物，其特征在于，为如下结构所示的化合物：The present invention provides a phosphorus-containing compound characterized by being a compound represented by the following structure:
Figure PCTCN2018087629-appb-000001
Figure PCTCN2018087629-appb-000001
R 1选自烷基、芳基、苄基及其衍生物； R 1 is selected from the group consisting of alkyl, aryl, benzyl and derivatives thereof;
R 1中烷基选自如：甲基、乙基、丙基、异丙基、丁基、异丁基、戊基、环戊基、己基、环己基等碳原子数为1-12的支链、支链或环烷基； The alkyl group in R 1 is selected from the group consisting of a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, a pentyl group, a cyclopentyl group, a hexyl group, a cyclohexyl group and the like having 1 to 12 carbon atoms. , branched or cyclic alkyl;
芳基选自如：苯基、萘基、喹啉基，或苯环上任一或任几个碳为碳原子数为1-3的烷基/苯基/卤素/硝基/氨基/磺酰基/氨基/碳原子数为1-3的烷氧基取代的苯基、萘基或喹啉基；The aryl group is selected from, for example, a phenyl group, a naphthyl group, a quinolyl group, or an alkyl group/phenyl group/halogen/nitro/amino/sulfonyl group having any one or more carbons of 1 to 3 carbon atoms. a phenyl, naphthyl or quinolyl group substituted with an alkoxy group having an amino group/carbon number of 1-3;
苄基选自如：苯环上任一或任几个碳为碳原子数为1-3的烷基/苯基/卤素/硝基/氨基/磺酰基/碳原子数为1-3的烷基取代的苄基；The benzyl group is selected from, for example, an alkyl group having at least one or several carbons having a carbon number of 1-3, a phenyl group/halogen/nitro group/amino group/sulfonyl group and an alkyl group having 1 to 3 carbon atoms. Benzyl;
R 2选自羟基、烷基、氢、烷氧基、胺基、烷胺基； R 2 is selected from the group consisting of a hydroxyl group, an alkyl group, a hydrogen, an alkoxy group, an amine group, and an alkylamine group;
R 2中烷基选自如：甲基、乙基、丙基、异丙基、环丙基、丁基、异丁基、戊基、环戊基、己基、环己基等碳原子数为1-12的支链、支链或环烷基； The alkyl group in R 2 is selected from, for example, methyl group, ethyl group, propyl group, isopropyl group, cyclopropyl group, butyl group, isobutyl group, pentyl group, cyclopentyl group, hexyl group, cyclohexyl group and the like, and the number of carbon atoms is 1- a branched, branched or cyclic alkyl group of 12;
烷氧基选自如：甲氧基、乙氧基等碳原子数为1-12的支链、支链或环烷氧基；The alkoxy group is selected from the group consisting of a branched, branched or cyclic alkoxy group having a carbon number of from 1 to 12 such as a methoxy group or an ethoxy group;
n选自0或1；n is selected from 0 or 1;
X选自碳、氧、氮；X is selected from the group consisting of carbon, oxygen, and nitrogen;
在X中，当其为碳时，此处可以为(-CH 2-)，或(-C(R1R2)-,其中，R1、R2可以为相同或不相同的任意取代基团，如：氢、甲基、乙基等烷基，苯基、苄基等芳香基团，羟基，烷氧基，卤素等；) In X, when it is carbon, it may be (-CH 2 -), or (-C(R1R2)-, wherein R1, R2 may be the same or different any substituent group, such as: hydrogen An alkyl group such as a methyl group or an ethyl group; an aromatic group such as a phenyl group or a benzyl group; a hydroxyl group, an alkoxy group, a halogen, etc.;
在X中，当其为氮时，此处可以为(-NH-)，或(-N(R N)-，其中，R N可以为任意取代基团，如：甲基、乙基等烷基，苯基、苄基等芳香基团等；) In X, when it is nitrogen, it may be (-NH-), or (-N(R N )-, wherein R N may be any substituent such as methyl, ethyl or the like. An aromatic group such as a phenyl group or a benzyl group;
Z选自碳、氧、硫、氮；Z is selected from the group consisting of carbon, oxygen, sulfur, and nitrogen;
在Z中，当其为碳时，此处可以为羰基(-(C＝O)-)、烷基(-C nH 2n-，其中，n为10以下的自然数)、或(-C(R1R2)-、支链烷基，其中，R1、R2可以为相同或不相同的任意取代基团，如：氢、甲基、乙基等烷基，苯基、苄 基等芳香基团，羟基，烷氧基，卤素等；) In Z, when it is carbon, it may be a carbonyl group (-(C=O)-), an alkyl group (-C n H 2n - wherein n is a natural number of 10 or less), or (-C ( R1R2)-, a branched alkyl group, wherein R1 and R2 may be the same or different optional substituent groups, such as an alkyl group such as hydrogen, methyl or ethyl, an aromatic group such as a phenyl group or a benzyl group, and a hydroxyl group. , alkoxy, halogen, etc. ;)
在Z中，当其为氮时，此处可以为(-NH-)，或(-N(R N)-，其中，R N可以为任意取代基团，如：甲基、乙基等烷基，苯基、苄基等芳香基团等；) In Z, when it is nitrogen, it may be (-NH-), or (-N(R N )-, wherein R N may be any substituent such as methyl, ethyl or the like. An aromatic group such as a phenyl group or a benzyl group;
R 3为苯环上任一或任几取代的氢、烷基、烷氧基、卤素、氨基、氰基、羟基、硝基、芳基； R 3 is any optionally substituted hydrogen, alkyl, alkoxy, halogen, amino, cyano, hydroxy, nitro, aryl;
Y选自碳、氧、氮；Y is selected from the group consisting of carbon, oxygen, and nitrogen;
在Y中，当其为碳时，此处可以为(-CH 2-)，或(-C(R1R2)-,其中，R1、R2可以为相同或不相同的任意取代基团，如：氢、甲基、乙基等烷基，苯基、苄基等芳香基团，卤素等；) In Y, when it is carbon, it may be (-CH 2 -), or (-C(R1R2)-, wherein R1, R2 may be the same or different any substituent group, such as: hydrogen , an alkyl group such as a methyl group or an ethyl group, an aromatic group such as a phenyl group or a benzyl group, a halogen or the like;
在Y中，当其为氮时，此处可以为(-NH-)，或(-N(R N)-，其中，R N可以为任意取代基团，如：甲基、乙基等烷基，苯基、苄基等芳香基团等)； In Y, when it is nitrogen, it may be (-NH-), or (-N(R N )-, wherein R N may be any substituent such as methyl, ethyl or the like. a group, an aromatic group such as a phenyl group or a benzyl group, etc.);
R 4选自烷基、烷氧基、芳基、苄基及其衍生物； R 4 is selected from the group consisting of alkyl, alkoxy, aryl, benzyl and derivatives thereof;
R 5选自氢、烷基、芳基、苄基及其衍生物； R 5 is selected from the group consisting of hydrogen, alkyl, aryl, benzyl and derivatives thereof;
G1和G2所代表的取代基团呈间位、对位或邻位的设置于苯环上。The substituent groups represented by G1 and G2 are disposed on the benzene ring in a meta, para or ortho position.
进一步地，本发明提供的一种含磷化合物，还具有这样的特点：即、上述芳基选自苯基及其衍生物、萘基及其衍生物、N杂或O杂环苯基及其衍生物、N杂或O杂环萘基及其衍生物；Further, the present invention provides a phosphorus-containing compound which is further characterized in that the above aryl group is selected from the group consisting of a phenyl group and a derivative thereof, a naphthyl group and a derivative thereof, an N or O heterocyclic phenyl group, and a derivative, an N or O heterocyclonaphthyl group and a derivative thereof;
其中，上述衍生物指在芳香环上任一或任几取代的氢、烷基、烷氧基、卤素、氨基、氰基、羟基、硝基、芳基、烷基磺酰基、苯基磺酰基。Here, the above derivative means a hydrogen, an alkyl group, an alkoxy group, a halogen group, an amino group, a cyano group, a hydroxyl group, a nitro group, an aryl group, an alkylsulfonyl group or a phenylsulfonyl group which is optionally substituted on the aromatic ring.
进一步地，本发明提供的一种含磷化合物，还具有这样的特点：即、上X选自氨(-NH-)、胺(-N(R3)-)；Further, the present invention provides a phosphorus-containing compound which is further characterized in that X is selected from the group consisting of ammonia (-NH-) and amine (-N(R3)-);
上述Y选自(-NH-)、胺(-N(R N)-)、铵(-N +(R4R5)-,其中，R N可以为任意取代基团，如：甲基、乙基等烷基，苯基、苄基等芳香基团等)，R4、 R5可以为相同或不相同的任意取代基团，如：甲基、乙基等烷基，苯基、苄基等芳香基团等，与N +配位的阴离子可选自卤素)。 The above Y is selected from (-NH-), amine (-N(R N )-), ammonium (-N + (R4R5)-, wherein R N may be any substituent group such as methyl, ethyl, etc. An alkyl group, a phenyl group, a benzyl group or the like, etc.), and R4 and R5 may be the same or different substituent groups, such as an alkyl group such as a methyl group or an ethyl group, or an aromatic group such as a phenyl group or a benzyl group. Alternatively, the anion coordinated to N + may be selected from halogens).
进一步地，本发明提供的一种含磷化合物，还具有这样的特点：即、Further, the phosphorus-containing compound provided by the present invention further has the following characteristics:
上述R 4选自如下结构所示的基团： The above R 4 is selected from the group consisting of the following structures:
Figure PCTCN2018087629-appb-000002
Figure PCTCN2018087629-appb-000002
n选自0-5的整数；n is selected from an integer from 0 to 5;
上述A选自硫、碳、氮、氧；The above A is selected from the group consisting of sulfur, carbon, nitrogen, and oxygen;
上述R 42选自芳基、烷基、烷基氨基、烷基磺酰胺基、环烷基、取代的环烷基、杂环烷基、取代的杂环烷基； The above R 42 is selected from the group consisting of aryl, alkyl, alkylamino, alkylsulfonylamino, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl;
其中，上述芳基选自6-12元的芳香基及其衍生物、5-12元的芳香环上，任一或任几个碳原子为氧、氮、硫替代的杂芳基；Wherein the above aryl group is selected from the group consisting of a 6-12 membered aromatic group and a derivative thereof, a 5-12 membered aromatic ring, and any or any of several carbon atoms is a heteroaryl group substituted with oxygen, nitrogen or sulfur;
其中，上述衍生物指在芳香环上任一或任几取代的氢、烷基、烷氧基、卤素、氨基、氰基、羟基、硝基、磺酰基、烷基磺酰基、苯基磺酰基。Here, the above derivative means a hydrogen, an alkyl group, an alkoxy group, a halogen group, an amino group, a cyano group, a hydroxyl group, a nitro group, a sulfonyl group, an alkylsulfonyl group or a phenylsulfonyl group which is optionally substituted on the aromatic ring.
上述杂芳基上还可以具有-N-R 422的结构； The above heteroaryl group may further have a structure of -NR 422 ;
上述R 422为磺酰基、烷基磺酰基、烷基、羟基； The above R 422 is a sulfonyl group, an alkylsulfonyl group, an alkyl group, or a hydroxyl group;
上述环烷基为3-12元的环烷基；The above cycloalkyl group is a 3-12 membered cycloalkyl group;
上述取代的环烷基为环上任一或任几取代的磺酰基、烷基磺酰基、烷基、烷氧基、羟基、氨基、硝基；The above substituted cycloalkyl group is a sulfonyl group, an alkylsulfonyl group, an alkyl group, an alkoxy group, a hydroxyl group, an amino group, a nitro group;
上述杂环烷基为3-12元的杂环烷基上，任一或任几个碳原子为氧、氮、硫替代；The above heterocycloalkyl group is a 3-12 membered heterocycloalkyl group, and any or any of several carbon atoms is replaced by oxygen, nitrogen or sulfur;
上述杂环烷基上的碳原子还可以为C＝O基和/或SO和/或SO 2基替代； The carbon atom on the above heterocycloalkyl group may also be a C=O group and/or a SO and/or SO 2 group;
上述取代的杂环烷基为环上任一或任几磺酰基、烷基磺酰基、烷基、烷氧基、羟基、氨基、硝基、羰基取代的氮杂、氧杂或硫杂的四元、五元、六元或七元环的环烷基；The above substituted heterocycloalkyl group is a quaternary ring of any or any of a plurality of sulfonyl groups, alkylsulfonyl groups, alkyl groups, alkoxy groups, hydroxyl groups, amino groups, nitro groups, carbonyl-substituted aza, oxa or thia groups. a five-, six- or seven-membered cycloalkyl group;
上述取代的杂环烷基上还可以具有-N-R 422的结构； The above substituted heterocycloalkyl group may further have a structure of -NR 422 ;
上述R 422为磺酰基、烷基磺酰基、烷基、羟基； The above R 422 is a sulfonyl group, an alkylsulfonyl group, an alkyl group, or a hydroxyl group;
上述R 42还可以选自如下结构的基团： The above R 42 may also be selected from the group consisting of the following structures:
Figure PCTCN2018087629-appb-000003
Figure PCTCN2018087629-appb-000003
上述R 43、R 44为相同或不相同的烷基、羟基、羟基取代的碳原子数不大于5的烷基； The above R 43 and R 44 are the same or different alkyl group, a hydroxyl group or a hydroxyl group substituted with an alkyl group having not more than 5 carbon atoms;
G3为3-12元的环；G3 is a ring of 3-12 yuan;
上述G3的环上的碳原子还可以部分的为氧、硫、氮、C＝O或SO 2替代； The carbon atom on the ring of G3 above may also be partially replaced by oxygen, sulfur, nitrogen, C=O or SO 2 ;
上述R 45为G3环上一个或几个取代的烷基、羟基、烷氧基、氨基。 R 45 above is an alkyl group, a hydroxyl group, an alkoxy group or an amino group substituted on the G 3 ring.
进一步地，本发明提供的一种含磷化合物，还具有这样的特点：即、为如下结构所示的化合物：Further, the phosphorus-containing compound provided by the present invention further has the following characteristics: that is, a compound represented by the following structure:
Figure PCTCN2018087629-appb-000004
Figure PCTCN2018087629-appb-000004
其中，上述C 1和C 2均为碳原子，其之间的碳键为单键、双键或三键。 Wherein C 1 and C 2 are each a carbon atom, and the carbon bond therebetween is a single bond, a double bond or a triple bond.
进一步地，本发明提供的一种含磷化合物，还具有这样的特点：即、为如下结构所示的化合物：Further, the phosphorus-containing compound provided by the present invention further has the following characteristics: that is, a compound represented by the following structure:
Figure PCTCN2018087629-appb-000005
Figure PCTCN2018087629-appb-000005
R 11选自苯环上任一或任几取代的氢、烷基、烷氧基、卤素、氨基、氰基、羟基、硝基； R 11 is selected from any one or more substituted hydrogen, alkyl, alkoxy, halogen, amino, cyano, hydroxy, nitro;
R 2选自羟基、烷基、烷氧基、卤素； R 2 is selected from the group consisting of hydroxyl, alkyl, alkoxy, halogen;
R 3为苯环上任一或任几取代的氢、烷基、烷氧基、卤素、氨基、氰基、羟基、硝基、芳基； R 3 is any optionally substituted hydrogen, alkyl, alkoxy, halogen, amino, cyano, hydroxy, nitro, aryl;
Y 1选自氢、烷基、芳基； Y 1 is selected from the group consisting of hydrogen, alkyl, and aryl;
R 41选自苯环上任一或任几取代的氢、烷基、烷氧基、烷基磺酰基、芳基磺酰基、卤素、氨基、氰基、羟基、硝基； R 41 is selected from any one or more substituted hydrogen, alkyl, alkoxy, alkylsulfonyl, arylsulfonyl, halogen, amino, cyano, hydroxy, nitro;
R 5选自氢、烷基、芳基、苄基及其衍生物。 R 5 is selected from the group consisting of hydrogen, alkyl, aryl, benzyl and derivatives thereof.
进一步地，本发明提供的一种含磷化合物，还具有这样的特点：即、为如下结构所示的化合物：Further, the phosphorus-containing compound provided by the present invention further has the following characteristics: that is, a compound represented by the following structure:
Figure PCTCN2018087629-appb-000006
Figure PCTCN2018087629-appb-000006
其中，X 1、X 2、X 3、X 4选自氢、烷基、卤素、羟基。 Wherein X 1 , X 2 , X 3 and X 4 are selected from the group consisting of hydrogen, alkyl, halogen and hydroxyl.
进一步地，本发明提供的一种含磷化合物，还具有这样的特点：即、上述含磷化合物选自如下化合物：Further, the phosphorus-containing compound provided by the present invention has the feature that the phosphorus-containing compound is selected from the following compounds:
(2s)-2-(2，6-二氯-4-(2-(羟基(苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸；(2s)-2-(2,6-Dichloro-4-(2-(hydroxy(phenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl)phenyl) Propionic acid
(2s)-2-(2，6-二氯-4-((羟基(3-羟基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸；(2s)-2-(2,6-Dichloro-4-((hydroxy(3-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl)phenyl) Propionic acid
(2s)-2-(2，6-二氯-4-(2-(羟基(间羟基苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸；(2s)-2-(2,6-Dichloro-4-(2-(hydroxy(m-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl)benzene Propionate
(2s)-2-(2，6-二氯-4-(2-(甲氧基(苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸；(2s)-2-(2,6-Dichloro-4-(2-(methoxy(phenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl)benzene Propionate
(2s)-2-(2，6-二氯-4-(甲氧基(苯基)磷酰基)乙炔基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸；(2s)-2-(2,6-Dichloro-4-(methoxy(phenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl)phenyl)propene acid;
(2s)-2-(2，6-二氯-4-(2-(乙氧基(间羟基苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸；(2s)-2-(2,6-Dichloro-4-(2-(ethoxy(m-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl) Phenyl) propionic acid;
(2s)-2-(2，6-二氯-4-((甲氧基(3-羟基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸；(2s)-2-(2,6-Dichloro-4-((methoxy(3-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl)benzene Propionate
(2s)-2-(2，6-二氯-4-(2-(甲氧基(3-羟基苯基)磷酰基)乙基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸；(2s)-2-(2,6-Dichloro-4-(2-(methoxy(3-hydroxyphenyl)phosphoryl)ethyl)benzylamino)-3-(3-(methylsulfonyl) Phenyl) propionic acid;
(2s)-2-(2，6-二氯-4-(甲基(苯基)磷酰基)乙炔基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸；(2s)-2-(2,6-Dichloro-4-(methyl(phenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl)phenyl)propanoic acid ;
(2s)-2-(2，6-二氯-4-(2-(甲基(苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸；(2s)-2-(2,6-Dichloro-4-(2-(methyl(phenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl)phenyl Propionic acid
(2s)-2-(2，6-二氯-4-((甲基(3-羟基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸；(2s)-2-(2,6-Dichloro-4-((methyl(3-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl)phenyl Propionic acid
(2s)-2-(2，6-二氯-4-((甲氧基(3-甲氧基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸；(2s)-2-(2,6-Dichloro-4-((methoxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl) Phenyl) propionic acid;
(2s)-2-(2，6-二氯-4-((羟基(3-甲氧基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸。(2s)-2-(2,6-Dichloro-4-((hydroxy(3-methoxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl)benzene Base) propionic acid.
另外，本发明还提供了上述含磷化合物的制备方法，其特征在于：In addition, the present invention also provides a method for preparing the above phosphorus-containing compound, which is characterized in that:
由化合物A和化合物C，依次与化合物B上的活性点位进行反应后获得；Obtained by reacting Compound A and Compound C with the active sites on Compound B in sequence;
其中，上述化合物A为如下结构所示的化合物：Wherein the above compound A is a compound represented by the following structure:
Figure PCTCN2018087629-appb-000007
Figure PCTCN2018087629-appb-000007
上述化合物C为如下结构所示的化合物：The above compound C is a compound represented by the following structure:
Figure PCTCN2018087629-appb-000008
Figure PCTCN2018087629-appb-000008
上述化合物B为如下结构所示的化合物：The above compound B is a compound represented by the following structure:
Figure PCTCN2018087629-appb-000009
Figure PCTCN2018087629-appb-000009
其中，L 1与L 1‘以及L 2与L 2‘分别为一对可相互发生反应的活性基团，在反应的过程中，通过L 1与L 1‘反应，L 2与L 2‘反应获得目标产物； Wherein, L 1 L 1 'and L 2 and L 2', respectively, a pair of reactive groups can react with each other, in the course of the reaction, by L 1 and L 1 'reaction, L 2 and L 2' is reacted with Obtaining the target product;
R 1选自烷基、芳基、苄基及其衍生物； R 1 is selected from the group consisting of alkyl, aryl, benzyl and derivatives thereof;
R 2选自羟基、烷基、烷氧基、卤素； R 2 is selected from the group consisting of hydroxyl, alkyl, alkoxy, halogen;
n选自1-3的自然数；n is selected from a natural number of 1-3;
R 3为苯环上任一或任几取代的氢、烷基、烷氧基、卤素、氨基、氰基、羟基、硝基、芳基； R 3 is any optionally substituted hydrogen, alkyl, alkoxy, halogen, amino, cyano, hydroxy, nitro, aryl;
R 4选自烷基、烷氧基、芳基、苄基及其衍生物； R 4 is selected from the group consisting of alkyl, alkoxy, aryl, benzyl and derivatives thereof;
R 5选自氢、烷基、芳基、苄基及其衍生物。 R 5 is selected from the group consisting of hydrogen, alkyl, aryl, benzyl and derivatives thereof.
进一步地，本发明提供的含磷化合物的制备方法，还具有这样的特点：即、上述L 1与L 1‘以及L 2与L 2‘之间可以发生，取代反应、加成反应、消除反应或置换反应，从而形成L 1与L 1‘以及L 2与L 2‘之间的连接键。 Further, the method for preparing a phosphorus-containing compound provided by the present invention further has the characteristics that the above L 1 and L 1 ' and between L 2 and L 2 ' can occur, and the substitution reaction, the addition reaction, and the elimination reaction are carried out. Or the displacement reaction, thereby forming a linkage between L 1 and L 1 'and L 2 and L 2 '.
进一步地，本发明提供的含磷化合物的制备方法，还具有这样的特点：即、上述L 1选自卤素、氨基、氰基、硫基、羟基、烷氧基； Further, the method for preparing a phosphorus-containing compound provided by the present invention further has the feature that the above L 1 is selected from the group consisting of halogen, amino, cyano, thio, hydroxy, alkoxy;
上述L 1‘选自卤素、炔基、羧基、氨基、氰基、酯基、烷氧基、磺酰氨基基、烷氧基磺酰基； The above L 1 ' is selected from the group consisting of halogen, alkynyl, carboxy, amino, cyano, ester, alkoxy, sulfonylamino, alkoxysulfonyl;
上述L 2选自卤素、羧基、氨基、氰基、酯基、烷氧基、磺酰氨基基、烷氧基磺酰基； The above L 2 is selected from the group consisting of halogen, carboxyl, amino, cyano, ester, alkoxy, sulfonylamino, alkoxysulfonyl;
上述L 2‘选自卤素、氨基、硫基、羟基、烷氧基。 The above L 2 ' is selected from the group consisting of halogen, amino, thio, hydroxy and alkoxy.
进一步地，本发明提供的含磷化合物的制备方法，还具有这样的特点：即、上述化合物A与化合物C的摩尔比为1:0.1-10；Further, the method for preparing a phosphorus-containing compound provided by the present invention has the characteristic that the molar ratio of the above compound A to the compound C is 1:0.1-10;
上述化合物C与化合物B的摩尔比为1:0.1-10。The molar ratio of the above compound C to the compound B is 1:0.1-10.
进一步地，本发明提供的含磷化合物的制备方法，还具有这样的特点：即、具体工艺步骤如下所示：Further, the method for preparing a phosphorus-containing compound provided by the present invention further has the following characteristics: that is, the specific process steps are as follows:
步骤一、于磷酸二酯的衍生物中加入卤化试剂，于50-100℃的温度下反应1-5小时后，直接旋干获得底物1；Step 1, adding a halogenating reagent to the derivative of the phosphoric acid diester, reacting at a temperature of 50-100 ° C for 1-5 hours, and directly spinning to obtain a substrate 1;
在本步骤中，旨在将磷酸二酯的衍生物制作成具有活性反应基团的底物，如直接选用具有活性基团L 1的化合物A的话，可省略该步骤一。 In this step, it is intended to prepare a derivative of a phosphoric acid diester into a substrate having an active reactive group, and if the compound A having a reactive group L 1 is directly selected, the first step can be omitted.
在本发明中，磷酸二酯的衍生物为如下结构所示的化合物：In the present invention, the derivative of the phosphodiester is a compound represented by the following structure:
Figure PCTCN2018087629-appb-000010
Figure PCTCN2018087629-appb-000010
R 111、R 112、R 113选自芳基(如：苯基、萘基、喹啉基等芳香基团)、烷基(如：甲基、乙基、丙基、异丙基、环己基、环戊基等烷基基团)； R 111 , R 112 , and R 113 are selected from an aryl group (e.g., an aromatic group such as a phenyl group, a naphthyl group, or a quinolyl group), an alkyl group (e.g., a methyl group, an ethyl group, a propyl group, an isopropyl group, or a cyclohexyl group). , an alkyl group such as a cyclopentyl group);
卤化试剂一般选用二氯亚砜、光气、溴等提供卤素的试剂；The halogenating reagent generally uses a reagent for providing halogen such as chlorosulfoxide, phosgene or bromine;
该反应优选在氮气、氩气、氦气等保护气的保护下进行。The reaction is preferably carried out under the protection of a shielding gas such as nitrogen, argon or helium.
该反应中，每100mg的磷酸二酯的衍生物，添加的卤化试剂的量为0.5-4ml。In the reaction, the amount of the halogenating agent added is 0.5 to 4 ml per 100 mg of the derivative of the phosphodiester.
步骤二、于0℃以下的温度下，于乙炔基苯甲酸甲酯的衍生物中，依次加入格氏试剂和底物1，反应0.1-2小时，用酸溶液淬灭反应，萃取有机相旋干，得到中间产物1；Step 2, in the derivative of methyl ethynylbenzoate at a temperature below 0 ° C, the Grignard reagent and the substrate 1 are sequentially added, the reaction is carried out for 0.1-2 hours, the reaction is quenched with an acid solution, and the organic phase is extracted. Dry to obtain intermediate product 1;
在本发明中，乙炔基苯甲酸甲酯的衍生物为如下结构所示的化合物：In the present invention, the derivative of methyl ethynylbenzoate is a compound represented by the following structure:
Figure PCTCN2018087629-appb-000011
Figure PCTCN2018087629-appb-000011
其中，R 311选自任一或任几取代的卤素、硝基、芳基(如：苯基、萘基、喹啉基等芳香基团)、烷基(如：甲基、乙基、丙基、异丙基、环己基、环戊基等烷基基团)等； Wherein R 311 is selected from any or a plurality of substituted halogen, nitro, aryl (eg, aryl, naphthyl, quinolyl, etc.), alkyl (eg, methyl, ethyl, propyl) An alkyl group such as an isopropyl group, a cyclohexyl group or a cyclopentyl group;
该乙炔基和甲酸甲酯基团可以为对位、邻位或间位；The ethynyl and methyl formate groups may be para, ortho or meta;
该乙炔基苯甲酸甲酯的衍生物与格氏试剂和底物1的质量比例为1:0.01-10:1:-10；The mass ratio of the derivative of methyl ethynylbenzoate to Grignard reagent and substrate 1 is 1:0.01-10:1:-10;
该反应优选在氮气、氩气、氦气等保护气的保护下进行；The reaction is preferably carried out under the protection of a protective gas such as nitrogen, argon or helium;
该反应优选在醚类溶剂中进行；The reaction is preferably carried out in an ether solvent;
用于淬灭反应的酸优选为无机酸，该酸的浓度优选为0.5-1.5mol/L，该酸的用量优选为反应物总量的0.01-10倍；The acid used for the quenching reaction is preferably a mineral acid, the concentration of the acid is preferably from 0.5 to 1.5 mol/L, and the amount of the acid is preferably from 0.01 to 10 times the total amount of the reactant;
该萃取用的试剂优选为酯溶剂。The reagent for extraction is preferably an ester solvent.
步骤三、将中间产物1和脱酯基试剂，在100-150摄氏度的温度下反应2-5小时，加入酸溶液，萃取有机相旋干，得到中间产物2；Step 3, the intermediate product 1 and the de-esterification reagent, reacted at a temperature of 100-150 degrees Celsius for 2-5 hours, adding an acid solution, extracting the organic phase to spin dry, to obtain an intermediate product 2;
该中间产物1和脱酯基试剂的质量比为1:0.5-3；The mass ratio of the intermediate product 1 and the deesterification reagent is 1:0.5-3;
该反应优选在氮气、氩气、氦气等保护气的保护下进行；The reaction is preferably carried out under the protection of a protective gas such as nitrogen, argon or helium;
用于淬灭反应的酸优选为无机酸，该酸的浓度优选为0.5-1.5mol/L，该酸的用量优选为反应物总量的0.01-10倍；The acid used for the quenching reaction is preferably a mineral acid, the concentration of the acid is preferably from 0.5 to 1.5 mol/L, and the amount of the acid is preferably from 0.01 to 10 times the total amount of the reactant;
该萃取用的试剂优选为酯溶剂。The reagent for extraction is preferably an ester solvent.
步骤四、于中间产物2中，依次加入L 2为氨基的化合物C，碱性催化剂，在20-50℃的温度下，反应1-10小时，用酸溶液淬灭反应，萃取有机相旋干，得到含炔基的含磷化合物。 Step 4, in the intermediate product 2, sequentially add L 2 as the amino group of the compound C, a basic catalyst, at a temperature of 20-50 ° C, the reaction is 1-10 hours, quench the reaction with an acid solution, extract the organic phase spin dry A phosphorus-containing compound containing an alkynyl group is obtained.
该中间产物2、化合物C和碱性催化剂的摩尔比为1:1-5:5-20；The molar ratio of the intermediate product 2, the compound C and the basic catalyst is 1:1-5:5-20;
用于淬灭反应的酸优选为无机酸，该酸的浓度优选为0.5-1.5mol/L，该酸的用量优选为反应物总量的0.01-10倍；The acid used for the quenching reaction is preferably a mineral acid, the concentration of the acid is preferably from 0.5 to 1.5 mol/L, and the amount of the acid is preferably from 0.01 to 10 times the total amount of the reactant;
该萃取用的试剂优选为酯溶剂。The reagent for extraction is preferably an ester solvent.
上述反应过程均适用于不提纯直接进行下一步骤的方案，其每步的产率约为50-95％，总产率约为50-80％。The above reaction procedures are all applicable to the scheme in which the next step is carried out without purification, and the yield per step is about 50 to 95%, and the total yield is about 50 to 80%.
上述过程的具体方程式如下所示：The specific equations for the above process are as follows:
Figure PCTCN2018087629-appb-000012
Figure PCTCN2018087629-appb-000012
进一步地，本发明提供的一种含磷化合物的制备方法，还具有这样的特点，即、上述含炔基的含磷化合物经还原反应、酯化反应、酰胺化反应、取代反应、加成反应中的一种或几种反应后,可得到相应的含磷产物。Further, the method for preparing a phosphorus-containing compound provided by the present invention has the feature that the alkynyl-containing phosphorus-containing compound undergoes a reduction reaction, an esterification reaction, an amidation reaction, a substitution reaction, and an addition reaction. After one or more of the reactions, the corresponding phosphorus-containing product can be obtained.
此外，本发明还提供了上述含磷化合物的应用，其特征在于：能作为一种免疫细胞迁徙抑制剂使用。Further, the present invention provides the use of the above phosphorus-containing compound, which is characterized in that it can be used as an immune cell migration inhibitor.
另外，本发明还提供了上述含磷化合物的应用，其特征在于：包含有上述含磷化合物的眼药水，可用于缓解和治疗干眼症。Further, the present invention provides the use of the above phosphorus-containing compound, characterized in that the eye drops containing the above phosphorus-containing compound are useful for alleviating and treating dry eye.
该眼药水制剂的制备方法可以为任何常规制剂法。The preparation method of the ophthalmic preparation can be any conventional preparation method.
例如：将上述化合物加入到10-200倍重量的无菌生理盐水中，加入0.01-1倍的碱溶液，搅拌得到透明溶液；再向以上得到的溶液中加入缓冲溶液中直至溶液的pH在6.5-7.5之间；再向得到水溶液加入无菌生理盐水，直到总体积达到原体积的1.5-20倍。再向以上溶液通氮气，鼓泡0.5-10小时，所得溶液密封，放在5℃避光保存。分装到一次性使用的滴眼液袋中待用。 其中，上述氢氧化钠 和NaH 2PO 4饱和水溶液可以被其他缓冲溶液替代。 For example, the above compound is added to 10-200 times by weight of sterile physiological saline, 0.01-1 times of alkali solution is added, and stirred to obtain a transparent solution; and the buffer solution is added to the solution obtained above until the pH of the solution is 6.5. Between -7.5; add sterile physiological saline to the obtained aqueous solution until the total volume reaches 1.5-20 times the original volume. The above solution was again purged with nitrogen, bubbling for 0.5-10 hours, and the resulting solution was sealed and stored at 5 ° C in the dark. Dispense into a disposable eye drop bag for use. Wherein the sodium hydroxide and a saturated aqueous solution of NaH 2 PO 4 may be replaced by other buffer solutions.
本发明的作用和效果：The effects and effects of the present invention:
在本发明中，合成了一类新的含磷化合物，为新型的免疫细胞迁徙抑制剂。其具有很好的亲水性，容易开发成眼药水，对免疫细胞迁徙有很强的抑制能力，可能会缓解大多数干眼症病人的症状。In the present invention, a new class of phosphorus-containing compounds is synthesized, which is a novel immune cell migration inhibitor. It has good hydrophilicity, is easy to develop into eye drops, has a strong inhibitory effect on immune cell migration, and may alleviate the symptoms of most dry eye patients.
具体实施方式Detailed ways
实施例1Example 1
Figure PCTCN2018087629-appb-000013
Figure PCTCN2018087629-appb-000013
(2s)-2-(2，6-二氯-4-(2-(羟基(苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(hydroxy(phenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl)phenyl) Propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000014
Figure PCTCN2018087629-appb-000014
步骤A：甲氧基苯基磷酰氯(化合物1.1)Step A: methoxyphenylphosphoryl chloride (Compound 1.1)
称取150mg苯基磷酸二甲酯，加入4ml二氯亚砜，氮气保护，在75度下反应2小时后直接旋干即可。150 mg of dimethyl phenyl phosphate was weighed, added with 4 ml of thionyl chloride, and protected with nitrogen. After reacting at 75 °C for 2 hours, it was directly dried.
步骤B：2,6-二氯-4-((苯基(甲氧基)磷酰基)乙炔基)苯甲酸甲酯(化合物1.2)Step B: Methyl 2,6-dichloro-4-((phenyl(methoxy)phosphoryl)ethynyl)benzoate (Compound 1.2)
将50mg的2,6-二氯-4-乙炔基苯甲酸甲酯溶于1ml四氢呋喃中，氮气保护，并在0度下加入0.2ml的2mol/L的异丙基氯化镁，搅拌20分钟；将化合物1.1溶于0.5ml的四氢呋喃并加入，反应20分钟。用1mol/L的稀盐酸溶液淬灭反应，用30mL乙酸乙酯分三次萃取，合并有机相，旋干，纯化得产物(50mg，60％)。LCMS ESI(+)m/z:382.6(M+1).50 mg of methyl 2,6-dichloro-4-ethynylbenzoate was dissolved in 1 ml of tetrahydrofuran, protected with nitrogen, and 0.2 ml of 2 mol/L of isopropylmagnesium chloride was added at 0 °C, and stirred for 20 minutes; Compound 1.1 was dissolved in 0.5 ml of tetrahydrofuran and added for 20 minutes. The reaction was quenched with 1 mol/L EtOAc (EtOAc)EtOAc. LCMS ESI (+) m/z: 382.6 (M+1).
步骤C：2,6-二氯-4-((羟基(苯基)磷酰基)乙炔基)苯甲酸(化合物1.3)Step C: 2,6-Dichloro-4-((hydroxy(phenyl)phosphoryl)ethynyl)benzoic acid (Compound 1.3)
将化合物1.2(50mg)和碘化锂(50mg)溶于1ml吡啶中氮气保护，在120度搅拌3个小时，冷却旋干，加入10ml的1mol/L的稀盐酸溶液，用30ml的乙酸乙酯分3次萃取，合并有机相，旋干无需纯化。LCMS ESI(+)m/z:354.6(M+1).步骤D：(2s)-2-(2，6-二氯-4-((羟基(苯基)磷酰基)乙炔基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸苄酯(化合物1.4)Compound 1.2 (50 mg) and lithium iodide (50 mg) were dissolved in 1 ml of pyridine, protected with nitrogen, stirred at 120 °C for 3 hours, cooled and dried, and 10 ml of a 1 mol/L dilute hydrochloric acid solution was added to 30 ml of ethyl acetate. The extraction was carried out in 3 portions, and the organic phases were combined and dried without further purification. LCMS ESI (+) m/z: 354.6 (M + 1). Step D: (2s)-2-(2,6-dichloro-4-((hydroxy(phenyl)phosphoryl)ethynyl) Benzyl)-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester (Compound 1.4)
将化合物1.3溶于DMF中，加入(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸苄酯盐酸盐(2eq)，后加入DIPEA(10eq)，HATU(2.5eq)。在常温下搅拌4h，加入10ml稀盐酸溶液，用EA萃取3次，合并有机相，旋干。用反相制备纯化，45度减压旋干得目标产物40mg。LCMS ESI(+)m/z:669.5(M+1).Compound 1.3 was dissolved in DMF and (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester hydrochloride (2 eq) was added followed by DIPEA (10 eq), HATU ( 2.5eq). After stirring at normal temperature for 4 h, 10 ml of dilute hydrochloric acid solution was added, and extracted with EA three times, and the organic phases were combined and dried. Purification by reverse phase preparation, spin-drying at 45 ° under reduced pressure afforded 40 mg. LCMS ESI (+) m/z: 669.5 (M+1).
步骤E：(2s)-2-(2，6-二氯-4-(2-(羟基(苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(化合物1)Step E: (2s)-2-(2,6-Dichloro-4-(2-(hydroxy(phenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl) Phenyl)propionic acid (compound 1)
将化合物1.4溶于1ml的甲醇中，加入Pd/C(10％，0.1eq)，常压氢化1h,过滤，旋干，反相制备，冻干的产物10mg。LCMS ESI(+)m/z:583.6(M+1)； 1H-NMR(400MHz,DMSO)δ9.02(d,J＝8Hz,1H),7.86(s,1H)，7.77(m,3H)，7.66(m,1H)，7.55(m,2H)，7.51(m,2H),7.29(s,2H),4.75(m,1H),3.29(dd,J＝15Hz,J＝4.4Hz,1H),3.15(s,3H),3.03(dd,J＝15.5Hz,J＝10.4Hz,1H),2.70(m,2H),2.11(m,2H)。 Compound 1.4 was dissolved in 1 ml of methanol, Pd/C (10%, 0.1 eq) was added, and then hydrogenated under normal pressure for 1 h, filtered, dried, and then evaporated. LCMS ESI (+) m/z: 58 </RTI> (M + 1); 1 H-NMR (400 MHz, DMSO) δ 9.02 (d, J=8 Hz, 1H), 7.86 (s, 1H), 7.77 (m, 3H) ), 7.66 (m, 1H), 7.55 (m, 2H), 7.51 (m, 2H), 7.29 (s, 2H), 4.75 (m, 1H), 3.29 (dd, J = 15 Hz, J = 4.4 Hz, 1H), 3.15 (s, 3H), 3.03 (dd, J = 15.5 Hz, J = 10.4 Hz, 1H), 2.70 (m, 2H), 2.11 (m, 2H).
实施例2Example 2
Figure PCTCN2018087629-appb-000015
Figure PCTCN2018087629-appb-000015
(2s)-2-(2，6-二氯-4-((羟基(3-羟基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-((hydroxy(3-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl)phenyl) Propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000016
Figure PCTCN2018087629-appb-000016
步骤A：(间甲氧基苯基)乙氧基磷酰氯(化合物2.1)Step A: (m-methoxyphenyl)ethoxyphosphoryl chloride (Compound 2.1)
称取200mg间甲氧基苯基磷酸二乙酯，加入4ml二氯亚砜，氮气保护，在75度下反应12小时后直接旋干即可。200 mg of m-methoxyphenylphosphoric acid diethyl ester was weighed, and 4 ml of thionyl chloride was added thereto, and the mixture was shielded by nitrogen gas, and reacted at 75 °C for 12 hours, and then directly dried.
步骤B：2,6-二氯-4-(((间甲氧基苯基)(乙氧基)磷酰基)乙炔基)苯甲酸甲酯(化合物2.2)Step B: Methyl 2,6-dichloro-4-((m-methoxyphenyl)(ethoxy)phosphoryl)ethynyl)benzoate (Compound 2.2)
将100mg的2,6-二氯-4-乙炔基苯甲酸甲酯溶于1.5ml四氢呋喃中，氮气保护，并在0度下加入0.7ml的2mol/L的异丙基氯化镁，搅拌20分钟；将化合物2.1溶于0.5ml的四氢呋喃并加入，反应20分钟。用1mol/L的稀盐酸溶液淬灭反应，用30mL乙酸乙酯分三次萃取，合并有机相，旋干，纯化得产物(100mg，60％)。LCMS ESI(+)m/z:426.6(M+1).100 mg of methyl 2,6-dichloro-4-ethynylbenzoate was dissolved in 1.5 ml of tetrahydrofuran, protected with nitrogen, and 0.7 ml of 2 mol/L of isopropylmagnesium chloride was added at 0 °C, and stirred for 20 minutes; Compound 2.1 was dissolved in 0.5 ml of tetrahydrofuran and added for 20 minutes. The reaction was quenched with 1 mol/L EtOAc (EtOAc)EtOAc. LCMS ESI (+) m/z: 426.6 (M+1).
步骤C：2,6-二氯-4-((羟基(间甲氧苯基)磷酰基)乙炔基)苯甲酸(化合物2.3)Step C: 2,6-Dichloro-4-((hydroxy(m-methoxyphenyl)phosphoryl)ethynyl)benzoic acid (Compound 2.3)
将化合物2.2(100mg)和碘化锂(100mg)溶于2ml吡啶中氮气保护，在120度搅拌3个小时，冷却旋干，加入10ml的1mol/L的稀盐酸溶液，用30ml的乙酸乙酯分3次萃取，合并有机相，旋干无需纯化。LCMS ESI(+)m/z:384.6(M+1).Compound 2.2 (100 mg) and lithium iodide (100 mg) were dissolved in 2 ml of pyridine in nitrogen, stirred at 120 °C for 3 hours, cooled and dried, and 10 ml of 1 mol/L dilute hydrochloric acid solution was added to 30 ml of ethyl acetate. The extraction was carried out in 3 portions, and the organic phases were combined and dried without further purification. LCMS ESI (+) m/z: 384.6 (M+1).
步骤D：(2s)-2-(2，6-二氯-4-((羟基(间甲氧苯基)磷酰基)乙炔基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸苄酯(化合物2.4)Step D: (2s)-2-(2,6-Dichloro-4-((hydroxy(m-methoxyphenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl) Phenyl) benzyl propionate (compound 2.4)
将化合物2.3溶于DMF中，加入(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸苄酯盐酸盐(2eq)，后加入DIPEA(10eq)，HATU(2.5eq)。在常温下搅拌4h，加入10ml稀盐酸溶液，用EA萃取3次，合并有机相，旋干。用反相制备纯化，45度减压旋干得目标产物80mg。LCMS ESI(+)m/z:699.5(M+1).Compound 2.3 was dissolved in DMF, and (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester hydrochloride (2 eq) was added followed by DIPEA (10 eq), HATU ( 2.5eq). After stirring at normal temperature for 4 h, 10 ml of dilute hydrochloric acid solution was added, and extracted with EA three times, and the organic phases were combined and dried. Purification by reverse phase preparation, spin-drying at 45 ° under reduced pressure afforded the desired product, 80 mg. LCMS ESI (+) m/z: 699.5 (M+1).
步骤E：(2s)-2-(2，6-二氯-4-((羟基(3-羟基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(化合物2)Step E: (2s)-2-(2,6-Dichloro-4-((hydroxy(3-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl) Phenyl) propionic acid (compound 2)
将化合物2.4(40mg)溶于1ml的DCM中，氮气保护，在-40度下加入0.2ml三溴化硼(1mol/L)，然后在0度下搅拌30分钟。在-40度下加入水淬灭，用30mlEA萃取，用无水硫酸钠干燥，旋干，纯化得15mg产物。Compound 2.4 (40 mg) was dissolved in 1 ml of DCM, and then filtered under nitrogen, and 0.2 ml of boron tribromide (1 mol/L) was added at -40 °, and then stirred at 0 ° for 30 minutes. It was quenched by the addition of water at -40 °, extracted with 30 mL of EA, dried over anhydrous sodium sulfate.
LCMS ESI(+)m/z:595.5(M+1).LCMS ESI (+) m/z: 595.5 (M+1).
1H-NMR(400MHz,DMSO),δ9.16(d,J＝8.4Hz,1H),7.86(s,1H),7.76(d,J＝7.6Hz,1H),7.67(d,J＝7.6Hz,1H),7.56(dd,J＝8Hz,J＝7.6Hz,1H),7.44(s,2H),7.28(m,1H),7.16(m,2H),6.77(m,1H),4.78(m,1H),3.29(m,1H),3.14(s,3H),3.01(dd,J＝14,J＝10.4,1H). 1 H-NMR (400 MHz, DMSO), δ 9.16 (d, J = 8.4 Hz, 1H), 7.86 (s, 1H), 7.76 (d, J = 7.6 Hz, 1H), 7.67 (d, J = 7.6) Hz, 1H), 7.56 (dd, J = 8 Hz, J = 7.6 Hz, 1H), 7.44 (s, 2H), 7.28 (m, 1H), 7.16 (m, 2H), 6.77 (m, 1H), 4.78 (m, 1H), 3.29 (m, 1H), 3.14 (s, 3H), 3.01 (dd, J = 14, J = 10.4, 1H).
实施例3Example 3
Figure PCTCN2018087629-appb-000017
Figure PCTCN2018087629-appb-000017
(2s)-2-(2，6-二氯-4-(2-(羟基(间羟基苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(hydroxy(m-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl)benzene Propionate
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000018
Figure PCTCN2018087629-appb-000018
将化合物2.5(10mg)溶于1ml甲醇，加入1mgPd/C(10％)，常压氢化1.5h后过滤。旋干纯化得3mg产物。LCMS ESI(+)m/z:599.6(M+1).Compound 2.5 (10 mg) was dissolved in 1 ml of methanol, and 1 mg of Pd/C (10%) was added thereto, and hydrogenated at normal pressure for 1.5 hr. The product was purified by spin-drying to give 3 mg. LCMS ESI (+) m/z: 599.6 (M+1).
1H-NMR(400MHz,DMSO)δ9.72(s,1H),9.04(d,J＝8.4Hz,1H),7.86(s,1H),7.77(d,J＝8Hz,1H,)，7.67(d,J＝7.6Hz,1H),7.56(t,J＝7.6Hz,1H),7.33(m,1H)，7.28(s,2H),7.15(m,2H),6.93(m,1H),4.75(m,1H),3.30(m,1H),3.15(s,3H),3.01(dd,J＝10.8Hz,J＝9.6Hz,1H),2.69(m,2H),2.04(m,2H). 1 H-NMR (400 MHz, DMSO) δ 9.72 (s, 1H), 9.04 (d, J = 8.4 Hz, 1H), 7.86 (s, 1H), 7.77 (d, J = 8 Hz, 1H,), 7.67 (d, J = 7.6 Hz, 1H), 7.56 (t, J = 7.6 Hz, 1H), 7.33 (m, 1H), 7.28 (s, 2H), 7.15 (m, 2H), 6.93 (m, 1H) , 4.75 (m, 1H), 3.30 (m, 1H), 3.15 (s, 3H), 3.01 (dd, J = 10.8 Hz, J = 9.6 Hz, 1H), 2.69 (m, 2H), 2.04 (m, 2H).
实施例4Example 4
Figure PCTCN2018087629-appb-000019
Figure PCTCN2018087629-appb-000019
(2s)-2-(2，6-二氯-4-(2-(甲氧基(苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(methoxy(phenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl)benzene Propionate
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000020
Figure PCTCN2018087629-appb-000020
步骤A：(2s)-2-(2，6-二氯-4-((甲氧基(苯基)磷酰基)乙炔基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸苄酯(化合物4.1)Step A: (2s)-2-(2,6-Dichloro-4-((methoxy(phenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl) Phenyl) benzyl propionate (Compound 4.1)
将化合物1.4(20mg)溶于0.5ml甲醇中，加入三甲基硅基重氮甲烷(3eq)，在常温下搅拌30分钟。加入适量的乙酸淬灭，旋干，加入5ml的稀盐酸溶液，用EA萃取3次，合并有机相旋干即可。LCMS ESI(+)m/z:683.6(M+1).Compound 1.4 (20 mg) was dissolved in 0.5 ml of methanol, trimethylsilyldiazomethane (3 eq) was added, and stirred at room temperature for 30 minutes. Quenched with an appropriate amount of acetic acid, spun dry, add 5 ml of dilute hydrochloric acid solution, extract 3 times with EA, and combine the organic phase to dry. LCMS ESI (+) m/z: 683.6 (M+1).
步骤B：(2s)-2-(2，6-二氯-4-(2-(甲氧基(苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(化合物4)Step B: (2s)-2-(2,6-Dichloro-4-(2-(methoxy(phenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonate) Acyl)phenyl)propionic acid (compound 4)
将化合物4.1溶于甲醇(1ml)中，加入1mgPd/C(10％)，常压氢化1h，过滤旋干，纯化得目标产物。LCMS ESI(+)m/z:597.6(M+1).The compound 4.1 was dissolved in methanol (1 ml), and 1 mg of Pd/C (10%) was added thereto, and the mixture was hydrogenated at normal pressure for 1 hour, and the mixture was evaporated to dryness to give the desired product. LCMS ESI (+) m/z: 597.6 (M+1).
1H-NMR(400MHz,DMSO)δ9.03(d,J＝8.4Hz,1H),7.86(s,1H),7.75(m,3H),7.66(m,2H),7.56(m,3H),7.32(s,2H),4.75(m,1H),3.51(d,J＝11.2Hz,3H),3.28(dd,J＝14.4Hz,J＝3.6Hz,1H),3.15(s,3H),3.01(dd,J＝14.4Hz,J＝10.8Hz,1H),2.72(m,2H),2.34(m,2H). 1 H-NMR (400 MHz, DMSO) δ 9.03 (d, J = 8.4 Hz, 1H), 7.86 (s, 1H), 7.75 (m, 3H), 7.66 (m, 2H), 7.56 (m, 3H) , 7.32 (s, 2H), 4.75 (m, 1H), 3.51 (d, J = 11.2 Hz, 3H), 3.28 (dd, J = 14.4 Hz, J = 3.6 Hz, 1H), 3.15 (s, 3H) , 3.01 (dd, J = 14.4 Hz, J = 10.8 Hz, 1H), 2.72 (m, 2H), 2.34 (m, 2H).
实施例5Example 5
Figure PCTCN2018087629-appb-000021
Figure PCTCN2018087629-appb-000021
(2s)-2-(2，6-二氯-4-(甲氧基(苯基)磷酰基)乙炔基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(methoxy(phenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl)phenyl)propene acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000022
Figure PCTCN2018087629-appb-000022
将化合物4.1溶于DCM中，在-40度下加入1mol/L的三溴化硼(10eq)，在0度下搅拌30分钟后在-40度下加水淬灭反应。用EA萃取3次，合并有机相， 干燥旋干，纯化得目标产物。LCMS ESI(+)m/z:593.6(M+1).Compound 4.1 was dissolved in DCM, and 1 mol/L of boron tribromide (10 eq) was added at -40 °C, stirred at 0 °C for 30 minutes and then quenched with water at -40 °C. It was extracted 3 times with EA, and the organic phases were combined, dried and dried to give the desired product. LCMS ESI (+) m/z: 593.6 (M+1).
1H-NMR(400MHz,DMSO)δ9.21(d,J＝8.4Hz,1H),7.88(m,5H),7.77(m,1H),7.72(m,1H),7.67(m,1H),7.63(m,2H),7.57(m,1H),3.83(d,J＝12.4Hz,3H),3.30(m,1H),3.15(s,3H),3.03(dd,J＝13.6Hz,J＝10.4Hz,1H).1H-NMR (400MHz, DMSO) δ 9.21 (d, J = 8.4 Hz, 1H), 7.88 (m, 5H), 7.77 (m, 1H), 7.72 (m, 1H), 7.67 (m, 1H), 7.63 (m, 2H), 7.57 (m, 1H), 3.83 (d, J = 12.4 Hz, 3H), 3.30 (m, 1H), 3.15 (s, 3H), 3.03 (dd, J = 13.6 Hz, J =10.4Hz, 1H).
实施例6Example 6
Figure PCTCN2018087629-appb-000023
Figure PCTCN2018087629-appb-000023
(2s)-2-(2，6-二氯-4-(2-(乙氧基(间羟基苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(ethoxy(m-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl) Phenyl) propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000024
Figure PCTCN2018087629-appb-000024
步骤A：2,6-二氯-4-(((间羟基苯基)(乙氧基)磷酰基)乙炔基)苯甲酸(化合物6.1)Step A: 2,6-Dichloro-4-(((hydroxyphenyl)(ethoxy)phosphoryl)ethynyl)benzoic acid (Compound 6.1)
将化合物2.2溶于DCM中在低温下加入1mol/L的三溴化硼(10eq)，在0度下搅拌30分钟，后在-40度下淬灭反应，EA萃取3次，合并有机相旋干即可。Compound 2.2 was dissolved in DCM, and 1 mol/L of boron tribromide (10 eq) was added at a low temperature, and stirred at 0 °C for 30 minutes, then quenched at -40 °C, extracted with EA three times, and combined with organic phase rotation. Just do it.
LCMS ESI(+)m/z:398.6(M+1).LCMS ESI (+) m/z: 398.6 (M+1).
步骤B：(2s)-2-(2，6-二氯-4-((乙氧基(间羟基苯基)磷酰基)乙炔基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸苄酯(化合物6.2)Step B: (2s)-2-(2,6-Dichloro-4-((ethoxy(m-hydroxyphenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonate) Benzyl)phenyl)propionic acid benzyl ester (Compound 6.2)
将化合物6.1溶于DMF中，加入(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙 酸苄酯盐酸盐(2eq)，后加入DIPEA(10eq)，HATU(2.5eq)。在常温下搅拌4h，加入10ml稀盐酸溶液，用EA萃取3次，合并有机相，旋干。用反相制备纯化，45度减压旋干得目标产物。LCMS ESI(+)m/z:713.5(M+1).Compound 6.1 was dissolved in DMF, and (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester hydrochloride (2 eq) was added, followed by DIPEA (10 eq), HATU ( 2.5eq). After stirring at normal temperature for 4 h, 10 ml of dilute hydrochloric acid solution was added, and extracted with EA three times, and the organic phases were combined and dried. Purification by reverse phase preparation, rotary drying at 45 ° to give the desired product. LCMS ESI (+) m/z: 713.5 (M+1).
步骤C：(2s)-2-(2，6-二氯-4-(2-(乙氧基(间羟基苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(化合物6)Step C: (2s)-2-(2,6-Dichloro-4-(2-(ethoxy(m-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3-( Methanesulfonyl)phenyl)propionic acid (compound 6)
将化合物6.2溶于1ml的甲醇中，加入Pd/C(10％，0.1eq)，常压氢化1h，过滤，旋干，纯化得产物。LCMS ESI(+)m/z:627.5(M+1).Compound 6.2 was dissolved in 1 ml of methanol, Pd/C (10%, 0.1 eq) was added, and the mixture was hydrogenated under normal pressure for 1 h, filtered, and dried to give the product. LCMS ESI (+) m/z: 627.5 (M+1).
1H-NMR(400MHz,DMSO)δ9.85(s,1H),9.05(d,J＝5.6Hz,1H),7.86(s,1H),7.76(d,J＝4.8Hz,1H),7.66(d,J＝4.8Hz,1H),7.57(dd,J＝5.2Hz,J＝5.2Hz,1H),7.35(m,1H),7.33(s,2H),7.16(m,2H),6.98(m,1H),4.75(m,1H),3.91(m,1H),3.78(m,1H),3.30(m,1H),3.15(s,3H),3.01(m,1H),2.70(m,2H),2.25(m,2H,)1.19(t,J＝4.8Hz,3H). 1 H-NMR (400 MHz, DMSO) δ 9.85 (s, 1H), 9.05 (d, J = 5.6 Hz, 1H), 7.86 (s, 1H), 7.76 (d, J = 4.8 Hz, 1H), 7.66 (d, J = 4.8 Hz, 1H), 7.57 (dd, J = 5.2 Hz, J = 5.2 Hz, 1H), 7.35 (m, 1H), 7.33 (s, 2H), 7.16 (m, 2H), 6.98 (m, 1H), 4.75 (m, 1H), 3.91 (m, 1H), 3.78 (m, 1H), 3.30 (m, 1H), 3.15 (s, 3H), 3.01 (m, 1H), 2.70 ( m, 2H), 2.25 (m, 2H,) 1.19 (t, J = 4.8 Hz, 3H).
实施例7Example 7
Figure PCTCN2018087629-appb-000025
Figure PCTCN2018087629-appb-000025
(2s)-2-(2，6-二氯-4-((甲氧基(3-羟基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-((methoxy(3-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl)benzene Propionate
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000026
Figure PCTCN2018087629-appb-000026
步骤A：(2s)-2-(2，6-二氯-4-((甲氧基(3-甲氧基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸苄酯(化合物7.1)Step A: (2s)-2-(2,6-Dichloro-4-((methoxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-( Methyl sulfonyl)phenyl)propanoate (Compound 7.1)
将化合物2.4(40mg)溶于1ml甲醇中，加入三甲基硅基重氮甲烷(3eq)，在常温下搅拌30分钟。加入适量的乙酸淬灭，旋干，加入5ml的稀盐酸溶液，用EA萃取3次，合并有机相旋干即可。LCMS ESI(+)m/z:713.5(M+1).Compound 2.4 (40 mg) was dissolved in 1 ml of methanol, and trimethylsilyldiazomethane (3 eq) was added, and the mixture was stirred at room temperature for 30 minutes. Quenched with an appropriate amount of acetic acid, spun dry, add 5 ml of dilute hydrochloric acid solution, extract 3 times with EA, and combine the organic phase to dry. LCMS ESI (+) m/z: 713.5 (M+1).
步骤B：(2s)-2-(2，6-二氯-4-((甲氧基(3-羟基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(化合物7)Step B: (2s)-2-(2,6-Dichloro-4-((methoxy(3-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonate) Acyl)phenyl)propionic acid (compound 7)
将化合物7.1(30mg)溶于DCM中，在-40度下加入1mol/L的三溴化硼(10eq)，在0度下搅拌30分钟后在-40度下加水淬灭反应。用EA萃取3次，合并有机相，干燥旋干，纯化得目标产物15mg。LCMS ESI(+)m/z:609.5(M+1).Compound 7.1 (30 mg) was dissolved in DCM, and 1 mol/L of boron tribromide (10 eq) was added at -40 °C, stirred at 0 °C for 30 minutes and then quenched with water at -40 °C. It was extracted 3 times with EA, and the organic phases were combined, dried and dried to give 15 mg of the desired product. LCMS ESI (+) m/z: 609.5 (M+1).
1H-NMR(400MHz,DMSO)δ10.05(s,1H),7.85(s,3H),7.76(d,J＝8Hz,1H),7.67(d,J＝7.6Hz,1H),7.56(dd,J＝8Hz,J＝7.6Hz,1H),7.42(m,1H),7.29(m,1H),7.25(m,1H),7.07(m,1H),4.75(m,1H),3.80(d,J＝12.4Hz,3H),3.30(m,1H),3.15(s,3H),3.04(m,1H). 1 H-NMR (400 MHz, DMSO) δ 10.05 (s, 1H), 7.85 (s, 3H), 7.76 (d, J = 8 Hz, 1H), 7.67 (d, J = 7.6 Hz, 1H), 7.56 ( Dd, J=8 Hz, J=7.6 Hz, 1H), 7.42 (m, 1H), 7.29 (m, 1H), 7.25 (m, 1H), 7.07 (m, 1H), 4.75 (m, 1H), 3.80 (d, J = 12.4 Hz, 3H), 3.30 (m, 1H), 3.15 (s, 3H), 3.04 (m, 1H).
实施例8Example 8
Figure PCTCN2018087629-appb-000027
Figure PCTCN2018087629-appb-000027
(2s)-2-(2，6-二氯-4-(2-(甲氧基(3-羟基苯基)磷酰基)乙基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(methoxy(3-hydroxyphenyl)phosphoryl)ethyl)benzylamino)-3-(3-(methylsulfonyl) Phenyl) propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000028
Figure PCTCN2018087629-appb-000028
将化合物7.2(10mg)溶于甲醇(1ml)中，加入1mgPd/C(10％)，常压氢化1h，过滤旋干，纯化得目标产物4mg。LCMS ESI(+)m/z:613.6(M+1).The compound 7.2 (10 mg) was dissolved in methanol (1 ml), 1 mg of Pd/C (10%) was added, and the mixture was hydrogenated under normal pressure for 1 hour, and the mixture was evaporated to dryness to give 4 mg of desired product. LCMS ESI (+) m/z: 613.6 (M+1).
1H-NMR(400MHz,DMSO)δ9.87(s,1H),9.04(d,J＝8.4Hz,1H),7.86(s,1H),7.77(d,J＝8Hz,1H),7.67(d,J＝7.6Hz,1H),7.56(dd,J＝8Hz,J＝7.6Hz,1H),7.37(m,1H),7.34(s,2H),7.15(m,2H),7.00(m,1H),4.75(m,1H),3.50(d,J＝11.6Hz,3H),3.27(m,1H),3.15(s,3H),3.01(m,1H),2.72(m,2H),2.30(m,2H). 1 H-NMR (400 MHz, DMSO) δ 9.87 (s, 1H), 9.04 (d, J = 8.4 Hz, 1H), 7.86 (s, 1H), 7.77 (d, J = 8 Hz, 1H), 7.67 ( d, J = 7.6 Hz, 1H), 7.56 (dd, J = 8 Hz, J = 7.6 Hz, 1H), 7.37 (m, 1H), 7.34 (s, 2H), 7.15 (m, 2H), 7.00 (m) , 1H), 4.75 (m, 1H), 3.50 (d, J = 11.6 Hz, 3H), 3.27 (m, 1H), 3.15 (s, 3H), 3.01 (m, 1H), 2.72 (m, 2H) , 2.30 (m, 2H).
实施例9Example 9
Figure PCTCN2018087629-appb-000029
Figure PCTCN2018087629-appb-000029
(2s)-2-(2，6-二氯-4-(甲基(苯基)磷酰基)乙炔基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(methyl(phenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl)phenyl)propanoic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000030
Figure PCTCN2018087629-appb-000030
步骤A：甲基苯基磷酰氯(化合物9.1)Step A: Methylphenylphosphoryl chloride (Compound 9.1)
称取500mg甲基苯基磷酸甲酯，加入10ml二氯亚砜，氮气保护，在75度下反应2小时后直接旋干即可。Weigh 500 mg of methyl phenyl phosphate, add 10 ml of thionyl chloride, protect with nitrogen, and react directly at 75 °C for 2 hours and then spin dry.
步骤B：2,6-二氯-4-((苯基(甲基)磷酰基)乙炔基)苯甲酸甲酯(化合物9.2)Step B: Methyl 2,6-dichloro-4-((phenyl(methyl)phosphoryl)ethynyl)benzoate (Compound 9.2)
将200mg的2,6-二氯-4-乙炔基苯甲酸甲酯溶于2ml四氢呋喃中，氮气保护，并在0度下加入0.66ml的2mol/L的异丙基氯化镁，搅拌20分钟；将化合物9.1溶于0.5ml的四氢呋喃并加入，反应20分钟。用1mol/L的稀盐酸溶液淬灭反应，用30mL乙酸乙酯分三次萃取，合并有机相，旋干，纯化得产物(200mg，60％)。LCMS ESI(+)m/z:366.6(M+1).200 mg of methyl 2,6-dichloro-4-ethynylbenzoate was dissolved in 2 ml of tetrahydrofuran, protected with nitrogen, and 0.66 ml of 2 mol/L isopropylmagnesium chloride was added at 0 °C, and stirred for 20 minutes; Compound 9.1 was dissolved in 0.5 ml of tetrahydrofuran and added for 20 minutes. The reaction was quenched with 1 mol/L EtOAc (EtOAc)EtOAc. LCMS ESI (+) m/z: 366.6 (M+1).
步骤C：2,6-二氯-4-((甲基(苯基)磷酰基)乙炔基)苯甲酸(化合物9.3)Step C: 2,6-Dichloro-4-((methyl(phenyl)phosphoryl)ethynyl)benzoic acid (Compound 9.3)
将化合物9.2(200mg)和碘化锂(200mg)溶于2ml吡啶中氮气保护，在120度搅拌3个小时，冷却旋干，加入10ml的1mol/L的稀盐酸溶液，用40ml的乙酸乙酯分3次萃取，合并有机相，旋干无需纯化(150mg)。LCMS ESI(+)m/z:352.6(M+1).Compound 9.2 (200 mg) and lithium iodide (200 mg) were dissolved in 2 ml of pyridine in nitrogen, stirred at 120 °C for 3 hours, cooled and dried, and 10 ml of 1 mol/L dilute hydrochloric acid solution was added to 40 ml of ethyl acetate. The extraction was carried out in 3 portions, and the organic phases were combined and evaporated to dryness (150 mg). LCMS ESI (+) m/z: 352.6 (M+1).
步骤D：(2s)-2-(2，6-二氯-4-((甲基(苯基)磷酰基)乙炔基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸苄酯(化合物9.4)Step D: (2s)-2-(2,6-Dichloro-4-((methyl(phenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl)benzene Base) benzyl propionate (compound 9.4)
将化合物9.3溶于DMF中，加入(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙 酸苄酯盐酸盐(2eq)，后加入DIPEA(10eq)，HATU(2.5eq)。在常温下搅拌4h，加入10ml稀盐酸溶液，用EA萃取3次，合并有机相，旋干。纯化，得目标产物150mg。LCMS ESI(+)m/z:667.5(M+1).Compound 9.3 was dissolved in DMF, and (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester hydrochloride (2 eq) was added, followed by DIPEA (10 eq), HATU ( 2.5eq). After stirring at normal temperature for 4 h, 10 ml of dilute hydrochloric acid solution was added, and extracted with EA three times, and the organic phases were combined and dried. Purification gave the target product 150 mg. LCMS ESI (+) m/z: 667.5 (M+1).
步骤E：(2s)-2-(2，6-二氯-4-((甲基(苯基)磷酰基)乙炔基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(化合物9)Step E: (2s)-2-(2,6-Dichloro-4-((methyl(phenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl)benzene Propionate (compound 9)
将化合物9.4(20mg)溶于DCM中在低温下加入1mol/L的三溴化硼(10eq)，在0度下搅拌30分钟，后在-40度下淬灭反应，EA萃取3次，合并有机相旋干纯化的目标产物10mg。LCMS ESI(+)m/z:577.6(M+1).Compound 9.4 (20 mg) was dissolved in DCM and 1 mol/L boron tribromide (10 eq) was added at low temperature, stirred at 0 °C for 30 minutes, then quenched at -40 °C, EA extracted 3 times, combined The target product of the organic phase was purified by spin-drying 10 mg. LCMS ESI (+) m/z: 577.6 (M+1).
1H-NMR(400MHz,DMSO)δ9.21(d,J＝8.4Hz,1H),7.91(m,2H),7.86(s,1H),7.80(s,2H),7.77(d,J＝4.4Hz,1H),7.67(m,2H),7.62(m,2H),7.57(m,1H),4.80(m,1H),3.29(m,1H),3.15(s,3H),3.03(m,1H),2.02(d,J＝14.8Hz,3H),2.11(m,2H). 1 H-NMR (400 MHz, DMSO) δ 9.21 (d, J = 8.4 Hz, 1H), 7.91 (m, 2H), 7.86 (s, 1H), 7.80 (s, 2H), 7.77 (d, J = 4.4 Hz, 1H), 7.67 (m, 2H), 7.62 (m, 2H), 7.57 (m, 1H), 4.80 (m, 1H), 3.29 (m, 1H), 3.15 (s, 3H), 3.03 ( m, 1H), 2.02 (d, J = 14.8 Hz, 3H), 2.11 (m, 2H).
实施例10Example 10
Figure PCTCN2018087629-appb-000031
Figure PCTCN2018087629-appb-000031
(2s)-2-(2，6-二氯-4-(2-(甲基(苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(methyl(phenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl)phenyl Propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000032
Figure PCTCN2018087629-appb-000032
将化合物9.4(10mg)溶于甲醇(1ml)中，加入1mgPd/C(10％)，常压氢化1h，过滤旋干，纯化得目标产物3mg。LCMS ESI(+)m/z:581.6(M+1).Compound 9.4 (10 mg) was dissolved in methanol (1 ml), and 1 mg of Pd/C (10%) was added, and the mixture was hydrogenated under normal pressure for 1 hour, and the mixture was evaporated to dryness to give the desired product. LCMS ESI (+) m/z: 581.6 (M+1).
1H-NMR(400MHz,DMSO)δ9.04(d,J＝8.4Hz,1H),7.86(s,1H),7.78(m,3H),7.67(d,J＝7.6Hz,1H),7.56(m,4H),7.31(s,2H),4.75(m,1H),3.27(m,1H),3.15(s,3H),3.01(dd,J＝14Hz,10.4Hz,1H),2.79(m,2H),2.29(m,2H),1.67(d,J＝13.4Hz,3H.)。 1 H-NMR (400 MHz, DMSO) δ 9.04 (d, J = 8.4 Hz, 1H), 7.86 (s, 1H), 7.78 (m, 3H), 7.67 (d, J = 7.6 Hz, 1H), 7.56 (m, 4H), 7.31 (s, 2H), 4.75 (m, 1H), 3.27 (m, 1H), 3.15 (s, 3H), 3.01 (dd, J = 14 Hz, 10.4 Hz, 1H), 2.79 ( m, 2H), 2.29 (m, 2H), 1.67 (d, J = 13.4 Hz, 3H.).
实施例11Example 11
Figure PCTCN2018087629-appb-000033
Figure PCTCN2018087629-appb-000033
(2s)-2-(2，6-二氯-4-((甲基(3-羟基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-((methyl(3-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl)phenyl Propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000034
Figure PCTCN2018087629-appb-000034
步骤A：(间甲氧基苯基)甲基磷酸乙酯(化合物11.1)Step A: (M-methoxyphenyl) methyl phosphate (Compound 11.1)
在化合物间甲氧基磷酸二乙酯(2g)加入二氯亚砜(10ml)，在75度下搅拌过夜，旋干后在0度下加入3mol/L的甲基氯化镁(5ml)，搅拌30分钟后在0度下用稀盐酸溶液淬灭反应，用EA萃取，干燥旋干，纯化得目标产物(1.2g，68％)。LCMS ESI(+)m/z:214.6(M+1).Diethyl sulfoxide (2 g) was added to the compound to dichlorosulfoxide (10 ml), and the mixture was stirred at 75 ° C overnight. After spinning, 3 mol/L methyl magnesium chloride (5 ml) was added at 0 ° C, and stirred 30 After a minute, the reaction was quenched with EtOAc (EtOAc)EtOAc. LCMS ESI (+) m/z: 214.6 (M+1).
步骤B:(间甲氧基苯基)甲基磷酰氯(化合物11.2)Step B: (m-methoxyphenyl)methylphosphoryl chloride (compound 11.2)
在化合物11.1(150mg)中加入二氯亚砜，在70度下搅拌3个小时，旋干即得目标产物。To the compound 11.1 (150 mg), thionyl chloride was added, and the mixture was stirred at 70 °C for 3 hours, and dried to give the desired product.
步骤C：2,6-二氯-4-(((间甲氧基苯基)(甲基)磷酰基)乙炔基)苯甲酸甲酯(化合物11.3)Step C: Methyl 2,6-dichloro-4-((m-methoxyphenyl)(methyl)phosphoryl)ethynyl)benzoate (Compound 11.3)
将100mg的2,6-二氯-4-乙炔基苯甲酸甲酯溶于1.5ml四氢呋喃中，氮气保护，并在0度下加入0.7ml的2mol/L的异丙基氯化镁，搅拌20分钟；将化合物11.2溶于0.5ml的四氢呋喃并加入，反应20分钟。用1mol/L的稀盐酸溶液淬灭反应，用30mL乙酸乙酯分三次萃取，合并有机相，旋干，纯化得产物(90mg，60％)。LCMS ESI(+)m/z:396.6(M+1).100 mg of methyl 2,6-dichloro-4-ethynylbenzoate was dissolved in 1.5 ml of tetrahydrofuran, protected with nitrogen, and 0.7 ml of 2 mol/L of isopropylmagnesium chloride was added at 0 °C, and stirred for 20 minutes; Compound 11.2 was dissolved in 0.5 ml of tetrahydrofuran and added for 20 minutes. The reaction was quenched with 1 mol/L EtOAc (EtOAc)EtOAc. LCMS ESI (+) m/z: 396.6 (M+1).
步骤D：2,6-二氯-4-((甲基(间羟基苯基)磷酰基)乙炔基)苯甲酸(化合物11.4)Step D: 2,6-Dichloro-4-((methyl(m-hydroxyphenyl)phosphoryl)ethynyl)benzoic acid (Compound 11.4)
将化合物11.3(90mg)溶于2ml的DCM中，氮气保护，在-40度下加入0.4ml三溴化硼(1mol/L)，然后在0度下搅拌30分钟。在-40度下加入水淬灭，用30mlEA萃取，用无水硫酸钠干燥，旋干，得80mg目标产物ESI(+)m/z:368.6(M+1).Compound 11.3 (90 mg) was dissolved in 2 ml of DCM, and then filtered under nitrogen, and 0.4 ml of boron tribromide (1 mol/L) was added at -40 degrees, and then stirred at 0 degree for 30 minutes. The mixture was quenched with EtOAc (3 mL), EtOAc (EtOAc)
步骤E:(2s)-2-(2，6-二氯-4-((甲基(3-羟基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸苄酯(化合物11.5)Step E: (2s)-2-(2,6-Dichloro-4-((methyl(3-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl) Phenyl) benzyl propionate (compound 11.5)
化合物11.4溶于DMF中，加入(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸苄酯盐酸盐(2eq)，后加入DIPEA(10eq)，HATU(2.5eq)。在常温下搅拌4h，加入10ml稀盐酸溶液，用EA萃取3次，合并有机相，旋干。用反相制备纯化，45度减压旋干得目标产物70mg。LCMS ESI(+)m/z:683.5(M+1).Compound 11.4 was dissolved in DMF and (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester hydrochloride (2 eq) was added followed by DIPEA (10 eq), HATU (2.5 Eq). After stirring at normal temperature for 4 h, 10 ml of dilute hydrochloric acid solution was added, and extracted with EA three times, and the organic phases were combined and dried. Purification by reverse phase preparation, and dried under reduced pressure at 45 ° to yield 70 mg of the desired product. LCMS ESI (+) m/z: 683.5 (M+1).
步骤F：(2s)-2-(2，6-二氯-4-((甲基(3-羟基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(化合物11)Step F: (2s)-2-(2,6-Dichloro-4-((methyl(3-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl) Phenyl) propionic acid (compound 11)
将化合物11.5(20mg)溶于1ml的DCM中，氮气保护，在-40度下加入0.5ml三溴化硼(1mol/L)，然后在0度下搅拌30分钟。在-40度下加入水淬灭，用30mlEA萃取，用无水硫酸钠干燥，旋干，纯化得8mg产物。Compound 11.5 (20 mg) was dissolved in 1 ml of DCM, and then filtered under nitrogen, and 0.5 ml of boron tribromide (1 mol/L) was added at -40 degree, and then stirred at 0 degree for 30 minutes. It was quenched by the addition of water at -40 °, extracted with 30 mL of EA, dried over anhydrous sodium sulfate.
LCMS ESI(+)m/z:593.5(M+1).LCMS ESI (+) m/z: 593.5 (M+1).
1H-NMR(400MHz,DMSO),δ9.98(s,1H),9.22(d,J＝8.4Hz,1H),7.86(s,1H),7.79(s,2H),7.77(m,1H),7.67(d,J＝8Hz,1H),7.58(dd,J＝8Hz,J＝7.6Hz,1H),7.40(m,1H),7.31(m,1H),7.26(m,1H),7.02(m,1H),4.80(m,1H),3.32(m,1H),3.15(s,3H),3.03(dd,J＝14,J＝10.8,1H),1.97(d,J＝14.8Hz,3H). 1 H-NMR (400MHz, DMSO ), δ9.98 (s, 1H), 9.22 (d, J = 8.4Hz, 1H), 7.86 (s, 1H), 7.79 (s, 2H), 7.77 (m, 1H ), 7.67 (d, J = 8 Hz, 1H), 7.58 (dd, J = 8 Hz, J = 7.6 Hz, 1H), 7.40 (m, 1H), 7.31 (m, 1H), 7.26 (m, 1H), 7.02 (m, 1H), 4.80 (m, 1H), 3.32 (m, 1H), 3.15 (s, 3H), 3.03 (dd, J = 14, J = 10.8, 1H), 1.97 (d, J = 14.8) Hz, 3H).
实施例12Example 12
Figure PCTCN2018087629-appb-000035
Figure PCTCN2018087629-appb-000035
(2s)-2-(2，6-二氯-4-((甲氧基(3-甲氧基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-((methoxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl) Phenyl) propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000036
Figure PCTCN2018087629-appb-000036
将化合物7.1(10mg)溶于1ml的DCM中，氮气保护，在-40度下加入0.5ml三溴化硼(1mol/L)，搅拌30分钟后加入水淬灭，用30mlEA萃取，用无水硫酸钠干燥，旋干，纯化得3mg产物。LCMS ESI(+)m/z:623.5(M+1).Compound 7.1 (10 mg) was dissolved in 1 ml of DCM, nitrogen was added, and 0.5 ml of boron tribromide (1 mol/L) was added at -40 °C, stirred for 30 minutes, then quenched with water, extracted with 30 ml of EA, with anhydrous Dry over sodium sulfate, spin dry and purify to give 3 mg. LCMS ESI (+) m/z: 623.5 (M+1).
1H-NMR(400MHz,DMSO)δ9.20(d,J＝7.6Hz,1H),7.87(s,2H),7.86(s,1H),7.77(d,J＝8Hz,1H),7.67(d,J＝7.6Hz,1H),7.55(m,2H),7.44(m,1H),7.30(m,2H), 4.80(m,1H),3.84(d,J＝12.4Hz,3H),3.83(s,3H),3.30(m,1H),3.15(s,3H),3.03(dd,J＝14,J＝9.4,1H).1H-NMR (400MHz, DMSO) δ 9.20 (d, J = 7.6 Hz, 1H), 7.87 (s, 2H), 7.86 (s, 1H), 7.77 (d, J = 8 Hz, 1H), 7.67 (d) , J=7.6Hz, 1H), 7.55 (m, 2H), 7.44 (m, 1H), 7.30 (m, 2H), 4.80 (m, 1H), 3.84 (d, J = 12.4 Hz, 3H), 3.83 (s, 3H), 3.30 (m, 1H), 3.15 (s, 3H), 3.03 (dd, J = 14, J = 9.4, 1H).
实施例13Example 13
Figure PCTCN2018087629-appb-000037
Figure PCTCN2018087629-appb-000037
(2s)-2-(2，6-二氯-4-((羟基(3-甲氧基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-((hydroxy(3-methoxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl)benzene Propionate
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000038
Figure PCTCN2018087629-appb-000038
将化合物7.1(10mg)溶于1ml的THF中，取氢氧化锂(20mg)溶于0.5ml水中，常温搅拌5分钟。浓盐酸调PH＝1旋干，纯化得4mg产物。Compound 7.1 (10 mg) was dissolved in 1 ml of THF, and lithium hydroxide (20 mg) was dissolved in 0.5 ml of water and stirred at room temperature for 5 minutes. Concentrated hydrochloric acid was adjusted to pH = 1 and dried to give 4 mg of product.
LCMS ESI(+)m/z:609.6(M+1).LCMS ESI (+) m/z: 609.6 (M+1).
1H-NMR(400MHz,DMSO)δ9.20(d,J＝8.4Hz,1H),7.85(s,1H),7.77(d,J＝8Hz,1H),7.66(d,J＝9.4Hz,1H),7.65(s,2H),7.57(dd,J＝8Hz,J＝7.6Hz,1H),7.43(m,1H),7.38(m,1H),7.28(m,1H),7.14(m,1H),4.79(m,1H),3.30(dd,J＝14.8Hz,J＝4.8Hz,1H),3.15(s,3H),3.03(dd,J＝14.4,J＝10.8,1H). 1 H-NMR (400 MHz, DMSO) δ 9.20 (d, J = 8.4 Hz, 1H), 7.85 (s, 1H), 7.77 (d, J = 8 Hz, 1H), 7.66 (d, J = 9.4 Hz, 1H), 7.65 (s, 2H), 7.57 (dd, J = 8 Hz, J = 7.6 Hz, 1H), 7.43 (m, 1H), 7.38 (m, 1H), 7.28 (m, 1H), 7.14 (m) , 1H), 4.79 (m, 1H), 3.30 (dd, J = 14.8 Hz, J = 4.8 Hz, 1H), 3.15 (s, 3H), 3.03 (dd, J = 14.4, J = 10.8, 1H).
实施例14Example 14
Figure PCTCN2018087629-appb-000039
Figure PCTCN2018087629-appb-000039
(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzylamino)-3-(3-(methylsulfonyl) Phenyl) propionic acid
Figure PCTCN2018087629-appb-000040
Figure PCTCN2018087629-appb-000040
将化合物11.5(10mg)溶于甲醇(1ml)中，加入1mgPd/C(10％)，常压氢化1h，过滤旋干，纯化得目标产物3mg。LCMS ESI(+)m/z:597.6(M+1).The compound 11.5 (10 mg) was dissolved in methanol (1 ml), and 1 mg of Pd/C (10%) was added, and the mixture was hydrogenated at normal pressure for 1 hour, and the mixture was evaporated to dryness to give 3 mg. LCMS ESI (+) m/z: 597.6 (M+1).
1H-NMR(400MHz,DMSO)δ9.79(s,1H),9.05(d,J＝8Hz,1H),7.86(s,1H),7.77(d,J＝8Hz,1H),7.67(d,J＝8Hz,1H),7.56(t,J＝7.6Hz,1H),7.34(s,2H),7.33(m,1H),7.17(m,2H),6.93(d,J＝7.6Hz,1H),4.75(m,1H),3.29(dd,J＝14Hz,J＝4.4Hz,1H),3.15(s,3H),3.00(dd,J＝14Hz,10.4Hz,1H),2.78(m,2H),2.23(m,2H),1.97(d,J＝13.2Hz,3H)。 1 H-NMR (400 MHz, DMSO) δ 9.79 (s, 1H), 9.05 (d, J = 8 Hz, 1H), 7.86 (s, 1H), 7.77 (d, J = 8 Hz, 1H), 7.67 (d) , J = 8 Hz, 1H), 7.56 (t, J = 7.6 Hz, 1H), 7.34 (s, 2H), 7.33 (m, 1H), 7.17 (m, 2H), 6.93 (d, J = 7.6 Hz, 1H), 4.75 (m, 1H), 3.29 (dd, J = 14 Hz, J = 4.4 Hz, 1H), 3.15 (s, 3H), 3.00 (dd, J = 14 Hz, 10.4 Hz, 1H), 2.78 (m) , 2H), 2.23 (m, 2H), 1.97 (d, J = 13.2 Hz, 3H).
实施例15Example 15
Figure PCTCN2018087629-appb-000041
Figure PCTCN2018087629-appb-000041
(2s)-2-(2，6-二氯-4-((甲基(4-羟基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-((methyl(4-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl)phenyl Propionic acid
采用实施例11中完全相同的步骤，将“间甲氧基磷酸二乙酯”替换为“对甲氧 基磷酸二乙酯”制备出实施例15。LCMS ESI(+)m/z:594.1(M+1).Example 15 was prepared by the same procedure as in Example 11 except that "dimethoxymethoxyphosphate" was replaced with "p-methoxyphosphoric acid diethyl ester". LCMS ESI (+) m/z: 594.1 (M+1).
1H-NMR(400MHz,DMSO)δ10.39(s,1H),9.21(d,J＝8Hz,1H),7.86(s,1H),7.77(s,2H),7.71-7.66(m,3H),7.57(t,J＝8Hz,1H),6.94(d,J＝7.6Hz,2H),4.79(m,1H),3.29(dd,J＝14Hz,J＝4.4Hz,1H),3.15(s,3H),3.02(dd,J＝14Hz,10.4Hz,1H),1.94(d,J＝13.2Hz,3H)。 1 H-NMR (400 MHz, DMSO) δ 10.39 (s, 1 H), 9.21. (d, J = 8 Hz, 1H), 7.86 (s, 1H), 7.77 (s, 2H), 7.71-7.66 (m, 3H) ), 7.57 (t, J = 8 Hz, 1H), 6.94 (d, J = 7.6 Hz, 2H), 4.79 (m, 1H), 3.29 (dd, J = 14 Hz, J = 4.4 Hz, 1H), 3.15 ( s, 3H), 3.02 (dd, J = 14 Hz, 10.4 Hz, 1H), 1.94 (d, J = 13.2 Hz, 3H).
实施例16Example 16
Figure PCTCN2018087629-appb-000042
Figure PCTCN2018087629-appb-000042
(2s)-2-(2，6-二氯-4-((羟基(苯基)磷酰基)乙炔基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸苄酯(2s)-2-(2,6-Dichloro-4-((hydroxy(phenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl)phenyl)propanoic acid Benzyl ester
采用实施例13中完全相同的步骤，将“化合物7.1”替换为“化合物1.4”制备出实施例16。LCMS ESI(+)m/z:580.1(M+1).Example 16 was prepared by the same procedure as in Example 13 except that "Compound 7.1" was replaced with "Compound 1.4". LCMS ESI (+) m/z: 580.1 (M+1).
1H-NMR(400MHz,DMSO)δ9.16(d,J＝8Hz,1H),7.84(s,1H),7.81-7.76(m,3H),7.66(d,J＝8Hz,1H),7.59(s,2H),7.57-7.47(m,5H),4.80(m,1H),3.29(dd,J＝14Hz,J＝4.4Hz,1H),3.14(s,3H),3.02(dd,J＝14Hz,10.4Hz,1H) 1 H-NMR (400 MHz, DMSO) δ 9.16 (d, J = 8 Hz, 1H), 7.84 (s, 1H), 7.81 - 7.76 (m, 3H), 7.66 (d, J = 8 Hz, 1H), 7.59 (s, 2H), 7.57-7.47 (m, 5H), 4.80 (m, 1H), 3.29 (dd, J = 14 Hz, J = 4.4 Hz, 1H), 3.14 (s, 3H), 3.02 (dd, J =14Hz, 10.4Hz, 1H)
实施例17Example 17
Figure PCTCN2018087629-appb-000043
Figure PCTCN2018087629-appb-000043
(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲酰氨基) -3-(2-噻吩酰胺基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzoylamino)-3-(2-thienylamino) Propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000044
Figure PCTCN2018087629-appb-000044
步骤A：3-(2-噻吩酰胺基)-N-[(1,1-二甲基乙氧基)羰基]-L-丙氨酸甲酯(化合物17.1)Step A: 3-(2-Thienylamino)-N-[(1,1-dimethylethoxy)carbonyl]-L-alanine methyl ester (Compound 17.1)
将((S)-3-氨基-2-((1,1-二甲基乙氧基)酰胺基)丙酸甲酯，HCl盐(2.55g,10mmol)溶解在水(30mL)中，放进冰浴并搅拌。向所得溶液加入THF(20mL)，NaOH(1.0M水溶液，25mL)，2-Thiophenecarbonyl chloride(11mmol)。反应搅拌10分钟后，向其中加入EtOAc(100mL)，水层分离掉，有机层用水(25mL),饱和NaCl水溶液洗，然后用无水MgSO4干燥，过滤，旋干得到纯的化合物17.1(3.3g,100％)。LCMS ESI(+)m/z:329(M+1).Methyl ((S)-3-amino-2-((1,1-dimethylethoxy)amido)propanoate, HCl salt (2.55 g, 10 mmol) was dissolved in water (30 mL) Into the ice bath and stirred. To the obtained solution was added THF (20 mL), EtOAc (EtOAc (EtOAc, EtOAc, EtOAc) The organic layer was washed with EtOAc (EtOAc) (EtOAc) 1).
步骤B：(S)-2-氨基-3-(2-噻吩酰胺基)丙酸甲酯(化合物17.2)Step B: (S)-2-Amino-3-(2-thienamido)propionic acid methyl ester (Compound 17.2)
将化合物17.1(1.0g)溶于DCM(20ml)中，加入HCl-dioxane溶液(4.0M，5mL)，反应搅拌2h，旋干，得到的产物(盐酸盐)直接用于下一步反应。LCMS ESI(+)m/z:229(M+1).The compound 17.1 (1.0 g) was dissolved in DCM (20 mL). EtOAc (EtOAc) LCMS ESI (+) m/z: 229 (M+1).
步骤C：2,6-二氯-4-(((间甲氧基苯基)(乙氧基)磷酰基)乙基)苯甲酸甲酯(化合物17.3)Step C: Methyl 2,6-dichloro-4-((m-methoxyphenyl)(ethoxy)phosphoryl)ethyl)benzoate (Compound 17.3)
将化合物2.2(2.0g)溶于甲醇(20ml)中，加入100mg Pd/C(5％)，常 压氢化2h，过滤旋干，纯化得目标产物2.0g.LCMS ESI(+)m/z:401.1(M+1)The compound 2.2 (2.0 g) was dissolved in methanol (20 ml), and 100 mg of Pd/C (5%) was added, and the mixture was hydrogenated under normal pressure for 2 h, filtered and dried to give the desired product: 2.0 g. LCMS ESI (+) m/z: 401.1 (M+1)
步骤D：2,6-二氯-4-(((间羟基苯基)(乙氧基)磷酰基)乙基)苯甲酸甲酯(化合物17.4)Step D: 2,6-Dichloro-4-(((hydroxyphenyl)(ethoxy)phosphoryl)ethyl)benzoate (Compound 17.4)
将化合物11.3(500mg)溶于10ml的DCM中，氮气保护，在-40度下加入3.0ml三溴化硼(1mol/L)，然后在0度下搅拌30分钟。在-40度下加入水淬灭，用30mlEA萃取3次，用无水硫酸钠干燥，旋干，得450mg目标产物ESI(+)m/z:373(M+1).Compound 11.3 (500 mg) was dissolved in 10 ml of DCM, and then filtered under nitrogen, and 3.0 ml of boron tribromide (1 mol/L) was added at -40 degrees, and then stirred at 0 degree for 30 minutes. After quenching with water at -40 °C, it was extracted with EtOAc (3 mL).
步骤E：(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲酰氨基)-3-(2-噻吩酰胺基)丙酸甲酯(化合物17.5)Step E: (2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzoylamino)-3-(2-thiophene) Amido) methyl propionate (compound 17.5)
化合物17.4(100mg)溶于DMF(3mL)中，加入化合物17.4(2eq)，后加入DIPEA(10eq)，HATU(2.5eq)。在常温下搅拌4h，加入10ml稀盐酸溶液，用EA萃取3次，合并有机相，旋干。用反相制备纯化，45度减压旋干得目标产物70mg。LCMS ESI(+)m/z:583(M+1).Compound 17.4 (100 mg) was dissolved in DMF (3 mL). Compound 17.4 (2 eq) was then added, then DIPEA (10 eq), HATU (2.5 eq). After stirring at normal temperature for 4 h, 10 ml of dilute hydrochloric acid solution was added, and extracted with EA three times, and the organic phases were combined and dried. Purification by reverse phase preparation, and dried under reduced pressure at 45 ° to yield 70 mg of the desired product. LCMS ESI (+) m/z: 583 (M+1).
步骤F:(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲酰氨基)-3-(2-噻吩酰胺基)丙酸(化合物17)Step F: (2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzoylamino)-3-(2-thiophene) Amido)propionic acid (compound 17)
将化合物17.5(10mg)溶于1ml的THF中，取氢氧化锂(20mg)溶于0.5ml水中，混合，常温搅拌5分钟。浓盐酸调PH＝1旋干，高压液相纯化得5.2mg产物。LCMS ESI(+)m/z:569(M+1)Compound 17.5 (10 mg) was dissolved in 1 ml of THF, and lithium hydroxide (20 mg) was dissolved in 0.5 ml of water, mixed, and stirred at room temperature for 5 minutes. Concentrated hydrochloric acid was adjusted to pH = 1 to dryness, and purified by high pressure liquid to give 5.2 mg of product. LCMS ESI(+) m/z: 569 (M+1)
1H NMR(400MHz,CD3OD):δ7.69-7.67(m,2H),7.42-7.37(m,1H),7.26(s,2H),7.23-7.12(m,3H),7.03(m,J＝8.4Hz,1H),4.98(m,1H),3.88-3.84(m,2H),2.93-2.86(m,1H),2.77-2.73(m,1H),2.42-2.32(m,2H),1.78(m,J＝13.2Hz,3H)。1H NMR (400MHz, CD3OD): δ 7.69-7.67 (m, 2H), 7.42-7.37 (m, 1H), 7.26 (s, 2H), 7.23 - 7.12 (m, 3H), 7.03 (m, J = 8.4 Hz, 1H), 4.98 (m, 1H), 3.88-3.84 (m, 2H), 2.93-2.86 (m, 1H), 2.77-2.73 (m, 1H), 2.42-2.32 (m, 2H), 1.78 (m, J = 13.2 Hz, 3H).
实施例18Example 18
Figure PCTCN2018087629-appb-000045
Figure PCTCN2018087629-appb-000045
(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲酰氨基)-3-(1-(甲磺酰胺)-1，2，5，6-四氢吡啶-3-基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzoylamino)-3-(1-(methanesulfonamide) )-1,2,5,6-tetrahydropyridin-3-yl)propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000046
Figure PCTCN2018087629-appb-000046
步骤A：(S)-2-((叔丁基氧羰基)氨基)-3-(1-甲基-1，2，5，6-四氢吡啶-3-基)丙酸甲酯)(化合物18.1)Step A: (S)-2-((tert-Butyloxycarbonyl)amino)-3-(1-methyl-1,2,5,6-tetrahydropyridin-3-yl)propanoic acid methyl ester) ( Compound 18.1)
化合物:Boc-3-(3-吡啶基)-L-丙氨酸甲酯(2.8g,10mmol)溶解在乙醇(40mL)中，向溶液中加入甲基碘(2mL),反应在40度搅拌，直到反应结束。反应温度降低到0度，向溶液中分批加入硼氢化钠(2g)。反应搅拌1小时后，向反应中加入5ml丙酮，加入EtOAc(100mL)。所得有机溶液用饱和NH4Cl水溶液(20mL)，水(30mL),饱和食盐水溶液洗。用无水MgSO4干燥，过滤，旋干。所得粗产品用硅胶柱纯化(0-10％的MeOH/DCM)得到化合物18.1.Compound: Boc-3-(3-pyridyl)-L-alanine methyl ester (2.8 g, 10 mmol) was dissolved in ethanol (40 mL), and methyl iodide (2 mL) was added to the solution, and the reaction was stirred at 40 degrees. Until the reaction is over. The reaction temperature was lowered to 0 degrees, and sodium borohydride (2 g) was added portionwise to the solution. After the reaction was stirred for 1 hour, 5 ml of acetone was added to the reaction, and EtOAc (100 mL) was added. The resulting organic solution was washed with aq. EtOAc (EtOAc) Dry over anhydrous MgSO.sub.4, filtered and dried. The crude product obtained was purified by silica gel column (0-10% MeOH / DCM) to afford compound 18.1.
步骤B:(S)-2-((叔丁基氧羰基)氨基)-3-(1-(甲磺酰胺)-1，2，5，6-四氢吡啶-3-基)丙酸甲酯(化合物18.2)Step B: (S)-2-((tert-Butyloxycarbonyl)amino)-3-(1-(methanesulfonamide)-1,2,5,6-tetrahydropyridin-3-yl)propanoic acid Ester (Compound 18.2)
化合物18.1(1.5g,5mmol)溶解在DCE(20mL)中，加入1-氯乙基氯甲酸酯(7.5mmol)，反应加热到40度2小时，旋干。加入干燥的甲醇(20mL),反应回流1小时，再旋干。向得到的中间产物中加入DCM(20ml)，TEA(15mmol)，降温到0度，向反应中加入甲磺酰氯(7.5mmol)。搅拌1小时候， 反应用EtOA(80ml)稀释，水洗两次，干燥，过滤，旋干得到化合物18.2Compound 18.1 (1.5 g, 5 mmol) was dissolved in DCC (20 mL). &lt;RTI ID=0.0&gt;&gt; Dry methanol (20 mL) was added and the mixture was refluxed for 1 hour and then dried. DCM (20 ml), TEA (15 mmol) was added to the obtained intermediate product, and the mixture was cooled to 0°, and methanesulfonyl chloride (7.5 mmol) was added to the reaction. After stirring for 1 hour, the reaction was diluted with EtOA (80 ml), washed twice with water, dried, filtered and dried to give compound 18.2.
步骤C：(S)-2-氨基-3-(1-(甲磺酰胺)-1，2，5，6-四氢吡啶-3-基)丙酸甲酯(化合物18.3)Step C: (S)-2-Amino-3-(1-(methylsulfonamide)-1,2,5,6-tetrahydropyridin-3-yl)propanoic acid methyl ester (Compound 18.3)
将化合物18.2(100mg)溶于DCM(5ml)中，加入HCl-dioxane溶液(4.0M，1mL)，反应搅拌2h，旋干，得到的产物(盐酸盐)直接用于下一步反应。LCMS ESI(+)m/z:229(M+1).The compound 18.2 (100 mg) was dissolved in DCM (5 mL). EtOAc (EtOAc) LCMS ESI (+) m/z: 229 (M+1).
步骤D：(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲酰氨基)-3-(1-(甲磺酰胺)-1，2，5，6-四氢吡啶-3-基)丙酸(化合物18)Step D: (2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzoylamino)-3-(1-( Methanesulfonamide)-1,2,5,6-tetrahydropyridin-3-yl)propionic acid (Compound 18)
从化合物17.4制备化合物17的同样的方法被用来从17.4制备化合物18，其中化合物17.3被化合物18.2置换掉。The same procedure for the preparation of compound 17 from compound 17.4 was used to prepare compound 18 from 17.4, wherein compound 17.3 was replaced by compound 18.2.
实施例19Example 19
Figure PCTCN2018087629-appb-000047
Figure PCTCN2018087629-appb-000047
(2s)-3-((R)-N’-氰基-3-羟基四氫吡咯-1-甲脒基)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲酰氨基)丙酸(2s)-3-((R)-N'-cyano-3-hydroxytetrahydropyrrole-1-carboxy)-2-(2,6-dichloro-4-(2-(methyl) 3-hydroxyphenyl)phosphoryl)ethyl)benzoylamino)propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000048
Figure PCTCN2018087629-appb-000048
步骤A：1-氰基-9，9-二甲基-2-(甲硫基)-7-羰基-8-氧基-1，3，6-三氮癸-1-烯-5-羧酸甲酯(化合物19.1)Step A: 1-Cyano-9,9-dimethyl-2-(methylthio)-7-carbonyl-8-oxy-1,3,6-triazin-1-ene-5-carboxylate Methyl ester (compound 19.1)
将3-氨基-N-叔丁氧羰基-L-丙氨酸甲酯，HCl salt(2.55g,10mmol)溶解在乙醇(30mL)中，加入N-氰亚胺基-S,S-二硫代碳酸二甲酯(10mmol),DIPEA(3mL)。反应搅拌6小时后，向其中加入EtOAc(100mL)，用水(25mL),饱和NaCl水溶液洗，然后用无水MgSO4干燥，过滤，旋干得到纯的化合物18.1，直接用于下一步反应。3-Amino-N-tert-butoxycarbonyl-L-alanine methyl ester, HCl salt (2.55 g, 10 mmol) was dissolved in ethanol (30 mL), and N-cyanamido-S, S-disulfide was added. Dimethyl carbonate (10 mmol), DIPEA (3 mL). After the reaction was stirred for 6 hr, EtOAc (EtOAc)EtOAc.
步骤B：1-氰基-2-((R)-3-羟基四氫吡咯-1-基)-9，9-二甲基-7-羰基-8-氧基-1，3，6-三氮癸-1-烯-5-羧酸甲酯。Step B: 1-cyano-2-((R)-3-hydroxytetrahydropyrrol-1-yl)-9,9-dimethyl-7-carbonyl-8-oxy-1,3,6- Methyl triazain-1-ene-5-carboxylate.
所得的化合物19.1溶解在乙腈(20ml)中，加入(R)-pyrrolidin-3-ol(20mmol)，硝酸银(20mmol)。反应回流8小时，用硅胶过滤，旋干。粗产品用硅胶柱分离，用100/5/1的DCM/MeOH/TEA分离得到产品。The obtained compound 19.1 was dissolved in acetonitrile (20 ml), and (R)-pyrrolidin-3-ol (20 mmol) and silver nitrate (20 mmol) were added. The reaction was refluxed for 8 hours, filtered over silica gel and dried. The crude product was separated on a silica gel column eluting with 100/5/1 DCM/MeOH/EtOAc.
步骤C：(2s)-2-氨基-3-((R)-N’-氰基-3-羟基四氫吡咯-1-甲脒基)苯甲酰氨基)丙酸(化合物19.3)Step C: (2s)-2-Amino-3-((R)-N'-cyano-3-hydroxytetrahydropyrrole-1-carbenyl)benzoylamino)propanoic acid (Compound 19.3)
将化合物19.2(100mg)溶于DCM(5ml)中，加入HCl-dioxane溶液(4.0M，1mL)，反应搅拌2h，旋干，得到的产物(盐酸盐)直接用于下一步反应。The compound 19.2 (100 mg) was dissolved in DCM (5 mL). EtOAc (EtOAc)
步骤D：Step D:
(2s)-3-((R)-N’-氰基-3-羟基四氫吡咯-1-甲脒基)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲酰氨基)丙酸(化合物19)(2s)-3-((R)-N'-cyano-3-hydroxytetrahydropyrrole-1-carboxy)-2-(2,6-dichloro-4-(2-(methyl) 3-hydroxyphenyl)phosphoryl)ethyl)benzoylamino)propionic acid (Compound 19)
从化合物17.4制备化合物17的同样的方法被用来从17.4制备化合物19，其中化合物17.3被化合物19.3置换掉。LC-MS:m/z 595.7(M+H)+.The same procedure for the preparation of compound 17 from compound 17.4 was used to prepare compound 19 from 17.4, wherein compound 17.3 was replaced by compound 19.3. LC-MS: m/z 595.7 (M+H)+.
1H NMR(400MHz,CD3OD):δ7.40-7.35(m,1H),7.28-7.27(m,2H),7.21-7.12(m,2H),7.05-7.02(m,1H),4.80-4.86(m,1H),4.60-4.29(m,4H),3.45-3.41(m,1H), 3.27-3.24(m,1H),3.15-2.71(m,3H),2.44-2.32(m,2H),2.05-1.74(m,5H)。 1 H NMR (400MHz, CD3OD) : δ7.40-7.35 (m, 1H), 7.28-7.27 (m, 2H), 7.21-7.12 (m, 2H), 7.05-7.02 (m, 1H), 4.80-4.86 (m, 1H), 4.60-4.29 (m, 4H), 3.45-3.41 (m, 1H), 3.27-3.24 (m, 1H), 3.15-2.71 (m, 3H), 2.44-2.32 (m, 2H) , 2.05-1.74 (m, 5H).
实施例20Example 20
Figure PCTCN2018087629-appb-000049
Figure PCTCN2018087629-appb-000049
(2s)-2-(2，6-二氯-4-(乙基(3-羟基苯基)磷酰基)乙炔基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(ethyl(3-hydroxyphenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl)phenyl Propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000050
Figure PCTCN2018087629-appb-000050
步骤A：乙基(3-甲氧基苯基)磷酸甲酯(化合物20.1)Step A: Ethyl (3-methoxyphenyl) phosphate (compound 20.1)
(3-甲氧基苯基)磷酸二甲酯(2.16g，10mmol)溶解在无水THF(30mL)中，溶液冷却到-78度，加入EtMgBr(1.0M，10.5ml)。反应渐渐升温到室温(升温过程约2小时)，反应用饱和NH4Cl水溶液淬灭，加入EtOAc(100mL)，水洗一次，加无水MgSO4干燥，过滤，旋干。粗产物硅胶柱分离得到目标产物。步骤B：乙基(3-甲氧基苯基)磷酰氯(化合物20.2)(3-Methoxyphenyl)phosphoric acid dimethyl ester (2.16 g, 10 mmol) was dissolved in dry THF (30 mL), and the solution was cooled to -78, and EtMgBr (1.0M, 10.5ml) was added. The reaction was gradually warmed to room temperature (2 hrs). EtOAc (EtOAc)EtOAc. The crude product was isolated on a silica gel column to give the desired product. Step B: Ethyl (3-methoxyphenyl)phosphoryl chloride (Compound 20.2)
化合物20.1溶解在DCE(10mL)中，加入草酰氯(5ml)，所得溶液回流5小时，反应溶液旋干，得到目标产物，直接使用在下一步。Compound 20.1 was dissolved in DCE (10 mL), oxalyl chloride (5 ml) was added, and the resulting solution was refluxed for 5 hours, and the reaction solution was dried to give the desired product.
步骤C：(2s)-2-(2，6-二氯-4-(乙基(3-羟基苯基)磷酰基)乙炔基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(化合物20)Step C: (2s)-2-(2,6-Dichloro-4-(ethyl(3-hydroxyphenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl) Phenyl) propionic acid (compound 20)
制备化合物2的完全相同的步骤用了制备化合物20，其中化合物20.2用来取代化合物2.1。The exact same procedure for the preparation of compound 2 was carried out by the preparation of compound 20, wherein compound 20.2 was used to replace compound 2.1.
实施例21Example 21
Figure PCTCN2018087629-appb-000051
Figure PCTCN2018087629-appb-000051
(2s)-2-(2，6-二氯-4-(乙基(3-羟基苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(ethyl(3-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl)phenyl Propionic acid
制备实施例3的完全相同的方法用来制备实施例21，其中化合物20取代化合物2.LCMS ESI(+)m/z:612.1(M+1). 1H-NMR(400MHz,DMSO)δ9.75(s,1H),9.03(d,J＝8Hz,1H),7.85(s,1H),7.78(d,J＝8Hz,1H),7.67(d,J＝8Hz,1H),7.56(t,J＝7.6Hz,1H),7.43(m,1H),7.41(s,2H),7.16-7.12(m,2H),6.93(d,J＝7.6Hz,1H),4.75(m,1H),3.29(dd,J＝14Hz,J＝4.4Hz,1H),3.15(s,3H),3.00(dd,J＝14Hz,10.4Hz,1H),2.78-2.50(m,2H),2.26(m,2H),1.86(m,2H),0.93(dt,J＝13.0Hz,7.2Hz,3H)。 The exact same procedure as in Preparation Example 3 was used to prepare Example 21, wherein Compound 20 was substituted for Compound 2. LCMS ESI (+) m/z: 612.1 (M+1). 1 H-NMR (400 MHz, DMSO) δ 9. 75(s,1H), 9.03 (d, J=8Hz, 1H), 7.85(s,1H), 7.78(d,J=8Hz,1H), 7.67(d,J=8Hz,1H),7.56(t , J=7.6 Hz, 1H), 7.43 (m, 1H), 7.41 (s, 2H), 7.16-7.12 (m, 2H), 6.93 (d, J = 7.6 Hz, 1H), 4.75 (m, 1H) , 3.29 (dd, J = 14 Hz, J = 4.4 Hz, 1H), 3.15 (s, 3H), 3.00 (dd, J = 14 Hz, 10.4 Hz, 1H), 2.78-2.50 (m, 2H), 2.26 (m) , 2H), 1.86 (m, 2H), 0.93 (dt, J = 13.0 Hz, 7.2 Hz, 3H).
实施例22Example 22
Figure PCTCN2018087629-appb-000052
Figure PCTCN2018087629-appb-000052
(2s)-2-(2，6-二氯-4-(环丙基(3-羟基苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(cyclopropyl(3-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl)benzene Propionate
制备实施例21的完全相同的方法用来制备实施例22，其中环丙基格氏试剂 取代乙基格氏试剂。LCMS ESI(+)m/z:624.1(M+1).The exact same procedure as in Preparation Example 21 was used to prepare Example 22, wherein the cyclopropyl Grignard reagent was substituted for the ethyl Grignard reagent. LCMS ESI (+) m/z: 624.1 (M+1).
1H-NMR(400MHz,DMSO)δ9.80(s,1H),9.03(d,J＝8Hz,1H),7.85(s,1H),7.78(d,J＝8Hz,1H),7.67(d,J＝8Hz,1H),7.56(t,J＝7.6Hz,1H),7.41(s,2H),7.40(m,1H),7.18-7.23(m,2H),6.93(d,J＝7.6Hz,1H),4.75(m,1H),3.29(dd,J＝14Hz,J＝4.4Hz,1H),3.15(s,3H),3.02(dd,J＝14Hz,10.4Hz,1H),2.80(m,1H),2.60(m,1H),2.26(m,2H),1.22(m,1H),0.82(m,2H),0.71(m,1H),0.52(m,1H)。 1 H-NMR (400 MHz, DMSO) δ 9.80 (s, 1H), 9.03 (d, J = 8 Hz, 1H), 7.85 (s, 1H), 7.78 (d, J = 8 Hz, 1H), 7.67 (d) , J=8Hz, 1H), 7.56 (t, J=7.6Hz, 1H), 7.41(s, 2H), 7.40(m, 1H), 7.18-7.23(m, 2H), 6.93(d, J=7.6 Hz, 1H), 4.75 (m, 1H), 3.29 (dd, J = 14 Hz, J = 4.4 Hz, 1H), 3.15 (s, 3H), 3.02 (dd, J = 14 Hz, 10.4 Hz, 1H), 2.80 (m, 1H), 2.60 (m, 1H), 2.26 (m, 2H), 1.22 (m, 1H), 0.82 (m, 2H), 0.71 (m, 1H), 0.52 (m, 1H).
实施例23Example 23
Figure PCTCN2018087629-appb-000053
Figure PCTCN2018087629-appb-000053
(2s)-2-(2，6-二氯-4-(丁基(3-羟基苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(butyl(3-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl)phenyl Propionic acid
制备实施例21的完全相同的方法用来制备实施例23，其中丁基格氏试剂取代乙基格氏试剂。LCMS ESI(+)m/z:630.1(M+1).The exact same procedure as in Preparation Example 21 was used to prepare Example 23, wherein the butyl Grignard reagent was substituted for the ethyl Grignard reagent. LCMS ESI (+) m/z: 630.1 (M+1).
1H-NMR(400MHz,DMSO)δ9.75(s,1H),9.03(d,J＝8Hz,1H),7.85(s,1H),7.78(d,J＝8Hz,1H),7.67(d,J＝8Hz,1H),7.56(t,J＝7.6Hz,1H),7.42(m,1H),7.41(s,2H),7.18-7.12(m,2H),6.93(d,J＝7.6Hz,1H),4.75(m,1H),3.29(dd,J＝14Hz,J＝4.4Hz,1H),3.15(s,3H),3.00(dd,J＝14Hz,10.4Hz,1H),2.78(m,1H),2.52(m,1H),2.26(m,2H),1.83(m,2H),1.45(m,2H),1.24-1.20(m,3H),0.80(t,J＝7.2Hz,3H)。 1 H-NMR (400 MHz, DMSO) δ 9.75 (s, 1H), 9.03 (d, J = 8 Hz, 1H), 7.85 (s, 1H), 7.78 (d, J = 8 Hz, 1H), 7.67 (d) , J = 8 Hz, 1H), 7.56 (t, J = 7.6 Hz, 1H), 7.42 (m, 1H), 7.41 (s, 2H), 7.18-7.12 (m, 2H), 6.93 (d, J = 7.6 Hz, 1H), 4.75 (m, 1H), 3.29 (dd, J = 14 Hz, J = 4.4 Hz, 1H), 3.15 (s, 3H), 3.00 (dd, J = 14 Hz, 10.4 Hz, 1H), 2.78 (m, 1H), 2.52 (m, 1H), 2.26 (m, 2H), 1.83 (m, 2H), 1.45 (m, 2H), 1.24-1.20 (m, 3H), 0.80 (t, J = 7.2) Hz, 3H).
实施例24Example 24
Figure PCTCN2018087629-appb-000054
Figure PCTCN2018087629-appb-000054
(2s)-2-(2，6-二氯-4-((甲基(苯并呋喃-6-基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-((methyl(benzofuran-6-yl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl) Phenyl) propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000055
Figure PCTCN2018087629-appb-000055
步骤A：苯并呋喃-6-基磷酰酸二乙酯(化合物24.1)Step A: Diethyl benzofuran-6-ylphosphoric acid (Compound 24.1)
化合物6-溴苯并呋喃(2.0g，10mmol)溶解亚磷酸二乙酯(6mL)，加入Pd(OAc)2(200mg),TEA(1ml)。反应在微波反应器上加热到200度，30分钟。加入EtOAc(80ml)，水洗两次，干燥，过滤，旋干。粗产物用硅胶柱分离，0-10％的MeOH/DCM为流动相，得到目标产物24.1。Compound 6-bromobenzofuran (2.0 g, 10 mmol) was dissolved in diethyl phosphite (6 mL). Pd(OAc) 2 (200 mg), TEA (1 ml). The reaction was heated to 200 degrees on a microwave reactor for 30 minutes. EtOAc (80 ml) was added, washed twice with water, dried and filtered. The crude product was separated on a silica gel column eluting EtOAc/EtOAc (EtOAc)
步骤B：甲基(苯并呋喃-6-基)磷酰酸乙酯(化合物24.2)Step B: Methyl (benzofuran-6-yl)phosphoric acid ethyl ester (Compound 24.2)
化合物24.1(1.27g)溶解在THF(20ml)中溶液冷却到-78度，加入MeMgBr(1.0M，5ml)。反应渐渐升温到室温(升温过程约2小时)，反应用饱和NH4Cl水溶液淬灭，加入EtOAc(100mL)，水洗一次，加无水MgSO4干燥，过滤，旋干。粗产物硅胶柱分离得到目标产物。Compound 24.1 (1.27 g) was dissolved in THF (20 mL). The solution was cooled to -78[deg.], and MeMgBr (1.0M, 5ml). The reaction was gradually warmed to room temperature (2 hrs). EtOAc (EtOAc)EtOAc. The crude product was isolated on a silica gel column to give the desired product.
步骤C：甲基(苯并呋喃-6-基)磷酰氯(化合物24.3)Step C: Methyl (benzofuran-6-yl)phosphoryl chloride (Compound 24.3)
化合物24.2溶解在二氯亚砜中(5ml)，所得溶液回流5小时，反应溶液旋干，得到目标产物，直接使用在下一步。The compound 24.2 was dissolved in thionyl chloride (5 ml), and the resulting solution was refluxed for 5 hours, and the reaction solution was dried to give the desired product, which was used directly in the next step.
步骤D：(2s)-2-(2，6-二氯-4-((甲基(苯并呋喃-6-基)磷酰基)乙炔基)苯 甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(化合物24)Step D: (2s)-2-(2,6-Dichloro-4-((methyl(benzofuran-6-yl)phosphoryl)ethynyl)benzylamino)-3-(3-(A) Sulfonyl)phenyl)propionic acid (compound 24)
制备化合物2的完全相同的步骤用来制备化合物24，其中化合物24.3用来取代化合物2.1。LCMS ESI(+)m/z:619.4(M+1).The exact same procedure for the preparation of compound 2 was used to prepare compound 24, wherein compound 24.3 was used to replace compound 2.1. LCMS ESI (+) m/z: 619.4 (M+1).
1H-NMR(400MHz,DMSO)δ9.20(d,J＝8.4Hz,1H),7.85-7.74(m,5H),7.70-7.63(m,2H),7.59-7.44(m,2H),7.40-7.33(m,1H),7.57-7.47(m,5H),4.80(m,1H),3.30(dd,J＝19Hz,J＝4.8Hz,1H),3.14(d,J＝3.2Hz,3H),3.02(m,1H),2.16(d,J＝16Hz,3H)。 1 H-NMR (400 MHz, DMSO) δ 9.20 (d, J = 8.4 Hz, 1H), 7.85-7.74 (m, 5H), 7.70-7.63 (m, 2H), 7.59-7.44 (m, 2H), 7.40-7.33 (m, 1H), 7.57-7.47 (m, 5H), 4.80 (m, 1H), 3.30 (dd, J = 19 Hz, J = 4.8 Hz, 1H), 3.14 (d, J = 3.2 Hz, 3H), 3.02 (m, 1H), 2.16 (d, J = 16 Hz, 3H).
实施例25Example 25
Figure PCTCN2018087629-appb-000056
Figure PCTCN2018087629-appb-000056
(2s)-2-(2，6-二氯-4-((甲基(苯并呋喃-6-基)磷酰基)乙基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-((methyl(benzofuran-6-yl)phosphoryl)ethyl)benzylamino)-3-(3-(methylsulfonyl) Phenyl) propionic acid
制备实施例3的完全相同的方法用来制备实施例2，其中化合物24取代化合物2。LCMS ESI(+)m/z:622.1(M+1).The exact same procedure as in Preparation Example 3 was used to prepare Example 2, in which Compound 24 was substituted for Compound 2. LCMS ESI (+) m/z: 6221. (M+1).
1H-NMR(400MHz,DMSO)δ9.04(d,J＝8Hz,1H),8.16(s,1H),8.03(d,J＝11Hz,1H)7.87(s,1H),7.83(d,J＝8Hz,1H),7.78(d,J＝8Hz,1H),7.67(d,J＝8Hz,1H),7.57(t,J＝7.6Hz,1H),7.42(s,1H),7.32(s,2H),7.07(s,1H),4.75(m,1H),3.29(dd,J＝14Hz,J＝4.4Hz,1H),3.16(s,3H),3.02(dd,J＝14Hz,10.4Hz,1H),2.88(m,1H),2.68(m,1H),2.34(m,2H),1.71(d,J＝13.2Hz,3H)。 1 H-NMR (400 MHz, DMSO) δ 9.04 (d, J = 8 Hz, 1H), 8.16 (s, 1H), 8.03 (d, J = 11 Hz, 1H) 7.78 (s, 1H), 7.83 (d, J=8 Hz, 1H), 7.78 (d, J=8 Hz, 1H), 7.67 (d, J=8 Hz, 1H), 7.57 (t, J=7.6 Hz, 1H), 7.42 (s, 1H), 7.32 ( s, 2H), 7.07 (s, 1H), 4.75 (m, 1H), 3.29 (dd, J = 14 Hz, J = 4.4 Hz, 1H), 3.16 (s, 3H), 3.02 (dd, J = 14 Hz, 10.4 Hz, 1H), 2.88 (m, 1H), 2.68 (m, 1H), 2.34 (m, 2H), 1.71 (d, J = 13.2 Hz, 3H).
实施例26Example 26
Figure PCTCN2018087629-appb-000057
Figure PCTCN2018087629-appb-000057
(2s)-2-(2，6-二氯-4-((甲基(苯并呋喃-2-基)磷酰基)乙基)苯甲酰氨基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-((methyl(benzofuran-2-yl)phosphoryl)ethyl)benzoylamino)-3-(3-(methylsulfonyl) Phenyl) propionic acid
制备化合物25的完全相同的步骤用来制备化合物26，其中2-苯并呋喃用来取代6-苯并呋喃。LCMS ESI(+)m/z:623.6(M+1).The exact same procedure for the preparation of compound 25 was used to prepare compound 26, wherein 2-benzofuran was used to replace 6-benzofuran. LCMS ESI (+) m/z: 623.6 (M+1).
1H-NMR(400MHz,DMSO)δ9.03(d,J＝8Hz,1H),7.86(s,1H),7.76(d,J＝8Hz,2H),7.67(t,2H),7.60(s,1H),7.56(t,1H),7.45(t,1H),7.38(s,2H),7.34(t,1H),4.75(m,1H),3.29(dd,J＝18.4Hz,J＝4.4Hz,1H),3.01(dd,J＝20.4Hz,J＝10.4Hz 1H),2.85(m2H),2.39(m,2H),1.84(d,J＝14Hz,3H)。 1 H-NMR (400 MHz, DMSO) δ 9.03 (d, J = 8 Hz, 1H), 7.86 (s, 1H), 7.76 (d, J = 8 Hz, 2H), 7.67 (t, 2H), 7.60 (s) , 1H), 7.56 (t, 1H), 7.45 (t, 1H), 7.38 (s, 2H), 7.34 (t, 1H), 4.75 (m, 1H), 3.29 (dd, J = 18.4 Hz, J = 4.4 Hz, 1H), 3.01 (dd, J = 20.4 Hz, J = 10.4 Hz 1H), 2.85 (m2H), 2.39 (m, 2H), 1.84 (d, J = 14 Hz, 3H).
实施例27Example 27
Figure PCTCN2018087629-appb-000058
Figure PCTCN2018087629-appb-000058
(2s)-2-(2，6-二氯-4-((甲基(1H-吲哚-5-基)磷酰基)乙基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-((methyl(1H-indol-5-yl)phosphoryl)ethyl)benzylamino)-3-(3-(methylsulfonyl) Phenyl) propionic acid
制备化合物25的完全相同的步骤用来制备化合物26，其中1-Ms-5-Br-indole用来取代6-苯并呋喃。The exact same procedure for the preparation of compound 25 was used to prepare compound 26, wherein 1-Ms-5-Br-indole was used to replace 6-benzofuran.
实施例28Example 28
Figure PCTCN2018087629-appb-000059
Figure PCTCN2018087629-appb-000059
(2S)-2-(2,6-二氯-4-(((甲基(苯基)磷酰基)氧基)甲基)苯甲酰氨基)-3-(3-(甲磺酰基)苯基)丙酸(2S)-2-(2,6-Dichloro-4-(((methyl(phenyl)phosphoryl)oxy)methyl)benzoylamino)-3-(3-(methylsulfonyl)) Phenyl) propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000060
Figure PCTCN2018087629-appb-000060
步骤A：2,6-二氯-(4-羟甲基)苯甲酸甲酯(化合物28.1)2,6-二氯对苯二甲酸二甲酯(2.63g，10mmol)溶解在THF(50ml)中，慢慢加入硼氢化锂(12mmml)，反应搅拌1小时后，加入丙酮(1ml),EtOAc(100ml)。所得溶液水洗二次，无水Na2SO4干燥，过滤，旋干，粗产物直接用于下一步。Step A: Methyl 2,6-dichloro-(4-hydroxymethyl)benzoate (Compound 28.1) Dimethyl 2,6-dichloroterephthalate (2.63 g, 10 mmol) dissolved in THF (50 mL) The lithium borohydride (12 mm ml) was slowly added, and the mixture was stirred for 1 hour, then acetone (1 ml) and EtOAc (100 ml). The resulting solution was washed twice with water, dried over anhydrous Na.sub.2SO4, filtered and evaporated.
步骤B：2,6-二氯-(4-羟甲基)苯甲酸(化合物28.2)Step B: 2,6-Dichloro-(4-hydroxymethyl)benzoic acid (Compound 28.2)
化合物28.1溶解在吡啶(20ml)中，加入碘化锂(15mmml)，反应回流搅拌5小时后，旋干，粗产物用硅胶柱纯化，流动相95/5/0.5(v/v/v)的DCM/MeOH/AcOH。The compound 28.1 was dissolved in pyridine (20 ml), lithium iodide (15 mmml) was added, and the reaction was stirred at reflux for 5 hours, then dried, and the crude product was purified using silica gel column, mobile phase 95/5/0.5 (v/v/v) DCM/MeOH/AcOH.
步骤C：(S)-2-(2,6-二氯-4-(羟甲基)苯甲酰胺基)-3-(3-(3-(3-(甲磺酰基)苯基)丙酸甲酯(化合物28.3)Step C: (S)-2-(2,6-Dichloro-4-(hydroxymethyl)benzamide)-3-(3-(3-(3-(methylsulfonyl)phenyl)propyl Methyl ester (compound 28.3)
化合物28.2溶于DMF中，加入(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸甲酯盐酸盐(2eq)，后加入DIPEA(10eq)，HATU(2.5eq)。在常温下搅拌4h，加入10ml稀盐酸溶液，用EA萃取3次，合并有机相，旋干。用反相制 备纯化，45度减压旋干得目标产物.Compound 28.2 was dissolved in DMF, and (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoic acid methyl ester hydrochloride (2 eq) was added followed by DIPEA (10 eq), HATU (2.5 Eq). After stirring at normal temperature for 4 h, 10 ml of dilute hydrochloric acid solution was added, and extracted with EA three times, and the organic phases were combined and dried. Purified by reverse phase preparation, and dried under reduced pressure at 45 ° to obtain the target product.
步骤D：(S)-2-(2,6-dichloro-4-(羟甲基)苯甲酰胺基)-3-(3-(3-(3-(甲磺酰基)苯基)丙酸(化合物28.4)Step D: (S)-2-(2,6-dichloro-4-(hydroxymethyl)benzamide)-3-(3-(3-(3-(methylsulfonyl)phenyl)propanoic acid (Compound 28.4)
化合物28.3溶于THF中，加入LiOH(2eq)。在常温下搅拌4h，加入稀盐酸溶液值得pH约2-3，用EA萃取3次，合并有机相，旋干。得到目标产物，该产物不经纯化，直接用于下一步。Compound 28.3 was dissolved in THF and LiOH (2 eq) was added. Stir at room temperature for 4 h, add dilute hydrochloric acid solution worth about 2-3, extract with EA 3 times, combine the organic phase, and spin dry. The desired product was obtained which was used in the next step without purification.
步骤E：(2S)-2-(2,6-二氯-4-(((甲基(苯基)磷酰基)氧基)甲基)苯甲酰氨基)-3-(3-(甲磺酰基)苯基)丙酸(化合物28)。Step E: (2S)-2-(2,6-Dichloro-4-(((methyl(phenyl)phosphoryl)oxy)methyl)benzoylamino)-3-(3-(A) Sulfonyl)phenyl)propionic acid (Compound 28).
化合物28.4溶解在DCM中，冷却的0度，加入TEA(10eq)，再加入甲基苯基膦酰氯(5eq)。反应在室温搅拌5小时，加水(20eq)淬灭，旋干。粗产物用反相制备HPLC纯化，得到目标产物。Compound 28.4 was dissolved in DCM, cooled to 0. EtOAc (10 EtOAc). The reaction was stirred at room temperature for 5 h, quenched with water (20 EtOAc) and evaporated. The crude product was purified by reverse phase preparative HPLC to give the desired product.
实施例29Example 29
Figure PCTCN2018087629-appb-000061
Figure PCTCN2018087629-appb-000061
(2S)-2-(2,6-二氯-4-(((3-羟基苯基)(甲基)磷酰基)甲基)氨基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2S)-2-(2,6-Dichloro-4-((3-hydroxyphenyl)(methyl)phosphoryl)methyl)amino)benzamide)-3-(3-(A Sulfonyl)phenyl)propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000062
Figure PCTCN2018087629-appb-000062
步骤A：(氯甲基)(3-甲氧基苯基)(甲基)氧化磷(化合物29.1)Step A: (Chloromethyl)(3-methoxyphenyl)(methyl)phosphorus oxide (Compound 29.1)
氯甲基(甲基)次磷酰氯(10mmol)溶于无水THF(30ml),冷却到-78度，加入(3-甲氧基)苯基锂(1.01eq)。反应搅拌1小时后，在-78度用稀盐酸淬灭。反应回到室温后，加入EtOAc(80ml)，水洗一次，干燥，过滤，旋干。粗产品用硅胶柱纯化，流动相是0-10％MeOH/DCM(v/v)。Chloromethyl (methyl)phosphoryl chloride (10 mmol) was dissolved in dry THF (30 mL), cooled to -78, and (3-methoxy)phenyl lithium (1.01 eq). After the reaction was stirred for 1 hour, it was quenched with dilute hydrochloric acid at -78. After the reaction was returned to room temperature, EtOAc (EtOAc) (EtOAc) The crude product was purified on a silica gel column eluting with 0-10% MeOH / DCM (v/v).
步骤B：2,6-二氯-4-((((3-甲氧基苯基)(甲基)磷酰基)甲基)氨基)苯甲酸甲酯(化合物29.2)Step B: Methyl 2,6-dichloro-4-(((3-methoxyphenyl)(methyl)phosphoryl)methyl)amino)benzoate (Compound 29.2)
化合物29.1(1eq)，4-氨基-2,6-二氯苯甲酸甲酯(1.5eq)溶于无水DMF(30ml),加入碘化钠(0.1eq)冷却到0度，加入钠氢(3eq)。反应在室温下搅拌值得化合物29.1消失，在-78度用饱和NH4Cl水溶液淬灭。反应回到室温后，加入EtOAc(80ml)，水洗三次，干燥，过滤，旋干。粗产品用硅胶柱纯化，流动相是0-10％MeOH/DCM(v/v)。Compound 29.1 (1 eq), methyl 4-amino-2,6-dichlorobenzoate (1.5 eq) was dissolved in anhydrous DMF (30 mL). 3eq). The reaction was stirred at room temperature and the compound 29.1 disappeared and was quenched with saturated aqueous NH.sub.4Cl. After the reaction was returned to room temperature, EtOAc (EtOAc) (EtOAc) The crude product was purified on a silica gel column eluting with 0-10% MeOH / DCM (v/v).
步骤C：2,6-二氯-4-((((3-甲氧基苯基)(甲基)磷酰基)甲基)氨基)苯甲酸(化合物29.3)Step C: 2,6-Dichloro-4-(((3-methoxyphenyl)(methyl)phosphoryl)methyl)amino)benzoic acid (Compound 29.3)
将化合物29.2溶于DCM中在0度下加入1mol/L的三溴化硼(10eq)，在25度下搅拌30分钟，后在-40度下淬灭反应，EA萃取3次，合并有机相旋干。得到的粗产物直接用于下一步。Compound 29.2 was dissolved in DCM and 1 mol/L of boron tribromide (10 eq) was added at 0 °C, stirred at 25 °C for 30 minutes, then quenched at -40 °C, extracted with EA 3 times, combined organic phase Spin dry. The crude product obtained was used directly in the next step.
步骤D：(2S)-2-(2,6-二氯-4-((((3-羟基苯基)(甲基)磷酰基)甲基)氨基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸甲酯(化合物29.4)Step D: (2S)-2-(2,6-Dichloro-4-(((3-hydroxyphenyl)(methyl)phosphoryl)methyl)amino)benzamide)-3-( Methyl 3-(methylsulfonyl)phenyl)propanoate (Compound 29.4)
化合物11.4溶于DMF中，加入(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸甲酯盐酸盐(2eq)，后加入DIPEA(10eq)，HATU(2.5eq)。在常温下搅拌4h，加入10ml稀盐酸溶液，用EA萃取3次，合并有机相，旋干。用反相制备纯化，45度减压旋干得目标产物.Compound 11.4 was dissolved in DMF and (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoic acid methyl ester hydrochloride (2 eq) was added followed by DIPEA (10 eq), HATU (2.5 Eq). After stirring at normal temperature for 4 h, 10 ml of dilute hydrochloric acid solution was added, and extracted with EA three times, and the organic phases were combined and dried. Purified by reverse phase preparation, and dried under reduced pressure at 45 degrees to obtain the target product.
步骤E：(2S)-2-(2,6-二氯-4-(((3-羟基苯基)(甲基)磷酰基)甲基)氨基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(化合物29)Step E: (2S)-2-(2,6-Dichloro-4-((3-hydroxyphenyl)(methyl)phosphoryl)methyl)amino)benzamide)-3-(3) -(Methanesulfonyl)phenyl)propionic acid (Compound 29)
化合物29.4溶于THF中，加入LiOH(3eq)。在常温下搅拌4h，加入稀盐酸溶液值得pH约2-3，用EA萃取3次，合并有机相，旋干。粗产物用反相HPLC纯化得到纯的目标产物。Compound 29.4 was dissolved in THF and LiOH (3 eq) was added. Stir at room temperature for 4 h, add dilute hydrochloric acid solution worth about 2-3, extract with EA 3 times, combine the organic phase, and spin dry. The crude product was purified by reverse phase HPLC to afford pure title product.
实施例30和实施例31Example 30 and Example 31
Figure PCTCN2018087629-appb-000063
Figure PCTCN2018087629-appb-000063
手性制备HPLC用来拆分实施例14所得的化合物。手性柱为Chiralcel OZ-H型号，流动相为Hexane/EtOH/TFA,比例60/40/0.1(V/V/V)。两个异构体得到很好的分离。LCMS与 1HNMR数据同化合物14. Chiral preparative HPLC was used to resolve the compound obtained in Example 14. The chiral column is Chiralcel OZ-H model, the mobile phase is Hexane/EtOH/TFA, and the ratio is 60/40/0.1 (V/V/V). The two isomers were well separated. LCMS and 1 H NMR data were the same as compound 14.
实施例32Example 32
(S,E)-2-(4-(2-(3-羟基苯基)膦酰基乙烯基)-2,6-二氯苯酰胺)-3-(3-(甲磺酰基)苯基)丙酸(S,E)-2-(4-(2-(3-hydroxyphenyl)phosphonovinyl)-2,6-dichlorobenzamide)-3-(3-(methylsulfonyl)phenyl) Propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000064
Figure PCTCN2018087629-appb-000064
步骤A：(E)-2,6-二氯-4-(2-(二乙氧基膦酰基)乙烯基)苯甲酸甲酯(化合物32.1)Step A: (E)-2,6-Dichloro-4-(2-(diethoxyphosphono)vinyl)benzoic acid methyl ester (Compound 32.1)
2,6-二氯-4-醛基苯甲酸甲酯1g和亚甲基二磷酸四乙酯2.1g，溶于20mlDMF，加入K2CO3固体2g，搅拌2h,除去溶剂，纯化得产物。1 g of 2,6-dichloro-4-aldehyde benzoic acid methyl ester and 2.1 g of tetramethyl methylene diphosphate were dissolved in 20 ml of DMF, and 2 g of K 2 CO 3 solid was added thereto, and the mixture was stirred for 2 hours to remove the solvent, and the product was purified.
步骤B:(E)-2,6-二氯-4-(2-(氯乙氧基膦酰基)乙烯基)苯甲酸甲酯(化合物32.2)Step B: (E)-2,6-Dichloro-4-(2-(chloroethoxyphosphonyl)vinyl)benzoic acid methyl ester (Compound 32.2)
1g化合物32.1溶于10mlSOCl2，加热至70度反应4h，除去溶剂，得到产物。步骤C:(E)-4-(2-(3-甲氧基苯基)膦酰基乙烯基)-2,6-二氯苯甲酸甲酯(化合物32.3)1 g of compound 32.1 was dissolved in 10 ml of SOCl2, heated to 70 degrees for 4 h, and the solvent was removed to give the product. Step C: (E)-4-(2-(3-Methoxyphenyl)phosphonovinyl)methyl-2,6-dichlorobenzoate (Compound 32.3)
1g化合物32.2，溶于20mlTHF，降温至0度，加入间甲氧基苯基溴化镁，室温搅拌4h，除去THF，纯化得产物。1 g of compound 32.2, dissolved in 20 ml of THF, cooled to 0 °, added m-methoxyphenylmagnesium bromide, stirred at room temperature for 4 h, THF was removed and purified.
步骤D：(E)-4-(2-(3-羟基苯基)乙氧膦酰基乙烯基)-2,6-二氯苯甲酸(化合物32.4)Step D: (E)-4-(2-(3-Hydroxyphenyl)ethoxyphosphonovinyl)-2,6-dichlorobenzoic acid (Compound 32.4)
200mg化合物32.3，溶于20mlCH2Cl2，降温至0度，加入BBr3，反应2h后，加入水，用EA萃取后干燥，旋干得产物。200 mg of compound 32.3, dissolved in 20 ml of CH 2 Cl 2 , cooled to 0 °, added BBr3, after 2 h of reaction, water was added, extracted with EA, dried, and dried to give the product.
步骤E:(S,E)-2-(4-(2-(3-羟基苯基)乙氧膦酰基乙烯基)-2,6-二氯苯酰 胺)-3-(3-(甲磺酰基)苯基)丙酸苄酯(化合物32.5)Step E: (S,E)-2-(4-(2-(3-hydroxyphenyl)ethoxyphosphonylvinyl)-2,6-dichlorobenzamide)-3-(3-(methylsulfonate) Benzyl)phenyl)propionic acid benzyl ester (compound 32.5)
50mg化合物32.4与(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸苄酯盐酸盐40mg和DIPEA 40mg溶于DMF 5ml，加入HATU 60mg，搅拌过夜，除去DMF，纯化得产物。50 mg of compound 32.4 and (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester hydrochloride 40 mg and DIPEA 40 mg dissolved in DMF 5 ml, HATU 60 mg was added, stirred overnight to remove DMF , purified product.
步骤F：(S,E)-2-(4-(2-(3-羟基苯基)乙氧膦酰基乙烯基)-2,6-二氯苯酰胺)-3-(3-(甲磺酰基)苯基)丙酸(化合物32)Step F: (S,E)-2-(4-(2-(3-hydroxyphenyl)ethoxyphosphonylvinyl)-2,6-dichlorobenzamide)-3-(3-(methylsulfonate) Acyl)phenyl)propionic acid (compound 32)
15mg化合物32.5溶于1mlTHF,加入LiOH水溶液0.2ml，搅拌5min，除去溶剂，纯化得产物。LCMS ESI(+)m/z:629.5(M+1).15 mg of the compound 32.5 was dissolved in 1 ml of THF, and 0.2 ml of a LiOH aqueous solution was added thereto, and the mixture was stirred for 5 minutes, and the solvent was removed to obtain a product. LCMS ESI (+) m/z: 629.5 (M+1).
1H-NMR(400MHz,DMSO)δ9.18(d,J＝8.4Hz,1H),7.85(s,1H),7.82(s,2H),7.78(d,J＝7.6Hz,1H),7.68(d,J＝7.2Hz,1H),7.58(t,1H),7.37(m,2H),7.18(m,3H)7.01(d,J＝6Hz,1H),4.80(m,1H),3.92(m,2H),3.3(m,2H),3.14(s,1H),1.24(t,J＝7.8Hz，3H)。 1 H-NMR (400 MHz, DMSO) δ 9.18 (d, J = 8.4 Hz, 1H), 7.85 (s, 1H), 7.82 (s, 2H), 7.78 (d, J = 7.6 Hz, 1H), 7.68 (d, J = 7.2 Hz, 1H), 7.58 (t, 1H), 7.37 (m, 2H), 7.18 (m, 3H) 7.01 (d, J = 6 Hz, 1H), 4.80 (m, 1H), 3.92 (m, 2H), 3.3 (m, 2H), 3.14 (s, 1H), 1.24 (t, J = 7.8 Hz, 3H).
实施例33Example 33
Figure PCTCN2018087629-appb-000065
Figure PCTCN2018087629-appb-000065
(2s)-2-(2，6-二氯-4-(2-(羟基(4-羟基苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(hydroxy(4-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl) Phenyl) propionic acid
采用实施例14中完全相同的步骤，将“实施例16”转变为实施例33。"Example 16" was converted to Example 33 using the exact same procedure as in Example 14.
LCMS ESI(+)m/z:597.6(M+1). 1H-NMR(400MHz,DMSO)δ10.05(s,1H),9.04(d,J＝8Hz,1H),7.86(s,1H),7.77(d,J＝8Hz,1H),7.67(d,J＝8Hz,1H),7.56(m,3H),7.31(s,2H),6.88(d,J＝7.6Hz,1H),4.75(m,1H),3.29(dd,J＝14Hz,J＝4.4Hz,1H), 3.15(s,3H),3.02(dd,J＝14Hz,10.4Hz,1H),2.78(m,1H),2.60(m,1H),2.18(m,2H),1.59(d,J＝13.2Hz,3H). LCMS ESI (+) m / z :. 597.6 (M + 1) 1 H-NMR (400MHz, DMSO) δ10.05 (s, 1H), 9.04 (d, J = 8Hz, 1H), 7.86 (s, 1H ), 7.77 (d, J = 8 Hz, 1H), 7.67 (d, J = 8 Hz, 1H), 7.56 (m, 3H), 7.31 (s, 2H), 6.88 (d, J = 7.6 Hz, 1H), 4.75 (m, 1H), 3.29 (dd, J = 14 Hz, J = 4.4 Hz, 1H), 3.15 (s, 3H), 3.02 (dd, J = 14 Hz, 10.4 Hz, 1H), 2.78 (m, 1H) , 2.60 (m, 1H), 2.18 (m, 2H), 1.59 (d, J = 13.2 Hz, 3H).
实施例34Example 34
Figure PCTCN2018087629-appb-000066
Figure PCTCN2018087629-appb-000066
(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲酰氨基)-3-(3-羟基苯甲酰胺基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzoylamino)-3-(3-hydroxybenzamide Propionate
实施例17中的2-噻吩羧酸置换成3-羟基苯甲酸制得实施例34。Example 34 was prepared by substituting 2-thiophenecarboxylic acid in Example 17 to 3-hydroxybenzoic acid.
LC-MS:m/z 579.2(M+H)+LC-MS: m/z 579.2 (M+H)+
1H NMR(400MHz,CD3OD):δ7.40-7.30(m,1H),7.26-7.20(m,5H),7.19-7.16(m,1H),7.30(d,J＝8.8,1H),7.00(d,J＝4.8Hz,1H),6.95-6.92(m,1H),4.95(t,J＝4.0Hz,1H),3.84(d,J＝4.4,2H),2.92-2.87(m,1H),2.74-2.71(m,1H),2.38-2.30(m,2H),1.75(d,J＝8.4,3H). 1 H NMR (400MHz, CD3OD) : δ7.40-7.30 (m, 1H), 7.26-7.20 (m, 5H), 7.19-7.16 (m, 1H), 7.30 (d, J = 8.8,1H), 7.00 (d, J = 4.8 Hz, 1H), 6.95-6.92 (m, 1H), 4.95 (t, J = 4.0 Hz, 1H), 3.84 (d, J = 4.4, 2H), 2.92 - 2.87 (m, 1H) ), 2.74-2.71 (m, 1H), 2.38-2.30 (m, 2H), 1.75 (d, J = 8.4, 3H).
实施例35Example 35
Figure PCTCN2018087629-appb-000067
Figure PCTCN2018087629-appb-000067
(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲酰氨基)-3-(3，5-二羟基苯甲酰胺基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzoylamino)-3-(3,5-dihydroxyl Benzoylamide)propionic acid
实施例17中的2-噻吩羧酸置换成3，5-二羟基苯甲酸制得实施例35。Example 35 was prepared by substituting the 2-thiophenecarboxylic acid of Example 17 with 3,5-dihydroxybenzoic acid.
LC-MS:m/z 595.2(M+H) + LC-MS: m/z 595.2 (M+H) +
1H NMR(400MHz,CD 3OD):δ7.41-7.37(m,1H),7.27(s,2H),7.22-7.14(m,2H),7.04-7.01(m,J＝5.2Hz,1H),6.73(d,J＝1.6Hz,2H),6.45(t,J＝1.6Hz,1H),4.97(m,1H),3.80-3.87(m,2H),2.95-2.88(m,1H),2.78-2.71(m,1H),2.43-2.29(m,2H)1.78(d,J＝8.8Hz,3H)。 1 H NMR (400 MHz, CD 3 OD): δ 7.41-7.37 (m, 1H), 7.27 (s, 2H), 7.22 - 7.14 (m, 2H), 7.04-7.01 (m, J = 5.2 Hz, 1H) ), 6.73 (d, J = 1.6 Hz, 2H), 6.45 (t, J = 1.6 Hz, 1H), 4.97 (m, 1H), 3.80-3.87 (m, 2H), 2.95-2.88 (m, 1H) , 2.78-2.71 (m, 1H), 2.43-2.29 (m, 2H) 1.78 (d, J = 8.8 Hz, 3H).
实施例36Example 36
Figure PCTCN2018087629-appb-000068
Figure PCTCN2018087629-appb-000068
2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲氨基)-3-(3，5-(二甲磺酰基)苯基)丙酸2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzylamino)-3-(3,5-(dimethylsulfonyl)benzene Propionate
将实施例14和11中的(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸苄酯盐酸盐替换为2-氨基-3-(3，5-(二甲磺酰基)苯基)丙酸苄酯盐酸盐制备得到实施例36。LCMS ESI(+)m/z:583.6(M+1)。Replacing the benzyl ester of (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoate in Examples 14 and 11 with 2-amino-3-(3,5-( Preparation of dimethyl sulfonyl)phenyl)propionate hydrochloride gave Example 36. LCMS ESI (+) m/z: 58.
1H-NMR(400MHz,DMSO)δ9.78(d,J＝8Hz,1H),8.25(d,J＝7.6Hz,2H),7.34(s,2H),7.18(m,2H)，6.95(m,2H)，4.89(m,1H),3.45(dd,J＝15Hz,J＝4.4Hz,1H),3.29(s,6H),3.22(dd,J＝15.5Hz,J＝10.4Hz,1H),2.70(m,2H),2.11(m,2H)，1.65(d，J＝13.2，3H)。 1 H-NMR (400 MHz, DMSO) δ 9.78 (d, J = 8 Hz, 1H), 8.25 (d, J = 7.6 Hz, 2H), 7.34 (s, 2H), 7.18 (m, 2H), 6.95 ( m, 2H), 4.89 (m, 1H), 3.45 (dd, J = 15 Hz, J = 4.4 Hz, 1H), 3.29 (s, 6H), 3.22 (dd, J = 15.5 Hz, J = 10.4 Hz, 1H) ), 2.70 (m, 2H), 2.11 (m, 2H), 1.65 (d, J = 13.2, 3H).
实施例37Example 37
Figure PCTCN2018087629-appb-000069
Figure PCTCN2018087629-appb-000069
(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲氨基)-6-(甲磺酰胺)己酸(2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzylamino)-6-(methanesulfonamide)hexanoic acid
将实施例14和11中的(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸苄酯盐酸盐替换为(2s)-2-氨基-6-(甲磺酰胺)己酸甲酯盐酸盐制备得到实施例37.LC-MS:m/z 578.1(M+H) +1H NMR(400MHz,CD 3OD):δ7.40(m,1H),7.29(s,2H),7.21-7.17(s,2H),7.22(m,1H),7.17(m,J＝8.8Hz,1H),7.02(d,J＝5.2Hz,1H),4.02(m,1H),3.41(t,J＝4.4Hz,2H),3.96-3.91(m,1H),2.97(s,3H),2.95-2.90(m,1H),2.82-2.72(m,1H),2.47-2.32(m,2H),1.92-1.90(m,1H),1.79(d,J＝8.8,3H),1.79(m,5H). Replacing benzyl (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoate in Examples 14 and 11 with (2s)-2-amino-6-(A Preparation of sulfonamide) hexanoic acid methyl ester hydrochloride Example 37. LC-MS: m/z 578.1 (M+H) +1 H NMR (400 MHz, CD 3 OD): δ 7.40 (m, 1H), 7.29(s,2H), 7.21-7.17(s,2H), 7.22(m,1H), 7.17(m,J=8.8Hz,1H),7.02(d,J=5.2Hz,1H),4.02(m , 1H), 3.41 (t, J = 4.4 Hz, 2H), 3.96-3.91 (m, 1H), 2.97 (s, 3H), 2.95-2.90 (m, 1H), 2.82-2.72 (m, 1H), 2.47-2.32 (m, 2H), 1.92-1.90 (m, 1H), 1.79 (d, J = 8.8, 3H), 1.79 (m, 5H).
实施例38Example 38
Figure PCTCN2018087629-appb-000070
Figure PCTCN2018087629-appb-000070
(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲氨基)-3-(5-(甲磺酰基)吡啶-3-基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzylamino)-3-(5-(methylsulfonyl) Pyridin-3-yl)propionic acid
将实施例14和11中的(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸苄酯盐酸盐替换为(2s)-2-氨基-3-(5-(甲磺酰基)吡啶-3-基)丙酸盐酸盐制备得到实施例38.LCMS ESI(+)m/z:600.4(M+1).Replacing the benzyl ester of (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoate in Examples 14 and 11 with (2s)-2-amino-3-(5 -(Methanesulfonyl)pyridin-3-yl)propanoic acid hydrochloride was prepared as Example 38. LCMS ESI (+) m/z: 600.4 (M+1).
1H-NMR(400MHz,MeOD)δ8.96(s,1H),8.83(s,1H),8.39(s,1H),7.38(m,H),7.24(s,2H),7.17(m,2H),7.02(d,J＝7.6Hz,1H),5.08(m,1H),3.52(m,1H),3.20(s,1H),2.88(m,1H),2.74(m,1H),2.33(m,2H),1.79(d,J＝13.2Hz,3H) 1 H-NMR (400MHz, MeOD ) δ8.96 (s, 1H), 8.83 (s, 1H), 8.39 (s, 1H), 7.38 (m, H), 7.24 (s, 2H), 7.17 (m, 2H), 7.02 (d, J = 7.6 Hz, 1H), 5.08 (m, 1H), 3.52 (m, 1H), 3.20 (s, 1H), 2.88 (m, 1H), 2.74 (m, 1H), 2.33 (m, 2H), 1.79 (d, J = 13.2 Hz, 3H)
实施例39Example 39
Figure PCTCN2018087629-appb-000071
Figure PCTCN2018087629-appb-000071
(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲氨基)-3-(1-(甲磺酰基)-1H-吡咯-3-基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzylamino)-3-(1-(methylsulfonyl) -1H-pyrrol-3-yl)propionic acid
将实施例14和11中的(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸苄酯盐酸盐替换为(2s)-2-氨基-3-(1-(甲磺酰基)-1H-吡咯-3-基)丙酸盐酸盐制备得到实施例39.Replacing the benzyl ester of (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoate in Examples 14 and 11 with (2s)-2-amino-3-(1) -(Methanesulfonyl)-1H-pyrrol-3-yl)propionic acid hydrochloride was prepared to give Example 39.
实施例40Example 40
Figure PCTCN2018087629-appb-000072
Figure PCTCN2018087629-appb-000072
(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲氨基)-3-(1-(甲磺酰基)-1H-吡唑-3-基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzylamino)-3-(1-(methylsulfonyl) -1H-pyrazol-3-yl)propionic acid
将实施例14和11中的(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸苄酯盐酸盐替换为(2s)-2-氨基-3-(1-(甲磺酰基)-1H-吡唑-3-基)丙酸盐酸盐制备得到实施例40.Replacing the benzyl ester of (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoate in Examples 14 and 11 with (2s)-2-amino-3-(1) -(Methanesulfonyl)-1H-pyrazol-3-yl)propionic acid hydrochloride was prepared to give Example 40.
实施例41Example 41
Figure PCTCN2018087629-appb-000073
Figure PCTCN2018087629-appb-000073
(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲氨基)-3-(1-(甲磺酰基)-1H-吡唑-4-基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzylamino)-3-(1-(methylsulfonyl) -1H-pyrazol-4-yl)propionic acid
将实施例14和11中的(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸苄酯盐酸盐替换为(2s)-2-氨基-3-(1-(甲磺酰基)-1H-吡唑-4-基)丙酸盐酸盐制备得到实施例41.Replacing the benzyl ester of (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoate in Examples 14 and 11 with (2s)-2-amino-3-(1) -(Methanesulfonyl)-1H-pyrazol-4-yl)propionic acid hydrochloride was prepared to give Example 41.
实施例42Example 42
Figure PCTCN2018087629-appb-000074
Figure PCTCN2018087629-appb-000074
(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲氨基)-3-(1-(甲磺酰基)吡咯烷-3-基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzylamino)-3-(1-(methylsulfonyl) Pyrrolidin-3-yl)propionic acid
将实施例14和11中的(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸苄酯盐酸盐替换为(2s)-2-氨基-3-(1-(甲磺酰基)吡咯烷-3-基)丙酸盐酸盐制备得到实施例42.LC-MS:m/z 591.1(M+H) +. Replacing the benzyl ester of (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoate in Examples 14 and 11 with (2s)-2-amino-3-(1) - propionic acid hydrochloride (methylsulfonyl) pyrrolidin-3-yl) obtained in Example 42.LC-MS: m / z 591.1 (m + H) +.
1H NMR(400MHz,CD 3OD):δ7.43-7.37(m,1H),7.30(s,2H),7.25-7.20(m,2H),7.04-7.01(m,1H),4.71-4.63(m,1H),3.68-3.55(m,1H),3.50-3.44(m,1H),3.30-3.28(m,1H),3.05-2.90(m,2H),2.89(d,J＝2.0Hz,3H),2.88-2.75(m,1H), 2.52-2.50(m,1H),2.50-2.10(m,3H),2.09-1.82(m,2H),1.79(d,J＝9.2Hz,3H),1.69(m,1H)。 1 H NMR (400 MHz, CD 3 OD): δ 7.43 - 7.37 (m, 1H), 7.30 (s, 2H), 7.25-7.20 (m, 2H), 7.04-7.01 (m, 1H), 4.71-4.63 (m, 1H), 3.68-3.55 (m, 1H), 3.50-3.44 (m, 1H), 3.30-3.28 (m, 1H), 3.05-2.90 (m, 2H), 2.89 (d, J = 2.0 Hz) , 3H), 2.88-2.75 (m, 1H), 2.52-2.50 (m, 1H), 2.50-2.10 (m, 3H), 2.09-1.82 (m, 2H), 1.79 (d, J = 9.2 Hz, 3H ), 1.69 (m, 1H).
实施例43Example 43
Figure PCTCN2018087629-appb-000075
Figure PCTCN2018087629-appb-000075
(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲酰氨基)-3-((S)-3-羟基吡咯烷-1-羧酰胺)丙酸(2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzoylamino)-3-((S)-3- Hydroxypyrrolidine-1-carboxamide)propionic acid
实施例17中的2-噻吩羧酸置换成(s)-3-羟基吡咯烷-1-羰酰氯制得实施例43。Example 43 was prepared by substituting 2-thiophenecarboxylic acid in Example 17 with (s)-3-hydroxypyrrolidine-1-carbonyl chloride.
LC-MS:m/z 595.7(M+H)+.LC-MS: m/z 595.7 (M+H)+.
1HNMR(400MHz,CD3OD):δ7.39-7.35(m,1H),7.26(s,2H),7.19-7.12(m,2H),7.01(d,J＝8.4,1H),4.78-4.74(m,1H),4.36-4.40(m,1H),3.66-3.64(m,2H),3.45-3.41(m,3H),3.27-3.25(m,1H),2.95-2.85(m,1H),2.79-2.69(m,1H),2.39-2.27(m,2H),2.05-1.92(m,2H),1.77(d,J＝12.4,1H)。 1 H NMR (400 MHz, CD 3 OD): δ 7.39-7.35 (m, 1H), 7.26 (s, 2H), 7.19-7.12 (m, 2H), 7.01 (d, J = 8.4, 1H), 4.78-4.74 ( m,1H), 4.36-4.40 (m,1H), 3.66-3.64 (m,2H), 3.45-3.41 (m,3H), 3.27-3.25 (m,1H), 2.95-2.85 (m,1H), 2.79-2.69 (m, 1H), 2.39-2.27 (m, 2H), 2.05-1.92 (m, 2H), 1.77 (d, J = 12.4, 1H).
实施例44Example 44
Figure PCTCN2018087629-appb-000076
Figure PCTCN2018087629-appb-000076
(2s)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲氨基)-5(甲磺酰胺)戊酸(2s)-2-(2,6-Dichloro-4-(2-(methyl(3-hydroxyphenyl)phosphoryl)ethyl)benzylamino)-5(methanesulfonamide)pentanoic acid
将实施例14和11中的(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸苄酯盐酸盐替换为(2s)-2-氨基-5(甲磺酰胺)戊酸甲酯盐酸盐制备得到实施例44Replacing benzyl (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoate in Examples 14 and 11 with (2s)-2-amino-5 (methanesulfonate) Preparation of amide) methyl valerate hydrochloride to give Example 44
LC-MS:m/z 565.1(M+H) +. LC-MS: m/z 565.1 (M+H) + .
1H NMR(400MHz,CD 3OD):δ7.39-7.36(m,1H),7.27(s,2H),7.21-7.17(m,1H),7.14(d,J＝8.4Hz,1H)7.00(d,J＝5.6Hz,1H),4.05(m,1H),3.42-3.40(m,2H),2.95(s,3H),2.91-2.87(m,1H),2.74-2.72(m,1H),2.39-2.28(m,2H),2.04-2.02(m,1H),1.81-1.74(m,6H)。 1 H NMR (400 MHz, CD 3 OD): δ 7.39-7.36 (m, 1H), 7.27 (s, 2H), 7.21-7.17 (m, 1H), 7.14 (d, J = 8.4 Hz, 1H) 7.00 (d, J = 5.6 Hz, 1H), 4.05 (m, 1H), 3.42-3.40 (m, 2H), 2.95 (s, 3H), 2.91-2.87 (m, 1H), 2.74-2.72 (m, 1H) ), 2.39-2.28 (m, 2H), 2.04-2.02 (m, 1H), 1.81-1.74 (m, 6H).
实施例45Example 45
Figure PCTCN2018087629-appb-000077
Figure PCTCN2018087629-appb-000077
(2s)-3-((反式)-N’-氰基-3，4-二羟基吡咯烷-1-甲脒基)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲酰氨基)丙酸(2s)-3-((trans)-N'-cyano-3,4-dihydroxypyrrolidine-1-carboxy)-2-(2,6-dichloro-4-(2-( Methyl (3-hydroxyphenyl)phosphoryl)ethyl)benzoylamino)propionic acid
实施例19中的(R)-3-吡咯烷醇置换成(反式)-3，4-吡咯烷二醇制得实施例45。Example 45 was prepared by substituting (R)-3-pyrrolidinol in Example 19 with (trans)-3,4-pyrrolidine diol.
LC-MS:m/z 611.7(M+H) +. LC-MS: m/z 611.7 (M+H) + .
1H NMR(400MHz,CD 3OD):δ7.41-7.39(m,1H),7.30-7.28(m,2H),7.24-7.20(m,2H),7.05-7.03(m,1H),5.11(dd,J＝10.4,4.4,1H),4.11-4.18(m,1H),4.04-4.01(m,1H),3.79-3.72(m,1H),3.51-3.56(m,1H),3.26-3.25(m,1H),3.23-3.22(m,1H),3.17-3.13(m,1H),3.07-3.03(m,1H),3.00-2.96(m,1H),2.94-2.86(m,1H),2.77-2.66(m,1H),2.45-2.33(m,2H),1.79-1.75(m,3H)。 1 H NMR (400 MHz, CD 3 OD): δ 7.41-7.39 (m, 1H), 7.30-7.28 (m, 2H), 7.24-7.20 (m, 2H), 7.05-7.03 (m, 1H), 5.11 (dd, J = 10.4, 4.4, 1H), 4.11-4.18 (m, 1H), 4.04-4.01 (m, 1H), 3.79-3.72 (m, 1H), 3.51-3.56 (m, 1H), 3.26- 3.25(m,1H),3.23-3.22(m,1H), 3.17-3.13(m,1H),3.07-3.03(m,1H), 3.00-2.96(m,1H),2.94-2.86(m,1H) ), 2.77-2.66 (m, 1H), 2.45-2.33 (m, 2H), 1.79-1.75 (m, 3H).
实施例46Example 46
Figure PCTCN2018087629-appb-000078
Figure PCTCN2018087629-appb-000078
(2s)-3-(2-氰基-3，3-二(2-羟基乙基)胍基)-2-(2，6-二氯-4-(2-(甲基(3-羟基苯基)磷酰基)乙基)苯甲酰氨基)丙酸(2s)-3-(2-cyano-3,3-bis(2-hydroxyethyl)indolyl)-2-(2,6-dichloro-4-(2-(methyl(3-hydroxy)) Phenyl)phosphoryl)ethyl)benzoylamino)propionic acid
实施例19中的(R)-3-吡咯烷醇置换成二乙醇胺制得实施例46。Example 46 was prepared by replacing (R)-3-pyrrolidinol in Example 19 with diethanolamine.
LC-MS:m/z 595.6(M-OH) +. LC-MS: m/z 595.6 (M-OH) + .
1H NMR(400MHz,CD 3OD):δ7.38-7.37(m,1H),7.30-7.27(m,2H),7.21-7.17(m,2H),7.01-6.99(m,1H),5.71(dd,J＝10.0,4.0,1H),4.07-4.04(m,1H),3.96-3.91(m,1H),3.79-3.43(m,8H),2.99-2.90(m,1H),2.82-2.72(m,1H),2.47-2.36(m,2H),1.75(d,J＝12.8,3H)。 1 H NMR (400 MHz, CD 3 OD): δ 7.38-7.37 (m, 1H), 7.30-7.27 (m, 2H), 7.21-7.17 (m, 2H), 7.01-6.99 (m, 1H), 5.71 (dd, J = 10.0, 4.0, 1H), 4.07-4.04 (m, 1H), 3.96-3.91 (m, 1H), 3.79-3.43 (m, 8H), 2.99-2.90 (m, 1H), 2.82 2.72 (m, 1H), 2.47-2.36 (m, 2H), 1.75 (d, J = 12.8, 3H).
实施例47Example 47
Figure PCTCN2018087629-appb-000079
Figure PCTCN2018087629-appb-000079
(2s)-2-(2，6-二氯-4-((甲氧基(2-羟基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-((methoxy(2-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl)benzene Propionate
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000080
Figure PCTCN2018087629-appb-000080
步骤A：2,6-二氯-4-(((邻甲氧基苯基)(乙氧基)磷酰基)乙炔基)苯甲酸甲酯(化合物47.2)Step A: Methyl 2,6-dichloro-4-((o-methoxyphenyl)(ethoxy)phosphoryl)ethynyl)benzoate (compound 47.2)
将100mg的2,6-二氯-4-乙炔基苯甲酸甲酯溶于1.5ml四氢呋喃中，氮气保护，并在0度下加入0.7ml的2mol/L的异丙基氯化镁，搅拌20分钟；将化合物47.1溶于0.5ml的四氢呋喃并加入，反应20分钟。用1mol/L的稀盐酸溶液淬灭反应，用30mL乙酸乙酯分三次萃取，合并有机相，旋干，纯化得产物(100mg，60％)。LCMS ESI(+)m/z:426.6(M+1).100 mg of methyl 2,6-dichloro-4-ethynylbenzoate was dissolved in 1.5 ml of tetrahydrofuran, protected with nitrogen, and 0.7 ml of 2 mol/L of isopropylmagnesium chloride was added at 0 °C, and stirred for 20 minutes; Compound 47.1 was dissolved in 0.5 ml of tetrahydrofuran and added for 20 minutes. The reaction was quenched with 1 mol/L EtOAc (EtOAc)EtOAc. LCMS ESI (+) m/z: 426.6 (M+1).
步骤B：2,6-二氯-4-((羟基(邻甲氧苯基)磷酰基)乙炔基)苯甲酸(化合物47.3)Step B: 2,6-Dichloro-4-((hydroxy(o-methoxyphenyl)phosphoryl)ethynyl)benzoic acid (Compound 47.3)
将化合物47.3(100mg)和碘化锂(100mg)溶于2ml吡啶中氮气保护，在120度搅拌3个小时，冷却旋干，加入10ml的1mol/L的稀盐酸溶液，用30ml的乙酸乙酯分3次萃取，合并有机相，旋干无需纯化。LCMS ESI(+)m/z:384.6(M+1).Compound 47.3 (100 mg) and lithium iodide (100 mg) were dissolved in 2 ml of pyridine in nitrogen, stirred at 120 °C for 3 hours, cooled and dried, and 10 ml of 1 mol/L dilute hydrochloric acid solution was added, and 30 ml of ethyl acetate was added. The extraction was carried out in 3 portions, and the organic phases were combined and dried without further purification. LCMS ESI (+) m/z: 384.6 (M+1).
步骤C：(2s)-2-(2，6-二氯-4-((羟基(邻甲氧苯基)磷酰基)乙炔基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸苄酯(化合物47.4)Step C: (2s)-2-(2,6-Dichloro-4-((hydroxy(o-methoxyphenyl)phosphoryl)ethynyl)benzamide)-3-(3-(methylsulfonyl) Phenyl) benzyl propionate (compound 47.4)
将化合物47.3溶于DMF中，加入(2s)-2-氨基-3-(3-(甲磺酰基)苯基) 丙酸苄酯盐酸盐(2eq)，后加入DIPEA(10eq)，HATU(2.5eq)。在常温下搅拌4h，加入10ml稀盐酸溶液，用EA萃取3次，合并有机相，旋干。用反相制备纯化，45度减压旋干得目标产物85mg。LCMS ESI(+)m/z:699.5(M+1).Compound 47.3 was dissolved in DMF and (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester hydrochloride (2 eq) was added followed by DIPEA (10 eq), HATU ( 2.5eq). After stirring at normal temperature for 4 h, 10 ml of dilute hydrochloric acid solution was added, and extracted with EA three times, and the organic phases were combined and dried. Purification by reverse phase preparation, spin-drying at 45 ° under reduced pressure to give the desired product, 85 mg. LCMS ESI (+) m/z: 699.5 (M+1).
步骤D：(2s)-2-(2，6-二氯-4-((甲氧基(2-甲氧基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸苄酯(化合物47.5)Step D: (2s)-2-(2,6-Dichloro-4-((methoxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-( Methyl sulfonyl)phenyl)propanoate (Compound 47.5)
将化合物47.4(40mg)溶于1ml甲醇中，加入三甲基硅基重氮甲烷(3eq)，在常温下搅拌30分钟。加入适量的乙酸淬灭，旋干，加入5ml的稀盐酸溶液，用EA萃取3次，合并有机相旋干即可。LCMS ESI(+)m/z:713.5(M+1).Compound 47.4 (40 mg) was dissolved in 1 ml of methanol, and trimethylsilyldiazomethane (3 eq) was added and stirred at room temperature for 30 minutes. Quenched with an appropriate amount of acetic acid, spun dry, add 5 ml of dilute hydrochloric acid solution, extract 3 times with EA, and combine the organic phase to dry. LCMS ESI (+) m/z: 713.5 (M+1).
步骤E：(2s)-2-(2，6-二氯-4-((甲氧基(2-羟基苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(化合物47)Step E: (2s)-2-(2,6-Dichloro-4-((methoxy(2-hydroxyphenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonate) Acyl)phenyl)propionic acid (compound 47)
将化合物47.5(30mg)溶于DCM中，在-40度下加入1mol/L的三溴化硼(10eq)，在0度下搅拌30分钟后在-40度下加水淬灭反应。用EA萃取3次，合并有机相，干燥旋干，纯化得目标产物15mg。LCMS ESI(+)m/z:609.5(M+1).Compound 47.5 (30 mg) was dissolved in DCM, and 1 mol/L of boron tribromide (10 eq) was added at -40 °C, stirred at 0 °C for 30 minutes and then quenched with water at -40 °C. It was extracted 3 times with EA, and the organic phases were combined, dried and dried to give 15 mg of the desired product. LCMS ESI (+) m/z: 609.5 (M+1).
1H-NMR(400MHz,DMSO),δ10.57(s,1H),9.21(d,J＝8.4Hz,1H),7.88(s,1H),7.78(s,2H),7.67(m,2H),7.57(t,J＝7.6Hz,1H),5.96(m,2H),4.80(m,1H),3.77(d,J＝12.4Hz,3H),3.30(m,1H),3.15(s,3H),3.03(dd,J＝14,J＝9.4,1H). 1 H-NMR (400 MHz, DMSO), δ 10.57 (s, 1H), 9.21. (d, J = 8.4 Hz, 1H), 7.78 (s, 1H), 7.78 (s, 2H), 7.67 (m, 2H) ), 7.57 (t, J = 7.6 Hz, 1H), 5.96 (m, 2H), 4.80 (m, 1H), 3.77 (d, J = 12.4 Hz, 3H), 3.30 (m, 1H), 3.15 (s) , 3H), 3.03 (dd, J=14, J=9.4, 1H).
实施例48Example 48
Figure PCTCN2018087629-appb-000081
Figure PCTCN2018087629-appb-000081
(2s)-2-(2，6-二氯-4-((甲基(4-羟基-3-氯苯基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-((methyl(4-hydroxy-3-chlorophenyl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonate) Acyl)phenyl)propionic acid
采用实施例11中完全相同的步骤，将“间甲氧基磷酸二乙酯”替换为“4-甲氧基-3-氯磷酸二乙酯”制备出实施例48。LCMS ESI(+)m/z:628.1(M+1).Example 48 was prepared by the same procedure as in Example 11 except that "dimethoxym-methoxyphosphate" was replaced with "4-methoxy-3-chlorophosphate diethyl ester". LCMS ESI (+) m/z: 628.1 (M+1).
1H-NMR(400MHz,DMSO)δ11.15(s,1H),9.21(d,J＝8Hz,1H),7.86(s,1H),7.79-7.82(m,4H),7.68(m,2H),7.57(t,J＝8Hz,1H),7.15(dd,J＝7.6Hz,4.0Hz,1H),4.79(m,1H),3.30(dd,J＝14Hz,J＝4.4Hz,1H),3.15(s,3H),3.03(dd,J＝14Hz,10.4Hz,1H),1.99(d,J＝13.2Hz,3H). 1 H-NMR (400 MHz, DMSO) δ 11.15 (s, 1H), 9.21. (d, J=8 Hz, 1H), 7.86 (s, 1H), 7.79-7.82 (m, 4H), 7.68 (m, 2H) ), 7.57 (t, J = 8 Hz, 1H), 7.15 (dd, J = 7.6 Hz, 4.0 Hz, 1H), 4.79 (m, 1H), 3.30 (dd, J = 14 Hz, J = 4.4 Hz, 1H) , 3.15 (s, 3H), 3.03 (dd, J = 14 Hz, 10.4 Hz, 1H), 1.99 (d, J = 13.2 Hz, 3H).
实施例49Example 49
Figure PCTCN2018087629-appb-000082
Figure PCTCN2018087629-appb-000082
(2s)-2-(2，6-二氯-4-((羟基(苯并呋喃-6-基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-((hydroxy(benzofuran-6-yl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonyl)benzene Propionate
Figure PCTCN2018087629-appb-000083
Figure PCTCN2018087629-appb-000083
步骤A：2,6-二氯-4-(((苯并呋喃-6-基)(乙氧基)磷酰基)乙炔基)苯甲酸甲酯(化合物49.1)Step A: 2,6-Dichloro-4-(((benzofuran-6-yl)(ethoxy)phosphoryl)ethynyl)benzoic acid methyl ester (Compound 49.1)
将100mg的2,6-二氯-4-乙炔基苯甲酸甲酯溶于1.5ml四氢呋喃中，氮气保护，并在0度下加入0.7ml的2mol/L的异丙基氯化镁，搅拌20分钟；将化合物24.3溶于0.5ml的四氢呋喃并加入，反应20分钟。用1mol/L的稀盐酸溶液淬灭 反应，用30mL乙酸乙酯分三次萃取，合并有机相，旋干，纯化得产物(100mg，60％)。LCMS ESI(+)m/z:437.1(M+1).100 mg of methyl 2,6-dichloro-4-ethynylbenzoate was dissolved in 1.5 ml of tetrahydrofuran, protected with nitrogen, and 0.7 ml of 2 mol/L of isopropylmagnesium chloride was added at 0 °C, and stirred for 20 minutes; Compound 26.3 was dissolved in 0.5 ml of tetrahydrofuran and added for 20 minutes. The reaction was quenched with 1 mol/L EtOAc (EtOAc)EtOAc. LCMS ESI (+) m/z: 437.1 (M+1).
步骤B：2,6-二氯-4-((羟基(苯并呋喃-6-基)磷酰基)乙炔基)苯甲酸(化合物49.2)Step B: 2,6-Dichloro-4-((hydroxy(benzofuran-6-yl)phosphoryl)ethynyl)benzoic acid (Compound 49.2)
将化合物49.1(100mg)和碘化锂(100mg)溶于2ml吡啶中氮气保护，在120度搅拌3个小时，冷却旋干，加入10ml的1mol/L的稀盐酸溶液，用30ml的乙酸乙酯分3次萃取，合并有机相，旋干无需纯化。Compound 49.1 (100 mg) and lithium iodide (100 mg) were dissolved in 2 ml of pyridine under nitrogen, stirred at 120 °C for 3 hours, cooled and dried, and 10 ml of 1 mol/L dilute hydrochloric acid solution was added to 30 ml of ethyl acetate. The extraction was carried out in 3 portions, and the organic phases were combined and dried without further purification.
步骤C：(2s)-2-(2，6-二氯-4-((羟基(苯并呋喃-6-基)磷酰基)乙炔基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸苄酯(化合物49.3)Step C: (2s)-2-(2,6-Dichloro-4-((hydroxy(benzofuran-6-yl)phosphoryl)ethynyl)benzamide)-3-(3-(A) Benzyl sulfonyl)phenyl)propionate (Compound 49.3)
将化合物49.2溶于DMF中，加入(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸苄酯盐酸盐(2eq)，后加入DIPEA(10eq)，HATU(2.5eq)。在常温下搅拌4h，加入10ml稀盐酸溶液，用EA萃取3次，合并有机相，旋干。用反相制备纯化，45度减压旋干得目标产物85mg。LCMS ESI(+)m/z:710.1(M+1).Compound 49.2 was dissolved in DMF, and (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester hydrochloride (2 eq) was added followed by DIPEA (10 eq), HATU ( 2.5eq). After stirring at normal temperature for 4 h, 10 ml of dilute hydrochloric acid solution was added, and extracted with EA three times, and the organic phases were combined and dried. Purification by reverse phase preparation, spin-drying at 45 ° under reduced pressure to give the desired product, 85 mg. LCMS ESI (+) m/z: 710.1 (M+1).
步骤D：(2s)-2-(2，6-二氯-4-((羟基(苯并呋喃-6-基)磷酰基)乙炔基)苯甲氨基)-3-(3-(甲磺酰基)苯基)丙酸(化合物49)Step D: (2s)-2-(2,6-Dichloro-4-((hydroxy(benzofuran-6-yl)phosphoryl)ethynyl)benzylamino)-3-(3-(methylsulfonate) Acyl)phenyl)propionic acid (compound 49)
将化合物49.3(30mg)溶于DCM中，在-40度下加入1mol/L的三溴化硼(10eq)，在0度下搅拌30分钟后在-40度下加水淬灭反应。用EA萃取3次，合并有机相，干燥旋干，纯化得目标产物15mg。LCMS ESI(+)m/z:620.0(M+1).Compound 49.3 (30 mg) was dissolved in DCM, and 1 mol/L of boron tribromide (10 eq) was added at -40 °C, stirred at 0 °C for 30 minutes and then quenched with water at -40 °C. It was extracted 3 times with EA, and the organic phases were combined, dried and dried to give 15 mg of the desired product. LCMS ESI (+) m/z: 620.0 (M+1).
1H-NMR(400MHz,DMSO)δ9.19(d,J＝8Hz,1H),8.17(s,1H),7.98(d,J＝248Hz,1H),7.86(s,1H),7.79-7.76(m,2H),7.55-7.66(m,3H),7.57(t,J＝8Hz,1H),7.07(s,1H),4.79(m,1H),3.30(dd,J＝14Hz,J＝4.4Hz,1H),3.15(s,3H),3.02(dd,J＝14Hz,10.4Hz,1H). 1 H-NMR (400 MHz, DMSO) δ 9.19 (d, J = 8 Hz, 1H), 8.17 (s, 1H), 7.78 (d, J = 248 Hz, 1H), 7.86 (s, 1H), 7.79-7.76 (m, 2H), 7.55-7.66 (m, 3H), 7.57 (t, J = 8 Hz, 1H), 7.07 (s, 1H), 4.79 (m, 1H), 3.30 (dd, J = 14 Hz, J = 4.4 Hz, 1H), 3.15 (s, 3H), 3.02 (dd, J = 14 Hz, 10.4 Hz, 1H).
实施例50Example 50
Figure PCTCN2018087629-appb-000084
Figure PCTCN2018087629-appb-000084
(2s)-2-(2，6-二氯-4-(1-羟基2-(羟基(3-羟基苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(1-hydroxy2-(hydroxy(3-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3-(A Sulfonyl)phenyl)propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000085
Figure PCTCN2018087629-appb-000085
步骤A：2，6-二氯-4-(1-羟基2-(甲氧基(3-甲氧基苯基)磷酰基)乙基)苯甲酸甲酯(化合物50.1)Step A: Methyl 2,6-dichloro-4-(1-hydroxy 2-(methoxy(3-methoxyphenyl)phosphoryl)ethyl)benzoate (Compound 50.1)
称取300mg甲基(3-甲氧基苯基)膦酸乙酯溶于10ml干燥四氢呋喃中，氮气保护，冰浴下，加入1.1mL(2M)的LDA，在冰浴下搅拌1小时后，加入419mg的2，6-二氯-4-甲酰基苯甲酸甲酯，在常温下搅拌2小时，饱和NH 4Cl在冰浴下淬灭反应，用30mL乙酸乙酯分三次萃取，合并有机相，旋干，柱纯化得到产物(100mg，23％)。LCMS ESI(+)m/z:432.8。 300 mg of ethyl methyl (3-methoxyphenyl)phosphonate was weighed and dissolved in 10 ml of dry tetrahydrofuran under nitrogen, and 1.1 mL (2M) of LDA was added thereto, and the mixture was stirred for 1 hour in an ice bath. Add 419 mg of methyl 2,6-dichloro-4-formylbenzoate, stir at room temperature for 2 hours, quench the saturated NH 4 Cl in an ice bath, extract three times with 30 mL of ethyl acetate, and combine the organic phases. The product was purified (100 mg, 23%). LCMS ESI (+) m/z: 432.8.
步骤B：2，6-二氯-4-(1-羟基2-(羟基(3-羟基苯基)磷酰基)乙基)苯甲酸(化合物50.2)Step B: 2,6-Dichloro-4-(1-hydroxy 2-(hydroxy(3-hydroxyphenyl)phosphoryl)ethyl)benzoic acid (Compound 50.2)
将化合物50.1(100mg)溶于8ml二氯甲烷中氮气保护，-40℃下，加入1mL的三溴化硼，-40℃下搅拌4个小时，加入10ml的水淬灭，用20ml的二 氯甲烷分3次萃取，合并有机相，旋干无需纯化，粗品70mg。LCMS ESI(+)m/z:390.8(M+1)。Compound 50.1 (100 mg) was dissolved in 8 ml of dichloromethane and protected with nitrogen. At -40 ° C, 1 mL of boron tribromide was added, stirred at -40 ° C for 4 hours, quenched with 10 ml of water, and 20 ml of dichloro The methane was extracted three times, and the organic phase was combined. LCMS ESI (+) m/z: 39.
步骤C：(2s)-2-(2，6-二氯-4-(1-羟基2-(羟基(3-羟基苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸苄酯(化合物50.3)Step C: (2s)-2-(2,6-Dichloro-4-(1-hydroxy2-(hydroxy(3-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3) -(Methanesulfonyl)phenyl)propanoic acid benzyl ester (Compound 50.3)
将化合物50.2溶于DMF中，加入(2s)-2-氨基-3-(3，5-(二甲磺酰基)苯基)丙酸苄酯盐酸盐(2eq)，后加入DIPEA(10eq)，HATU(2.5eq)。在常温下搅拌4h，加入10ml稀盐酸溶液，用EA萃取3次，合并有机相，旋干。用反相制备纯化，45度减压旋干得目标产物40mg。LCMS ESI(+)m/z:705.7(M+1).Compound 50.2 was dissolved in DMF, and (2s)-2-amino-3-(3,5-(dimethylsulfonyl)phenyl)propanoic acid benzyl ester hydrochloride (2 eq) was added, followed by DIPEA (10 eq) , HATU (2.5 eq). After stirring at normal temperature for 4 h, 10 ml of dilute hydrochloric acid solution was added, and extracted with EA three times, and the organic phases were combined and dried. Purification by reverse phase preparation, spin-drying at 45 ° under reduced pressure afforded 40 mg. LCMS ESI (+) m/z: 705.7 (M+1).
步骤D：(2s)-2-(2，6-二氯-4-(1-羟基2-(羟基(3-羟基苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(化合物50)Step D: (2s)-2-(2,6-Dichloro-4-(1-hydroxy2-(hydroxy(3-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3) -(Methanesulfonyl)phenyl)propionic acid (Compound 50)
将化合物1.3溶于2ml的甲醇和0.3mL的水中，冰浴下加入2eq的一水合氢氧化锂，室温下搅拌1h,1N的盐酸调节pH＝6，20mL乙酸乙酯旋分3次萃取，水洗，干燥，旋蒸，反相制备，冻干的产物5mg。LCMS ESI(+)m/z:583.6(M+1)；The compound 1.3 was dissolved in 2 ml of methanol and 0.3 mL of water, 2 eq of lithium hydroxide monohydrate was added under ice bath, stirred at room temperature for 1 h, 1N hydrochloric acid was adjusted to pH=6, and 20 mL of ethyl acetate was spun and extracted three times, and washed with water. , dry, rotary steaming, reversed phase preparation, lyophilized product 5 mg. LCMS ESI (+) m/z: 58.
1H-NMR(400MHz,CD 3OD-d 4)δ7.95(s,1H),7.86(d,J＝5.6Hz,1H),7.73(d,J＝4.8Hz,1H)，7.62(t,J＝5.2Hz,1H)，7.31(m,2H),7.26(s,2H)，7.21(dd,J＝5.2Hz,J＝5.2Hz,1H)，7.08(d,J＝8.4Hz,1H)，6.96(t,J＝5.2Hz,1H)，5.10(m,1H),4.91(m,1H),3.50(dd,J＝3.2Hz,J＝3.2Hz,1H),3.22(dd,J＝2.8Hz,J＝3.2Hz,1H),3.29(s,3H),2.39(m,2H)。 1 H-NMR (400 MHz, CD 3 OD-d 4 ) δ 7.95 (s, 1H), 7.86 (d, J = 5.6 Hz, 1H), 7.73 (d, J = 4.8 Hz, 1H), 7.62 (t , J = 5.2 Hz, 1H), 7.31 (m, 2H), 7.26 (s, 2H), 7.21 (dd, J = 5.2 Hz, J = 5.2 Hz, 1H), 7.08 (d, J = 8.4 Hz, 1H) ), 6.96 (t, J = 5.2 Hz, 1H), 5.10 (m, 1H), 4.91 (m, 1H), 3.50 (dd, J = 3.2 Hz, J = 3.2 Hz, 1H), 3.22 (dd, J = 2.8 Hz, J = 3.2 Hz, 1H), 3.29 (s, 3H), 2.39 (m, 2H).
实施例51Example 51
Figure PCTCN2018087629-appb-000086
Figure PCTCN2018087629-appb-000086
(2s)-2-(2，6-二氯-4-(2-(羟基(间羟基苯基)磷酰基)乙烯基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(2-(hydroxy(m-hydroxyphenyl)phosphoryl)vinyl)benzamide)-3-(3-(methylsulfonyl)benzene Propionate
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000087
Figure PCTCN2018087629-appb-000087
将化合物32溶于5ml的四氢呋喃中，加入LiOH水溶液1ml，室温搅拌5h，旋干，纯化得产物。LCMS ESI(+)m/z:599.4(M+1).The compound 32 was dissolved in 5 ml of tetrahydrofuran, and 1 ml of a LiOH aqueous solution was added thereto, and the mixture was stirred at room temperature for 5 hr. LCMS ESI (+) m/z: 599.4 (M+1).
1H-NMR(400MHz,DMSO)δ9.73(s,1H),9.13(d,J＝8.8Hz,1H),7.87(s,1H),7.77(d,J＝8Hz,2H),7.69(d,J＝7.6Hz,1H),7.58(t,J＝7.6Hz,1H),7.31(m,2H),7.16(m,2H),7.02(m,2H),6.98(m,1H),4.78(m,1H),3.15(s,3H),3.03(m,1H). 1 H-NMR (400 MHz, DMSO) δ 9.73 (s, 1H), 9.13 (d, J = 8.8 Hz, 1H), 7.78 (s, 1H), 7.77 (d, J = 8 Hz, 2H), 7.69 ( d, J = 7.6 Hz, 1H), 7.58 (t, J = 7.6 Hz, 1H), 7.31 (m, 2H), 7.16 (m, 2H), 7.02 (m, 2H), 6.98 (m, 1H), 4.78 (m, 1H), 3.15 (s, 3H), 3.03 (m, 1H).
实施例52Example 52
Figure PCTCN2018087629-appb-000088
Figure PCTCN2018087629-appb-000088
(S,E)-2-(4-(2-(二(3-羟基苯基)膦酰基)乙烯基)-2,6-二氯苯酰胺)-3-(3-(甲磺酰基)苯基)丙酸(S,E)-2-(4-(2-(bis(3-hydroxyphenyl)phosphono)vinyl)-2,6-dichlorobenzamide)-3-(3-(methylsulfonyl) Phenyl) propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000089
Figure PCTCN2018087629-appb-000089
步骤A:(E)-4-(2-(二(3-甲氧基苯基)膦酰基)乙烯基)-2,6-二氯苯甲酸甲酯(化合物52.1)Step A: (E)-4-(2-(bis(3-methoxyphenyl)phosphono)vinyl)-2,6-dichlorobenzoic acid methyl ester (compound 52.1)
1g化合物32.2，溶于20mlTHF，降温至0度，加入5当量间甲氧基苯基溴化镁溶液，室温搅拌4h，除去THF，纯化得产物。1 g of compound 32.2, dissolved in 20 ml of THF, cooled to 0 °, added 5 equivalents of m-methoxyphenylmagnesium bromide solution, stirred at room temperature for 4 h, THF was removed and purified.
步骤B：(E)-4-(2-(二(3-羟基苯基)膦酰基)乙烯基)-2,6-二氯苯甲酸(化合物52.2)Step B: (E)-4-(2-(bis(3-hydroxyphenyl)phosphono)vinyl)-2,6-dichlorobenzoic acid (Compound 52.2)
160mg化合物52.1，溶于10ml CH2Cl2，降温至0度，加入BBr3，反应2h后，加入水，用EA萃取后干燥，旋干得产物。160 mg of compound 52.1, dissolved in 10 ml of CH2Cl2, cooled to 0 °, added BBr3, after 2 h of reaction, water was added, extracted with EA, dried, and dried to give the product.
步骤C:(S,E)-2-(4-(2-(二(3-羟基苯基)膦酰基)乙烯基)-2,6-二氯苯酰胺)-3-(3-(甲磺酰基)苯基)丙酸苄酯(化合物52.3)Step C: (S,E)-2-(4-(2-(bis(3-hydroxyphenyl)phosphono)vinyl)-2,6-dichlorobenzamide)-3-(3-(A Benzyl sulfonyl)phenyl)propionate (compound 52.3)
50mg化合物52.2，(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸苄酯盐酸盐40mg和DIPEA 40mg溶于DMF 5ml，加入HATU 60mg，搅拌过夜，除去DMF，纯化得产物。50 mg of compound 52.2, (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester hydrochloride 40 mg and DIPEA 40 mg dissolved in DMF 5 ml, HATU 60 mg was added, stirred overnight to remove DMF , purified product.
步骤D：(S,E)-2-(4-(2-(二(3-羟基苯基)膦酰基)乙烯基)-2,6-二氯苯酰胺)-3-(3-(甲磺酰基)苯基)丙酸(化合物52)Step D: (S,E)-2-(4-(2-(bis(3-hydroxyphenyl)phosphono)vinyl)-2,6-dichlorobenzamide)-3-(3-(A Sulfonyl)phenyl)propionic acid (compound 52)
15mg化合物52.3溶于1mlTHF,加入LiOH水溶液0.2ml，搅拌0.5h，除去溶剂，纯化得产物。LCMS ESI(+)m/z:675.5(M+1).15 mg of the compound 52.3 was dissolved in 1 ml of THF, and 0.2 ml of a LiOH aqueous solution was added thereto, and the mixture was stirred for 0.5 hour, and the solvent was removed to obtain a product. LCMS ESI (+) m/z: 675.5 (M+1).
1H-NMR(400MHz,DMSO)δ9.81(s,2H),7.87(s,1H),7.89(s,2H),7.86(s,1H),7.58(t,1H),7.76(d,J＝4.6Hz,2H),7.66(m,2H),7.51(t,1H),7.34(m,3H),7.13(m,4H),6.94(d,J＝4.4Hz,1H),3.29(dd,J＝14Hz,J＝4.4Hz,1H),3.15(s,3H),3.01(dd,J＝14Hz,10.4Hz,1H)。 1 H-NMR (400 MHz, DMSO) δ 9.81 (s, 2H), 7.78 (s, 1H), 7.89 (s, 2H), 7.86 (s, 1H), 7.58 (t, 1H), 7.76 (d, J = 4.6 Hz, 2H), 7.66 (m, 2H), 7.51 (t, 1H), 7.34 (m, 3H), 7.13 (m, 4H), 6.94 (d, J = 4.4 Hz, 1H), 3.29 ( Dd, J = 14 Hz, J = 4.4 Hz, 1H), 3.15 (s, 3H), 3.01 (dd, J = 14 Hz, 10.4 Hz, 1H).
实施例53Example 53
Figure PCTCN2018087629-appb-000090
Figure PCTCN2018087629-appb-000090
(S,E)-2-(4-(2-(二(3-羟基苯基)膦酰基)乙基)-2,6-二氯苯酰胺)-3-(3-(甲磺酰基)苯基)丙酸(S,E)-2-(4-(2-(bis(3-hydroxyphenyl)phosphonyl)ethyl)-2,6-dichlorobenzamide)-3-(3-(methylsulfonyl) Phenyl) propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000091
Figure PCTCN2018087629-appb-000091
12mg化合物52溶于0.5mlMeOH,加入10％钯碳1mg，通入H2，反应3h，过滤，纯化得产物。LCMS ESI(+)m/z:677.5(M+1).12 mg of compound 52 was dissolved in 0.5 ml of MeOH, and 1 mg of 10% palladium on carbon was added thereto, and H2 was added thereto, and the mixture was reacted for 3 hours. LCMS ESI (+) m/z: 677.5 (M+1).
1H-NMR(400MHz,DMSO)δ10.63(s,2H),8.67(s,1H),8.59(d,J＝8.4Hz,1H),8.50(d,J＝8Hz,1H),8.40(t,1H),8.18(s,2H),8.16(m,3H)，8.01(m,5H),7.75(dd,J＝10Hz,2Hz,2H),3.29(dd,J＝14Hz,J＝4.4Hz,1H),3.15(s,3H),3.01(dd,J＝14Hz,10.4Hz,1H),2.81(m,2H),2.11(m,2H). 1 H-NMR (400MHz, DMSO ) δ10.63 (s, 2H), 8.67 (s, 1H), 8.59 (d, J = 8.4Hz, 1H), 8.50 (d, J = 8Hz, 1H), 8.40 ( t, 1H), 8.18 (s, 2H), 8.16 (m, 3H), 8.01 (m, 5H), 7.75 (dd, J = 10 Hz, 2 Hz, 2H), 3.29 (dd, J = 14 Hz, J = 4.4 Hz, 1H), 3.15 (s, 3H), 3.01 (dd, J = 14 Hz, 10.4 Hz, 1H), 2.81 (m, 2H), 2.11 (m, 2H).
实施例54Example 54
Figure PCTCN2018087629-appb-000092
Figure PCTCN2018087629-appb-000092
(S,E)-2-(4-(2-(4-羟基苯基)膦酰基乙烯基)-2,6-二氯苯酰胺)-3-(3-(甲磺酰基)苯基)丙酸(S,E)-2-(4-(2-(4-hydroxyphenyl)phosphonovinyl)-2,6-dichlorobenzamide)-3-(3-(methylsulfonyl)phenyl) Propionic acid
制备实施例32的完全相同的方法用来制备实施例54，其中对甲氧基溴化镁取代间甲氧基溴化镁。LCMS ESI(+)m/z:629.5(M+1).The exact same procedure as in Preparation Example 32 was used to prepare Example 54, wherein p-methoxymagnesium bromide was substituted for m-methoxymagnesium bromide. LCMS ESI (+) m/z: 629.5 (M+1).
1H-NMR(400MHz,DMSO)δ9.15(d,J＝8Hz,1H),7.87(s,1H),7.73(t,3H),7.60(d,J＝6Hz,1H),7.58(d,J＝7.2Hz,1H),7.58(t,1H),7.18(m,3H),6.91(dd,J＝10.4Hz,2Hz,2H),4.80(m,1H),3.92(m,2H),3.3(m,2H),3.12(s,1H),1.26(t,J＝7.8Hz,3H)。 1 H-NMR (400 MHz, DMSO) δ 9.15 (d, J = 8 Hz, 1H), 7.78 (s, 1H), 7.73 (t, 3H), 7.60 (d, J = 6 Hz, 1H), 7.58 (d) , J=7.2 Hz, 1H), 7.58 (t, 1H), 7.18 (m, 3H), 6.91 (dd, J = 10.4 Hz, 2 Hz, 2H), 4.80 (m, 1H), 3.92 (m, 2H) , 3.3 (m, 2H), 3.12 (s, 1H), 1.26 (t, J = 7.8 Hz, 3H).
实施例55Example 55
Figure PCTCN2018087629-appb-000093
Figure PCTCN2018087629-appb-000093
(2s)-2-(2，6-二氯-4-(1-羟基2-(甲基(苯并呋喃-3-基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(1-hydroxy2-(methyl(benzofuran-3-yl)phosphoryl)ethyl)benzamide)-3-(3) -(methylsulfonyl)phenyl)propionic acid
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000094
Figure PCTCN2018087629-appb-000094
步骤A：二甲基(苯并呋喃-3-基)氧膦(化合物55.1)Step A: dimethyl (benzofuran-3-yl) phosphine oxide (compound 55.1)
将100mg的甲基(苯并呋喃-3-基)膦酰氯溶于2ml四氢呋喃中，氮气保护，并在0度下加入0.7ml的3mol/L的甲基基溴化镁，搅拌20分钟。用1mol/L的 稀盐酸溶液淬灭反应，用30mL乙酸乙酯分三次萃取，合并有机相，旋干，纯化得产物(100mg，60％)。LCMS ESI(+)m/z:195.1(M+1).100 mg of methyl (benzofuran-3-yl)phosphonoyl chloride was dissolved in 2 ml of tetrahydrofuran, protected with nitrogen, and 0.7 ml of 3 mol/L methyl magnesium bromide was added at 0 ° C and stirred for 20 minutes. The reaction was quenched with 1 mol/L EtOAc EtOAc (EtOAc)EtOAc. LCMS ESI (+) m/z: 195.1 (M+1).
步骤B：(2s)-2-(2，6-二氯-4-(1-羟基2-(甲基(苯并呋喃-3-基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(化合物55)Step B: (2s)-2-(2,6-Dichloro-4-(1-hydroxy2-(methyl(benzofuran-3-yl)phosphoryl)ethyl)benzamide)-3 -(3-(methylsulfonyl)phenyl)propionic acid (Compound 55)
采用实施例50中完全相同的步骤，将“甲基(3-甲氧基苯基)膦酸乙酯”替换为“二甲基(苯并呋喃-3-基)氧膦”制备出化合物55。Using the exact same procedure as in Example 50, replacing "methyl (3-methoxyphenyl)phosphonate" with "dimethyl(benzofuran-3-yl)phosphine oxide" to prepare compound 55 .
LCMS ESI(+)m/z:638.1(M+1).LCMS ESI (+) m/z: 638.1 (M+1).
1H-NMR(400MHz,DMSO)δ9.09(d,J＝8Hz,1H),8.16(s,1H),8.01(t,J＝8Hz,1H)，7.87(s,1H),7.77-7.82(m,2H),7.68(m,2H),7.57(t,J＝8Hz,1H),7.38(s,2H),7.05(dd,J＝7.6Hz,4.0Hz,1H),4.90(m,1H),4.77(m,1H),3.30(dd,J＝14Hz,J＝4.4Hz,1H),3.15(s,3H),3.03(dd,J＝14Hz,10.4Hz,1H),2.40(m,2H),1.80(d,J＝13.2Hz,3H). 1 H-NMR (400 MHz, DMSO) δ 9.09 (d, J = 8 Hz, 1H), 8.16 (s, 1H), 8.1 (t, J = 8 Hz, 1H), 7.78 (s, 1H), 7.77-7.82 (m, 2H), 7.68 (m, 2H), 7.57 (t, J = 8 Hz, 1H), 7.38 (s, 2H), 7.05 (dd, J = 7.6 Hz, 4.0 Hz, 1H), 4.90 (m, 1H), 4.77 (m, 1H), 3.30 (dd, J = 14 Hz, J = 4.4 Hz, 1H), 3.15 (s, 3H), 3.03 (dd, J = 14 Hz, 10.4 Hz, 1H), 2.40 (m) , 2H), 1.80 (d, J = 13.2 Hz, 3H).
实施例56Example 56
Figure PCTCN2018087629-appb-000095
Figure PCTCN2018087629-appb-000095
(2s)-2-(2，6-二氯-4-(异丙基(3-羟基苯基)磷酰基)乙基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(isopropyl(3-hydroxyphenyl)phosphoryl)ethyl)benzamide)-3-(3-(methylsulfonyl)benzene Propionate
制备实施例21的完全相同的方法用来制备实施例56，其中异丙基格氏试剂取代乙基格氏试剂。LCMS ESI(+)m/z:626.1(M+1).The exact same procedure as in Preparation Example 21 was used to prepare Example 56, wherein the isopropyl Grignard reagent was substituted for the ethyl Grignard reagent. LCMS ESI (+) m/z: 626.1 (M+1).
1H-NMR(400MHz,DMSO)δ9.76(s,1H),9.03(d,J＝8Hz,1H),7.86(s,1H),7.77(d,J＝8Hz,1H),7.67(d,J＝8Hz,1H),7.57(t,J＝7.6Hz,1H),7.31-7.35(m,3H),7.12-7.17(m,2H),6.94(d,J＝7.6Hz,1H),4.75(m,1H),3.29(dd,J＝14Hz,J＝4.4Hz,1H), 3.15(s,3H),3.01(dd,J＝14Hz,10.4Hz,1H),2.74(m,1H),2.50(m,1H),2.29(m,2H),2.07(m,1H),1.10(dd,J＝16Hz,7.0Hz,3H),0.88(dd,J＝16Hz,7.0Hz,3H). 1 H-NMR (400 MHz, DMSO) δ 9.76 (s, 1H), 9.03 (d, J = 8 Hz, 1H), 7.86 (s, 1H), 7.77 (d, J = 8 Hz, 1H), 7.67 (d) , J = 8 Hz, 1H), 7.57 (t, J = 7.6 Hz, 1H), 7.31 - 7.35 (m, 3H), 7.12 - 7.17 (m, 2H), 6.94 (d, J = 7.6 Hz, 1H), 4.75 (m, 1H), 3.29 (dd, J = 14 Hz, J = 4.4 Hz, 1H), 3.15 (s, 3H), 3.01 (dd, J = 14 Hz, 10.4 Hz, 1H), 2.74 (m, 1H) , 2.50 (m, 1H), 2.29 (m, 2H), 2.07 (m, 1H), 1.10 (dd, J = 16 Hz, 7.0 Hz, 3H), 0.88 (dd, J = 16 Hz, 7.0 Hz, 3H).
实施例57Example 57
Figure PCTCN2018087629-appb-000096
Figure PCTCN2018087629-appb-000096
(2s)-2-(2，6-二氯-4-(环丙基(3-羟基苯基)磷酰基)乙烯基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(cyclopropyl(3-hydroxyphenyl)phosphoryl)vinyl)benzamide)-3-(3-(methylsulfonyl)benzene Propionate
具体反应方程式如下所示：The specific reaction equation is as follows:
Figure PCTCN2018087629-appb-000097
Figure PCTCN2018087629-appb-000097
步骤A:(E)-4-(2-(3-甲氧基苯基)氯膦酰基乙烯基)-2,6-二氯苯甲酸甲酯(化合物57.1)Step A: (E)-4-(2-(3-Methoxyphenyl)chlorophosphonovinyl)methyl-2,6-dichlorobenzoate (Compound 57.1)
1g化合物32.3，溶于20mlSOCl2，80度加热3小时，浓缩得产物。1 g of compound 32.3 was dissolved in 20 ml of SOCl2, heated at 80 °C for 3 hours, and concentrated to give the product.
步骤B：(E)-4-(2-(3-甲氧基苯基)环丙基膦酰基乙烯基)-2,6-二氯苯甲酸甲酯(化合物57.2)Step B: (E)-4-(2-(3-Methoxyphenyl)cyclopropylphosphonovinyl)methyl-2,6-dichlorobenzoate (Compound 57.2)
10毫升1M环丙基氯化镁的THF溶液，加入0.5g氯化铈，再加入1g化合物57.1，室温反应1h后加氯化铵溶液淬灭，萃取纯化后得产物。10 ml of 1 M solution of cyclopropylmagnesium chloride in THF, 0.5 g of ruthenium chloride, and 1 g of compound 57.1 were added, and reacted at room temperature for 1 hour, and then quenched with ammonium chloride solution to obtain a product after extraction and purification.
步骤C：(E)-4-(2-((3-羟基苯基)环丙基膦酰基)乙烯基)-2,6-二氯苯甲酸(化合物57.3)Step C: (E)-4-(2-((3-Hydroxyphenyl)cyclopropylphosphono)vinyl)-2,6-dichlorobenzoic acid (Compound 57.3)
160mg化合物57.2，溶于10ml CH2Cl2，降温至0度，加入BBr3，反应2h后，加入水，用EA萃取后干燥，旋干得产物。160 mg of compound 57.2, dissolved in 10 ml of CH2Cl2, cooled to 0 degree, BBr3 was added, and after 2 h of reaction, water was added, extracted with EA, dried, and dried to give a product.
步骤D:(S,E)-2-(4-(2-(3-羟基苯基)环丙基膦酰基)乙烯基)-2,6-二氯苯酰胺)-3-(3-(甲磺酰基)苯基)丙酸苄酯(化合物57.4)Step D: (S,E)-2-(4-(2-(3-hydroxyphenyl)cyclopropylphosphono)vinyl)-2,6-dichlorobenzamide)-3-(3-( Methyl sulfonyl)phenyl)propanoate (compound 57.4)
50mg化合物57.3，(2s)-2-氨基-3-(3-(甲磺酰基)苯基)丙酸苄酯盐酸盐40mg和DIPEA 40mg溶于DMF 5ml，加入HATU 60mg，搅拌过夜，除去DMF，纯化得产物。50 mg of compound 57.3, (2s)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester hydrochloride 40 mg and DIPEA 40 mg dissolved in DMF 5 ml, added HATU 60 mg, stirred overnight to remove DMF , purified product.
步骤E：(S,E)-2-(4-(2-(3-羟基苯基)环丙基膦酰基)乙烯基)-2,6-二氯苯酰胺)-3-(3-(甲磺酰基)苯基)丙酸(化合物57)Step E: (S,E)-2-(4-(2-(3-hydroxyphenyl)cyclopropylphosphono)vinyl)-2,6-dichlorobenzamide)-3-(3-( Methanesulfonyl)phenyl)propionic acid (compound 57)
15mg化合物57.4溶于1mlTHF,加入LiOH水溶液0.2ml，搅拌0.5h，除去溶剂，纯化得产物。LCMS ESI(+)m/z:622.1(M+1).15 mg of the compound 57.4 was dissolved in 1 ml of THF, and 0.2 ml of a LiOH aqueous solution was added thereto, and the mixture was stirred for 0.5 hour, and the solvent was removed to obtain a product. LCMS ESI (+) m/z: 6221. (M+1).
1H-NMR(400MHz,DMSO)δ9.80(s,1H),9.13(d,J＝8.8Hz,1H),7.87(s,1H),7.82(s,2H),7.78(d,J＝8Hz,2H),7.69(d,J＝7.6Hz,1H),7.58(t,J＝7.6Hz,1H),7.17-7.37(m,5H),6.96(m,1H),4.79(m,1H),3.30(dd,J＝14Hz,J＝4.4Hz,1H),3.15(s,3H),3.04(dd,J＝14Hz,10.4Hz,1H),1.25(m,1H),0.66-0.89(m,4H). 1 H-NMR (400MHz, DMSO ) δ9.80 (s, 1H), 9.13 (d, J = 8.8Hz, 1H), 7.87 (s, 1H), 7.82 (s, 2H), 7.78 (d, J = 8 Hz, 2H), 7.69 (d, J = 7.6 Hz, 1H), 7.58 (t, J = 7.6 Hz, 1H), 7.17-7.37 (m, 5H), 6.96 (m, 1H), 4.79 (m, 1H) ), 3.30 (dd, J = 14 Hz, J = 4.4 Hz, 1H), 3.15 (s, 3H), 3.04 (dd, J = 14 Hz, 10.4 Hz, 1H), 1.25 (m, 1H), 0.66-0.89 ( m, 4H).
实施例58Example 58
Figure PCTCN2018087629-appb-000098
Figure PCTCN2018087629-appb-000098
(2s)-2-(2，6-二氯-4-(甲基(3-羟基苯基)磷酰基)乙烯基)苯甲酰胺基)-3-(3-(甲磺酰基)苯基)丙酸(2s)-2-(2,6-Dichloro-4-(methyl(3-hydroxyphenyl)phosphoryl)vinyl)benzamide)-3-(3-(methylsulfonyl)phenyl Propionic acid
制备实施例57的完全相同的方法用来制备实施例58，其中甲基格氏试剂取代环丙基格氏试剂。LCMS ESI(+)m/z:596.1(M+1).The exact same procedure as in Preparation Example 57 was used to prepare Example 58 wherein the methyl Grignard reagent was substituted for the cyclopropyl Grignard reagent. LCMS ESI (+) m/z: 596.1 (M+1).
1H-NMR(400MHz,DMSO)δ9.80(s,1H),9.14(d,J＝8.8Hz,1H),7.87(s,1H),7.78(m,3H),7.69(d,J＝7.6Hz,1H),7.58(t,J＝7.6Hz,1H),7.38(m,1H),7.25(d,J＝20Hz,2H),7.15-7.18(m,2H),6.96(m,1H),4.79(m,1H),3.30(dd,J＝14Hz,J＝4.4Hz,1H),3.15(s,3H),3.04(dd,J＝14Hz,10.4Hz,1H),1.73(d,,J＝13.2Hz 3H). 1 H-NMR (400 MHz, DMSO) δ 9.80 (s, 1H), 9.14 (d, J = 8.8 Hz, 1H), 7.78 (s, 1H), 7.78 (m, 3H), 7.69 (d, J = 7.6 Hz, 1H), 7.58 (t, J = 7.6 Hz, 1H), 7.38 (m, 1H), 7.25 (d, J = 20 Hz, 2H), 7.15-7.18 (m, 2H), 6.96 (m, 1H) ), 4.79 (m, 1H), 3.30 (dd, J = 14 Hz, J = 4.4 Hz, 1H), 3.15 (s, 3H), 3.04 (dd, J = 14 Hz, 10.4 Hz, 1H), 1.73 (d, , J=13.2Hz 3H).
细胞粘和抑制实验：Cell adhesion and inhibition experiments:
使用人类T淋巴细胞株Jurkat(ATCC TIB-152)进行T-细胞粘附试验：在PBS中将山羊Anti-Human IgG(Fc specific)(Sigma I8885)稀释至10μg/ml，在4℃每孔100uL/96孔板培育12小时.倾倒掉孔板中的液体，用200uL 1％的BSA在37℃封闭90分钟，再用用PBS洗板3次。每孔加入50uL的1ug/mL的ICAM-1(含0.1％BSA，0.01％Tween20)，在37℃培育3小时。用assay的缓冲液(20mM HEPES pH 7.6,140mM NaCl,1mM MgCl 2,1mM MnCl 2,0.2％glucose)洗板3次。 T-cell adhesion assay using human T lymphocyte strain Jurkat (ATCC TIB-152): goat anti-Human IgG (Fc specific) (Sigma I8885) was diluted to 10 μg/ml in PBS at 100 °L per well at 4 °C The cells were incubated for 12 hours in a /96 well plate. The liquid in the well plate was poured out, blocked with 200 uL of 1% BSA at 37 ° C for 90 minutes, and washed three times with PBS. 50 uL of 1 ug/mL ICAM-1 (containing 0.1% BSA, 0.01% Tween 20) was added to each well and incubated at 37 ° C for 3 hours. The plate was washed 3 times with assay buffer (20 mM HEPES pH 7.6, 140 mM NaCl, 1 mM MgCl 2 , 1 mM MnCl 2 , 0.2% glucose).
Jurkat细胞仪100-G离心处理，重新将细胞混悬入37℃的assay的缓冲液(20mM HEPES pH 7.6,140mM NaCl,1mM MgCl 2,1mM MnCl2,0.2％ glucose)， The Jurkat cytometer was centrifuged at 100-G, and the cells were resuspended in an assay buffer (20 mM HEPES pH 7.6, 140 mM NaCl, 1 mM MgCl 2 , 1 mM MnCl 2 , 0.2% glucose) at 37 ° C.
每mL的细胞悬浮液中加入2μl 1mM of BCECF-AM。在37℃培育30分钟，培育过程中，每10分钟搅拌一次。培育完后，细胞用37℃的assay缓冲液洗一下。将细胞悬浮到6x10 6/mL的浓度。 2 μl of 1 mM of BCECF-AM was added per mL of the cell suspension. Incubate at 37 ° C for 30 minutes, and stir every 10 minutes during the incubation. After the incubation, the cells were washed with an assay buffer at 37 °C. The cells were suspended to a concentration of 6 x 10 6 /mL.
在测定缓冲液中将抑制剂稀释至2X终浓度，并在室温下取50uL的化合物溶液和60uL的Jurkat细胞混合，在37℃培育30分钟。将100μL/孔的细胞和抑制剂加入到板中并在室温下培养1小时。用荧光仪测量总荧光度:ex:485；em:530；cutoff:530来测量总荧光度。将板用assay缓冲液洗一次，再用荧光仪测量荧光度:ex:485；em:530；cutoff:。得出结果绘成抑制-浓度关系图，并用标准方法计算出EC 50。表一显示了用此办法测得的部分EC 50值。 The inhibitor was diluted to a final concentration of 2X in assay buffer, and 50 uL of the compound solution and 60 uL of Jurkat cells were mixed at room temperature, and incubated at 37 ° C for 30 minutes. 100 μL/well of cells and inhibitor were added to the plate and incubated for 1 hour at room temperature. The total fluorescence was measured by a fluorometer: ex: 485; em: 530; cutoff: 530 to measure the total fluorescence. The plate was washed once with the assay buffer and the fluorescence was measured with a fluorometer: ex: 485; em: 530; cutoff:. Results obtained are plotted inhibition - concentration diagram, and the EC 50 calculated by standard methods. Table 1 shows the partial EC 50 values measured by this method.
表一：细胞粘和抑制的EC 50 Table 1: EC 50 of cell adhesion and inhibition
实施例Example EC50(nM)EC50(nM) 实施例Example EC50(nM)EC50(nM)
11 3030 21twenty one 7.37.3
22 9.49.4 22twenty two 22twenty two
33 8.58.5 23twenty three 6161
44 1111 24twenty four 10.210.2
55 22twenty two 2525 6363
66 1717 2626 23twenty three
77 7.27.2 2727 NA * NA *
88 6.26.2 2828 NA * NA *
99 11.811.8 2929 NA * NA *
1010 7878 3030 7.27.2
1111 13.513.5 3131 7.17.1
1212 2929 3232 3.73.7
1313 2929 3333 10.810.8
1414 8.58.5 3434 12.512.5
1515 1.81.8 3535 3.83.8
1616 7.27.2 3636 3030
1717 1515 3737 >1000>1000
1818 NA * NA * 3838 24twenty four
1919 230230 3939 NA * NA *
2020 NA * NA * 4040 NA * NA *
表一(继续)Table 1 (continued)
实施例Example EC50(nM)EC50(nM) 实施例Example EC50(nM)EC50(nM)
4141 NANA 5151 4.24.2
4242 150150 5252 7474
4343 6969 5353 22twenty two
4444 NA * NA * 5454 9.69.6
4545 >1000>1000 5555 1919
4646 >1000>1000 5656 6363
4747 340340 5757 5.35.3
4848 6.86.8 5858 1.91.9
4949 10.810.8
5050 1616
*NA:没有数据 * NA: No data
实施例：制剂Example: Preparation
实施例11所得到的化合物，5.0g加入90mL的无菌生理盐水中，加入0.7g的NaOH，搅拌得到透明溶液；向以上得到的溶液中加入NaH 2PO 4饱和水溶液，直到溶液的pH在6.75-7.25之间。向得到水溶液加入无菌生理盐水，直到总体积达到100.0mL。向以上溶液通氮气，鼓泡1小时，所得溶液密封，放在5℃避光保存。分装到一次性使用的滴眼液袋中，每个含60mL的制剂溶液。根据具体化合物性质和使用要求的需要，该制剂的方法和具体比例还可按需调整。 The compound obtained in Example 11 was added to 90 mL of sterile physiological saline, and 0.7 g of NaOH was added thereto, followed by stirring to obtain a transparent solution; and a saturated aqueous solution of NaH 2 PO 4 was added to the solution obtained above until the pH of the solution was 6.75. Between -7.25. Sterile physiological saline was added to the obtained aqueous solution until the total volume reached 100.0 mL. The above solution was purged with nitrogen, and bubbled for 1 hour, and the resulting solution was sealed and stored at 5 ° C in the dark. Dispense into disposable eye drops bags, each containing 60 mL of formulation solution. The method and specific ratio of the formulation can also be adjusted as needed, depending on the nature of the particular compound and the requirements of the application.

Claims (15)

  1. 一种含磷化合物，其特征在于，为如下结构所示的化合物：A phosphorus-containing compound characterized by being a compound represented by the following structure:
    Figure PCTCN2018087629-appb-100001
    Figure PCTCN2018087629-appb-100001
    R 1选自烷基、芳基、苄基及其衍生物； R 1 is selected from the group consisting of alkyl, aryl, benzyl and derivatives thereof;
    R 2选自羟基、烷基、氢、烷氧基、胺基，烷胺基； R 2 is selected from the group consisting of a hydroxyl group, an alkyl group, a hydrogen, an alkoxy group, an amine group, and an alkylamine group;
    n选自0和1；n is selected from 0 and 1;
    X选自碳、氧、氮；X is selected from the group consisting of carbon, oxygen, and nitrogen;
    Z选自羰基、烷基、支链烷基、磺酰基、氮、氧、硫；Z is selected from the group consisting of a carbonyl group, an alkyl group, a branched alkyl group, a sulfonyl group, a nitrogen, an oxygen, and a sulfur;
    R 3为苯环上任一或任几取代的氢、烷基、烷氧基、卤素、氨基、氰基、羟基、硝基、芳基； R 3 is any optionally substituted hydrogen, alkyl, alkoxy, halogen, amino, cyano, hydroxy, nitro, aryl;
    Y选自碳、氧、氮；Y is selected from the group consisting of carbon, oxygen, and nitrogen;
    R 4选自烷基、芳基、苄基及其衍生物； R 4 is selected from the group consisting of alkyl, aryl, benzyl and derivatives thereof;
    R 5选自氢、烷基、芳基、苄基及其衍生物； R 5 is selected from the group consisting of hydrogen, alkyl, aryl, benzyl and derivatives thereof;
    G1和G2所代表的取代基团呈间位、对位或邻位的设置于苯环上。The substituent groups represented by G1 and G2 are disposed on the benzene ring in a meta, para or ortho position.
  2. 如权利要求1所述的一种含磷化合物，其特征在于：A phosphorus-containing compound according to claim 1 wherein:
    所述芳基选自苯基及其衍生物、萘基及其衍生物、N杂或O杂环苯基及其衍生物、 N杂或O杂环萘基及其衍生物；The aryl group is selected from the group consisting of phenyl and its derivatives, naphthyl and its derivatives, N or O heterocyclic phenyl and its derivatives, N or O heterocyclonaphthyl and derivatives thereof;
    其中，所述衍生物指在苯环上任一或任几取代的氢、烷基、烷氧基、卤素、氨基、氰基、羟基、硝基、芳基、烷基磺酰基、苯基磺酰基。Wherein the derivative refers to any or any of a substituted hydrogen, alkyl, alkoxy, halogen, amino, cyano, hydroxy, nitro, aryl, alkylsulfonyl, phenylsulfonyl group on the phenyl ring. .
  3. 如权利要求1所述的一种含磷化合物，其特征在于：A phosphorus-containing compound according to claim 1 wherein:
    所述R 4选自如下结构所示的基团： The R 4 is selected from the group consisting of the following structures:
    Figure PCTCN2018087629-appb-100002
    Figure PCTCN2018087629-appb-100002
    n选自0-5的整数；n is selected from an integer from 0 to 5;
    所述A选自硫、碳、氮、氧；The A is selected from the group consisting of sulfur, carbon, nitrogen, and oxygen;
    所述R 42选自芳基、烷基、烷基氨基、烷基磺酰胺基、环烷基、取代的环烷基、杂环烷基、取代的杂环烷基； The R 42 is selected from the group consisting of aryl, alkyl, alkylamino, alkylsulfonylamino, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl;
    其中，所述芳基选自6-12元的芳香基及其衍生物、5-12元的芳香环上，任一或任几个碳原子为氧、氮、硫替代的杂芳基；Wherein the aryl group is selected from the group consisting of a 6-12 membered aromatic group and a derivative thereof, a 5-12 membered aromatic ring, and any or any of several carbon atoms is a heteroaryl group substituted with oxygen, nitrogen or sulfur;
    其中，所述衍生物指在芳香环上任一或任几取代的氢、烷基、烷氧基、卤素、氨基、氰基、羟基、硝基、磺酰基、烷基磺酰基、苯基磺酰基。Wherein said derivative refers to any or any of a substituted hydrogen, alkyl, alkoxy, halogen, amino, cyano, hydroxy, nitro, sulfonyl, alkylsulfonyl, phenylsulfonyl group on the aromatic ring. .
    所述杂芳基上还可以具有-N-R 422的结构； The heteroaryl group may also have a structure of -NR 422 ;
    所述R 422为磺酰基、烷基磺酰基、烷基、羟基； The R 422 is a sulfonyl group, an alkylsulfonyl group, an alkyl group, or a hydroxyl group;
    所述环烷基为3-12元的环烷基；The cycloalkyl group is a 3-12 membered cycloalkyl group;
    所述取代的环烷基为环上任一或任几取代的磺酰基、烷基磺酰基、烷基、烷氧基、羟基、氨基、硝基；The substituted cycloalkyl group is a sulfonyl group, an alkylsulfonyl group, an alkyl group, an alkoxy group, a hydroxyl group, an amino group, a nitro group;
    所述杂环烷基为3-12元的杂环烷基上，任一或任几个碳原子为氧、氮、硫替代；The heterocycloalkyl group is a 3-12 membered heterocycloalkyl group, and any or any of the carbon atoms is replaced by oxygen, nitrogen, or sulfur;
    所述杂环烷基上的碳原子还可以为C＝O基和/或SO和/或SO 2基替代； The carbon atom on the heterocycloalkyl group may also be a C=O group and/or a SO and/or SO 2 group substitution;
    所述取代的杂环烷基为环上任一或任几磺酰基、烷基磺酰基、烷基、烷氧基、羟基、氨基、硝基、羰基取代的氮杂、氧杂或硫杂的四元、五元、六元或七元环的环烷基；The substituted heterocycloalkyl group is a ring of any or any of a sulfonyl group, an alkylsulfonyl group, an alkyl group, an alkoxy group, a hydroxyl group, an amino group, a nitro group, a carbonyl-substituted aza, an oxa or a thia. a cycloalkyl group of a meta, five, six or seven membered ring;
    所述取代的杂环烷基上还可以具有-N-R 422的结构； The substituted heterocycloalkyl group may further have a structure of -NR 422 ;
    所述R 422为磺酰基、烷基磺酰基、烷基、羟基； The R 422 is a sulfonyl group, an alkylsulfonyl group, an alkyl group, or a hydroxyl group;
    所述R 42还可以选自如下结构的基团： The R 42 may also be selected from the group consisting of:
    Figure PCTCN2018087629-appb-100003
    Figure PCTCN2018087629-appb-100003
    所述R 43、R 44为相同或不相同的烷基、羟基、羟基取代的碳原子数不大于5的烷基； The R 43 and R 44 are the same or different alkyl group, a hydroxyl group, and a hydroxyl group substituted with an alkyl group having not more than 5 carbon atoms;
    G3为3-12元的环；G3 is a ring of 3-12 yuan;
    所述G3的环上的碳原子还可以部分的为氧、硫、氮、C＝O或SO 2替代； The carbon atom on the ring of G3 may also be partially replaced by oxygen, sulfur, nitrogen, C=O or SO 2 ;
    所述R 45为G3环上一个或几个取代的烷基、羟基、烷氧基、氨基。 The R 45 is one or more substituted alkyl groups, hydroxyl groups, alkoxy groups, amino groups on the G3 ring.
  4. 如权利要求1所述的一种含磷化合物，其特征在于，为如下结构所示的化合物：A phosphorus-containing compound according to claim 1, which is a compound represented by the following structure:
    Figure PCTCN2018087629-appb-100004
    Figure PCTCN2018087629-appb-100004
    其中，所述C 1和C 2之间的碳键为单键、双键或三键。 Wherein, the carbon bond between C 1 and C 2 is a single bond, a double bond or a triple bond.
  5. 如权利要求4所述的一种含磷化合物，其特征在于，为如下结构所示的化合物：A phosphorus-containing compound according to claim 4, which is a compound represented by the following structure:
    Figure PCTCN2018087629-appb-100005
    Figure PCTCN2018087629-appb-100005
    R 11选自苯环上任一或任几取代的氢、烷基、烷氧基、卤素、氨基、氰基、羟基、硝基； R 11 is selected from any one or more substituted hydrogen, alkyl, alkoxy, halogen, amino, cyano, hydroxy, nitro;
    R 2选自羟基、烷基、烷氧基； R 2 is selected from the group consisting of hydroxyl, alkyl, alkoxy;
    R 3为苯环上任一或任几取代的氢、烷基、烷氧基、卤素、氨基、氰基、羟基、硝基、芳基； R 3 is any optionally substituted hydrogen, alkyl, alkoxy, halogen, amino, cyano, hydroxy, nitro, aryl;
    Y 1选自氢、烷基； Y 1 is selected from the group consisting of hydrogen and alkyl;
    R 41选自苯环上任一或任几取代的氢、烷基、烷氧基、烷基磺酰基、芳基磺酰基、卤素、氨基、氰基、羟基、硝基； R 41 is selected from any one or more substituted hydrogen, alkyl, alkoxy, alkylsulfonyl, arylsulfonyl, halogen, amino, cyano, hydroxy, nitro;
    R 5选自氢、烷基、芳基、苄基及其衍生物。 R 5 is selected from the group consisting of hydrogen, alkyl, aryl, benzyl and derivatives thereof.
  6. 如权利要求5所述的一种含磷化合物，其特征在于，为如下结构所示的化合物：A phosphorus-containing compound according to claim 5, which is a compound represented by the following structure:
    Figure PCTCN2018087629-appb-100006
    Figure PCTCN2018087629-appb-100006
    其中，X 1、X 2、X 3、X 4选自氢、烷基、卤素、羟基、烷氧基。 Wherein X 1 , X 2 , X 3 and X 4 are selected from the group consisting of hydrogen, alkyl, halogen, hydroxy and alkoxy.
  7. 一种含磷化合物的制备方法，其特征在于：A method for preparing a phosphorus-containing compound, characterized in that:
    由化合物A和化合物C，依次与化合物B上的活性点位进行反应后获得；Obtained by reacting Compound A and Compound C with the active sites on Compound B in sequence;
    其中，所述化合物A为如下结构所示的化合物：Wherein the compound A is a compound represented by the following structure:
    Figure PCTCN2018087629-appb-100007
    Figure PCTCN2018087629-appb-100007
    所述化合物C为如下结构所示的化合物：The compound C is a compound represented by the following structure:
    Figure PCTCN2018087629-appb-100008
    Figure PCTCN2018087629-appb-100008
    所述化合物B为如下结构所示的化合物：The compound B is a compound represented by the following structure:
    Figure PCTCN2018087629-appb-100009
    Figure PCTCN2018087629-appb-100009
    其中，L 1与L 1‘以及L 2与L 2‘分别为一对可相互发生反应的活性基团，在反应的过程中，通过L 1与L 1‘反应，L 2与L 2‘反应获得目标产物； Wherein, L 1 L 1 'and L 2 and L 2', respectively, a pair of reactive groups can react with each other, in the course of the reaction, by L 1 and L 1 'reaction, L 2 and L 2' is reacted with Obtaining the target product;
    R 1选自烷基、芳基、苄基及其衍生物； R 1 is selected from the group consisting of alkyl, aryl, benzyl and derivatives thereof;
    R 2选自羟基、烷基、烷氧基、卤素； R 2 is selected from the group consisting of hydroxyl, alkyl, alkoxy, halogen;
    n选自1-3的自然数；n is selected from a natural number of 1-3;
    R 3为苯环上任一或任几取代的氢、烷基、烷氧基、卤素、氨基、氰基、羟基、硝基、芳基； R 3 is any optionally substituted hydrogen, alkyl, alkoxy, halogen, amino, cyano, hydroxy, nitro, aryl;
    R 4选自烷基、烷氧基、芳基、苄基及其衍生物； R 4 is selected from the group consisting of alkyl, alkoxy, aryl, benzyl and derivatives thereof;
    R 5选自氢、烷基、芳基、苄基及其衍生物。 R 5 is selected from the group consisting of hydrogen, alkyl, aryl, benzyl and derivatives thereof.
  8. 如权利要求7所述的一种含磷化合物的制备方法，其特征在于：A method of preparing a phosphorus-containing compound according to claim 7, wherein:
    所述L 1与L 1‘以及L 2与L 2‘之间发生，取代反应、加成反应、消除反应或置换反应。 The L 1 and L 1 ' and L 2 and L 2 ' occur between the substitution reaction, the addition reaction, the elimination reaction or the displacement reaction.
  9. 如权利要求7所述的一种含磷化合物的制备方法，其特征在于：A method of preparing a phosphorus-containing compound according to claim 7, wherein:
    所述L 1选自卤素、氨基、氰基、硫基、羟基、烷氧基； The L 1 is selected from the group consisting of halogen, amino, cyano, thio, hydroxy, alkoxy;
    所述L 1‘选自卤素、炔基、羧基、氨基、氰基、酯基、烷氧基、磺酰氨基基、烷氧基磺酰基； The L 1 ' is selected from the group consisting of halogen, alkynyl, carboxy, amino, cyano, ester, alkoxy, sulfonylamino, alkoxysulfonyl;
    所述L 2选自卤素、羧基、氨基、氰基、酯基、烷氧基、磺酰氨基基、烷氧基磺酰基； The L 2 is selected from the group consisting of halogen, carboxyl, amino, cyano, ester, alkoxy, sulfonylamino, alkoxysulfonyl;
    所述L 2‘选自卤素、氨基、硫基、羟基、烷氧基。 The L 2 ' is selected from the group consisting of halogen, amino, thio, hydroxy, alkoxy.
  10. 如权利要求7所述的一种含磷化合物的制备方法，其特征在于：A method of preparing a phosphorus-containing compound according to claim 7, wherein:
    所述化合物A与化合物C的摩尔比为1:0.1-10；The molar ratio of the compound A to the compound C is 1:0.1-10;
    所述化合物C与化合物B的摩尔比为1:0.1-10。The molar ratio of the compound C to the compound B is 1:0.1-10.
  11. 如权利要求7所述的一种含磷化合物的制备方法，其特征在于，具体工艺步骤如下所示：A method for preparing a phosphorus-containing compound according to claim 7, wherein the specific process steps are as follows:
    步骤一、于磷酸二酯的衍生物中加入卤化试剂，于50-100℃的温度下反应1-5小时后，直接旋干获得底物1；Step 1, adding a halogenating reagent to the derivative of the phosphoric acid diester, reacting at a temperature of 50-100 ° C for 1-5 hours, and directly spinning to obtain a substrate 1;
    步骤二、于0℃以下的温度下，于乙炔基苯甲酸甲酯的衍生物中，依次加入格氏试剂和底物1，反应0.1-2小时，用酸溶液淬灭反应，萃取有机相旋干，得到中间产物1；Step 2, in the derivative of methyl ethynylbenzoate at a temperature below 0 ° C, the Grignard reagent and the substrate 1 are sequentially added, the reaction is carried out for 0.1-2 hours, the reaction is quenched with an acid solution, and the organic phase is extracted. Dry to obtain intermediate product 1;
    步骤三、将中间产物1和脱酯基试剂，在100-150摄氏度的温度下反应2-5小时，加入酸溶液，萃取有机相旋干，得到中间产物2；Step 3, the intermediate product 1 and the de-esterification reagent, reacted at a temperature of 100-150 degrees Celsius for 2-5 hours, adding an acid solution, extracting the organic phase to spin dry, to obtain an intermediate product 2;
    步骤四、于中间产物2中，依次加入L 2为氨基的化合物C，碱性催化剂，在20-50℃的温度下，反应1-10小时，用酸溶液淬灭反应，萃取有机相旋干，得到含炔基的含磷化合物。 Step 4, in the intermediate product 2, sequentially add L 2 as the amino group of the compound C, a basic catalyst, at a temperature of 20-50 ° C, the reaction is 1-10 hours, quench the reaction with an acid solution, extract the organic phase spin dry A phosphorus-containing compound containing an alkynyl group is obtained.
  12. 如权利要求11所述的一种含磷化合物的制备方法，其特征在于：A method for preparing a phosphorus-containing compound according to claim 11, wherein:
    所述含炔基的含磷化合物经还原反应、酯化反应、酰胺化反应、取代反应、加成反应中的一种或几种反应后,可得到相应的含磷产物。After the alkynyl-containing phosphorus-containing compound is subjected to one or more reactions of a reduction reaction, an esterification reaction, an amidation reaction, a substitution reaction, and an addition reaction, a corresponding phosphorus-containing product can be obtained.
  13. 一种含磷化合物的应用，其特征在于：如权利要求1-12任一所述的含磷化合物，能作为一种免疫细胞迁徙抑制剂使用。Use of a phosphorus-containing compound, characterized in that the phosphorus-containing compound according to any one of claims 1 to 12 can be used as an immune cell migration inhibitor.
  14. 如权利要求13所述的一种含磷化合物的应用，其特征在于：包含有如权利要求1-12任一所述的含磷化合物的眼药水，用于缓解和治疗干眼症。The use of a phosphorus-containing compound according to claim 13, comprising an eyedrop containing a phosphorus-containing compound according to any one of claims 1 to 12 for relieving and treating dry eye.
  15. 如权利要求14所述的一种含磷化合物的眼药水，其特征在于：A eye drop containing a phosphorus compound according to claim 14, wherein:
    将如权利要求1-12任一所述的含磷化合物加入无菌生理盐水中后，加入氢氧化钠搅拌得到透明溶液；向以上得到的溶液中加入NaH 2PO 4饱和水溶液，直到溶液的pH在6.75-7.25之间，用无菌生理盐水定容后，向以上溶液通氮气，鼓泡0.1-5小时，所得溶液密封，放在5℃避光保存待用； After adding the phosphorus-containing compound according to any one of claims 1 to 12 to a sterile physiological saline, stirring with sodium hydroxide to obtain a transparent solution; adding a saturated aqueous solution of NaH 2 PO 4 to the solution obtained above until the pH of the solution Between 6.75-7.25, after constant volume with sterile physiological saline, nitrogen gas is bubbled through the above solution, bubbling for 0.1-5 hours, and the resulting solution is sealed and stored at 5 ° C in the dark;
    其中，氢氧化钠和NaH 2PO 4饱和水溶液可以被其他缓冲溶液替代。 Among them, sodium hydroxide and a saturated aqueous solution of NaH 2 PO 4 can be replaced by other buffer solutions.
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US11111259B2 (en) 2015-12-18 2021-09-07 Unity Biotechnology, Inc. Acylsulfonamide derivatives for treating senescence-associated diseases and disorders
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