WO2020177128A1 - 2,6-diazaspiro[3.4]octane pyrimidine-hydroxamic acid compound and use thereof - Google Patents

2,6-diazaspiro[3.4]octane pyrimidine-hydroxamic acid compound and use thereof Download PDF

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WO2020177128A1
WO2020177128A1 PCT/CN2019/077356 CN2019077356W WO2020177128A1 WO 2020177128 A1 WO2020177128 A1 WO 2020177128A1 CN 2019077356 W CN2019077356 W CN 2019077356W WO 2020177128 A1 WO2020177128 A1 WO 2020177128A1
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compound
group
unsubstituted
substituted
diazaspiro
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PCT/CN2019/077356
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French (fr)
Chinese (zh)
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李剑
江陆斌
王熳炯
李若曦
汤同柯
凌大正
黄正辉
朱进
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华东理工大学
中国科学院上海巴斯德研究所
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Priority to PCT/CN2019/077356 priority Critical patent/WO2020177128A1/en
Publication of WO2020177128A1 publication Critical patent/WO2020177128A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00—Antiparasitic agents
    • A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
    • A61P33/06—Antimalarials
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing three or more hetero rings
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present invention relates to the fields of medicinal chemistry and pharmacotherapy. Specifically, the present invention provides a class of small molecule compounds with 2,6-diazaspiro[3.4]octane-like pyrimidine-hydroxamic acid structure and their pharmacy Acceptable salt, the compound has excellent malaria parasite killing activity in vivo and in vitro. Such compounds are expected to be developed into new anti-malaria drugs.
  • Malaria is one of the most prominent problems in public health in the world today, and it is the most widespread and harmful parasitic disease in the world. According to the World Health Organization (WHO) World Malaria Report in 2018, there were 219 million malaria cases worldwide in 2017, including 435,000 deaths; most of the malaria cases occurred in the WHO African region (200 million cases, 92%), among which children under 5 years old are the most vulnerable population. In addition to endangering human health and life safety, the malaria epidemic has severely weakened the health systems of many African countries, profoundly affected the socio-economic development of developing countries, and made poverty a vicious circle that cannot be escaped. Coupled with factors such as global warming, frequent international exchanges, and a slowdown in global funding for malaria control, global malaria control will face enormous challenges.
  • Malaria is caused by a parasite called Plasmodium, which is spread by the bite of an infected mosquito. This parasite reproduces in the human liver and then infects red blood cells. Symptoms of malaria include fever, headache, and vomiting, which usually appear 10-15 days after a mosquito bite. If left untreated, malaria may interrupt the blood supply to vital organs that sustain life, thereby quickly threatening life.
  • the purpose of the present invention is to provide a pharmaceutical compound that can effectively kill malaria parasites, especially drug-resistant malaria parasites.
  • the first aspect of the present invention provides a 2,6-diazaspiro[3.4]octane pyrimidine-hydroxamic acid compound represented by the following formula I, or a pharmaceutically acceptable salt or optical isoform thereof Structure,
  • R 1 is selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C15 monocyclic, bicyclic or tricyclic aryl, substituted Or unsubstituted 5-15 membered monocyclic, bicyclic or tricyclic heterocyclic group (including saturated, partially unsaturated or aromatic heterocyclic group); wherein, the heteroaryl group includes one or more selected from Group of heteroatoms as the ring skeleton: N, O or S;
  • R 2 is selected from the group consisting of NHOH, or OR 3 ;
  • R 3 is selected from the following group: hydrogen, substituted or unsubstituted C1-C3 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 5-15 membered heterocyclic group;
  • R 2 is selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, or substituted or unsubstituted Groups of the following groups:
  • X is selected from N, O, and S.
  • the aryl group is an aryl group containing 1 to 4 bicyclic rings.
  • the heterocyclic group is a heteroaryl group containing 1-4 bicyclic rings, wherein each bicyclic ring is independently a 5- to 6-membered aromatic ring or aromatic heterocyclic ring.
  • the C1-C6 alkyl group is methyl or ethyl.
  • the R 1 is selected from the group consisting of substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C15 aryl , Substituted or unsubstituted 5-15 membered heteroaryl; wherein, the C3-C8 cycloalkyl group is selected from the following group: cyclopentyl, cyclohexyl;
  • the aryl group is selected from the following group: phenyl, naphthyl, phenanthryl;
  • the 5-15 membered heteroaryl group is selected from the following group:
  • the substituents are selected from the group consisting of halogen, OH, NH 2 , CN, C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl, C1-C6 alkoxy Group, C1-C6 alkylamino.
  • the R 1 is NHOH.
  • the compound of formula I can be selected from the following group:
  • the second aspect of the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising (a) a therapeutically effective amount of the compound as described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, Hydrate or solvate; and (b) a pharmaceutically acceptable carrier.
  • the pharmaceutical composition is used for:
  • the pharmaceutical composition is used to regulate the activity or expression of HDAC.
  • the third aspect of the present invention provides a use of the compound of formula I as described in the first aspect of the present invention for preparing a pharmaceutical composition for treating or preventing diseases or disorders caused by malaria parasites.
  • the disease or condition is malaria.
  • the Plasmodium is resistant or non-drug resistant.
  • the plasmodium is a plasmodium in a stage selected from the group consisting of hepatic stage, gametophyte stage, and intraerythrocytic stage.
  • the drug-resistant plasmodium is a plasmodium that has developed resistance to drugs selected from the group consisting of artemisinin, dihydroartemisinin, artemether, artesunate, and fluorene Alcohol, sulfadoxine, pyrimethamine, pyronaridine, atovaquinone, quinine, chloroquine, piperquine, mefloquine, amodiaquine, primaquine, and tafenoquine.
  • drugs selected from the group consisting of artemisinin, dihydroartemisinin, artemether, artesunate, and fluorene Alcohol, sulfadoxine, pyrimethamine, pyronaridine, atovaquinone, quinine, chloroquine, piperquine, mefloquine, amodiaquine, primaquine, and tafenoquine.
  • the fourth aspect of the present invention provides a use of the compound of formula I as described in the first aspect of the present invention for preparing a pharmaceutical composition for treating or preventing diseases or disorders related to the activity or expression of HDAC.
  • the disease or condition is a tumor.
  • the tumor is selected from the group consisting of lung cancer, colon cancer, prostate cancer, breast cancer, ovarian cancer, and lymphatic system tumors.
  • Figure 1 is a Western Blot experimental result diagram of the compound of the present invention.
  • Figure 2 is a broken line diagram of the protozoan rate in mice in the in vivo drug efficacy experiment in mice;
  • Figure 3 is a broken line graph of the survival ratio of mice in the results of in vivo drug efficacy experiments in mice.
  • the inventor prepared a compound with HDAC inhibitory activity, which can kill drug-resistant and non-drug-resistant plasmodium activity in various periods, and has low toxicity to human cells, so it can For the treatment of diseases caused by malaria parasites such as malaria. Based on the above findings, the inventor completed the present invention.
  • the present invention provides a compound having the structure of the following general formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:
  • R 1 is selected from H, straight or branched chain alkyl or II:
  • Ring A in formula II is an aliphatic cycloalkyl group, a ring aryl group, and a heteroatom-containing ring aryl group.
  • the compound according to general formula I is characterized in that the A ring in II is a 3-8 membered alicyclic alkyl group.
  • the compound of general formula I is characterized in that the ring A in II is a ring aryl group with or without heteroatoms, wherein the number of heteroatoms is 0-4, and the type of heteroatoms is selected from one of nitrogen, oxygen, and sulfur. Two or three.
  • the compound in accordance with formula I is characterized in that the cyclic aryl group described by ring A is an aryl group containing 1 to 4 parallel rings, wherein the aromatic ring is composed of a 5- to 6-membered aromatic ring or aromatic heterocyclic ring, each The number of heteroatoms constituting an aromatic ring is 0-2, and the heteroatom types are selected from one or two of nitrogen, oxygen, and sulfur.
  • R 2 and R 3 are independently selected from H, halogen, OH, NH 2 , CN, linear or branched alkyl, cycloalkyl, aryl, alkoxy, alkyl Amino.
  • X is selected from N, O, S.
  • the compound according to the general formula I is characterized in that it is used to prepare a pharmaceutical composition or preparation for inhibiting the activity of histone deacetylase (HDAC), or for preparing a pharmaceutical composition or preparation for inhibiting Plasmodium or treating malaria.
  • HDAC histone deacetylase
  • the pharmaceutical composition contains 0.001 to 99% by weight, preferably 0.1 to 90% by weight, and more preferably 1 to 80% by weight of the compound of general formula I or a pharmaceutically acceptable salt thereof, based on the total weight of the composition.
  • the dosage form of the medicine or pharmaceutical composition is an oral dosage form or an injection. Oral dosage forms include tablets, capsules, films, granules, etc., and also include sustained-release or non-sustained-release dosage forms.
  • the pharmaceutical composition may also contain other pharmaceutical ingredients with anti-malarial activity, including but not limited to artemisinin, dihydroartemisinin, artemether, artesunate, benfluorenol, sulfadoxine, pyrimethamine, Pyrolidine, atovaquinone, quinine, chloroquine, piperaquine, mefloquine, amodiaquine, primaquine, talfinoquine, etc.
  • other pharmaceutical ingredients with anti-malarial activity including but not limited to artemisinin, dihydroartemisinin, artemether, artesunate, benfluorenol, sulfadoxine, pyrimethamine, Pyrolidine, atovaquinone, quinine, chloroquine, piperaquine, mefloquine, amodiaquine, primaquine, talfinoquine, etc.
  • the present invention also provides a preparation method of 2,6-diazaspiro[3.4]octane pyrimidine-hydroxamic acid and its intermediate compounds with the structure of general formula I.
  • the specific synthesis method is as follows:
  • step 2 1) Dissolve the compound obtained in step 1 and 2,6-di-tert-butylpyridine in dichloromethane, cool to 0 ⁇ 5°C in an ice water bath, slowly add trimethylsilyl trifluoromethanesulfonate dropwise, and the addition is complete Keep the ice water bath, stir for 15 min, remove the ice water bath, and stir at room temperature until the reaction is complete (TLC detection). Then cool the reaction liquid in an ice-water bath, slowly add anhydrous methanol (the addition amount is the same as trimethylsilyl trifluoromethanesulfonate) to the system under vigorous stirring. After the methanol is added, remove the ice-water bath and wait for the system to rise.
  • anhydrous methanol the addition amount is the same as trimethylsilyl trifluoromethanesulfonate
  • Step 3 The compound obtained in Step 2 was dissolved in 1,2-dichloroethane is added an aldehyde substituted with R 1, glacial acetic acid, triacetoxy sodium borohydride, stirred at room temperature overnight. After the reaction is completed, a saturated aqueous sodium bicarbonate solution is added, the liquid is separated by shaking, the organic phase is distilled under reduced pressure to remove the solvent, and the remaining mixture is separated and purified by silica gel column chromatography to obtain compound Id.
  • step 4 Dissolve the compound obtained in step 3 with a mixed solution of methanol and water, add solid potassium carbonate, and heat to react at 65-70°C for 5-6 hours. After the reaction was completed, the solvent was distilled off under reduced pressure, the residue was acidified with 2M hydrochloric acid to a pH of about 1, and water was distilled off under reduced pressure to obtain compound I-e mixed with inorganic salts and used directly in the next reaction.
  • step 5 Dissolve the mixture obtained in step 4 with N,N-dimethylformamide solution, add 1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide in sequence Amine hydrochloride, after stirring for 30 min at room temperature, O-(tetrahydro-2H-pyran-2-yl)hydroxylamine and triethylamine were added in sequence, and stirred at room temperature for 48 hours.
  • step 6 Dissolve the compound obtained in step 5 with dichloromethane, add 4M hydrogen chloride-dioxane solution, and stir for 30 min at room temperature. Filtration with suction and washing the solid with a large amount of dichloromethane to obtain compound I.
  • the compound of the present invention Since the compound of the present invention has excellent HDAC activation activity, the compound of the present invention and its various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and containing the compound of the present invention as the main active ingredient
  • the pharmaceutical composition can be used to treat, prevent and alleviate diseases caused by the activity or expression of HDAC.
  • the compounds of the present invention can be used to treat the following diseases: lung cancer, colon cancer, prostate cancer, breast cancer, ovarian cancer, lymphatic system tumors, and diseases caused by malaria parasites (such as malaria).
  • the pharmaceutical composition of the present invention contains the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier within a safe and effective amount.
  • the "safe and effective amount” refers to: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects.
  • the pharmaceutical composition contains 0.1-1000 mg of the compound of the present invention per agent, more preferably, 0.5-500 mg of the compound of the present invention per agent.
  • the "one dose" is a capsule or tablet.
  • “Pharmaceutically acceptable carrier” refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use, and must have sufficient purity and sufficiently low toxicity. "Compatibility” here means that the components in the composition can be blended with the compound of the present invention and between them without significantly reducing the efficacy of the compound.
  • pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, and solid lubricants (such as stearic acid).
  • Magnesium stearate calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as Tween), wetting Agents (such as sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.
  • vegetable oils such as soybean oil, sesame oil, peanut oil, olive oil, etc.
  • polyols such as propylene glycol, glycerin, mannitol, sorbitol, etc.
  • emulsifiers such as Tween
  • wetting Agents such as sodium lauryl sulfate
  • coloring agents such as sodium lauryl sulfate
  • flavoring agents such as pepperminophen, sorbitol, etc.
  • the administration method of the compound or pharmaceutical composition of the present invention is not particularly limited.
  • Representative administration methods include (but are not limited to): oral, rectal, parenteral (intravenous, intramuscular, or subcutaneous), and topical administration.
  • a particularly preferred mode of administration is oral.
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules.
  • the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with the following ingredients: (a) fillers or compatibilizers, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) humectant, For example, glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) Absorption accelerators, such as quaternary amine compounds; (g) wetting agents, such as cetyl alcohol and gly
  • Solid dosage forms such as tablets, sugar pills, capsules, pills and granules can be prepared with coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the active compound or the release of the compound in such a composition may be released in a certain part of the digestive tract in a delayed manner. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microcapsules with one or more of the above-mentioned excipients.
  • Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures.
  • the liquid dosage form may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-Butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.
  • composition may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.
  • adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.
  • the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.
  • suspending agents for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.
  • composition for parenteral injection may contain physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions.
  • Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.
  • the dosage form of the compound of the present invention for topical administration includes ointment, powder, patch, spray and inhalant.
  • the active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required if necessary.
  • the compound of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds.
  • a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage is the pharmaceutically effective dosage considered to be administered.
  • the daily dose administered is usually 0.2 to 1000 mg, preferably 0.5 to 500 mg.
  • the specific dosage should also consider factors such as the route of administration, the patient's health status, etc., which are within the skill range of a skilled physician.
  • Example 4 The solid obtained in Example 4 was dissolved in 10 mL of N,N-dimethylformamide solution, and 0.16g 1-hydroxybenzotriazole and 0.23g 1-(3-dimethylaminopropyl)-3-ethyl were added in sequence. After stirring for 30 min at room temperature, 0.35 g of O-(tetrahydro-2H-pyran-2-yl) hydroxylamine and 0.42 mL of triethylamine were added in sequence, and stirred at room temperature for 48 hours.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 2-furaldehyde, and the remaining raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-2d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 3-furaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-3d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 2-thiophenecarbaldehyde, and the remaining raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-4d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 3-thiophenecarbaldehyde, and the remaining raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-5d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 2-pyridinecarboxaldehyde, and the remaining raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-6d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 3-pyridinecarboxaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-7d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 4-pyridinecarboxaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-8d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 1-naphthaldehyde, and the remaining raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-9d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 2-naphthaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-10d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with biphenyl-4-carbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-11d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with benzaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-12d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with cyclopentylcarbaldehyde, and the remaining raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-13d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with cyclohexylcarbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-14d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 1-benzothiophene-2-carboxaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-15d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 3-carboxaldehyde benzothiophene, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-16d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with benzo[b]furan-2-carbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-17d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 4-indolecarbaldehyde, and the remaining raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-18d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 1-methylindole-2-carbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-19d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 3-quinolinecarboxaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-20d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with quinoline-2-carboxaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-21d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with quinoline-6-carbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-22d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with quinoline-8-carbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-23d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 8-isoquinolinecarboxaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-24d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with isoquinoline-5-carbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-25d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 4-quinolinecarboxaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-26d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with isoquinoline-4-carbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-27d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with imidazo[1,2-a]pyridine-3-carbaldehyde, and the remaining raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I- 28d.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 1-methyl-2-formylbenzimidazole, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-29d.
  • Example 3 Replace the N-methylindole-3-carbaldehyde in Example 3 with 1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde.
  • the remaining raw materials, reagents and preparation methods are the same In Example 3, I-30d was obtained.
  • Example 3 Replace the N-methylindole-3-carboxaldehyde in Example 3 with 1-methyl-1H-pyrrolo[2,3-c]pyridine-3-carboxylic acid, and other required raw materials, reagents and preparation methods Same as Example 3 to obtain I-31d.
  • Example 3 Replace the N-methylindole-3-carbaldehyde in Example 3 with 1-methyl-1H-pyrrolo[3,2-b]pyridine-3-carbaldehyde, and the other required raw materials, reagents and preparation methods are the same In Example 3, I-32d was obtained.
  • Example 3 Replace the N-methylindole-3-carbaldehyde in Example 3 with 1-methyl-1H-indazole-3-carbaldehyde, and the other required raw materials, reagents and preparation methods are the same as those in Example 3 to obtain I-33d .
  • Example 3 Replace the N-methylindole-3-carbaldehyde in Example 3 with 1-methyl-1H-pyrrolo[3,2-c]pyridine-3-carbaldehyde, and the other required raw materials, reagents and preparation methods are the same In Example 3, I-34d was obtained.
  • Example 3 The N-methylindole-3-carbaldehyde in Example 3 was replaced with 9-anthracene aldehyde, and the remaining raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-35d.
  • Example 3 Replace the N-methylindole-3-carbaldehyde in Example 3 with 6-methoxy-1-methyl-1H-indole-3-carbaldehyde, and the other required raw materials, reagents and preparation methods are the same as those in the example 3. Get I-36d.
  • Example 3 Replace the N-methylindole-3-carbaldehyde in Example 3 with 1,6-dimethyl-1H-indole-3-carbaldehyde, and the remaining raw materials, reagents and preparation methods are the same as those in Example 3. I-37d.
  • Example 3 Replace the N-methylindole-3-carbaldehyde in Example 3 with 6-cyano-1-methyl-1H-indole-3-carbaldehyde, and the other required raw materials, reagents and preparation methods are the same as in Example 3. , Get I-38d.
  • Example 3 Replace the N-methylindole-3-carbaldehyde in Example 3 with 6-bromo-1-methyl-1H-indole-3-carbaldehyde, and the remaining raw materials, reagents and preparation methods are the same as in Example 3. Get I-39d.
  • Example 4 Replace I-1d in Example 4 with I-2d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-2 is obtained through a three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-3d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-3 is obtained through a three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-4d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-4 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-5d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-5 is obtained through a three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-6d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-6 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-7d, and the remaining raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-7 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-8d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-8 is obtained through a three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-9d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-9 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-10d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-10 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-11d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-11 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-12d, and the remaining raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-12 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-13d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-13 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-14d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-14 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-15d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-15 is obtained through a three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-16d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-16 is obtained through a three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-17d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-17 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-18d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-18 is obtained through a three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-19d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-19 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-20d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-20 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-21d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-21 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-22d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-22 is obtained through a three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-23d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-23 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-24d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-24 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-25d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-25 is obtained through a three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-26d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-26 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-27d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-27 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-28d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-28 is obtained through a three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-29d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-29 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-30d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-30 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-31d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-31 is obtained through a three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-32d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-32 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-33d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-33 is obtained through a three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-34d, and the remaining raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-34 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-35d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-35 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-36d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-36 is obtained through a three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-37d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-37 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-38d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-38 is obtained through three-step reaction.
  • Example 4 Replace I-1d in Example 4 with I-39d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-39 is obtained through three-step reaction.
  • Plasmodium culture Use PRMI (containing NaHCO 3 , HEPES, Albumax I, Hypoxanthine, Genaotamicin) complete medium (complete medium) for plasmodium culture, in a 37°C incubator (5% CO 2 , 5% O 2 ) In the cultivation.
  • PRMI containing NaHCO 3 , HEPES, Albumax I, Hypoxanthine, Genaotamicin
  • 3D7 is a wild-type strain and has no obvious resistance to drugs; Dd2 is resistant to chloroquine, quinine, sulfadoxine, pyrimethamine, and amodiaquine.
  • Table 1 shows that the compounds have strong in vitro insecticidal activity, and the IC 50 values of some compounds against 3D7 and Dd2 are comparable to DHA.
  • Inhibition rate (%) [A(0)-A(dosing)]/[A(0)-A(blank)] ⁇ 100%
  • the experiment uses mouse liver microsomes (0.5mg/mL), purchased from Corning.
  • the positive control is ketanserin.
  • the test compound is first prepared into a 10 mM DMSO solution and diluted to 0.5 mM with acetonitrile; the above 0.5 mM solution is added to the buffer containing liver microsomes to make the compound concentration 1.5 ⁇ M; Take 30 ⁇ L of the 1.5 ⁇ M compound/liver microsome mixture and add 15 ⁇ L of 6 mM NADPH solution to make the final concentration of the compound 1.5 ⁇ M and the final concentration of NADPH 2 mM.
  • the compound/liver microsome test solution was placed on the test plate, incubated in a 37°C water bath, and quenched by adding 135 ⁇ L of acetonitrile at each time point (0, 5, 15, 30, 45 min). After all the samples are quenched, shake the samples with a shaker (IKA, MTS 2/4) for 10 minutes (600 rpm/min), and then centrifuge at 5594g for 15 minutes (Thermo Multifuge ⁇ 3R). Take the supernatant, dilute it with distilled water 1:1, and analyze by LC-MS. The peak area response ratio (PARR) of the compound at 5, 15, 30, and 45 min was compared with the PARR at time 0 to determine the percentage of test compound retained at each time point.
  • PARR peak area response ratio
  • GB4 is resistant to chloroquine; C2A is resistant to quinine; CP286 is resistant to sulfadoxine, pyrimethamine, and mefloquine; 6218 and 6320 have time-dependent resistance to artemisinin drugs, only in It will be displayed within 6h after the ring phase is synchronized.
  • Table 1 and Table 4 the 72h IC 50 values of the compounds are equivalent to DHA, indicating that the compounds have the potential to treat malaria that is resistant to current first- and second-line antimalarial drugs and cope with malaria resistance.
  • Plasmodium culture uses RPMI (containing NaHCO3, HEPES, Albumax I, Hypoxanthine Genaotamicin) complete medium (Complete Medium), cultivated in a 37°C incubator
  • the drug was dissolved in DMSO to prepare an initial concentration of 200*20*IC50.
  • SDS-PAGE gel electrophoresis Load 10 ⁇ L in each well of the precast gel, run at 80V for 30 minutes, adjust the voltage to 120V, and then run until the loading is close to the bottom edge of the separation gel.
  • Transfer membrane Cut out the PVDF membrane with corresponding coverage area, adopt wet transfer method, fast transfer membrane buffer, 400mA constant current transfer for 35 minutes, and take out the PVDF membrane after finishing.
  • Sealing Put the membrane in the sealing solution (add 5% skimmed milk powder in TBST), shake and seal for 2h on a shaker.
  • Incubate the primary antibody use histone histone H3 antibody and H3K9 acetylated antibody, dilute with 5% skimmed milk powder at a ratio of 1:2000, incubate the PVDF membrane on a shaker for 2 hours, discard the incubation solution, add TBST to wash the membrane three times, each time for 10 minutes.
  • Incubate the secondary antibody Dilute the secondary antibody at 1:5000, incubate the PVDF membrane on a shaker for 1 hour and discard the incubation solution.
  • Color development and exposure Temporarily prepare color development solution and spread it evenly on the PVDF film. The exposure time can be adjusted according to the brightness of the strip.
  • JL01 is the positive control compound. Comparing the acetylation bands of Plasmodium histone H3 after treatment with compound and DMSO for 4 hours, it can be seen that the compound up-regulated the level of acetylation, that is, inhibited deacetylation. The activity of the enzyme indirectly proves that the compound is a pan pfHDAC inhibitor.
  • Protozoa rate number of red blood cells infected by plasmodium/total number of red blood cells ⁇ 100%
  • the results are shown in Figures 2 and 3.
  • the protozoan rate curve ( Figure 2) shows that compared with the blank group, compound I-39 has better insecticidal activity at 60 mg/kg, and the protozoan rate of the surviving mice is 0 on the 30th day. The protozoan has been eliminated.
  • the protozoan rate curve and survival curve ( Figure 3) comprehensively show that I-39 has a good balance of efficacy and toxicity.
  • (1) hHDAC1-3,6 test method add 250nL DMSO or compound solution to the OptiPlate TM-384F black assay plate via Echo, and add 15 ⁇ L enzyme solution and 10 ⁇ L GL-8 substrate solution to the assay plate in turn. Incubate at 25°C for 60 minutes and read the value using the setting of Ex350-360/Em450-465 (sensitive 60). The inhibition rate was calculated, and the IC 50 value was calculated with GraphPad Prism.
  • (2) hHDAC8 test method add 250nL DMSO or compound solution to the OptiPlate TM-384F black assay plate via Echo, add 15 ⁇ L enzyme solution, 10 ⁇ L corresponding substrate solution, and react at 25°C for 4 hours. Add 10 ⁇ L stop solution to stop the reaction, and use the setting of Ex350-360/Em450-465 to read the value. The inhibition rate was calculated, and the IC 50 value was calculated with GraphPad Prism.
  • (3) hSirt2 test method add 800nL DMSO or compound solution to the OptiPlate TM-384F black assay plate via Echo, add 10 ⁇ L enzyme solution and 10 ⁇ L corresponding substrate solution in sequence, and react at 25°C for 4h. Add 20 ⁇ L stop solution to stop the reaction, and use the setting of Ex350-360/Em450-465 to read the value. The inhibition rate was calculated, and the IC 50 value was calculated with GraphPad Prism.
  • the 2,6-diazaspiro[3.4]octane pyrimidine-hydroxamic acid compound of the present invention has a relatively simple molecular structure and a simple preparation process. It is useful in HDAC enzyme inhibition experiments and experiments that are closely related to the survival and reproduction of plasmodium. Both internal and external insecticidal efficacy experiments have shown strong inhibitory activity, and the cytotoxicity is weak, and the selectivity index can reach thousands. Therefore, such compounds are not only expected to be developed into a new type of single-drug antimalarial drugs, but also can be developed into antimalarial drugs for combined administration with existing antimalarial drugs.

Abstract

Disclosed are a 2,6-diazaspiro[3.4]octane pyrimidine-hydroxamic acid compound and the use thereof. Specifically, the present invention relates to a compound as represented by formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and a preparation method therefor, and the use thereof in the preparation of HDAC inhibitor anti-malarial drugs.

Description

2,6-二氮杂螺[3.4]辛烷类嘧啶-异羟肟酸化合物及其用途2,6-diazaspiro[3.4]octane pyrimidine-hydroxamic acid compound and its use 技术领域Technical field
本发明涉及药物化学和药物治疗学领域，具体地，本发明提供了一类具有2,6-二氮杂螺[3.4]辛烷类嘧啶-异羟肟酸结构的小分子化合物及其药学上可接受的盐，所述化合物具有优异的疟原虫体内外杀疟原虫虫活性。该类化合物有望开发成为新型抗疟疾药物。The present invention relates to the fields of medicinal chemistry and pharmacotherapy. Specifically, the present invention provides a class of small molecule compounds with 2,6-diazaspiro[3.4]octane-like pyrimidine-hydroxamic acid structure and their pharmacy Acceptable salt, the compound has excellent malaria parasite killing activity in vivo and in vitro. Such compounds are expected to be developed into new anti-malaria drugs.
背景技术Background technique
疟疾是当今世界公共卫生中最突出的问题之一，是全球流行最广、危害最大的寄生虫病。据世界卫生组织(World health organization，WHO)2018年世界疟疾报告统计，2017年全球共发生2.19亿疟疾病例，其中43.5万例死亡；大部分疟疾病例发生在世卫组织非洲区域(2亿病例，92％)，其中5岁以下的儿童为最易感染人群。疟疾的流行除了危害人类身体健康和生命安全，还严重削弱着许多非洲国家的卫生系统，深刻影响着发展中国家的社会经济发展，使贫穷成为无法摆脱的恶性循环。加上全球气候变暖、国际交流频繁、全球疟疾防治资金投入放缓等因素，全球疟疾防治工作将面临巨大挑战。Malaria is one of the most prominent problems in public health in the world today, and it is the most widespread and harmful parasitic disease in the world. According to the World Health Organization (WHO) World Malaria Report in 2018, there were 219 million malaria cases worldwide in 2017, including 435,000 deaths; most of the malaria cases occurred in the WHO African region (200 million cases, 92%), among which children under 5 years old are the most vulnerable population. In addition to endangering human health and life safety, the malaria epidemic has severely weakened the health systems of many African countries, profoundly affected the socio-economic development of developing countries, and made poverty a vicious circle that cannot be escaped. Coupled with factors such as global warming, frequent international exchanges, and a slowdown in global funding for malaria control, global malaria control will face enormous challenges.
疟疾由一种叫作疟原虫的寄生虫引起，通过受感染蚊子的叮咬传播。这种寄生虫在人体的肝脏中繁殖，然后感染血红细胞。疟疾的症状包括发烧、头痛和呕吐，通常在蚊子叮咬后10-15天显现。如不治疗，疟疾可能中断对维持生命的重要器官的供血，从而迅速威胁生命。Malaria is caused by a parasite called Plasmodium, which is spread by the bite of an infected mosquito. This parasite reproduces in the human liver and then infects red blood cells. Symptoms of malaria include fever, headache, and vomiting, which usually appear 10-15 days after a mosquito bite. If left untreated, malaria may interrupt the blood supply to vital organs that sustain life, thereby quickly threatening life.
在人类与疟疾的长期抗争中，化学药治疗自始至终是疟疾防治至关重要的一个环节。人们长期依赖的天然(奎宁)或人工合成(氯喹、甲氟喹、伯胺喹等)氨基喹啉类或抗生素类(磺胺多辛、乙胺嘧啶等)等前几代抗疟药物的耐药性在20世纪五六十年代广泛出现，使疗效大打折扣破坏了疟疾控制的努力并颠覆了在儿童存活方面取得的成果。随后青蒿素及青蒿素联合疗法(artemisinin combination therapy，ACT)改变了抗疟用药的困境，成为当代疟疾治疗的中坚力量。但是，10年前在柬埔寨地区出现了ACT疗法的抗性报道，恶性疟原虫对青蒿素衍生物的敏感性降低，其中单药治疗后患者的清除时间明显延长，并且进一步降低了临床疗效，加速潜在伴药耐药性的出现，这已经成为对ACT持续功效的潜在威胁。近年来青蒿素耐受的疟疾在东南亚湄公河流域广泛传播，甚至有散布至非洲的风险。In the long-term struggle between humans and malaria, chemical treatment has always been a crucial part of malaria prevention and treatment. People have long relied on natural (quinine) or synthetic (chloroquine, mefloquine, primaquine, etc.)aminoquinolines or antibiotics (sulfadoxine, pyrimethamine, etc.) and other antimalarial drugs of previous generations. The medicinal properties appeared widely in the 1950s and 1960s, which greatly reduced the efficacy, undermined the efforts of malaria control and overturned the achievements in child survival. Subsequently, artemisinin and artemisinin combination therapy (artemisinin combination therapy, ACT) changed the predicament of antimalarial drugs and became the backbone of contemporary malaria treatment. However, 10 years ago, there was a report of resistance to ACT therapy in Cambodia. The sensitivity of Plasmodium falciparum to artemisinin derivatives was reduced, and the clearance time of patients after monotherapy was significantly prolonged, and the clinical efficacy was further reduced. Accelerate the emergence of potential drug resistance, which has become a potential threat to the continued efficacy of ACT. In recent years, artemisinin-resistant malaria has spread widely in the Mekong River basin of Southeast Asia, and even has the risk of spreading to Africa.
随着时间推移，抗疟药耐药性反复出现和扩散以及蚊媒对杀虫剂产生抗性并逐渐扩散，使得传统疟疾防治方法面临严峻挑战，迫切需要寻找新的靶标和方法，为疟疾的研究、防御、治疗和控制提供新的技术和策略。With the passage of time, the repeated emergence and spread of antimalarial drug resistance and the resistance of mosquito vectors to insecticides and the gradual spread have made traditional malaria control methods face severe challenges, and there is an urgent need to find new targets and methods for malaria. Research, defense, treatment and control provide new technologies and strategies.
因此，本领域迫切需要一种能够有效杀灭疟原虫，特别是耐药性疟原虫的药物化合物。Therefore, there is an urgent need in the art for a pharmaceutical compound that can effectively kill malaria parasites, especially drug-resistant malaria parasites.
发明内容Summary of the invention
本发明的目的是提供一种能够有效杀灭疟原虫，特别是耐药性疟原虫的药物化合物。The purpose of the present invention is to provide a pharmaceutical compound that can effectively kill malaria parasites, especially drug-resistant malaria parasites.
本发明的第一方面，提供了一种如下式I所示的2,6-二氮杂螺[3.4]辛烷类嘧啶-异羟肟酸化合物，或其药学上可接受的盐或光学异构体，The first aspect of the present invention provides a 2,6-diazaspiro[3.4]octane pyrimidine-hydroxamic acid compound represented by the following formula I, or a pharmaceutically acceptable salt or optical isoform thereof Structure,
Figure PCTCN2019077356-appb-000001
Figure PCTCN2019077356-appb-000001
其中，among them,
R 1选自下组：取代或未取代的C1～C6烷基、取代或未取代的C3～C8环烷基、取代或未取代的C6-C15单环、二环或三环芳基、取代或未取代的5-15元单环、二环或三环杂环基(包括饱和、部分不饱和或芳香性杂环基)；其中，所述的杂芳基包括一个或多个选自下组的杂原子作为环骨架：N、O或S； R 1 is selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C15 monocyclic, bicyclic or tricyclic aryl, substituted Or unsubstituted 5-15 membered monocyclic, bicyclic or tricyclic heterocyclic group (including saturated, partially unsaturated or aromatic heterocyclic group); wherein, the heteroaryl group includes one or more selected from Group of heteroatoms as the ring skeleton: N, O or S;
R 2选自下组：NHOH，或OR 3； R 2 is selected from the group consisting of NHOH, or OR 3 ;
R 3选自下组：氢，取代或未取代的C1～C3烷基，取代或未取代的C3～C8环烷基、取代或未取代的5-15元杂环基； R 3 is selected from the following group: hydrogen, substituted or unsubstituted C1-C3 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 5-15 membered heterocyclic group;
其中，所述的取代指基团上的氢原子被一个或者多个(例如2个、3个、4个等)选自下组的取代基所取代：卤素、氘代、C1-C6烷氧基、卤代的C1-C6烷氧基、卤代的C3-C8环烷基、甲基砜基、-S(＝O) 2NH 2、氧代(＝O)、-CN、羟基、-NH 2、羧基、或取代或未取代的选自下组的基团：C1-C6烷基、C3-C8环烷基、C1-C6胺基、C6-C10芳基、具有1-3个选自N、S和O的杂原子的5-10元杂芳基、具有1-3个选自N、S和O的杂原子的5-12元杂环基，且所述的取代基选自下组：卤素、C1-C6烷基、C1-C6烷氧基、氧代、-CN、-NH 2、-OH、C6-C10芳基、C1-C6胺基、C1-C6酰胺基、具有1-3个选自N、S和O的杂原子的5-10元杂芳基。 Wherein, the substitution means that the hydrogen atom on the group is replaced by one or more (for example, 2, 3, 4, etc.) substituents selected from the following group: halogen, deuterated, C1-C6 alkoxy Group, halogenated C1-C6 alkoxy, halogenated C3-C8 cycloalkyl, methyl sulfone, -S(=O) 2 NH 2 , oxo(=O), -CN, hydroxyl,- NH 2 , carboxyl group, or substituted or unsubstituted group selected from the following group: C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 amino, C6-C10 aryl, with 1-3 selected A 5-10 membered heteroaryl group from heteroatoms of N, S and O, a 5-12 membered heterocyclic group with 1-3 heteroatoms selected from N, S and O, and the substituents are selected from The following group: halogen, C1-C6 alkyl, C1-C6 alkoxy, oxo, -CN, -NH 2 , -OH, C6-C10 aryl, C1-C6 amine, C1-C6 amide, with A 5-10 membered heteroaryl group with 1-3 heteroatoms selected from N, S and O.
在另一优选例中，所述的R 2选自下组：H、取代或未取代的C1～C6烷基，取代或未取代的C3～C8环烷基，或取代或未取代的选自下组的基团： In another preferred example, the R 2 is selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, or substituted or unsubstituted Groups of the following groups:
Figure PCTCN2019077356-appb-000002
Figure PCTCN2019077356-appb-000002
上述各式中，X选自N、O、S。In the above formulas, X is selected from N, O, and S.
在另一优选例中，所述的芳基为包含1～4个并环的芳基。In another preferred example, the aryl group is an aryl group containing 1 to 4 bicyclic rings.
在另一优选例中，所述的杂环基为包含1-4个并环的杂芳基，其中，各个并环各自独立地为5～6元的芳环或芳杂环。In another preferred example, the heterocyclic group is a heteroaryl group containing 1-4 bicyclic rings, wherein each bicyclic ring is independently a 5- to 6-membered aromatic ring or aromatic heterocyclic ring.
在另一优选例中，所述的C1-C6烷基为甲基或乙基。In another preferred embodiment, the C1-C6 alkyl group is methyl or ethyl.
在另一优选例中，所述的R 1选自下组：取代或未取代的C1～C6烷基、取代或未取代的C3～C8环烷基，取代或未取代的C6-C15芳基，取代或未取代的5-15元杂芳基；其中，所述的C3～C8环烷基选自下组：环戊基，环己基； In another preferred example, the R 1 is selected from the group consisting of substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C15 aryl , Substituted or unsubstituted 5-15 membered heteroaryl; wherein, the C3-C8 cycloalkyl group is selected from the following group: cyclopentyl, cyclohexyl;
所述的芳基选自下组：苯基、萘基、菲基；The aryl group is selected from the following group: phenyl, naphthyl, phenanthryl;
所述的5-15元杂芳基选自下组：
Figure PCTCN2019077356-appb-000003
Figure PCTCN2019077356-appb-000004
The 5-15 membered heteroaryl group is selected from the following group:
Figure PCTCN2019077356-appb-000003
Figure PCTCN2019077356-appb-000004
在另一优选例中，所述的取代基选自下组：卤素、OH、NH 2、CN、C1-C6烷基、C3～C8环烷基、C6-C10芳基、C1-C6烷氧基、C1-C6烷氨基。 In another preferred embodiment, the substituents are selected from the group consisting of halogen, OH, NH 2 , CN, C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl, C1-C6 alkoxy Group, C1-C6 alkylamino.
在另一优选例中，所述的R 1为NHOH。 In another preferred embodiment, the R 1 is NHOH.
在另一优选例中，所述的式I化合物可选自下组：In another preferred embodiment, the compound of formula I can be selected from the following group:
Figure PCTCN2019077356-appb-000005
Figure PCTCN2019077356-appb-000005
本发明的第二方面，提供了一种药物组合物，所述的药物组合物包括(a)治疗有效量的如本发明第一方面中所述的化合物、或其药学上可接受的盐、水合物或溶剂化物；和(b)药学上可接受的载体。The second aspect of the present invention provides a pharmaceutical composition comprising (a) a therapeutically effective amount of the compound as described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, Hydrate or solvate; and (b) a pharmaceutically acceptable carrier.
在另一优选例中，所述的药物组合物用于：In another preferred embodiment, the pharmaceutical composition is used for:
(a)治疗或预防疟原虫导致的疾病或病症；或(a) Treatment or prevention of diseases or conditions caused by malaria parasites; or
(b)治疗或预防与HDAC活性或表达量相关的疾病或病症。(b) Treating or preventing diseases or disorders related to the activity or expression of HDAC.
在另一优选例中，所述的药物组合物用于调节HDAC活性或表达量。In another preferred embodiment, the pharmaceutical composition is used to regulate the activity or expression of HDAC.
本发明的第三方面，提供了一种如本发明第一方面所述的式I化合物的用途，其用于制备治疗或预防疟原虫导致的疾病或病症的药物组合物。The third aspect of the present invention provides a use of the compound of formula I as described in the first aspect of the present invention for preparing a pharmaceutical composition for treating or preventing diseases or disorders caused by malaria parasites.
在另一优选例中，所述的疾病或病症为疟疾。In another preferred embodiment, the disease or condition is malaria.
在另一优选例中，所述疟原虫为耐药疟原虫或非耐药疟原虫。In another preferred embodiment, the Plasmodium is resistant or non-drug resistant.
在另一优选例中，所述的疟原虫为处于选自下组时期的疟原虫：肝期、配子体期、红细胞内期。In another preferred embodiment, the plasmodium is a plasmodium in a stage selected from the group consisting of hepatic stage, gametophyte stage, and intraerythrocytic stage.
在另一优选例中，所述的耐药疟原虫为对选自下组的药物已经产生抗性的疟原虫：青蒿素、二氢青蒿素、蒿甲醚、蒿琥酯、本芴醇、磺胺多辛、乙胺嘧啶、咯萘啶、阿托伐醌、奎宁、氯喹、哌喹、甲氟喹、阿莫地喹、伯胺喹、和他非诺喹。In another preferred embodiment, the drug-resistant plasmodium is a plasmodium that has developed resistance to drugs selected from the group consisting of artemisinin, dihydroartemisinin, artemether, artesunate, and fluorene Alcohol, sulfadoxine, pyrimethamine, pyronaridine, atovaquinone, quinine, chloroquine, piperquine, mefloquine, amodiaquine, primaquine, and tafenoquine.
本发明的第四方面，提供了一种如本发明第一方面所述的式I化合物的用途，其用于制备治疗或预防HDAC活性或表达量相关的疾病或病症的药物组合物。The fourth aspect of the present invention provides a use of the compound of formula I as described in the first aspect of the present invention for preparing a pharmaceutical composition for treating or preventing diseases or disorders related to the activity or expression of HDAC.
在另一优选例中，所述的疾病或病症为肿瘤，较佳地，所述的肿瘤选自下组：肺癌、结肠癌、前列腺癌、乳腺癌、卵巢癌、淋巴系统肿瘤。In another preferred example, the disease or condition is a tumor. Preferably, the tumor is selected from the group consisting of lung cancer, colon cancer, prostate cancer, breast cancer, ovarian cancer, and lymphatic system tumors.
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described in the following (such as the embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, I will not repeat them here.
附图说明Description of the drawings
图1为本发明化合物的Western Blot实验结果图；Figure 1 is a Western Blot experimental result diagram of the compound of the present invention;
图2为小鼠体内药效实验中小鼠体内原虫率折线图；Figure 2 is a broken line diagram of the protozoan rate in mice in the in vivo drug efficacy experiment in mice;
图3为小鼠体内药效实验结果中小鼠存活比例折线图。Figure 3 is a broken line graph of the survival ratio of mice in the results of in vivo drug efficacy experiments in mice.
具体实施方式detailed description
本发明人经过长期而深入的研究，制备了一种具有HDAC抑制活性的化合物，其可以杀灭各个时期的耐药和非耐药疟原虫活性，且对人源细胞具有较低毒性，因此可以用于疟原虫导致的疾病例如疟疾的治疗。基于上述发现，发明人完成了本发明。After long-term and in-depth research, the inventor prepared a compound with HDAC inhibitory activity, which can kill drug-resistant and non-drug-resistant plasmodium activity in various periods, and has low toxicity to human cells, so it can For the treatment of diseases caused by malaria parasites such as malaria. Based on the above findings, the inventor completed the present invention.
式I化合物及其制备Formula I compound and its preparation
本发明提供具有如下通式I结构的化合物、或其药学上可接受的盐、或其立体异构体：The present invention provides a compound having the structure of the following general formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:
Figure PCTCN2019077356-appb-000006
Figure PCTCN2019077356-appb-000006
通式I中：R 1选H、直链或支链烷基或II： In the general formula I: R 1 is selected from H, straight or branched chain alkyl or II:
Figure PCTCN2019077356-appb-000007
Figure PCTCN2019077356-appb-000007
式II中A环为脂肪环烷基、环芳基、含杂原子的环芳基。Ring A in formula II is an aliphatic cycloalkyl group, a ring aryl group, and a heteroatom-containing ring aryl group.
符合通式I的化合物，其特征在于，II中A环为3～8元脂肪环烷基。The compound according to general formula I is characterized in that the A ring in II is a 3-8 membered alicyclic alkyl group.
符合通式I的化合物，其特征在于，II中A环为含有或不含有杂原子的环芳基，其中杂原子数目为0～4，杂原子类型取自氮、氧、硫中一种、二种或三种。The compound of general formula I is characterized in that the ring A in II is a ring aryl group with or without heteroatoms, wherein the number of heteroatoms is 0-4, and the type of heteroatoms is selected from one of nitrogen, oxygen, and sulfur. Two or three.
符合通式I的化合物，其特征在于，A环描述的环芳基为包含1～4个并环的芳基，其中的组成芳环取自5～6元的芳环或芳杂环，每一个组成芳环的杂原子数目为0～2，杂原子类型取自氮、氧、硫中一种或两种。The compound in accordance with formula I is characterized in that the cyclic aryl group described by ring A is an aryl group containing 1 to 4 parallel rings, wherein the aromatic ring is composed of a 5- to 6-membered aromatic ring or aromatic heterocyclic ring, each The number of heteroatoms constituting an aromatic ring is 0-2, and the heteroatom types are selected from one or two of nitrogen, oxygen, and sulfur.
符合通式I的化合物，其特征在于，R 2与R 3独立选自H、卤素、OH、NH 2、CN、直链或支链烷基、环烷基、芳基、烷氧基、烷氨基。 A compound in accordance with the general formula I, characterized in that R 2 and R 3 are independently selected from H, halogen, OH, NH 2 , CN, linear or branched alkyl, cycloalkyl, aryl, alkoxy, alkyl Amino.
符合通式I的化合物，其特征在于，A环描述的环芳基为：The compound in accordance with general formula I is characterized in that the ring aryl group described by ring A is:
Figure PCTCN2019077356-appb-000008
Figure PCTCN2019077356-appb-000008
X选自N、O、S。X is selected from N, O, S.
符合通式I的化合物，其特征在于，用于制备抑制组蛋白去乙酰化酶(HDAC)活性的药物组合物或制剂，亦或用于制备抑制疟原虫或治疗疟疾的药物组合物或制剂。The compound according to the general formula I is characterized in that it is used to prepare a pharmaceutical composition or preparation for inhibiting the activity of histone deacetylase (HDAC), or for preparing a pharmaceutical composition or preparation for inhibiting Plasmodium or treating malaria.
符合通式I的化合物作为HDAC抑制剂以单一用药或组合用药在预防或治疗疟疾中的应用。所述药物组合物中含有0.001～99wt％，较佳地0.1～90wt％，更佳地1～80wt％的通式I化合物或其药学上可接受的盐，按组合物的总重量计。药物或药物组合物的剂型为口服剂型、或注射剂。口服剂型包括片剂、胶囊剂、膜剂、颗粒剂等，还包括缓释型或非缓释型剂型。药物组合物还可含有其它具有抗疟疾活性的药物成分，其中包括但不限于青蒿素、二氢青蒿素、蒿甲醚、蒿琥酯、本芴醇、磺胺多辛、乙胺嘧啶、咯萘啶、阿托伐醌、奎宁、氯喹、哌喹、甲氟喹、阿莫地喹、伯胺喹、他非诺喹等。The application of a compound conforming to the general formula I as an HDAC inhibitor in the prevention or treatment of malaria in single or combined medication. The pharmaceutical composition contains 0.001 to 99% by weight, preferably 0.1 to 90% by weight, and more preferably 1 to 80% by weight of the compound of general formula I or a pharmaceutically acceptable salt thereof, based on the total weight of the composition. The dosage form of the medicine or pharmaceutical composition is an oral dosage form or an injection. Oral dosage forms include tablets, capsules, films, granules, etc., and also include sustained-release or non-sustained-release dosage forms. The pharmaceutical composition may also contain other pharmaceutical ingredients with anti-malarial activity, including but not limited to artemisinin, dihydroartemisinin, artemether, artesunate, benfluorenol, sulfadoxine, pyrimethamine, Pyrolidine, atovaquinone, quinine, chloroquine, piperaquine, mefloquine, amodiaquine, primaquine, talfinoquine, etc.
式I化合物的制备Preparation of compound of formula I
本发明还提供通式I结构的2,6-二氮杂螺[3.4]辛烷类嘧啶-异羟肟酸及其中间体化合 物的制备方法，具体合成方法如下：The present invention also provides a preparation method of 2,6-diazaspiro[3.4]octane pyrimidine-hydroxamic acid and its intermediate compounds with the structure of general formula I. The specific synthesis method is as follows:
Figure PCTCN2019077356-appb-000009
Figure PCTCN2019077356-appb-000009
1)将2,6-二氮杂螺[3.4]辛烷-2-羧酸叔丁酯(化合物I-a)和2-氯-嘧啶-5-羧酸乙酯溶于二氯甲烷中，冰浴冷却至0～5℃，加入N,N-二异丙基乙胺，加毕，撤去冰浴，室温搅拌5～6h。向反应液中加入水，振荡分液，有机相减压蒸馏除去溶剂，剩余混合物通过硅胶柱层析分离纯化，得淡黄色固体6-(5-(乙氧基羰基)嘧啶-2-基)-2,6-二氮杂螺[3.4]辛烷-2-羧酸叔丁酯(化合物I-b)。1) Dissolve 2,6-diazaspiro[3.4]octane-2-carboxylic acid tert-butyl ester (Compound Ia) and 2-chloro-pyrimidine-5-carboxylic acid ethyl ester in dichloromethane, ice bath Cool to 0～5℃, add N,N-diisopropylethylamine, after addition, remove the ice bath, and stir at room temperature for 5～6h. Water was added to the reaction solution, the liquid was separated by shaking, the organic phase was distilled under reduced pressure to remove the solvent, and the remaining mixture was separated and purified by silica gel column chromatography to obtain a pale yellow solid 6-(5-(ethoxycarbonyl)pyrimidin-2-yl) -2,6-diazaspiro[3.4]octane-2-carboxylic acid tert-butyl ester (Compound Ib).
2)将步骤1得到的化合物与2,6-二叔丁基吡啶溶于二氯甲烷，冰水浴冷却至0～5℃，缓慢滴加三氟甲磺酸三甲基硅酯，滴加完毕，保持冰水浴，搅拌15min，撤去冰水浴，室温下搅拌至反应完毕(TLC检测)。再将反应液冷置于冰水浴中，剧烈搅拌下向体系缓慢滴加无水甲醇(加入量与三氟甲磺酸三甲基硅酯相同)滴加甲醇完毕，撤去冰水浴，待体系升至室温，将体系倒入二氯甲烷，抽滤，将固体用二氯甲烷洗至白色，烘干，得白色固体2-(2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯三氟甲磺酸盐(化合物I-c)。2) Dissolve the compound obtained in step 1 and 2,6-di-tert-butylpyridine in dichloromethane, cool to 0～5°C in an ice water bath, slowly add trimethylsilyl trifluoromethanesulfonate dropwise, and the addition is complete Keep the ice water bath, stir for 15 min, remove the ice water bath, and stir at room temperature until the reaction is complete (TLC detection). Then cool the reaction liquid in an ice-water bath, slowly add anhydrous methanol (the addition amount is the same as trimethylsilyl trifluoromethanesulfonate) to the system under vigorous stirring. After the methanol is added, remove the ice-water bath and wait for the system to rise. To room temperature, pour the system into dichloromethane, filter with suction, wash the solid with dichloromethane until white, and dry to obtain a white solid 2-(2,6-diazaspiro[3.4]oct-6-yl) Pyrimidine-5-carboxylic acid ethyl ester triflate (Compound Ic).
3)将步骤2得到的化合物溶于1,2-二氯乙烷中，加入R 1取代的醛，冰醋酸，三乙酰氧基硼氢化钠，室温搅拌过夜。反应毕，加入饱和碳酸氢钠水溶液，振荡分液，有机相减压蒸馏除去溶剂，剩余混合物用硅胶柱层析分离纯化，得化合物I-d。 3) The compound obtained in Step 2 was dissolved in 1,2-dichloroethane is added an aldehyde substituted with R 1, glacial acetic acid, triacetoxy sodium borohydride, stirred at room temperature overnight. After the reaction is completed, a saturated aqueous sodium bicarbonate solution is added, the liquid is separated by shaking, the organic phase is distilled under reduced pressure to remove the solvent, and the remaining mixture is separated and purified by silica gel column chromatography to obtain compound Id.
4)将步骤3得到的化合物用甲醇和水的混合溶液溶解，加入碳酸钾固体，65～70℃加热反应5-6h。反应毕，减压蒸馏除去溶剂，残留物用2M盐酸酸化至pH约为1，减压蒸馏除去水，得混有无机盐的化合物I-e直接用于下步反应。4) Dissolve the compound obtained in step 3 with a mixed solution of methanol and water, add solid potassium carbonate, and heat to react at 65-70°C for 5-6 hours. After the reaction was completed, the solvent was distilled off under reduced pressure, the residue was acidified with 2M hydrochloric acid to a pH of about 1, and water was distilled off under reduced pressure to obtain compound I-e mixed with inorganic salts and used directly in the next reaction.
5)将步骤4得到的混合物用N,N-二甲基甲酰胺溶液溶解，依次加入1-羟基苯并三唑和1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐，室温下搅拌30min后，依次加入O-(四氢-2H-吡喃-2-基)羟胺和三乙胺，室温下搅拌48h。反应毕，加入饱和碳酸氢钠水溶液和二氯甲烷，振荡分液，有机相用水萃取三次，有机相减压蒸馏除去溶剂，剩余混合物用硅胶柱层析分离纯化，得化合物I-f。5) Dissolve the mixture obtained in step 4 with N,N-dimethylformamide solution, add 1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide in sequence Amine hydrochloride, after stirring for 30 min at room temperature, O-(tetrahydro-2H-pyran-2-yl)hydroxylamine and triethylamine were added in sequence, and stirred at room temperature for 48 hours. After the reaction is complete, add saturated sodium bicarbonate aqueous solution and dichloromethane, shake and separate, the organic phase is extracted three times with water, the organic phase is distilled under reduced pressure to remove the solvent, and the remaining mixture is separated and purified by silica gel column chromatography to obtain compound I-f.
6)将步骤5得到的化合物用二氯甲烷溶解，加入4M氯化氢-二氧六环溶液，室温下搅拌30min。抽滤，以大量二氯甲烷洗涤固体，得到化合物I。6) Dissolve the compound obtained in step 5 with dichloromethane, add 4M hydrogen chloride-dioxane solution, and stir for 30 min at room temperature. Filtration with suction and washing the solid with a large amount of dichloromethane to obtain compound I.
根据上述制备方法的教导，本领域普通技术人员无需创造性劳动，即可获得式I所包含的所有化合物。According to the teaching of the above preparation method, a person of ordinary skill in the art can obtain all the compounds contained in Formula I without creative work.
药物组合物和施用方法Pharmaceutical composition and method of administration
由于本发明化合物具有优异的对HDAC的激活活性，因此本发明化合物及其各种晶型，药学上可接受的无机或有机盐，水合物或溶剂合物，以及含有本发明化合物为主要活性成分的药物组合物可用于治疗、预防以及缓解由HDAC的活性或表达量所导致的疾病。根据现有技术，本发明化合物可用于治疗以下疾病：肺癌、结肠癌、前列腺癌、乳 腺癌、卵巢癌、淋巴系统肿瘤、以及疟原虫导致的疾病(如疟疾)。Since the compound of the present invention has excellent HDAC activation activity, the compound of the present invention and its various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and containing the compound of the present invention as the main active ingredient The pharmaceutical composition can be used to treat, prevent and alleviate diseases caused by the activity or expression of HDAC. According to the prior art, the compounds of the present invention can be used to treat the following diseases: lung cancer, colon cancer, prostate cancer, breast cancer, ovarian cancer, lymphatic system tumors, and diseases caused by malaria parasites (such as malaria).
本发明的药物组合物包含安全有效量范围内的本发明化合物或其药理上可接受的盐及药理上可以接受的赋形剂或载体。其中“安全有效量”指的是：化合物的量足以明显改善病情，而不至于产生严重的副作用。通常，药物组合物含有0.1-1000mg本发明化合物/剂，更佳地，含有0.5-500mg本发明化合物/剂。较佳地，所述的“一剂”为一个胶囊或药片。The pharmaceutical composition of the present invention contains the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier within a safe and effective amount. The "safe and effective amount" refers to: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Generally, the pharmaceutical composition contains 0.1-1000 mg of the compound of the present invention per agent, more preferably, 0.5-500 mg of the compound of the present invention per agent. Preferably, the "one dose" is a capsule or tablet.
“药学上可以接受的载体”指的是：一种或多种相容性固体或液体填料或凝胶物质，它们适合于人使用，而且必须有足够的纯度和足够低的毒性。“相容性”在此指的是组合物中各组份能和本发明的化合物以及它们之间相互掺和，而不明显降低化合物的药效。药学上可以接受的载体部分例子有纤维素及其衍生物(如羧甲基纤维素钠、乙基纤维素钠、纤维素乙酸酯等)、明胶、滑石、固体润滑剂(如硬脂酸、硬脂酸镁)、硫酸钙、植物油(如豆油、芝麻油、花生油、橄榄油等)、多元醇(如丙二醇、甘油、甘露醇、山梨醇等)、乳化剂(如吐温)、润湿剂(如十二烷基硫酸钠)、着色剂、调味剂、稳定剂、抗氧化剂、防腐剂、无热原水等。"Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use, and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be blended with the compound of the present invention and between them without significantly reducing the efficacy of the compound. Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, and solid lubricants (such as stearic acid). , Magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as Tween), wetting Agents (such as sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.
本发明化合物或药物组合物的施用方式没有特别限制，代表性的施用方式包括(但并不限于)：口服、直肠、肠胃外(静脉内、肌肉内或皮下)、和局部给药。特别优选的施用方式是口服。The administration method of the compound or pharmaceutical composition of the present invention is not particularly limited. Representative administration methods include (but are not limited to): oral, rectal, parenteral (intravenous, intramuscular, or subcutaneous), and topical administration. A particularly preferred mode of administration is oral.
用于口服给药的固体剂型包括胶囊剂、片剂、丸剂、散剂和颗粒剂。在这些固体剂型中，活性化合物与至少一种常规惰性赋形剂(或载体)混合，如柠檬酸钠或磷酸二钙，或与下述成分混合：(a)填料或增容剂，例如，淀粉、乳糖、蔗糖、葡萄糖、甘露醇和硅酸；(b)粘合剂，例如，羟甲基纤维素、藻酸盐、明胶、聚乙烯基吡咯烷酮、蔗糖和阿拉伯胶；(c)保湿剂，例如，甘油；(d)崩解剂，例如，琼脂、碳酸钙、马铃薯淀粉或木薯淀粉、藻酸、某些复合硅酸盐、和碳酸钠；(e)缓溶剂，例如石蜡；(f)吸收加速剂，例如，季胺化合物；(g)润湿剂，例如鲸蜡醇和单硬脂酸甘油酯；(h)吸附剂，例如，高岭土；和(i)润滑剂，例如，滑石、硬脂酸钙、硬脂酸镁、固体聚乙二醇、十二烷基硫酸钠，或其混合物。胶囊剂、片剂和丸剂中，剂型也可包含缓冲剂。Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with the following ingredients: (a) fillers or compatibilizers, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) humectant, For example, glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) Absorption accelerators, such as quaternary amine compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, hard Calcium fatty acid, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain buffering agents.
固体剂型如片剂、糖丸、胶囊剂、丸剂和颗粒剂可采用包衣和壳材制备，如肠衣和其它本领域公知的材料。它们可包含不透明剂，并且，这种组合物中活性化合物或化合物的释放可以延迟的方式在消化道内的某一部分中释放。可采用的包埋组分的实例是聚合物质和蜡类物质。必要时，活性化合物也可与上述赋形剂中的一种或多种形成微胶囊形式。Solid dosage forms such as tablets, sugar pills, capsules, pills and granules can be prepared with coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the active compound or the release of the compound in such a composition may be released in a certain part of the digestive tract in a delayed manner. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microcapsules with one or more of the above-mentioned excipients.
用于口服给药的液体剂型包括药学上可接受的乳液、溶液、悬浮液、糖浆或酊剂。除了活性化合物外，液体剂型可包含本领域中常规采用的惰性稀释剂，如水或其它溶剂，增溶剂和乳化剂，例知，乙醇、异丙醇、碳酸乙酯、乙酸乙酯、丙二醇、1,3-丁二醇、二甲基甲酰胺以及油，特别是棉籽油、花生油、玉米胚油、橄榄油、蓖麻油和芝麻油或这些物质的混合物等。Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-Butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.
除了这些惰性稀释剂外，组合物也可包含助剂，如润湿剂、乳化剂和悬浮剂、甜味剂、矫味剂和香料。In addition to these inert diluents, the composition may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.
除了活性化合物外，悬浮液可包含悬浮剂，例如，乙氧基化异十八烷醇、聚氧乙烯山梨醇和脱水山梨醇酯、微晶纤维素、甲醇铝和琼脂或这些物质的混合物等。In addition to the active compound, the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.
用于肠胃外注射的组合物可包含生理上可接受的无菌含水或无水溶液、分散液、悬浮液或乳液，和用于重新溶解成无菌的可注射溶液或分散液的无菌粉末。适宜的含水和 非水载体、稀释剂、溶剂或赋形剂包括水、乙醇、多元醇及其适宜的混合物。The composition for parenteral injection may contain physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.
用于局部给药的本发明化合物的剂型包括软膏剂、散剂、贴剂、喷射剂和吸入剂。活性成分在无菌条件下与生理上可接受的载体及任何防腐剂、缓冲剂，或必要时可能需要的推进剂一起混合。The dosage form of the compound of the present invention for topical administration includes ointment, powder, patch, spray and inhalant. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required if necessary.
本发明化合物可以单独给药，或者与其他药学上可接受的化合物联合给药。The compound of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds.
使用药物组合物时，是将安全有效量的本发明化合物适用于需要治疗的哺乳动物(如人)，其中施用时剂量为药学上认为的有效给药剂量，对于60kg体重的人而言，日给药剂量通常为0.2～1000mg，优选0.5～500mg。当然，具体剂量还应考虑给药途径、病人健康状况等因素，这些都是熟练医师技能范围之内的。When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage is the pharmaceutically effective dosage considered to be administered. For a 60kg body weight, the daily The dose administered is usually 0.2 to 1000 mg, preferably 0.5 to 500 mg. Of course, the specific dosage should also consider factors such as the route of administration, the patient's health status, etc., which are within the skill range of a skilled physician.
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数按重量计算。实施例中的所有参数以及其余的说明，除另有说明外，都是以质量(g)为单位。The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods that do not indicate specific conditions in the following examples usually follow the conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight. All parameters in the embodiment and the rest of the description, unless otherwise specified, are based on mass (g).
实施例1Example 1
6-(5-(乙氧基羰基)嘧啶-2-基)-2,6-二氮杂螺[3.4]辛烷-2-羧酸叔丁酯(中间体I-b)的制备Preparation of 6-(5-(ethoxycarbonyl)pyrimidin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylic acid tert-butyl ester (Intermediate I-b)
Figure PCTCN2019077356-appb-000010
Figure PCTCN2019077356-appb-000010
将21.2g 2,6-二氮杂螺[3.4]辛烷-2-羧酸叔丁酯和18.7g 2-氯-嘧啶-5-羧酸乙酯溶于500mL二氯甲烷中，冰浴冷却至0～5℃，加入18.2mL N,N-二异丙基乙胺，加毕，然后撤去冰浴，待反应液温度恢复至室温，搅拌6h。反应毕，向反应液中加入水，振荡分液，减压蒸除溶剂，剩余固体用硅胶柱层析分离纯化，得淡黄色固体。Dissolve 21.2g of 2,6-diazaspiro[3.4]octane-2-carboxylic acid tert-butyl ester and 18.7g of 2-chloro-pyrimidine-5-carboxylic acid ethyl ester in 500mL of dichloromethane and cool in an ice bath To 0～5℃, add 18.2mL N,N-diisopropylethylamine, after the addition, remove the ice bath, wait until the temperature of the reaction liquid returns to room temperature, and stir for 6h. After the reaction was completed, water was added to the reaction solution, the liquid was separated by shaking, the solvent was evaporated under reduced pressure, and the remaining solid was separated and purified by silica gel column chromatography to obtain a pale yellow solid.
1H NMR(400MHz,CDCl 3)δ8.86(s,2H),4.34(q,J＝7.1Hz,2H),3.94(d,J＝8.6Hz,2H),3.88(d,J＝8.6Hz,2H),3.80(s,2H),3.70(t,J＝6.9Hz,2H),2.22(t,J＝6.9Hz,2H),1.45(s,10H),1.36(t,J＝7.1Hz,3H). 1 H NMR(400MHz, CDCl 3 )δ8.86(s,2H), 4.34(q,J=7.1Hz,2H), 3.94(d,J=8.6Hz,2H), 3.88(d,J=8.6Hz ,2H),3.80(s,2H),3.70(t,J＝6.9Hz,2H),2.22(t,J＝6.9Hz,2H),1.45(s,10H),1.36(t,J＝7.1Hz ,3H).
实施例2Example 2
2-(2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯三氟甲磺酸盐(中间体I-c)的制备Preparation of 2-(2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester triflate (Intermediate I-c)
Figure PCTCN2019077356-appb-000011
Figure PCTCN2019077356-appb-000011
将33.3g 6-(5-(乙氧基羰基)嘧啶-2-基)-2,6-二氮杂螺[3.4]辛烷-2-羧酸叔丁酯与25.8g的2,6-二叔丁基吡啶溶于500mL二氯甲烷，冰水浴冷却至0～5℃，缓慢滴加24.4mL三氟甲磺酸三甲基硅酯，滴加完毕，保持冰水浴，搅拌15min，然后撤去冰水浴，室温下搅拌至TLC显示底物消失，反应完毕，冷却至冰水浴，剧烈搅拌下向体系缓慢滴加25mL无水甲醇，滴加完毕，撤去冰水浴，待体系升至室温，将体系倒入2500mL二氯甲烷，抽滤，将固体用二氯甲烷洗至白色，烘干，得到白色固体。Combine 33.3g 6-(5-(ethoxycarbonyl)pyrimidin-2-yl)-2,6-diazaspiro[3.4]octane-2-carboxylic acid tert-butyl ester and 25.8g of 2,6- Dissolve di-tert-butylpyridine in 500mL of dichloromethane, cool to 0～5℃ in an ice-water bath, slowly add 24.4mL of trimethylsilyl trifluoromethanesulfonate dropwise, after the addition, keep the ice-water bath, stir for 15min, then remove In an ice water bath, stir at room temperature until TLC shows that the substrate disappears. After the reaction is complete, cool to an ice water bath. Under vigorous stirring, slowly add 25 mL of anhydrous methanol to the system. After the addition is complete, remove the ice water bath. Pour into 2500 mL of dichloromethane, filter with suction, wash the solid with dichloromethane until white, and dry to obtain a white solid.
1H NMR(400MHz,DMSO-d 6)δ8.84(s,2H),4.31(q,J＝7.1Hz,2H),4.07(d,J＝10.8Hz,2H),3.95(d,J＝10.7Hz,2H),3.85(s,2H),3.64(t,J＝6.9Hz,2H),2.29(t,J＝6.9Hz, 2H),1.33(t,J＝7.1Hz,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ 8.84 (s, 2H), 4.31 (q, J = 7.1 Hz, 2H), 4.07 (d, J = 10.8 Hz, 2H), 3.95 (d, J = 10.7Hz, 2H), 3.85 (s, 2H), 3.64 (t, J = 6.9 Hz, 2H), 2.29 (t, J = 6.9 Hz, 2H), 1.33 (t, J = 7.1 Hz, 3H).
实施例3Example 3
2-(2-((1-甲基-1H-吲哚-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-1d)的制备2-(2-((1-methyl-1H-indol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (Compound I-1d) Preparation
Figure PCTCN2019077356-appb-000012
Figure PCTCN2019077356-appb-000012
将0.41g 2-(2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯三氟甲磺酸盐(中间体I-c)和0.19g N-甲基吲哚-3-甲醛，溶于15mL二氯甲烷中，加入0.27mL冰醋酸，0.76g三乙酰氧基硼氢化钠，室温搅拌过夜。反应毕，加入饱和碳酸氢钠水溶液，振荡分液，有机相减压蒸馏除去溶剂，剩余混合物用硅胶柱层析分离纯化，得淡黄色固体。0.41g 2-(2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester triflate (Intermediate Ic) and 0.19g N-methylindole Dole-3-carbaldehyde was dissolved in 15 mL of dichloromethane, 0.27 mL of glacial acetic acid and 0.76 g of sodium triacetoxyborohydride were added, and the mixture was stirred overnight at room temperature. After the reaction is completed, a saturated sodium bicarbonate aqueous solution is added, the liquid is separated by shaking, the organic phase is distilled under reduced pressure to remove the solvent, and the remaining mixture is separated and purified by silica gel column chromatography to obtain a light yellow solid.
1H NMR(400MHz,CDCl 3)δ8.83(s,2H),7.66(d,J＝7.9Hz,1H),7.31(d,J＝8.2Hz,1H),7.23(d,J＝7.2Hz,1H),7.16–7.12(m,1H),7.09(s,1H),4.33(q,J＝7.1Hz,2H),3.93(s,2H),3.78(s,3H),3.72(s,2H),3.64(t,J＝6.9Hz,2H),3.46(d,J＝17.7Hz,2H),3.37(d,J＝7.3Hz,2H),2.27(t,J＝6.9Hz,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR(400MHz,CDCl 3 )δ8.83(s,2H), 7.66(d,J=7.9Hz,1H), 7.31(d,J=8.2Hz,1H), 7.23(d,J=7.2Hz ,1H),7.16–7.12(m,1H),7.09(s,1H),4.33(q,J=7.1Hz,2H),3.93(s,2H),3.78(s,3H),3.72(s, 2H), 3.64 (t, J = 6.9 Hz, 2H), 3.46 (d, J = 17.7 Hz, 2H), 3.37 (d, J = 7.3 Hz, 2H), 2.27 (t, J = 6.9 Hz, 2H) ,1.36(t,J=7.1Hz,3H).
实施例4Example 4
2-(2-((1-甲基-1H-吲哚-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸(化合物I-1e)的制备2-(2-((1-methyl-1H-indol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid (compound I-1e) Preparation
Figure PCTCN2019077356-appb-000013
Figure PCTCN2019077356-appb-000013
将0.24g 2-(2-((1-甲基-1H-吲哚-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-1d)溶于甲醇—水的混合溶液(1:1)10mL，加入0.25g碳酸钾固体，65～70℃加热反应6h。反应毕，减压蒸除溶剂，残留物用2M盐酸酸化至pH约为1，减压蒸馏除去水，得淡粉色固体(含有无机盐)。0.24g 2-(2-((1-methyl-1H-indol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxy Ethyl acid (compound I-1d) was dissolved in 10 mL of methanol-water mixed solution (1:1), 0.25 g of potassium carbonate solid was added, and the reaction was heated at 65-70°C for 6 hours. After the reaction was completed, the solvent was evaporated under reduced pressure, the residue was acidified with 2M hydrochloric acid to a pH of about 1, and water was distilled off under reduced pressure to obtain a pale pink solid (containing inorganic salt).
1H NMR(400MHz,DMSO-d 6)δ11.12(s,1H),8.83–8.77(m,2H),7.90(dd,J＝7.8,4.6Hz,1H),7.66(d,J＝4.6Hz,1H),7.52(d,J＝8.2Hz,1H),7.26(t,J＝7.6Hz,1H),7.17(td,J＝7.1,2.8Hz,1H),4.58(t,J＝5.1Hz,2H),4.25–4.14(m,2H),4.07–3.94(m,2H),3.84(t,J＝3.0Hz,3H),3.64–3.57(m,2H),3.20(s,2H),2.31(dt,J＝31.9,6.9Hz,2H). 1 H NMR(400MHz,DMSO-d 6 )δ11.12(s,1H),8.83-8.77(m,2H),7.90(dd,J=7.8,4.6Hz,1H),7.66(d,J=4.6 Hz, 1H), 7.52 (d, J = 8.2 Hz, 1H), 7.26 (t, J = 7.6 Hz, 1H), 7.17 (td, J = 7.1, 2.8 Hz, 1H), 4.58 (t, J = 5.1 Hz, 2H), 4.25–4.14(m, 2H), 4.07–3.94(m, 2H), 3.84(t, J=3.0Hz, 3H), 3.64–3.57(m, 2H), 3.20(s, 2H) , 2.31 (dt, J = 31.9, 6.9 Hz, 2H).
实施例5Example 5
2-(2-((1-甲基-1H-吲哚-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)-N-((四氢-2H-吡喃-2-基)氧基)嘧啶-5-甲酰胺(化合物I-1f)的制备2-(2-((1-methyl-1H-indol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)-N-((tetrahydro- Preparation of 2H-pyran-2-yl)oxy)pyrimidine-5-carboxamide (Compound I-1f)
Figure PCTCN2019077356-appb-000014
Figure PCTCN2019077356-appb-000014
将实施例4得到的固体溶于N,N-二甲基甲酰胺溶液10mL，依次加入0.16g 1-羟基 苯并三唑和0.23g 1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐，室温下搅拌30min后，依次加入0.35g O-(四氢-2H-吡喃-2-基)羟胺和0.42mL三乙胺，室温下搅拌48h。反应毕，加入饱和碳酸氢钠水溶液和二氯甲烷，振荡分液，有机相用水萃取三次，有机相减压蒸馏除去溶剂，剩余混合物用硅胶柱层析分离纯化，得浅黄色固体。The solid obtained in Example 4 was dissolved in 10 mL of N,N-dimethylformamide solution, and 0.16g 1-hydroxybenzotriazole and 0.23g 1-(3-dimethylaminopropyl)-3-ethyl were added in sequence. After stirring for 30 min at room temperature, 0.35 g of O-(tetrahydro-2H-pyran-2-yl) hydroxylamine and 0.42 mL of triethylamine were added in sequence, and stirred at room temperature for 48 hours. After the reaction is completed, add saturated sodium bicarbonate aqueous solution and dichloromethane, shake and separate, the organic phase is extracted three times with water, the organic phase is distilled under reduced pressure to remove the solvent, and the remaining mixture is separated and purified by silica gel column chromatography to obtain a pale yellow solid.
1H NMR(400MHz,CDCl 3)δ8.68(s,1H),8.47(d,J＝7.2Hz,1H),7.66(d,J＝7.9Hz,1H),7.30(d,J＝8.2Hz,1H),7.25–7.21(m,1H),7.13(t,J＝7.4Hz,1H),7.04(s,1H),5.03(s,1H),3.98(dd,J＝19.4,8.3Hz,1H),3.87(s,2H),3.77(s,3H),3.73–3.65(m,3H),3.61(td,J＝6.9,2.8Hz,2H),3.35(d,J＝27.3Hz,4H),2.29–2.20(m,2H),1.83(d,J＝12.0Hz,3H),1.62(s,3H). 1 H NMR(400MHz,CDCl 3 )δ8.68(s,1H), 8.47(d,J=7.2Hz,1H), 7.66(d,J=7.9Hz,1H), 7.30(d,J=8.2Hz ,1H),7.25–7.21(m,1H),7.13(t,J=7.4Hz,1H),7.04(s,1H),5.03(s,1H),3.98(dd,J=19.4,8.3Hz, 1H), 3.87(s, 2H), 3.77(s, 3H), 3.73–3.65(m,3H), 3.61(td,J=6.9,2.8Hz,2H), 3.35(d,J=27.3Hz,4H ), 2.29–2.20 (m, 2H), 1.83 (d, J = 12.0 Hz, 3H), 1.62 (s, 3H).
实施例6Example 6
2-(2-((1-甲基-1H-吲哚-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-1)的制备2-(2-((1-methyl-1H-indol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamic acid Preparation of Hydrochloride (Compound I-1)
Figure PCTCN2019077356-appb-000015
Figure PCTCN2019077356-appb-000015
将0.14g 2-(2-((1-甲基-1H-吲哚-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)-N-((四氢-2H-吡喃-2-基)氧基)嘧啶-5-甲酰胺(化合物I-1f)用10mL二氯甲烷溶解，滴加0.5mL 4M氯化氢-二氧六环溶液，室温下搅拌30min。抽滤，以大量二氯甲烷洗涤固体，烘干，得浅黄色固体。0.14g 2-(2-((1-methyl-1H-indol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)-N-(( Tetrahydro-2H-pyran-2-yl)oxy)pyrimidine-5-carboxamide (compound I-1f) was dissolved in 10mL of dichloromethane, 0.5mL of 4M hydrogen chloride-dioxane solution was added dropwise, and stirred at room temperature 30min. Suction filtration, washing the solid with a large amount of dichloromethane, and drying to obtain a light yellow solid.
1H NMR(400MHz,DMSO-d 6)δ11.04(d,J＝30.2Hz,2H),8.72(d,J＝8.8Hz,2H),7.90(dd,J＝7.8,4.6Hz,1H),7.66(d,J＝4.5Hz,1H),7.52(d,J＝8.2Hz,1H),7.26(t,J＝7.6Hz,1H),7.21–7.14(m,1H),4.58(t,J＝4.9Hz,2H),4.25–4.14(m,2H),4.07–3.95(m,2H),3.84(t,J＝18.6Hz,5H),3.63–3.53(m,2H),2.30(dt,J＝31.1,7.0Hz,2H)；HRMS(ESI)m/z calcd for C 23H 29N 6O 3(M+H)+393.2039,found 393.2040. 1 H NMR(400MHz,DMSO-d 6 )δ11.04(d,J=30.2Hz,2H), 8.72(d,J=8.8Hz,2H), 7.90(dd,J=7.8,4.6Hz,1H) ,7.66(d,J=4.5Hz,1H),7.52(d,J=8.2Hz,1H),7.26(t,J=7.6Hz,1H),7.21-7.14(m,1H),4.58(t, J = 4.9Hz, 2H), 4.25–4.14 (m, 2H), 4.07–3.95 (m, 2H), 3.84 (t, J = 18.6 Hz, 5H), 3.63–3.53 (m, 2H), 2.30 (dt ,J=31.1,7.0Hz,2H); HRMS(ESI)m/z calcd for C 23 H 29 N 6 O 3 (M+H)+393.2039,found 393.2040.
实施例7Example 7
2-(2-(呋喃-2-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-2d)的制备Preparation of 2-(2-(furan-2-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-2d)
Figure PCTCN2019077356-appb-000016
Figure PCTCN2019077356-appb-000016
将实施例3中的N-甲基吲哚-3-甲醛替换成2-呋喃甲醛，其余所需原料，试剂及制备方法同实施例3，得I-2d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 2-furaldehyde, and the remaining raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-2d.
1H NMR(400MHz,CDCl 3)δ8.84(s,2H),7.38(d,J＝1.1Hz,1H),6.33(dd,J＝3.1,1.9Hz,1H),6.23(d,J＝3.1Hz,1H),4.35(dd,J＝7.1,3.0Hz,2H),3.71(s,2H),3.70(s,2H),3.64(d,J＝6.9Hz,2H),3.44(d,J＝8.2Hz,2H),3.36(t,J＝5.2Hz,2H),2.23(t,J＝7.0Hz,3H),1.38–1.36(m,3H). 1 H NMR(400MHz,CDCl 3 )δ8.84(s,2H), 7.38(d,J=1.1Hz,1H), 6.33(dd,J=3.1,1.9Hz,1H), 6.23(d,J= 3.1Hz, 1H), 4.35 (dd, J = 7.1, 3.0 Hz, 2H), 3.71 (s, 2H), 3.70 (s, 2H), 3.64 (d, J = 6.9 Hz, 2H), 3.44 (d, J = 8.2Hz, 2H), 3.36 (t, J = 5.2Hz, 2H), 2.23 (t, J = 7.0Hz, 3H), 1.38-1.36 (m, 3H).
实施例8Example 8
2-(2-(呋喃-3-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-3d)的制备Preparation of 2-(2-(furan-3-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-3d)
Figure PCTCN2019077356-appb-000017
Figure PCTCN2019077356-appb-000017
将实施例3中的N-甲基吲哚-3-甲醛替换成3-呋喃甲醛，其余所需原料，试剂及制备方法同实施例3，得I-3d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 3-furaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-3d.
1H NMR(400MHz,CDCl 3)δ8.83(s,2H),7.36(t,J＝1.6Hz,1H),7.32(s,1H),6.35(d,J＝0.9Hz,1H),4.32(q,J＝7.1Hz,2H),3.72(s,2H),3.64(t,J＝7.0Hz,2H),3.48(s,2H),3.32–3.24(m,2H),3.18(d,J＝7.8Hz,2H),2.21(t,J＝7.0Hz,2H),1.35(t,J＝7.1Hz,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.83 (s, 2H), 7.36 (t, J = 1.6 Hz, 1H), 7.32 (s, 1H), 6.35 (d, J = 0.9 Hz, 1H), 4.32 (q,J=7.1Hz,2H),3.72(s,2H), 3.64(t,J=7.0Hz,2H), 3.48(s,2H), 3.32–3.24(m,2H), 3.18(d, J=7.8Hz,2H),2.21(t,J=7.0Hz,2H),1.35(t,J=7.1Hz,3H).
实施例9Example 9
2-(2-(噻吩-2-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-4d)的制备Preparation of 2-(2-(thiophen-2-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-4d)
Figure PCTCN2019077356-appb-000018
Figure PCTCN2019077356-appb-000018
将实施例3中的N-甲基吲哚-3-甲醛替换成2-噻吩甲醛，其余所需原料，试剂及制备方法同实施例3，得I-4d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 2-thiophenecarbaldehyde, and the remaining raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-4d.
1H NMR(400MHz,CDCl 3)δ8.84(s,2H),7.22(dd,J＝5.0,1.1Hz,1H),6.94(dd,J＝5.0,3.5Hz,1H),6.91(d,J＝2.8Hz,1H),4.33(q,J＝7.1Hz,2H),3.83(s,2H),3.74(s,2H),3.65(t,J＝7.0Hz,2H),3.32(d,J＝7.6Hz,2H),3.24(d,J＝7.7Hz,2H),2.23(t,J＝7.0Hz,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR(400MHz, CDCl 3 )δ8.84(s,2H), 7.22(dd,J=5.0,1.1Hz,1H), 6.94(dd,J=5.0,3.5Hz,1H), 6.91(d, J = 2.8Hz, 1H), 4.33 (q, J = 7.1Hz, 2H), 3.83 (s, 2H), 3.74 (s, 2H), 3.65 (t, J = 7.0 Hz, 2H), 3.32 (d, J = 7.6 Hz, 2H), 3.24 (d, J = 7.7 Hz, 2H), 2.23 (t, J = 7.0 Hz, 2H), 1.36 (t, J = 7.1 Hz, 3H).
实施例10Example 10
2-(2-(噻吩-3-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-5d)的制备Preparation of 2-(2-(thiophen-3-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-5d)
Figure PCTCN2019077356-appb-000019
Figure PCTCN2019077356-appb-000019
将实施例3中的N-甲基吲哚-3-甲醛替换成3-噻吩甲醛，其余所需原料，试剂及制备方法同实施例3，得I-5d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 3-thiophenecarbaldehyde, and the remaining raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-5d.
1H NMR(400MHz,CDCl 3)δ8.84(s,2H),7.29–7.26(m,1H),7.13(s,1H),7.03(d,J＝4.9Hz,1H),4.33(q,J＝7.0Hz,2H),3.74(s,2H),3.69(s,2H),3.65(t,J＝6.9Hz,2H),3.34(d,J＝7.5Hz,2H),3.25(d,J＝7.5Hz,2H),2.23(t,J＝6.9Hz,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.84 (s, 2H), 7.29-7.26 (m, 1H), 7.13 (s, 1H), 7.03 (d, J = 4.9 Hz, 1H), 4.33 (q, J = 7.0Hz, 2H), 3.74 (s, 2H), 3.69 (s, 2H), 3.65 (t, J = 6.9 Hz, 2H), 3.34 (d, J = 7.5 Hz, 2H), 3.25 (d, J = 7.5Hz, 2H), 2.23 (t, J = 6.9Hz, 2H), 1.36 (t, J = 7.1Hz, 3H).
实施例11Example 11
2-(2-(吡啶-2-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-6d)的制备Preparation of 2-(2-(pyridin-2-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-6d)
Figure PCTCN2019077356-appb-000020
Figure PCTCN2019077356-appb-000020
将实施例3中的N-甲基吲哚-3-甲醛替换成2-吡啶甲醛，其余所需原料，试剂及制备方法同实施例3，得I-6d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 2-pyridinecarboxaldehyde, and the remaining raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-6d.
1H NMR(400MHz,CDCl 3)δ8.83(s,2H),8.54(d,J＝4.5Hz,1H),7.68(t,J＝7.3Hz,1H),7.36(d,J＝7.8Hz,1H),7.23–7.16(m,1H),4.31(q,J＝7.1Hz,2H),3.86(d,J＝8.3Hz,2H),3.76(s,2H),3.65(t,J＝7.0Hz,2H),3.47(d,J＝9.5Hz,2H),3.41(d,J＝7.5Hz,2H),2.27(t,J＝6.9Hz,2H),1.34(t,J＝7.1Hz,3H). 1 H NMR(400MHz,CDCl 3 )δ8.83(s,2H), 8.54(d,J=4.5Hz,1H), 7.68(t,J=7.3Hz,1H), 7.36(d,J=7.8Hz ,1H),7.23–7.16(m,1H),4.31(q,J=7.1Hz,2H), 3.86(d,J=8.3Hz,2H), 3.76(s,2H), 3.65(t,J= 7.0Hz, 2H), 3.47 (d, J = 9.5 Hz, 2H), 3.41 (d, J = 7.5 Hz, 2H), 2.27 (t, J = 6.9 Hz, 2H), 1.34 (t, J = 7.1 Hz ,3H).
实施例12Example 12
2-(2-(吡啶-3-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-7d)的制备Preparation of 2-(2-(pyridin-3-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-7d)
Figure PCTCN2019077356-appb-000021
Figure PCTCN2019077356-appb-000021
将实施例3中的N-甲基吲哚-3-甲醛替换成3-吡啶甲醛，其余所需原料，试剂及制备方法同实施例3，得I-7d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 3-pyridinecarboxaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-7d.
1H NMR(400MHz,CDCl 3)δ8.85(s,2H),8.55(s,2H),7.78(s,1H),7.29(dd,J＝7.7,5.0Hz,1H),4.34(dt,J＝14.2,5.4Hz,2H),3.79(s,4H),3.67(t,J＝7.0Hz,2H),3.44(d,J＝38.1Hz,4H),2.30(t,J＝6.7Hz,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.85 (s, 2H), 8.55 (s, 2H), 7.78 (s, 1H), 7.29 (dd, J = 7.7, 5.0 Hz, 1H), 4.34 (dt, J = 14.2, 5.4 Hz, 2H), 3.79 (s, 4H), 3.67 (t, J = 7.0 Hz, 2H), 3.44 (d, J = 38.1 Hz, 4H), 2.30 (t, J = 6.7 Hz, 2H), 1.36 (t, J=7.1Hz, 3H).
实施例13Example 13
2-(2-(吡啶-4-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-8d)的制备Preparation of 2-(2-(pyridin-4-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-8d)
Figure PCTCN2019077356-appb-000022
Figure PCTCN2019077356-appb-000022
将实施例3中的N-甲基吲哚-3-甲醛替换成4-吡啶甲醛，其余所需原料，试剂及制备方法同实施例3，得I-8d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 4-pyridinecarboxaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-8d.
1H NMR(400MHz,CDCl 3)δ8.85(s,2H),8.54(d,J＝5.9Hz,2H),7.25(s,2H),4.34(dt,J＝11.3,5.1Hz,2H),3.77(s,2H),3.70(s,2H),3.66(t,J＝7.0Hz,2H),3.37(d,J＝7.4Hz,2H),3.29(d,J＝7.6Hz,2H),2.25(t,J＝7.0Hz,2H),1.36(dd,J＝9.2,5.1Hz,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.85 (s, 2H), 8.54 (d, J = 5.9 Hz, 2H), 7.25 (s, 2H), 4.34 (dt, J = 11.3, 5.1 Hz, 2H) ,3.77(s,2H),3.70(s,2H),3.66(t,J=7.0Hz,2H), 3.37(d,J=7.4Hz,2H), 3.29(d,J=7.6Hz,2H) ,2.25(t,J=7.0Hz,2H),1.36(dd,J=9.2,5.1Hz,3H).
实施例14Example 14
2-(2-(萘-1-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-9d)的制备Preparation of 2-(2-(naphthalen-1-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-9d)
Figure PCTCN2019077356-appb-000023
Figure PCTCN2019077356-appb-000023
将实施例3中的N-甲基吲哚-3-甲醛替换成1-萘甲醛，其余所需原料，试剂及制备 方法同实施例3，得I-9d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 1-naphthaldehyde, and the remaining raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-9d.
1H NMR(400MHz,CDCl 3)δ8.85(d,J＝9.8Hz,2H),8.15(d,J＝8.2Hz,1H),7.89–7.84(m,1H),7.79(d,J＝7.9Hz,1H),7.57–7.49(m,2H),7.49–7.41(m,2H),4.35–4.29(m,2H),4.19(s,2H),3.76(d,J＝8.2Hz,2H),3.65(t,J＝7.0Hz,2H),3.46(d,J＝7.6Hz,2H),3.38(d,J＝7.5Hz,2H),2.26(t,J＝6.9Hz,2H),1.39–1.35(m,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.85 (d, J = 9.8 Hz, 2H), 8.15 (d, J = 8.2 Hz, 1H), 7.89-7.84 (m, 1H), 7.79 (d, J = 7.9Hz, 1H), 7.57–7.49(m, 2H), 7.49–7.41(m, 2H), 4.35–4.29(m, 2H), 4.19(s, 2H), 3.76(d, J=8.2Hz, 2H ), 3.65(t,J=7.0Hz,2H), 3.46(d,J=7.6Hz,2H), 3.38(d,J=7.5Hz,2H), 2.26(t,J=6.9Hz,2H), 1.39-1.35(m,3H).
实施例15Example 15
2-(2-(萘-2-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-10d)的制备Preparation of 2-(2-(naphthalen-2-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-10d)
Figure PCTCN2019077356-appb-000024
Figure PCTCN2019077356-appb-000024
将实施例3中的N-甲基吲哚-3-甲醛替换成2-萘甲醛，其余所需原料，试剂及制备方法同实施例3，得I-10d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 2-naphthaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-10d.
1H NMR(400MHz,CDCl 3)δ8.84(s,2H),7.82(d,J＝8.5Hz,3H),7.76(s,1H),7.47(dd,J＝8.1,5.7Hz,3H),4.32(dd,J＝7.1,3.2Hz,2H),3.94(s,2H),3.77(d,J＝4.2Hz,2H),3.66–3.63(m,2H),3.51(d,J＝8.2Hz,2H),3.41(d,J＝8.2Hz,2H),2.28(t,J＝6.9Hz,2H),1.36–1.33(m,3H). 1 H NMR(400MHz,CDCl 3 )δ8.84(s,2H),7.82(d,J=8.5Hz,3H),7.76(s,1H),7.47(dd,J=8.1,5.7Hz,3H) , 4.32(dd,J=7.1,3.2Hz,2H),3.94(s,2H),3.77(d,J=4.2Hz,2H),3.66-3.63(m,2H),3.51(d,J=8.2 Hz, 2H), 3.41 (d, J = 8.2 Hz, 2H), 2.28 (t, J = 6.9 Hz, 2H), 1.36-1.33 (m, 3H).
实施例16Example 16
2-(2-([1,1'-联苯基]-4-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-11d)的制备2-(2-([1,1'-biphenyl]-4-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester ( Preparation of compound I-11d)
Figure PCTCN2019077356-appb-000025
Figure PCTCN2019077356-appb-000025
将实施例3中的N-甲基吲哚-3-甲醛替换成联苯-4-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-11d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with biphenyl-4-carbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-11d.
1H NMR(400MHz,CDCl 3)δ8.85(s,2H),7.56(dd,J＝12.4,7.8Hz,4H),7.43(t,J＝7.6Hz,2H),7.34(dd,J＝16.2,7.7Hz,3H),4.33(q,J＝7.1Hz,2H),3.74(d,J＝19.8Hz,4H),3.66(t,J＝6.9Hz,2H),3.36(d,J＝7.5Hz,2H),3.27(d,J＝7.6Hz,2H),2.25(t,J＝7.0Hz,2H),1.40–1.33(m,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.85 (s, 2H), 7.56 (dd, J = 12.4, 7.8 Hz, 4H), 7.43 (t, J = 7.6 Hz, 2H), 7.34 (dd, J = 16.2, 7.7 Hz, 3H), 4.33 (q, J = 7.1 Hz, 2H), 3.74 (d, J = 19.8 Hz, 4H), 3.66 (t, J = 6.9 Hz, 2H), 3.36 (d, J = 7.5Hz, 2H), 3.27 (d, J = 7.6Hz, 2H), 2.25 (t, J = 7.0Hz, 2H), 1.40-1.33 (m, 3H).
实施例17Example 17
2-(2-苄基-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-12d)的制备Preparation of 2-(2-benzyl-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-12d)
Figure PCTCN2019077356-appb-000026
Figure PCTCN2019077356-appb-000026
将实施例3中的N-甲基吲哚-3-甲醛替换成苯甲醛，其余所需原料，试剂及制备方法同实施例3，得I-12d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with benzaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-12d.
1H NMR(400MHz,CDCl 3)δ8.84(s,2H),7.36–7.30(m,1H),7.30–7.26(m,3H),7.26–7.21(m,1H),4.32(q,J＝7.1Hz,2H),3.74(s,2H),3.68–3.61(m,4H),3.28(d,J＝7.6 Hz,2H),3.20(d,J＝7.6Hz,2H),2.22(t,J＝7.0Hz,2H),1.35(t,J＝7.1Hz,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.84 (s, 2H), 7.36-7.30 (m, 1H), 7.30-7.26 (m, 3H), 7.26-7.21 (m, 1H), 4.32 (q, J ＝7.1Hz,2H),3.74(s,2H),3.68–3.61(m,4H), 3.28(d,J＝7.6 Hz,2H), 3.20(d,J＝7.6Hz,2H), 2.22(t ,J=7.0Hz,2H),1.35(t,J=7.1Hz,3H).
实施例18Example 18
2-(2-(环戊基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-13d)的制备Preparation of 2-(2-(cyclopentylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-13d)
Figure PCTCN2019077356-appb-000027
Figure PCTCN2019077356-appb-000027
将实施例3中的N-甲基吲哚-3-甲醛替换成环戊基甲醛，其余所需原料，试剂及制备方法同实施例3，得I-13d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with cyclopentylcarbaldehyde, and the remaining raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-13d.
1H NMR(400MHz,DMSO-d 6)δ8.85(s,2H),4.34(q,J＝7.1Hz,2H),4.16(s,2H),3.80(d,J＝47.0Hz,4H),3.70(t,J＝7.0Hz,2H),3.00(d,J＝7.1Hz,2H),2.48(s,2H),2.14(dd,J＝15.5,7.6Hz,1H),1.90(dd,J＝11.0,7.1Hz,2H),1.69–1.55(m,4H),1.37(t,J＝7.1Hz,3H),1.26–1.19(m,2H). 1 H NMR(400MHz,DMSO-d 6 )δ8.85(s,2H), 4.34(q,J=7.1Hz,2H), 4.16(s,2H), 3.80(d,J=47.0Hz,4H) ,3.70(t,J=7.0Hz,2H),3.00(d,J=7.1Hz,2H), 2.48(s,2H), 2.14(dd,J=15.5,7.6Hz,1H),1.90(dd, J = 11.0, 7.1Hz, 2H), 1.69–1.55 (m, 4H), 1.37 (t, J = 7.1Hz, 3H), 1.26–1.19 (m, 2H).
实施例19Example 19
2-(2-(环己基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-14d)的制备Preparation of 2-(2-(cyclohexylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-14d)
Figure PCTCN2019077356-appb-000028
Figure PCTCN2019077356-appb-000028
将实施例3中的N-甲基吲哚-3-甲醛替换成环己基甲醛，其余所需原料，试剂及制备方法同实施例3，得I-14d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with cyclohexylcarbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-14d.
1H NMR(400MHz,CDCl 3)δ8.84(s,2H),4.33(q,J＝7.1Hz,2H),3.72(s,2H),3.64(t,J＝7.0Hz,2H),3.33(d,J＝7.3Hz,2H),3.18(d,J＝7.6Hz,2H),2.33(d,J＝6.9Hz,2H),2.23(t,J＝7.0Hz,2H),1.68(dd,J＝29.9,15.2Hz,5H),1.36(t,J＝7.1Hz,3H),1.28–1.06(m,4H),0.87(dd,J＝22.3,10.7Hz,2H). 1 H NMR (400MHz, CDCl 3 ) δ 8.84 (s, 2H), 4.33 (q, J = 7.1 Hz, 2H), 3.72 (s, 2H), 3.64 (t, J = 7.0 Hz, 2H), 3.33 (d,J=7.3Hz,2H), 3.18(d,J=7.6Hz,2H), 2.33(d,J=6.9Hz,2H), 2.23(t,J=7.0Hz,2H), 1.68(dd ,J=29.9,15.2Hz,5H),1.36(t,J=7.1Hz,3H),1.28-1.06(m,4H),0.87(dd,J=22.3,10.7Hz,2H).
实施例20Example 20
2-(2-(苯并[b]噻吩-2-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-15d)的制备2-(2-(Benzo[b]thiophen-2-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (Compound I-15d ) Preparation
Figure PCTCN2019077356-appb-000029
Figure PCTCN2019077356-appb-000029
将实施例3中的N-甲基吲哚-3-甲醛替换成1-苯并噻酚-2-羧醛，其余所需原料，试剂及制备方法同实施例3，得I-15d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 1-benzothiophene-2-carboxaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-15d.
1H NMR(400MHz,CDCl 3)δ8.85(s,2H),7.79(d,J＝7.7Hz,1H),7.69(d,J＝7.5Hz,1H),7.31(dd,J＝15.3,7.6Hz,2H),7.15(s,1H),4.33(q,J＝7.1Hz,2H),3.92(s,2H),3.76(s,2H),3.66(t,J＝7.0Hz,2H),3.39(d,J＝7.4Hz,2H),3.31(d,J＝7.4Hz,2H),2.25(t,J＝7.0Hz,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.85 (s, 2H), 7.79 (d, J = 7.7 Hz, 1H), 7.69 (d, J = 7.5 Hz, 1H), 7.31 (dd, J = 15.3, 7.6Hz, 2H), 7.15 (s, 1H), 4.33 (q, J = 7.1 Hz, 2H), 3.92 (s, 2H), 3.76 (s, 2H), 3.66 (t, J = 7.0 Hz, 2H) , 3.39 (d, J = 7.4 Hz, 2H), 3.31 (d, J = 7.4 Hz, 2H), 2.25 (t, J = 7.0 Hz, 2H), 1.36 (t, J = 7.1 Hz, 3H).
实施例21Example 21
2-(2-(苯并[b]噻吩-3-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-16d)的制备2-(2-(Benzo[b]thiophen-3-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (Compound I-16d ) Preparation
Figure PCTCN2019077356-appb-000030
Figure PCTCN2019077356-appb-000030
将实施例3中的N-甲基吲哚-3-甲醛替换成3-甲醛苯并噻吩，其余所需原料，试剂及制备方法同实施例3，得I-16d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 3-carboxaldehyde benzothiophene, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-16d.
1H NMR(400MHz,CDCl 3)δ8.85(d,J＝5.0Hz,2H),7.88–7.83(m,2H),7.42–7.32(m,2H),7.28(d,J＝9.2Hz,1H),4.33(q,J＝7.1Hz,2H),3.90(s,2H),3.76(s,2H),3.66(t,J＝7.0Hz,2H),3.36(d,J＝7.3Hz,2H),3.29(d,J＝7.3Hz,2H),2.24(t,J＝7.0Hz,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.85 (d, J = 5.0 Hz, 2H), 7.88-7.83 (m, 2H), 7.42-7.32 (m, 2H), 7.28 (d, J = 9.2 Hz, 1H),4.33(q,J=7.1Hz,2H),3.90(s,2H),3.76(s,2H), 3.66(t,J=7.0Hz,2H), 3.36(d,J=7.3Hz, 2H), 3.29 (d, J = 7.3 Hz, 2H), 2.24 (t, J = 7.0 Hz, 2H), 1.36 (t, J = 7.1 Hz, 3H).
实施例22Example 22
2-(2-(苯并呋喃-2-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-17d)的制备Preparation of 2-(2-(benzofuran-2-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-17d)
Figure PCTCN2019077356-appb-000031
Figure PCTCN2019077356-appb-000031
将实施例3中的N-甲基吲哚-3-甲醛替换成苯并[b]呋喃-2-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-17d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with benzo[b]furan-2-carbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-17d.
1H NMR(400MHz,CDCl 3)δ8.84(s,2H),7.53(d,J＝7.1Hz,1H),7.46(d,J＝8.3Hz,1H),7.30–7.26(m,1H),7.25–7.19(m,1H),6.63(s,1H),4.33(dt,J＝10.1,5.7Hz,2H),3.85(s,2H),3.75(s,2H),3.66(t,J＝6.9Hz,2H),3.58–3.35(m,4H),2.28(t,J＝6.9Hz,2H),1.40–1.33(m,3H). 1 H NMR(400MHz, CDCl 3 )δ8.84(s,2H), 7.53(d,J=7.1Hz,1H), 7.46(d,J=8.3Hz,1H), 7.30-7.26(m,1H) ,7.25–7.19(m,1H),6.63(s,1H),4.33(dt,J=10.1,5.7Hz,2H),3.85(s,2H),3.75(s,2H),3.66(t,J =6.9Hz,2H),3.58–3.35(m,4H),2.28(t,J=6.9Hz,2H),1.40–1.33(m,3H).
实施例23Example 23
2-(2-((1H-吲哚-4-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-18d)的制备2-(2-((1H-Indol-4-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (Compound I-18d ) Preparation
Figure PCTCN2019077356-appb-000032
Figure PCTCN2019077356-appb-000032
将实施例3中的N-甲基吲哚-3-甲醛替换成4-吲哚甲醛，其余所需原料，试剂及制备方法同实施例3，得I-18d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 4-indolecarbaldehyde, and the remaining raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-18d.
1H NMR(400MHz,CDCl 3)δ8.84(d,J＝13.8Hz,2H),8.45(s,1H),7.44(d,J＝7.8Hz,1H),7.30(s,1H),7.20(dd,J＝15.1,7.5Hz,2H),6.67(s,1H),4.33(q,J＝7.1Hz,2H),4.24(s,2H),3.83–3.72(m,2H),3.71–3.58(m,6H),2.37(s,2H),1.36(dd,J＝8.9,5.3Hz,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.84 (d, J = 13.8 Hz, 2H), 8.45 (s, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.30 (s, 1H), 7.20 (dd,J=15.1,7.5Hz,2H),6.67(s,1H),4.33(q,J=7.1Hz,2H),4.24(s,2H),3.83-3.72(m,2H),3.71- 3.58(m,6H),2.37(s,2H),1.36(dd,J=8.9,5.3Hz,3H).
实施例24Example 24
2-(2-((1-甲基-1H-吲哚-2-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-19d)的制备2-(2-((1-methyl-1H-indol-2-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (Compound I-19d) Preparation
Figure PCTCN2019077356-appb-000033
Figure PCTCN2019077356-appb-000033
将实施例3中的N-甲基吲哚-3-甲醛替换成1-甲基吲哚-2-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-19d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 1-methylindole-2-carbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-19d.
1H NMR(400MHz,CDCl 3)δ8.84(s,2H),7.55(d,J＝7.8Hz,1H),7.30(d,J＝8.3Hz,1H),7.20(t,J＝7.3Hz,1H),7.08(t,J＝7.2Hz,1H),6.41(s,1H),4.33(q,J＝7.1Hz,2H),3.84(s,2H),3.79(s,3H),3.76(s,2H),3.65(t,J＝6.9Hz,2H),3.32(s,4H),2.24(s,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR(400MHz,CDCl 3 )δ8.84(s,2H), 7.55(d,J=7.8Hz,1H), 7.30(d,J=8.3Hz,1H), 7.20(t,J=7.3Hz ,1H),7.08(t,J=7.2Hz,1H),6.41(s,1H),4.33(q,J=7.1Hz,2H),3.84(s,2H),3.79(s,3H),3.76 (s, 2H), 3.65 (t, J = 6.9 Hz, 2H), 3.32 (s, 4H), 2.24 (s, 2H), 1.36 (t, J = 7.1 Hz, 3H).
实施例25Example 25
2-(2-(喹啉-3-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-20d)的制备Preparation of 2-(2-(quinolin-3-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-20d)
Figure PCTCN2019077356-appb-000034
Figure PCTCN2019077356-appb-000034
将实施例3中的N-甲基吲哚-3-甲醛替换成3-喹啉甲醛，其余所需原料，试剂及制备方法同实施例3，得I-20d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 3-quinolinecarboxaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-20d.
1H NMR(400MHz,CDCl 3)δ8.87–8.84(m,2H),8.11–8.04(m,1H),7.80(d,J＝7.7Hz,1H),7.75–7.64(m,1H),7.61–7.51(m,1H),7.02–6.91(m,1H),6.64–6.45(m,1H),4.33(q,J＝7.1Hz,2H),3.77(d,J＝7.8Hz,2H),3.66(t,J＝7.0Hz,2H),3.38–3.32(m,2H),3.29(d,J＝7.5Hz,2H),2.25(dd,J＝8.9,5.0Hz,2H),1.41–1.33(m,3H). 1 H NMR (400MHz, CDCl 3 ) δ8.87–8.84(m,2H), 8.11–8.04(m,1H), 7.80(d,J=7.7Hz,1H), 7.75–7.64(m,1H), 7.61–7.51(m,1H), 7.02–6.91(m,1H), 6.64–6.45(m,1H), 4.33(q,J=7.1Hz,2H), 3.77(d,J=7.8Hz,2H) ,3.66(t,J=7.0Hz,2H),3.38–3.32(m,2H), 3.29(d,J=7.5Hz,2H), 2.25(dd,J=8.9,5.0Hz,2H),1.41– 1.33(m,3H).
实施例26Example 26
2-(2-(喹啉-2-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-21d)的制备Preparation of 2-(2-(quinolin-2-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-21d)
Figure PCTCN2019077356-appb-000035
Figure PCTCN2019077356-appb-000035
将实施例3中的N-甲基吲哚-3-甲醛替换成喹啉-2-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-21d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with quinoline-2-carboxaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-21d.
1H NMR(400MHz,CDCl 3)δ8.85(s,2H),8.14(d,J＝8.5Hz,1H),8.06(d,J＝8.4Hz,1H),7.80(d,J＝8.2Hz,1H),7.70(t,J＝7.7Hz,1H),7.53(dd,J＝16.5,8.0Hz,2H),4.33(q,J＝7.1Hz,2H),4.06(s,2H),3.80(s,2H),3.66(t,J＝7.0Hz,2H),3.50(d,J＝9.3Hz,5H),2.28(t,J＝6.9Hz,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR(400MHz,CDCl 3 )δ8.85(s,2H), 8.14(d,J=8.5Hz,1H), 8.06(d,J=8.4Hz,1H), 7.80(d,J=8.2Hz ,1H),7.70(t,J=7.7Hz,1H),7.53(dd,J=16.5,8.0Hz,2H),4.33(q,J=7.1Hz,2H),4.06(s,2H),3.80 (s, 2H), 3.66 (t, J = 7.0 Hz, 2H), 3.50 (d, J = 9.3 Hz, 5H), 2.28 (t, J = 6.9 Hz, 2H), 1.36 (t, J = 7.1 Hz ,3H).
实施例27Example 27
2-(2-(喹啉-6-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-22d)的制备Preparation of 2-(2-(quinolin-6-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-22d)
Figure PCTCN2019077356-appb-000036
Figure PCTCN2019077356-appb-000036
将实施例3中的N-甲基吲哚-3-甲醛替换成喹啉-6-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-22d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with quinoline-6-carbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-22d.
1H NMR(400MHz,CDCl 3)δ8.89(d,J＝2.7Hz,1H),8.85(s,2H),8.10(dd,J＝30.1,8.3Hz,2H),7.78–7.64(m,2H),7.40(dd,J＝8.1,4.1Hz,1H),4.33(q,J＝7.0Hz,2H),3.87(s,2H),3.78(s,2H),3.66(t,J＝6.9Hz,2H),3.35(dd,J＝27.7,7.0Hz,4H),2.26(t,J＝6.8Hz,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.89 (d, J = 2.7Hz, 1H), 8.85 (s, 2H), 8.10 (dd, J = 30.1, 8.3 Hz, 2H), 7.78–7.64 (m, 2H), 7.40 (dd, J = 8.1, 4.1 Hz, 1H), 4.33 (q, J = 7.0 Hz, 2H), 3.87 (s, 2H), 3.78 (s, 2H), 3.66 (t, J = 6.9 Hz, 2H), 3.35 (dd, J = 27.7, 7.0 Hz, 4H), 2.26 (t, J = 6.8 Hz, 2H), 1.36 (t, J = 7.1 Hz, 3H).
实施例28Example 28
2-(2-(喹啉-8-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-23d)的制备Preparation of 2-(2-(quinolin-8-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-23d)
Figure PCTCN2019077356-appb-000037
Figure PCTCN2019077356-appb-000037
将实施例3中的N-甲基吲哚-3-甲醛替换成喹啉-8-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-23d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with quinoline-8-carbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-23d.
1H NMR(400MHz,CDCl 3)δ8.84(s,2H),8.25–8.13(m,2H),7.90(d,J＝8.1Hz,1H),7.64(t,J＝7.7Hz,1H),7.49(dd,J＝8.3,4.2Hz,1H),4.99(s,2H),4.34(q,J＝7.1Hz,2H),4.09(s,4H),3.87(s,2H),3.66(t,J＝7.0Hz,2H),2.50(s,2H),1.37(t,J＝7.1Hz,3H). 1 H NMR(400MHz,CDCl 3 )δ8.84(s,2H),8.25-8.13(m,2H),7.90(d,J=8.1Hz,1H),7.64(t,J=7.7Hz,1H) ,7.49(dd,J=8.3,4.2Hz,1H), 4.99(s,2H), 4.34(q,J=7.1Hz,2H), 4.09(s,4H), 3.87(s,2H), 3.66( t,J=7.0Hz,2H), 2.50(s,2H), 1.37(t,J=7.1Hz,3H).
实施例29Example 29
2-(2-(异喹啉-8-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-24d)的制备Preparation of 2-(2-(isoquinolin-8-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-24d)
Figure PCTCN2019077356-appb-000038
Figure PCTCN2019077356-appb-000038
将实施例3中的N-甲基吲哚-3-甲醛替换成8-异喹啉甲醛，其余所需原料，试剂及制备方法同实施例3，得I-24d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 8-isoquinolinecarboxaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-24d.
1H NMR(400MHz,CDCl 3)δ9.67(s,1H),8.84(s,2H),8.56(d,J＝5.7Hz,1H),7.76(d,J＝8.1Hz,1H),7.63(dt,J＝10.9,6.3Hz,3H),4.33(q,J＝7.1Hz,2H),4.25(s,2H),3.77(s,2H),3.65(t,J＝7.0Hz,2H),3.44(s,2H),3.38(d,J＝6.8Hz,2H),2.27(t,J＝7.0Hz,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR(400MHz,CDCl 3 )δ9.67(s,1H),8.84(s,2H),8.56(d,J=5.7Hz,1H),7.76(d,J=8.1Hz,1H), 7.63 (dt,J=10.9,6.3Hz,3H),4.33(q,J=7.1Hz,2H),4.25(s,2H),3.77(s,2H),3.65(t,J=7.0Hz,2H) ,3.44(s,2H), 3.38(d,J=6.8Hz,2H), 2.27(t,J=7.0Hz,2H),1.36(t,J=7.1Hz,3H).
实施例30Example 30
2-(2-(异喹啉-5-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-25d)的制备Preparation of 2-(2-(isoquinolin-5-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-25d)
Figure PCTCN2019077356-appb-000039
Figure PCTCN2019077356-appb-000039
将实施例3中的N-甲基吲哚-3-甲醛替换成异喹啉-5-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-25d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with isoquinoline-5-carbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-25d.
1H NMR(400MHz,CDCl 3)δ9.28(s,1H),8.84(s,2H),8.61(d,J＝5.8Hz,1H),7.95(d,J＝5.3Hz,2H),7.81(s,1H),7.60(t,J＝7.4Hz,1H),4.33(q,J＝7.1Hz,2H),4.25(s,2H),3.78(s,2H),3.66(t,J＝6.9Hz,2H),3.47(s,4H),2.33(s,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR(400MHz,CDCl 3 )δ9.28(s,1H),8.84(s,2H),8.61(d,J=5.8Hz,1H),7.95(d,J=5.3Hz,2H),7.81 (s, 1H), 7.60 (t, J = 7.4 Hz, 1H), 4.33 (q, J = 7.1 Hz, 2H), 4.25 (s, 2H), 3.78 (s, 2H), 3.66 (t, J = 6.9Hz, 2H), 3.47 (s, 4H), 2.33 (s, 2H), 1.36 (t, J = 7.1 Hz, 3H).
实施例31Example 31
2-(2-(喹啉-4-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-26d)的制 备Preparation of 2-(2-(quinolin-4-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-26d)
Figure PCTCN2019077356-appb-000040
Figure PCTCN2019077356-appb-000040
将实施例3中的N-甲基吲哚-3-甲醛替换成4-喹啉甲醛，其余所需原料，试剂及制备方法同实施例3，得I-26d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 4-quinolinecarboxaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-26d.
1H NMR(400MHz,CDCl 3)δ8.87(d,J＝4.3Hz,1H),8.85(s,2H),8.11(dd,J＝12.1,8.6Hz,2H),7.73(t,J＝7.6Hz,1H),7.58(t,J＝7.6Hz,1H),7.45(d,J＝3.7Hz,1H),4.34(d,J＝7.1Hz,2H),4.16(s,2H),3.81(s,2H),3.68(t,J＝6.9Hz,2H),3.44(s,2H),3.37(d,J＝6.8Hz,2H),2.28(t,J＝6.9Hz,2H),1.36(t,J＝6.4Hz,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.87 (d, J = 4.3 Hz, 1H), 8.85 (s, 2H), 8.11 (dd, J = 12.1, 8.6 Hz, 2H), 7.73 (t, J = 7.6Hz,1H),7.58(t,J=7.6Hz,1H),7.45(d,J=3.7Hz,1H), 4.34(d,J=7.1Hz,2H), 4.16(s,2H), 3.81 (s, 2H), 3.68 (t, J = 6.9 Hz, 2H), 3.44 (s, 2H), 3.37 (d, J = 6.8 Hz, 2H), 2.28 (t, J = 6.9 Hz, 2H), 1.36 (t,J=6.4Hz,3H).
实施例32Example 32
2-(2-(异喹啉-4-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-27d)的制备Preparation of 2-(2-(isoquinolin-4-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-27d)
Figure PCTCN2019077356-appb-000041
Figure PCTCN2019077356-appb-000041
将实施例3中的N-甲基吲哚-3-甲醛替换成异喹啉-4-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-27d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with isoquinoline-4-carbaldehyde, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-27d.
1H NMR(400MHz,CDCl 3)δ9.19(s,1H),8.84(s,2H),8.44(s,1H),8.22(d,J＝8.4Hz,1H),7.98(d,J＝8.1Hz,1H),7.76(t,J＝7.6Hz,1H),7.63(t,J＝7.5Hz,1H),4.33(q,J＝7.1Hz,2H),4.06(s,2H),3.76(s,2H),3.64(t,J＝6.9Hz,2H),3.32(dd,J＝19.9,6.8Hz,4H),2.23(t,J＝6.9Hz,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR(400MHz, CDCl 3 )δ9.19(s,1H),8.84(s,2H),8.44(s,1H), 8.22(d,J=8.4Hz,1H),7.98(d,J= 8.1Hz, 1H), 7.76 (t, J = 7.6 Hz, 1H), 7.63 (t, J = 7.5 Hz, 1H), 4.33 (q, J = 7.1 Hz, 2H), 4.06 (s, 2H), 3.76 (s, 2H), 3.64 (t, J = 6.9 Hz, 2H), 3.32 (dd, J = 19.9, 6.8 Hz, 4H), 2.23 (t, J = 6.9 Hz, 2H), 1.36 (t, J = 7.1Hz, 3H).
实施例33Example 33
2-(2-(咪唑并[1,2-a]吡啶-3-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-28d)的制备2-(2-(imidazo[1,2-a]pyridin-3-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester ( Preparation of compound I-28d)
Figure PCTCN2019077356-appb-000042
Figure PCTCN2019077356-appb-000042
将实施例3中的N-甲基吲哚-3-甲醛替换成咪唑并[1,2-a]吡啶-3-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-28d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with imidazo[1,2-a]pyridine-3-carbaldehyde, and the remaining raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I- 28d.
1H NMR(400MHz,CDCl 3)δ8.84(s,2H),8.38(d,J＝6.8Hz,1H),7.64(d,J＝9.0Hz,1H),7.52(s,1H),7.25–7.20(m,1H),6.86(t,J＝6.8Hz,1H),4.33(q,J＝7.1Hz,2H),3.94(s,2H),3.76(s,2H),3.64(t,J＝7.0Hz,2H),3.23(s,4H),2.18(t,J＝7.0Hz,2H),1.39–1.31(m,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.84 (s, 2H), 8.38 (d, J = 6.8 Hz, 1H), 7.64 (d, J = 9.0 Hz, 1H), 7.52 (s, 1H), 7.25 –7.20(m,1H),6.86(t,J=6.8Hz,1H),4.33(q,J=7.1Hz,2H),3.94(s,2H),3.76(s,2H),3.64(t, J = 7.0Hz, 2H), 3.23 (s, 4H), 2.18 (t, J = 7.0Hz, 2H), 1.39-1.31 (m, 3H).
实施例34Example 34
2-(2-((1-甲基-1H-苯并[d]咪唑-2-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-29d)的制备2-(2-((1-methyl-1H-benzo[d]imidazol-2-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5- Preparation of ethyl carboxylate (compound I-29d)
Figure PCTCN2019077356-appb-000043
Figure PCTCN2019077356-appb-000043
将实施例3中的N-甲基吲哚-3-甲醛替换成1-甲基-2-甲酰苯并咪唑，其余所需原料，试剂及制备方法同实施例3，得I-29d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 1-methyl-2-formylbenzimidazole, and the remaining required raw materials, reagents and preparation methods were the same as in Example 3 to obtain I-29d.
1H NMR(400MHz,CDCl 3)δ8.84(s,2H),7.74–7.70(m,1H),7.35–7.31(m,1H),7.29(dd,J＝7.0,1.2Hz,1H),7.23(dd,J＝7.1,1.4Hz,1H),4.33(q,J＝7.1Hz,2H),3.95(s,2H),3.87(s,3H),3.77(s,2H),3.64(t,J＝7.0Hz,2H),3.36(s,4H),2.20(t,J＝7.0Hz,2H),1.38–1.32(m,3H). 1 H NMR(400MHz, CDCl 3 )δ8.84(s,2H), 7.74–7.70(m,1H), 7.35–7.31(m,1H), 7.29(dd,J=7.0,1.2Hz,1H), 7.23(dd,J=7.1,1.4Hz,1H),4.33(q,J=7.1Hz,2H),3.95(s,2H),3.87(s,3H),3.77(s,2H), 3.64(t ,J=7.0Hz,2H), 3.36(s,4H), 2.20(t,J=7.0Hz,2H), 1.38-1.32(m,3H).
实施例35Example 35
2-(2-((1-甲基-1H-吡咯并[2,3-b]吡啶-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-30d)的制备2-(2-((1-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl) Preparation of pyrimidine-5-carboxylic acid ethyl ester (compound I-30d)
Figure PCTCN2019077356-appb-000044
Figure PCTCN2019077356-appb-000044
将实施例3中的N-甲基吲哚-3-甲醛替换成1-甲基-1H-吡咯并[2,3-b]吡啶-3-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-30d。Replace the N-methylindole-3-carbaldehyde in Example 3 with 1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde. The remaining raw materials, reagents and preparation methods are the same In Example 3, I-30d was obtained.
1H NMR(400MHz,CDCl 3)δ8.84(s,2H),8.38(dd,J＝4.7,1.4Hz,1H),8.02(dd,J＝7.9,1.4Hz,1H),7.40(s,1H),7.12(dd,J＝7.9,4.7Hz,1H),4.34(q,J＝7.1Hz,2H),4.10(s,2H),3.90(s,3H),3.74(d,J＝12.9Hz,4H),3.66(t,J＝6.9Hz,2H),3.58(s,2H),2.36(s,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR(400MHz, CDCl 3 )δ8.84(s,2H), 8.38(dd,J=4.7,1.4Hz,1H), 8.02(dd,J=7.9,1.4Hz,1H), 7.40(s, 1H), 7.12 (dd, J = 7.9, 4.7 Hz, 1H), 4.34 (q, J = 7.1 Hz, 2H), 4.10 (s, 2H), 3.90 (s, 3H), 3.74 (d, J = 12.9 Hz, 4H), 3.66 (t, J = 6.9 Hz, 2H), 3.58 (s, 2H), 2.36 (s, 2H), 1.36 (t, J = 7.1 Hz, 3H).
实施例36Example 36
2-(2-((1-甲基-1H-吡咯并[2,3-c]吡啶-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-31d)的制备2-(2-((1-methyl-1H-pyrrolo[2,3-c]pyridin-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl) Preparation of pyrimidine-5-carboxylic acid ethyl ester (Compound I-31d)
Figure PCTCN2019077356-appb-000045
Figure PCTCN2019077356-appb-000045
将实施例3中的N-甲基吲哚-3-甲醛替换成1-甲基-1H-吡咯并[2,3-c]吡啶-3-羧酸，其余所需原料，试剂及制备方法同实施例3，得I-31d。Replace the N-methylindole-3-carboxaldehyde in Example 3 with 1-methyl-1H-pyrrolo[2,3-c]pyridine-3-carboxylic acid, and other required raw materials, reagents and preparation methods Same as Example 3 to obtain I-31d.
1H NMR(400MHz,CDCl 3)δ8.84(s,2H),8.74(s,1H),8.26(d,J＝5.5Hz,1H),7.58(dd,J＝5.5,0.9Hz,1H),7.22(s,1H),4.33(q,J＝7.1Hz,2H),3.87(d,J＝3.8Hz,3H),3.86(s,2H),3.74(s,2H),3.65(t,J＝7.0Hz,2H),3.38(d,J＝7.8Hz,2H),3.30(d,J＝7.9Hz,2H),2.25(t,J＝6.9Hz,2H),1.38–1.32(m,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.84 (s, 2H), 8.74 (s, 1H), 8.26 (d, J = 5.5 Hz, 1H), 7.58 (dd, J = 5.5, 0.9 Hz, 1H) ,7.22(s,1H),4.33(q,J=7.1Hz,2H), 3.87(d,J=3.8Hz,3H), 3.86(s,2H), 3.74(s,2H), 3.65(t, J = 7.0Hz, 2H), 3.38 (d, J = 7.8Hz, 2H), 3.30 (d, J = 7.9Hz, 2H), 2.25 (t, J = 6.9Hz, 2H), 1.38-1.32 (m, 3H).
实施例37Example 37
2-(2-((1-甲基-1H-吡咯并[3,2-b]吡啶-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-32d)的制备2-(2-((1-methyl-1H-pyrrolo[3,2-b]pyridin-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl) Preparation of pyrimidine-5-carboxylic acid ethyl ester (Compound I-32d)
Figure PCTCN2019077356-appb-000046
Figure PCTCN2019077356-appb-000046
将实施例3中的N-甲基吲哚-3-甲醛替换成1-甲基-1H-吡咯并[3,2-b]吡啶-3-甲醛，其 余所需原料，试剂及制备方法同实施例3，得I-32d。Replace the N-methylindole-3-carbaldehyde in Example 3 with 1-methyl-1H-pyrrolo[3,2-b]pyridine-3-carbaldehyde, and the other required raw materials, reagents and preparation methods are the same In Example 3, I-32d was obtained.
1H NMR(400MHz,CDCl 3)δ8.83(s,2H),8.47(dd,J＝4.6,1.1Hz,1H),7.76(s,1H),7.65(dd,J＝8.3,1.1Hz,1H),7.19(dd,J＝8.3,4.6Hz,1H),4.47(s,2H),4.33(q,J＝7.1Hz,2H),4.01(dd,J＝23.1,9.5Hz,4H),3.83(s,5H),3.66(t,J＝6.9Hz,2H),2.45(t,J＝6.5Hz,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR(400MHz,CDCl 3 )δ8.83(s,2H), 8.47(dd,J=4.6,1.1Hz,1H),7.76(s,1H),7.65(dd,J=8.3,1.1Hz, 1H), 7.19(dd,J=8.3,4.6Hz,1H), 4.47(s,2H), 4.33(q,J=7.1Hz,2H), 4.01(dd,J=23.1,9.5Hz,4H), 3.83 (s, 5H), 3.66 (t, J = 6.9 Hz, 2H), 2.45 (t, J = 6.5 Hz, 2H), 1.36 (t, J = 7.1 Hz, 3H).
实施例38Example 38
2-(2-((1-甲基-1H-吲唑-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-33d)的制备2-(2-((1-methyl-1H-indazol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (Compound I-33d) Preparation
Figure PCTCN2019077356-appb-000047
Figure PCTCN2019077356-appb-000047
将实施例3中的N-甲基吲哚-3-甲醛替换成1-甲基-1H-吲唑-3-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-33d。Replace the N-methylindole-3-carbaldehyde in Example 3 with 1-methyl-1H-indazole-3-carbaldehyde, and the other required raw materials, reagents and preparation methods are the same as those in Example 3 to obtain I-33d .
1H NMR(400MHz,CDCl 3)δ8.83(s,2H),7.84(d,J＝8.2Hz,1H),7.44–7.36(m,2H),7.21–7.15(m,1H),4.33(q,J＝7.1Hz,2H),4.16(s,2H),4.06(s,3H),3.68(s,2H),3.62(dd,J＝13.5,6.6Hz,4H),3.56(d,J＝8.2Hz,2H),2.27(t,J＝6.9Hz,2H),2.06(s,1H),1.36(t,J＝7.1Hz,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.83 (s, 2H), 7.84 (d, J = 8.2 Hz, 1H), 7.44-7.36 (m, 2H), 7.21-7.15 (m, 1H), 4.33 ( q,J=7.1Hz,2H),4.16(s,2H),4.06(s,3H),3.68(s,2H),3.62(dd,J=13.5,6.6Hz,4H),3.56(d,J ＝8.2Hz,2H), 2.27(t,J＝6.9Hz,2H),2.06(s,1H),1.36(t,J＝7.1Hz,3H).
实施例39Example 39
2-(2-((1-甲基-1H-吡咯并[3,2-c]吡啶-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-34d)的制备2-(2-((1-methyl-1H-pyrrolo[3,2-c]pyridin-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl) Preparation of pyrimidine-5-carboxylic acid ethyl ester (Compound I-34d)
Figure PCTCN2019077356-appb-000048
Figure PCTCN2019077356-appb-000048
将实施例3中的N-甲基吲哚-3-甲醛替换成1-甲基-1H-吡咯并[3,2-c]吡啶-3-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-34d。Replace the N-methylindole-3-carbaldehyde in Example 3 with 1-methyl-1H-pyrrolo[3,2-c]pyridine-3-carbaldehyde, and the other required raw materials, reagents and preparation methods are the same In Example 3, I-34d was obtained.
1H NMR(400MHz,DMSO-d 6)δ11.53(s,1H),11.24(s,1H),8.73(s,2H),8.06(d,J＝8.1Hz,1H),7.73(d,J＝8.5Hz,1H),7.55–7.46(m,1H),7.27(t,J＝7.5Hz,1H),4.85(d,J＝5.3Hz,2H),4.37–4.24(m,2H),4.19–4.06(m,5H),3.86(d,J＝47.4Hz,2H),3.61(s,1H),3.59–3.54(m,1H),2.32(dt,J＝37.4,6.9Hz,2H). 1 H NMR (400MHz, DMSO-d 6 ) δ 11.53 (s, 1H), 11.24 (s, 1H), 8.73 (s, 2H), 8.06 (d, J = 8.1 Hz, 1H), 7.73 (d, J=8.5Hz,1H), 7.55–7.46(m,1H), 7.27(t,J=7.5Hz,1H), 4.85(d,J=5.3Hz,2H), 4.37–4.24(m,2H), 4.19–4.06(m,5H), 3.86(d,J=47.4Hz,2H),3.61(s,1H),3.59–3.54(m,1H),2.32(dt,J=37.4,6.9Hz,2H) .
实施例40Example 40
2-(2-(蒽-9-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-35d)的制备Preparation of 2-(2-(anthracene-9-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxylic acid ethyl ester (compound I-35d)
Figure PCTCN2019077356-appb-000049
Figure PCTCN2019077356-appb-000049
将实施例3中的N-甲基吲哚-3-甲醛替换成9-蒽甲醛，其余所需原料，试剂及制备方法同实施例3，得I-35d。The N-methylindole-3-carbaldehyde in Example 3 was replaced with 9-anthracene aldehyde, and the remaining raw materials, reagents and preparation methods were the same as those in Example 3 to obtain I-35d.
1H NMR(400MHz,CDCl 3)δ8.79(d,J＝5.0Hz,2H),8.48(s,1H),8.42(d,J＝8.8Hz,2H),8.03(d,J＝8.3Hz,2H),7.56(dd,J＝15.1,6.8Hz,2H),7.52–7.47(m,2H),4.78(s,2H),4.31(q,J＝7.1Hz,2H),3.59(dd,J＝18.0,11.1Hz,4H),3.42(s,4H),2.25(d,J＝8.8Hz,2H),1.34(t,J＝7.1Hz,3H). 1 H NMR(400MHz,CDCl 3 )δ8.79(d,J=5.0Hz,2H), 8.48(s,1H), 8.42(d,J=8.8Hz,2H), 8.03(d,J=8.3Hz , 2H), 7.56 (dd, J = 15.1, 6.8 Hz, 2H), 7.52–7.47 (m, 2H), 4.78 (s, 2H), 4.31 (q, J = 7.1 Hz, 2H), 3.59 (dd, J = 18.0, 11.1 Hz, 4H), 3.42 (s, 4H), 2.25 (d, J = 8.8 Hz, 2H), 1.34 (t, J = 7.1 Hz, 3H).
实施例41Example 41
2-(2-((6-甲氧基-1-甲基-1H-吲哚-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-36d)的制备2-(2-((6-Methoxy-1-methyl-1H-indol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine- Preparation of ethyl 5-carboxylate (Compound I-36d)
Figure PCTCN2019077356-appb-000050
Figure PCTCN2019077356-appb-000050
将实施例3中的N-甲基吲哚-3-甲醛替换成6-甲氧基-1-甲基-1H-吲哚-3-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-36d。Replace the N-methylindole-3-carbaldehyde in Example 3 with 6-methoxy-1-methyl-1H-indole-3-carbaldehyde, and the other required raw materials, reagents and preparation methods are the same as those in the example 3. Get I-36d.
1H NMR(400MHz,CDCl 3)δ8.83(s,2H),7.51(d,J＝8.7Hz,1H),7.05(s,1H),6.83(dd,J＝8.7,2.2Hz,1H),6.76(d,J＝2.1Hz,1H),4.33(q,J＝7.1Hz,2H),4.00(s,2H),3.89(d,J＝5.4Hz,3H),3.73(s,3H),3.72(s,2H),3.64(t,J＝7.0Hz,2H),3.62–3.45(m,4H),2.30(t,J＝6.8Hz,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR(400MHz,CDCl 3 )δ8.83(s,2H),7.51(d,J=8.7Hz,1H),7.05(s,1H),6.83(dd,J=8.7,2.2Hz,1H) ,6.76(d,J=2.1Hz,1H),4.33(q,J=7.1Hz,2H), 4.00(s,2H), 3.89(d,J=5.4Hz,3H), 3.73(s,3H) ,3.72(s,2H),3.64(t,J=7.0Hz,2H),3.62–3.45(m,4H),2.30(t,J=6.8Hz,2H),1.36(t,J=7.1Hz, 3H).
实施例42Example 42
2-(2-((1,6-二甲基-1H-吲哚-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-37d)的制备2-(2-((1,6-Dimethyl-1H-indol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-carboxy Preparation of Ethyl Acetate (Compound I-37d)
Figure PCTCN2019077356-appb-000051
Figure PCTCN2019077356-appb-000051
将实施例3中的N-甲基吲哚-3-甲醛替换成1,6-二甲基-1H-吲哚-3-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-37d。Replace the N-methylindole-3-carbaldehyde in Example 3 with 1,6-dimethyl-1H-indole-3-carbaldehyde, and the remaining raw materials, reagents and preparation methods are the same as those in Example 3. I-37d.
1H NMR(400MHz,CDCl 3)δ8.83(s,2H),7.48(d,J＝8.1Hz,1H),7.31(s,1H),7.17(s,1H),7.05(d,J＝7.4Hz,1H),4.33(dd,J＝13.2,6.1Hz,4H),4.04(d,J＝10.6Hz,2H),3.92(d,J＝10.6Hz,2H),3.80(s,3H),3.73(d,J＝4.2Hz,2H),3.65(t,J＝7.0Hz,2H),2.50(s,3H),2.38(t,J＝6.9Hz,2H),1.40–1.33(m,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.83 (s, 2H), 7.48 (d, J = 8.1 Hz, 1H), 7.31 (s, 1H), 7.17 (s, 1H), 7.05 (d, J = 7.4Hz, 1H), 4.33 (dd, J = 13.2, 6.1 Hz, 4H), 4.04 (d, J = 10.6 Hz, 2H), 3.92 (d, J = 10.6 Hz, 2H), 3.80 (s, 3H) ,3.73(d,J=4.2Hz,2H), 3.65(t,J=7.0Hz,2H), 2.50(s,3H), 2.38(t,J=6.9Hz,2H),1.40–1.33(m, 3H).
实施例43Example 43
2-(2-((6-氰基-1-甲基-1H-吲哚-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-38d)的制备2-(2-((6-cyano-1-methyl-1H-indol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5 -Preparation of ethyl carboxylate (compound I-38d)
Figure PCTCN2019077356-appb-000052
Figure PCTCN2019077356-appb-000052
将实施例3中的N-甲基吲哚-3-甲醛替换成6-氰基-1-甲基-1H-吲哚-3-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-38d。Replace the N-methylindole-3-carbaldehyde in Example 3 with 6-cyano-1-methyl-1H-indole-3-carbaldehyde, and the other required raw materials, reagents and preparation methods are the same as in Example 3. , Get I-38d.
1H NMR(400MHz,CDCl 3)δ8.84(s,2H),7.75(d,J＝8.2Hz,1H),7.64(s,1H),7.36(d,J＝8.2Hz,1H),4.34(q,J＝7.1Hz,2H),3.80(d,J＝11.4Hz,5H),3.75(s,2H),3.65(t,J＝6.9Hz,2H),3.30(s,4H),2.26(s,2H),1.61(s,3H),1.36(t,J＝7.1Hz,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.84 (s, 2H), 7.75 (d, J = 8.2 Hz, 1H), 7.64 (s, 1H), 7.36 (d, J = 8.2 Hz, 1H), 4.34 (q,J=7.1Hz,2H), 3.80(d,J=11.4Hz,5H), 3.75(s,2H), 3.65(t,J=6.9Hz,2H), 3.30(s,4H), 2.26 (s,2H),1.61(s,3H),1.36(t,J=7.1Hz,3H).
实施例44Example 44
2-(2-((6-溴-1-甲基-1H-吲哚-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-羧酸乙酯(化合物I-39d)的制备2-(2-((6-Bromo-1-methyl-1H-indol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5- Preparation of ethyl carboxylate (compound I-39d)
Figure PCTCN2019077356-appb-000053
Figure PCTCN2019077356-appb-000053
将实施例3中的N-甲基吲哚-3-甲醛替换成6-溴-1-甲基-1H-吲哚-3-甲醛，其余所需原料，试剂及制备方法同实施例3，得I-39d。Replace the N-methylindole-3-carbaldehyde in Example 3 with 6-bromo-1-methyl-1H-indole-3-carbaldehyde, and the remaining raw materials, reagents and preparation methods are the same as in Example 3. Get I-39d.
1H NMR(400MHz,CDCl 3)δ8.83(s,2H),7.53(d,J＝8.4Hz,1H),7.23(d,J＝8.5Hz,1H),7.07(s,1H),4.33(dd,J＝14.1,7.1Hz,2H),3.89(s,2H),3.74(s,5H),3.64(t,J＝6.7Hz,2H),3.43(s,2H),3.35(d,J＝7.7Hz,2H),2.26(t,J＝6.8Hz,2H),1.36(t,J＝7.1Hz,3H). 1 H NMR (400MHz, CDCl 3 ) δ 8.83 (s, 2H), 7.53 (d, J = 8.4 Hz, 1H), 7.23 (d, J = 8.5 Hz, 1H), 7.07 (s, 1H), 4.33 (dd, J = 14.1, 7.1 Hz, 2H), 3.89 (s, 2H), 3.74 (s, 5H), 3.64 (t, J = 6.7 Hz, 2H), 3.43 (s, 2H), 3.35 (d, J = 7.7Hz, 2H), 2.26 (t, J = 6.8Hz, 2H), 1.36 (t, J = 7.1Hz, 3H).
实施例45Example 45
2-(2-(呋喃-2-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-2)的制备2-(2-(furan-2-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (Compound I-2) preparation
Figure PCTCN2019077356-appb-000054
Figure PCTCN2019077356-appb-000054
将实施例4中的I-1d替换成I-2d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-2。Replace I-1d in Example 4 with I-2d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-2 is obtained through a three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.77(s,1H),11.21(s,1H),8.73(s,2H),7.85–7.79(m,1H),6.74(dd,J＝5.9,3.3Hz,1H),6.58(dd,J＝4.9,2.9Hz,1H),4.57–4.48(m,2H),4.13(dd,J＝13.6,6.4Hz,2H),4.03(d,J＝11.2Hz,2H),3.91(s,1H),3.76(s,1H),3.60(dd,J＝12.9,6.1Hz,2H),2.37(t,J＝6.9Hz,1H),2.21(t,J＝7.1Hz,1H)；HRMS(ESI)m/z clcd for C 16H 20N 5O 3(M+H)+330.1566,found 330.1565. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.77 (s, 1H), 11.21 (s, 1H), 8.73 (s, 2H), 7.85-7.79 (m, 1H), 6.74 (dd, J = 5.9 ,3.3Hz,1H),6.58(dd,J=4.9,2.9Hz,1H),4.57–4.48(m,2H),4.13(dd,J=13.6,6.4Hz,2H),4.03(d,J= 11.2Hz, 2H), 3.91 (s, 1H), 3.76 (s, 1H), 3.60 (dd, J = 12.9, 6.1 Hz, 2H), 2.37 (t, J = 6.9 Hz, 1H), 2.21 (t, J = 7.1Hz, 1H); HRMS (ESI) m/z clcd for C 16 H 20 N 5 O 3 (M+H)+330.1566, found 330.1565.
实施例46Example 46
2-(2-(呋喃-3-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-3)的制备2-(2-(furan-3-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (compound I-3) preparation
Figure PCTCN2019077356-appb-000055
Figure PCTCN2019077356-appb-000055
将实施例4中的I-1d替换成I-3d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-3。Replace I-1d in Example 4 with I-3d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-3 is obtained through a three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.59(s,1H),11.18(s,1H),8.62(d,J＝81.9Hz,2H),7.91(d,J＝4.1Hz,1H),7.77(s,1H),6.74(s,1H),4.28(d,J＝5.4Hz,2H),4.13(d,J＝4.1Hz,4H), 3.91(s,1H),3.79(s,1H),3.63–3.55(m,2H),2.31(dt,J＝14.2,6.9Hz,2H)；HRMS(ESI)m/z calcd for C 16H 20N 5O 3(M+H)+330.1566,found 330.1567. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.59 (s, 1H), 11.18 (s, 1H), 8.62 (d, J = 81.9 Hz, 2H), 7.91 (d, J = 4.1 Hz, 1H) ,7.77(s,1H),6.74(s,1H), 4.28(d,J=5.4Hz,2H), 4.13(d,J=4.1Hz,4H), 3.91(s,1H), 3.79(s, 1H),3.63–3.55(m,2H),2.31(dt,J＝14.2,6.9Hz,2H); HRMS(ESI)m/z calcd for C 16 H 20 N 5 O 3 (M+H)+330.1566 ,found 330.1567.
实施例47Example 47
2-(2-(噻吩-2-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-4)的制备2-(2-(Thien-2-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (Compound I-4) preparation
Figure PCTCN2019077356-appb-000056
Figure PCTCN2019077356-appb-000056
将实施例4中的I-1d替换成I-4d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-4。Replace I-1d in Example 4 with I-4d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-4 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.48(s,1H),11.15(s,1H),8.72(s,2H),7.70(d,J＝5.1Hz,1H),7.40(s,1H),7.15(t,J＝4.3Hz,1H),4.67(t,J＝5.2Hz,2H),4.25–4.16(m,2H),3.99(d,J＝10.8Hz,2H),3.91(s,1H),3.81(s,1H),3.60(dd,J＝14.2,7.0Hz,2H),2.31(dt,J＝37.4,6.8Hz,2H)；HRMS(ESI)m/z calcd for C 16H 20N 5O 2S(M+H)+346.1338,found 346.1337. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.48 (s, 1H), 11.15 (s, 1H), 8.72 (s, 2H), 7.70 (d, J = 5.1 Hz, 1H), 7.40 (s, 1H), 7.15 (t, J = 4.3 Hz, 1H), 4.67 (t, J = 5.2 Hz, 2H), 4.25-4.16 (m, 2H), 3.99 (d, J = 10.8 Hz, 2H), 3.91 ( s, 1H), 3.81 (s, 1H), 3.60 (dd, J = 14.2, 7.0 Hz, 2H), 2.31 (dt, J = 37.4, 6.8 Hz, 2H); HRMS (ESI) m/z calcd for C 16 H 20 N 5 O 2 S(M+H)+346.1338,found 346.1337.
实施例48Example 48
2-(2-(噻吩-3-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-5)的制备2-(2-(Thien-3-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (compound I-5) preparation
Figure PCTCN2019077356-appb-000057
Figure PCTCN2019077356-appb-000057
将实施例4中的I-1d替换成I-5d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-5。Replace I-1d in Example 4 with I-5d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-5 is obtained through a three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.78(s,1H),11.26(s,1H),8.73(s,2H),7.81(d,J＝2.7Hz,1H),7.66(dd,J＝4.8,2.7Hz,1H),7.36(d,J＝4.9Hz,1H),4.44(t,J＝5.5Hz,2H),4.08–4.03(m,2H),3.99(dd,J＝14.2,7.8Hz,2H),3.92(s,1H),3.80(s,1H),3.58(dd,J＝15.3,7.1Hz,2H),2.32(dt,J＝46.7,6.8Hz,2H)；HRMS(ESI)m/z calcd for C 16H 20N 5O 2S(M+H)+346.1338,found 346.1339. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.78 (s, 1H), 11.26 (s, 1H), 8.73 (s, 2H), 7.81 (d, J = 2.7 Hz, 1H), 7.66 (dd, J = 4.8, 2.7 Hz, 1H), 7.36 (d, J = 4.9 Hz, 1H), 4.44 (t, J = 5.5 Hz, 2H), 4.08-4.03 (m, 2H), 3.99 (dd, J = 14.2 ,7.8Hz,2H),3.92(s,1H),3.80(s,1H),3.58(dd,J＝15.3,7.1Hz,2H),2.32(dt,J＝46.7,6.8Hz,2H); HRMS (ESI)m/z calcd for C 16 H 20 N 5 O 2 S(M+H)+346.1338,found 346.1339.
实施例49Example 49
2-(2-(吡啶-2-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-6)的制备2-(2-(Pyridin-2-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (compound I-6) preparation
Figure PCTCN2019077356-appb-000058
Figure PCTCN2019077356-appb-000058
将实施例4中的I-1d替换成I-6d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-6。Replace I-1d in Example 4 with I-6d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-6 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.60(s,2H),8.74(s,2H),8.67(d,J＝4.4Hz,1H),7.99(t,J＝7.7Hz,1H),7.63(d,J＝7.8Hz,1H),7.56–7.49(m,1H),4.69(s,2H),4.25(dd,J＝26.0,9.7 Hz,4H),3.88(s,2H),3.60(t,J＝6.3Hz,2H),2.37(s,2H)；HRMS(ESI)m/z calcd forC 17H 21N 6O 2(M+H)+341.1726,found 341.1727. 1 H NMR(400MHz,DMSO-d 6 )δ11.60(s,2H),8.74(s,2H),8.67(d,J=4.4Hz,1H),7.99(t,J=7.7Hz,1H) ,7.63(d,J=7.8Hz,1H),7.56–7.49(m,1H), 4.69(s,2H), 4.25(dd,J=26.0,9.7 Hz,4H), 3.88(s,2H), 3.60(t,J=6.3Hz,2H),2.37(s,2H); HRMS(ESI)m/z calcd for C 17 H 21 N 6 O 2 (M+H)+341.1726,found 341.1727.
实施例50Example 50
2-(2-(吡啶-3-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-7)的制备2-(2-(Pyridin-3-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (compound I-7) preparation
Figure PCTCN2019077356-appb-000059
Figure PCTCN2019077356-appb-000059
将实施例4中的I-1d替换成I-7d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-7。Replace I-1d in Example 4 with I-7d, and the remaining raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-7 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ12.58(s,1H),11.28(s,1H),9.18(s,1H),8.99(d,J＝5.2Hz,1H),8.81(d,J＝7.7Hz,1H),8.74(s,1H),8.53(s,1H),8.16–8.07(m,1H),4.70(d,J＝17.6Hz,2H),4.34(s,2H),4.12–3.99(m,2H),3.89(d,J＝36.4Hz,2H),3.60(s,2H),2.36(d,J＝28.4Hz,2H)；HRMS(ESI)m/z calcd for C 17H 21N 6O 2(M+H)+341.1726,found 341.1725. 1 H NMR (400MHz, DMSO-d 6 ) δ 12.58 (s, 1H), 11.28 (s, 1H), 9.18 (s, 1H), 8.99 (d, J = 5.2 Hz, 1H), 8.81 (d, J=7.7Hz, 1H), 8.74 (s, 1H), 8.53 (s, 1H), 8.16-8.07 (m, 1H), 4.70 (d, J = 17.6 Hz, 2H), 4.34 (s, 2H), 4.12–3.99(m,2H), 3.89(d,J=36.4Hz,2H), 3.60(s,2H), 2.36(d,J=28.4Hz,2H); HRMS(ESI)m/z calcd for C 17 H 21 N 6 O 2 (M+H)+341.1726,found 341.1725.
实施例51Example 51
2-(2-(吡啶-4-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-8)的制备2-(2-(Pyridin-4-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (compound I-8) preparation
Figure PCTCN2019077356-appb-000060
Figure PCTCN2019077356-appb-000060
将实施例4中的I-1d替换成I-8d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-8。Replace I-1d in Example 4 with I-8d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-8 is obtained through a three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ12.68(s,1H),10.33(s,1H),8.99(d,J＝6.3Hz,2H),8.52(s,2H),8.18(d,J＝6.0Hz,2H),4.80(s,2H),4.28(s,2H),4.17(d,J＝17.6Hz,2H),3.97(s,1H),3.83(s,1H),3.62–3.56(m,2H),2.43(s,1H),2.31(s,1H)；HRMS(ESI)m/z calcd for C 17H 21N 6O 2(M+H)+341.1726,found 341.1727. 1 H NMR (400MHz, DMSO-d 6 ) δ 12.68 (s, 1H), 10.33 (s, 1H), 8.99 (d, J = 6.3 Hz, 2H), 8.52 (s, 2H), 8.18 (d, J = 6.0Hz, 2H), 4.80 (s, 2H), 4.28 (s, 2H), 4.17 (d, J = 17.6 Hz, 2H), 3.97 (s, 1H), 3.83 (s, 1H), 3.62- 3.56(m,2H),2.43(s,1H),2.31(s,1H); HRMS(ESI)m/z calcd for C 17 H 21 N 6 O 2 (M+H)+341.1726,found 341.1727.
实施例52Example 52
2-(2-(萘-1-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-9)的制备2-(2-(Naphthalene-1-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (compound I-9) preparation
Figure PCTCN2019077356-appb-000061
Figure PCTCN2019077356-appb-000061
将实施例4中的I-1d替换成I-9d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-9。Replace I-1d in Example 4 with I-9d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-9 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.25(d,J＝36.0Hz,2H),8.73(d,J＝10.8Hz,2H),8.36(t,J＝9.0Hz,1H),8.06(d,J＝8.8Hz,2H),7.85(t,J＝6.8Hz,1H),7.65(ddd,J＝17.8,14.5,7.1Hz,3H),5.01(t,J＝5.4Hz,2H),4.21–4.14(m,2H),4.14–4.08(m,2H),3.95–3.85(m,2H),3.59 (dd,J＝15.2,7.3Hz,2H),2.42–2.31(m,2H)；HRMS(ESI)m/z calcd for C 22H 24N 5O 2(M+H)+390.1930,found 390.1931. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.25 (d, J = 36.0 Hz, 2H), 8.73 (d, J = 10.8 Hz, 2H), 8.36 (t, J = 9.0 Hz, 1H), 8.06 (d,J=8.8Hz,2H),7.85(t,J=6.8Hz,1H),7.65(ddd,J=17.8,14.5,7.1Hz,3H),5.01(t,J=5.4Hz,2H) ,4.21–4.14(m,2H),4.14–4.08(m,2H),3.95–3.85(m,2H),3.59 (dd,J=15.2,7.3Hz,2H),2.42–2.31(m,2H) ；HRMS(ESI)m/z calcd for C 22 H 24 N 5 O 2 (M+H)+390.1930,found 390.1931.
实施例53Example 53
2-(2-(萘-2-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-10)的制备2-(2-(Naphth-2-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (compound I-10) preparation
Figure PCTCN2019077356-appb-000062
Figure PCTCN2019077356-appb-000062
将实施例4中的I-1d替换成I-10d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-10。Replace I-1d in Example 4 with I-10d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-10 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.41–11.27(m,1H),11.14(s,1H),8.72(d,J＝9.3Hz,2H),8.13(s,1H),8.06–7.94(m,3H),7.71(d,J＝8.6Hz,1H),7.63(dd,J＝6.3,3.2Hz,2H),4.62(t,J＝5.9Hz,2H),4.30(d,J＝8.1Hz,2H),4.02(d,J＝6.5Hz,2H),3.91(s,1H),3.85(s,1H),3.59(dd,J＝15.6,7.3Hz,2H),2.33(dd,J＝15.9,8.3Hz,2H)；HRMS(ESI)m/z calcd for C 22H 24N 5O 2(M+H)+390.1930,found 390.1929. 1 H NMR(400MHz,DMSO-d 6 )δ11.41–11.27(m,1H),11.14(s,1H),8.72(d,J=9.3Hz,2H),8.13(s,1H),8.06– 7.94(m,3H),7.71(d,J=8.6Hz,1H), 7.63(dd,J=6.3,3.2Hz,2H), 4.62(t,J=5.9Hz,2H), 4.30(d,J ＝8.1Hz,2H),4.02(d,J＝6.5Hz,2H),3.91(s,1H),3.85(s,1H),3.59(dd,J＝15.6,7.3Hz,2H),2.33(dd ,J＝15.9,8.3Hz,2H); HRMS(ESI)m/z calcd for C 22 H 24 N 5 O 2 (M+H)+390.1930,found 390.1929.
实施例54Example 54
2-(2-([1,1'-联苯基]-4-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-11)的制备2-(2-([1,1'-biphenyl]-4-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate Preparation of acid salt (compound I-11)
Figure PCTCN2019077356-appb-000063
Figure PCTCN2019077356-appb-000063
将实施例4中的I-1d替换成I-11d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-11。Replace I-1d in Example 4 with I-11d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-11 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.84(s,1H),11.18(s,1H),8.73(s,2H),7.80–7.75(m,2H),7.72(t,J＝6.4Hz,4H),7.52(t,J＝7.6Hz,2H),7.43(t,J＝7.3Hz,1H),4.47(d,J＝6.2Hz,2H),4.22(dd,J＝16.8,10.3Hz,2H),4.08–3.99(m,2H),3.89(d,J＝41.8Hz,2H),3.64–3.54(m,2H),2.35(dt,J＝36.8,6.8Hz,2H)；HRMS(ESI)m/z calcd for C 24H 26N 5O 2(M+H)+416.2087,found 416.2086. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.84 (s, 1H), 11.18 (s, 1H), 8.73 (s, 2H), 7.80-7.75 (m, 2H), 7.72 (t, J = 6.4 Hz, 4H), 7.52 (t, J = 7.6 Hz, 2H), 7.43 (t, J = 7.3 Hz, 1H), 4.47 (d, J = 6.2 Hz, 2H), 4.22 (dd, J = 16.8, 10.3 Hz, 2H), 4.08–3.99 (m, 2H), 3.89 (d, J = 41.8 Hz, 2H), 3.64–3.54 (m, 2H), 2.35 (dt, J = 36.8, 6.8 Hz, 2H); HRMS (ESI)m/z calcd for C 24 H 26 N 5 O 2 (M+H)+416.2087,found 416.2086.
实施例55Example 55
2-(2-苄基-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-12)的制备Preparation of 2-(2-benzyl-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (compound I-12)
Figure PCTCN2019077356-appb-000064
Figure PCTCN2019077356-appb-000064
将实施例4中的I-1d替换成I-12d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-12。Replace I-1d in Example 4 with I-12d, and the remaining raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-12 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.97(s,1H),11.14(s,1H),8.74(s,2H),7.63(d,J＝3.9Hz,2H),7.46(d,J＝4.2Hz,3H),4.44(t,J＝6.1Hz,2H),4.24–4.12(m,2H),4.02–3.78(m,4H), 3.58(dt,J＝13.2,6.8Hz,2H),2.34(dt,J＝43.2,6.7Hz,2H)；HRMS(ESI)m/z calcd forC 18H 22N 5O 2(M+H)+340.1773,found 340.1774. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.97 (s, 1H), 11.14 (s, 1H), 8.74 (s, 2H), 7.63 (d, J = 3.9 Hz, 2H), 7.46 (d, J = 4.2Hz, 3H), 4.44 (t, J = 6.1Hz, 2H), 4.24-4.12 (m, 2H), 4.02-3.78 (m, 4H), 3.58 (dt, J = 13.2, 6.8 Hz, 2H ), 2.34(dt,J=43.2,6.7Hz,2H); HRMS(ESI)m/z calcd for C 18 H 22 N 5 O 2 (M+H)+340.1773,found 340.1774.
实施例56Example 56
2-(2-(环戊基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-13)的制备Preparation of 2-(2-(cyclopentylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (compound I-13)
Figure PCTCN2019077356-appb-000065
Figure PCTCN2019077356-appb-000065
将实施例4中的I-1d替换成I-13d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-13。Replace I-1d in Example 4 with I-13d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-13 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.15(d,J＝31.5Hz,2H),8.73(s,2H),4.18(dd,J＝15.5,8.9Hz,2H),4.14(d,J＝6.3Hz,2H),3.95(s,1H),3.78(s,1H),3.58(dt,J＝14.0,6.8Hz,2H),3.24–3.13(m,2H),2.34(dt,J＝66.3,6.8Hz,2H),2.08(d,J＝7.8Hz,1H),1.78(d,J＝6.8Hz,2H),1.69–1.48(m,4H),1.31–1.17(m,3H)；HRMS(ESI)m/z calcd for C 17H 26N 5O 2(M+H)+332.2087,found 332.2088. 1 H NMR (400MHz, DMSO-d 6 ) δ11.15 (d, J = 31.5Hz, 2H), 8.73 (s, 2H), 4.18 (dd, J = 15.5, 8.9 Hz, 2H), 4.14 (d, J = 6.3Hz, 2H), 3.95 (s, 1H), 3.78 (s, 1H), 3.58 (dt, J = 14.0, 6.8 Hz, 2H), 3.24-3.13 (m, 2H), 2.34 (dt, J ＝66.3,6.8Hz,2H),2.08(d,J＝7.8Hz,1H),1.78(d,J＝6.8Hz,2H),1.69–1.48(m,4H),1.31–1.17(m,3H) ；HRMS(ESI)m/z calcd for C 17 H 26 N 5 O 2 (M+H)+332.2087,found 332.2088.
实施例57Example 57
2-(2-(环己基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-14)的制备Preparation of 2-(2-(cyclohexylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (Compound I-14)
Figure PCTCN2019077356-appb-000066
Figure PCTCN2019077356-appb-000066
将实施例4中的I-1d替换成I-14d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-14。Replace I-1d in Example 4 with I-14d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-14 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.06(d,J＝29.5Hz,2H),8.73(s,2H),4.21–4.08(m,4H),3.95(s,1H),3.78(s,1H),3.58(dt,J＝20.4,6.7Hz,2H),3.10(d,J＝5.2Hz,2H),2.34(dt,J＝63.0,6.6Hz,2H),1.98(d,J＝31.6Hz,1H),1.84–1.55(m,6H),1.21(t,J＝8.5Hz,2H),1.03–0.89(m,2H)；HRMS(ESI)m/z calcd for C 18H 28N 5O 2(M+H)+346.2243,found 346.2244. 1 H NMR(400MHz,DMSO-d 6 )δ11.06(d,J=29.5Hz,2H), 8.73(s,2H), 4.21-4.08(m,4H), 3.95(s,1H), 3.78( s, 1H), 3.58 (dt, J = 20.4, 6.7 Hz, 2H), 3.10 (d, J = 5.2 Hz, 2H), 2.34 (dt, J = 63.0, 6.6 Hz, 2H), 1.98 (d, J ＝31.6Hz,1H),1.84–1.55(m,6H),1.21(t,J＝8.5Hz,2H),1.03–0.89(m,2H); HRMS(ESI)m/z calcd for C 18 H 28 N 5 O 2 (M+H)+346.2243,found 346.2244.
实施例58Example 58
2-(2-(苯并[b]噻吩-2-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-15)的制备2-(2-(Benzo[b]thiophen-2-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (compound I-15) Preparation
Figure PCTCN2019077356-appb-000067
Figure PCTCN2019077356-appb-000067
将实施例4中的I-1d替换成I-15d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-15。Replace I-1d in Example 4 with I-15d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-15 is obtained through a three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.88(s,1H),11.18(s,1H),8.72(s,2H),8.11–8.03(m,1H),7.97–7.89(m,1H),7.72(s,1H),7.53–7.40(m,2H),4.81(t,J＝5.5Hz,2H),4.34–4.20(m, 2H),4.17–4.04(m,2H),3.89(d,J＝39.5Hz,2H),3.60(dt,J＝17.6,6.9Hz,2H),2.34(dt,J＝34.7,7.0Hz,2H)；HRMS(ESI)m/z calcd for C 20H 22N 5O 2S(M+H)+396.1494,found 396.1495. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.88 (s, 1H), 11.18 (s, 1H), 8.72 (s, 2H), 8.11-8.03 (m, 1H), 7.97-7.89 (m, 1H) ),7.72(s,1H),7.53-7.40(m,2H),4.81(t,J=5.5Hz,2H),4.34-4.20(m, 2H),4.17-4.04(m,2H),3.89( d,J=39.5Hz,2H),3.60(dt,J=17.6,6.9Hz,2H),2.34(dt,J=34.7,7.0Hz,2H); HRMS(ESI)m/z calcd for C 20 H 22 N 5 O 2 S(M+H)+396.1494,found 396.1495.
实施例59Example 59
2-(2-(苯并[b]噻吩-3-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-16)的制备2-(2-(Benzo[b]thiophen-3-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (compound I-16) Preparation
Figure PCTCN2019077356-appb-000068
Figure PCTCN2019077356-appb-000068
将实施例4中的I-1d替换成I-16d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-16。Replace I-1d in Example 4 with I-16d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-16 is obtained through a three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.84–10.83(m,2H),8.77(dd,J＝27.8,4.4Hz,2H),8.17(d,J＝9.7Hz,2H),8.09(d,J＝7.8Hz,1H),7.50(dt,J＝14.9,7.2Hz,2H),4.78(t,J＝5.5Hz,2H),4.17(d,J＝6.3Hz,2H),4.12–4.07(m,2H),3.94(s,1H),3.85(s,1H),3.65–3.54(m,2H),2.35(dt,J＝31.8,6.7Hz,2H)；HRMS(ESI)m/z calcd for C 20H 22N 5O 2S(M+H)+396.1494,found 396.1493. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.84-10.83 (m, 2H), 8.77 (dd, J = 27.8, 4.4 Hz, 2H), 8.17 (d, J = 9.7 Hz, 2H), 8.09 ( d,J=7.8Hz,1H), 7.50(dt,J=14.9,7.2Hz,2H), 4.78(t,J=5.5Hz,2H), 4.17(d,J=6.3Hz,2H), 4.12– 4.07 (m, 2H), 3.94 (s, 1H), 3.85 (s, 1H), 3.65-3.54 (m, 2H), 2.35 (dt, J = 31.8, 6.7 Hz, 2H); HRMS (ESI) m/ z calcd for C 20 H 22 N 5 O 2 S(M+H)+396.1494,found 396.1493.
实施例60Example 60
2-(2-(苯并呋喃-2-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-17)的制备2-(2-(benzofuran-2-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (Compound I-17 ) Preparation
Figure PCTCN2019077356-appb-000069
Figure PCTCN2019077356-appb-000069
将实施例4中的I-1d替换成I-17d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-17。Replace I-1d in Example 4 with I-17d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-17 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.80(s,1H),11.18(s,1H),8.72(s,2H),7.74(d,J＝7.6Hz,1H),7.64(d,J＝8.2Hz,1H),7.42(t,J＝7.8Hz,1H),7.34(t,J＝7.5Hz,1H),7.20(d,J＝2.8Hz,1H),4.75(t,J＝5.0Hz,2H),4.39–4.27(m,2H),4.25–4.11(m,2H),3.87(d,J＝44.2Hz,2H),3.67–3.54(m,2H),2.33(dt,J＝42.0,6.9Hz,2H)；HRMS(ESI)m/z calcd for C 20H 22N 5O 3(M+H)+380.1723,found 380.1724. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.80 (s, 1H), 11.18 (s, 1H), 8.72 (s, 2H), 7.74 (d, J = 7.6 Hz, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.42 (t, J = 7.8 Hz, 1H), 7.34 (t, J = 7.5 Hz, 1H), 7.20 (d, J = 2.8 Hz, 1H), 4.75 (t, J = 5.0Hz, 2H), 4.39–4.27(m, 2H), 4.25–4.11(m, 2H), 3.87(d, J=44.2Hz, 2H), 3.67–3.54(m, 2H), 2.33(dt, J ＝42.0,6.9Hz,2H); HRMS(ESI)m/z calcd for C 20 H 22 N 5 O 3 (M+H)+380.1723,found 380.1724.
实施例61Example 61
2-(2-((1H-吲哚-4-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-18)的制备2-(2-((1H-Indol-4-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (compound I-18) Preparation
Figure PCTCN2019077356-appb-000070
Figure PCTCN2019077356-appb-000070
将实施例4中的I-1d替换成I-18d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-18。Replace I-1d in Example 4 with I-18d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-18 is obtained through a three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.35(d,J＝64.6Hz,2H),10.29(s,1H),8.71(s,1H),8.51(d,J＝5.0Hz,1H),7.58–7.46(m,2H),7.33–7.12(m,3H),4.71(s,2H),4.33–4.20(m,2H),4.07 (d,J＝24.3Hz,2H),3.86(d,J＝29.0Hz,4H),2.39–2.25(m,2H)；HRMS(ESI)m/z calcd for C 20H 23N 6O 2(M+H)+379.1882,found 379.1883. 1 H NMR(400MHz,DMSO-d 6 )δ11.35(d,J=64.6Hz,2H), 10.29(s,1H), 8.71(s,1H), 8.51(d,J=5.0Hz,1H) ,7.58–7.46(m,2H),7.33–7.12(m,3H),4.71(s,2H),4.33–4.20(m,2H),4.07 (d,J=24.3Hz,2H),3.86(d ,J=29.0Hz,4H),2.39–2.25(m,2H); HRMS(ESI)m/z calcd for C 20 H 23 N 6 O 2 (M+H)+379.1882,found 379.1883.
实施例62Example 62
2-(2-((1-甲基-1H-吲哚-2-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-19)的制备2-(2-((1-methyl-1H-indol-2-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamic acid Preparation of Hydrochloride (Compound I-19)
Figure PCTCN2019077356-appb-000071
Figure PCTCN2019077356-appb-000071
将实施例4中的I-1d替换成I-19d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-19。Replace I-1d in Example 4 with I-19d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-19 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.09(d,J＝74.3Hz,2H),8.72(s,2H),7.63–7.56(m,1H),7.53(d,J＝8.4Hz,1H),7.28–7.21(m,1H),7.13–7.07(m,1H),6.81(s,1H),4.74(d,J＝5.5Hz,2H),4.25(d,J＝6.2Hz,2H),4.15–4.10(m,2H),3.94(s,3H),3.87(d,J＝6.9Hz,2H),3.65–3.56(m,2H),2.47–2.29(m,2H)；HRMS(ESI)m/z calcd for C 21H 25N 6O 2(M+H)+393.2039,found 393.2040. 1 H NMR(400MHz,DMSO-d 6 )δ11.09(d,J=74.3Hz,2H), 8.72(s,2H), 7.63–7.56(m,1H), 7.53(d,J=8.4Hz, 1H), 7.28–7.21(m,1H), 7.13–7.07(m,1H), 6.81(s,1H), 4.74(d,J=5.5Hz,2H), 4.25(d,J=6.2Hz,2H ), 4.15–4.10 (m, 2H), 3.94 (s, 3H), 3.87 (d, J = 6.9 Hz, 2H), 3.65–3.56 (m, 2H), 2.47–2.29 (m, 2H); HRMS( ESI)m/z calcd for C 21 H 25 N 6 O 2 (M+H)+393.2039,found 393.2040.
实施例63Example 63
2-(2-(喹啉-3-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-20)的制备2-(2-(quinolin-3-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (Compound I-20) Preparation
Figure PCTCN2019077356-appb-000072
Figure PCTCN2019077356-appb-000072
将实施例4中的I-1d替换成I-20d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-20。Replace I-1d in Example 4 with I-20d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-20 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ12.01(s,1H),11.15(s,1H),9.26(d,J＝1.7Hz,1H),8.89(s,1H),8.73(d,J＝11.1Hz,2H),8.22(d,J＝8.5Hz,1H),8.16(d,J＝8.4Hz,1H),8.00(t,J＝7.1Hz,1H),7.83(t,J＝7.5Hz,1H),4.74(d,J＝5.6Hz,2H),4.37(d,J＝6.3Hz,2H),4.08(dd,J＝18.5,12.2Hz,2H),3.89(d,J＝24.9Hz,2H),3.59(dd,J＝13.3,6.7Hz,2H),2.40–2.30(m,2H)；HRMS(ESI)m/z calcd for C 21H 23N 6O 2(M+H)+391.1882,found 391.1883. 1 H NMR (400MHz, DMSO-d 6 ) δ 12.01 (s, 1H), 11.15 (s, 1H), 9.26 (d, J = 1.7 Hz, 1H), 8.89 (s, 1H), 8.73 (d, J = 11.1 Hz, 2H), 8.22 (d, J = 8.5 Hz, 1H), 8.16 (d, J = 8.4 Hz, 1H), 8.00 (t, J = 7.1 Hz, 1H), 7.83 (t, J = 7.5Hz, 1H), 4.74 (d, J = 5.6 Hz, 2H), 4.37 (d, J = 6.3 Hz, 2H), 4.08 (dd, J = 18.5, 12.2 Hz, 2H), 3.89 (d, J = 24.9Hz,2H),3.59(dd,J＝13.3,6.7Hz,2H),2.40–2.30(m,2H); HRMS(ESI)m/z calcd for C 21 H 23 N 6 O 2 (M+H )+391.1882,found 391.1883.
实施例64Example 64
2-(2-(喹啉-2-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-21)的制备2-(2-(Quinolin-2-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (Compound I-21) Preparation
Figure PCTCN2019077356-appb-000073
Figure PCTCN2019077356-appb-000073
将实施例4中的I-1d替换成I-21d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-21。Replace I-1d in Example 4 with I-21d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-21 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.57(s,1H),11.22(s,1H),8.75(s,2H),8.52(d,J＝8.5Hz,1H),8.09(t,J＝9.3Hz,2H),7.87(t,J＝7.2Hz,1H),7.69(dd,J＝18.1,8.1Hz,2H),4.93(s,2H),4.37(d,J＝5.4Hz,4H),3.91(s,2H),3.62(d,J＝11.1Hz,2H),2.41(s,2H)；HRMS(ESI)m/z calcd for C 21H 23N 6O 2(M+H)+391.1882,found 391.1833. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.57 (s, 1H), 11.22 (s, 1H), 8.75 (s, 2H), 8.52 (d, J = 8.5 Hz, 1H), 8.09 (t, J = 9.3Hz, 2H), 7.87 (t, J = 7.2Hz, 1H), 7.69 (dd, J = 18.1, 8.1Hz, 2H), 4.93 (s, 2H), 4.37 (d, J = 5.4Hz, 4H),3.91(s,2H),3.62(d,J=11.1Hz,2H),2.41(s,2H); HRMS(ESI)m/z calcd for C 21 H 23 N 6 O 2 (M+H )+391.1882,found 391.1833.
实施例65Example 65
2-(2-(喹啉-6-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-22)的制备2-(2-(Quinolin-6-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (Compound I-22) Preparation
Figure PCTCN2019077356-appb-000074
Figure PCTCN2019077356-appb-000074
将实施例4中的I-1d替换成I-22d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-22。Replace I-1d in Example 4 with I-22d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-22 is obtained through a three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ12.38(s,1H),11.24(s,1H),9.33(t,J＝8.6Hz,1H),9.10(d,J＝8.3Hz,1H),8.82–8.70(m,2H),8.57–8.43(m,2H),8.35(d,J＝8.8Hz,1H),8.08(dt,J＝18.6,9.3Hz,1H),4.76(d,J＝6.1Hz,2H),4.31(dd,J＝18.2,12.9Hz,2H),4.14–4.00(m,2H),3.95(s,1H),3.85(s,1H),3.58(dd,J＝15.3,7.0Hz,2H),2.37(dt,J＝33.9,6.8Hz,2H)；HRMS(ESI)m/z calcd for C 21H 23N 6O 2(M+H)+391.1882,found 391.1883. 1 H NMR (400MHz, DMSO-d 6 ) δ 12.38 (s, 1H), 11.24 (s, 1H), 9.33 (t, J = 8.6 Hz, 1H), 9.10 (d, J = 8.3 Hz, 1H) ,8.82–8.70(m,2H),8.57–8.43(m,2H),8.35(d,J＝8.8Hz,1H), 8.08(dt,J＝18.6,9.3Hz,1H),4.76(d,J =6.1Hz,2H),4.31(dd,J=18.2,12.9Hz,2H), 4.14–4.00(m,2H), 3.95(s,1H), 3.85(s,1H), 3.58(dd,J= 15.3,7.0Hz,2H),2.37(dt,J=33.9,6.8Hz,2H); HRMS(ESI)m/z calcd for C 21 H 23 N 6 O 2 (M+H)+391.1882,found 391.1883.
实施例66Example 66
2-(2-(喹啉-8-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-23)的制备2-(2-(Quinolin-8-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (Compound I-23) Preparation
Figure PCTCN2019077356-appb-000075
Figure PCTCN2019077356-appb-000075
将实施例4中的I-1d替换成I-23d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-23。Replace I-1d in Example 4 with I-23d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-23 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.15(s,1H),10.95(s,1H),9.09(d,J＝2.7Hz,1H),8.71(s,2H),8.56(d,J＝8.3Hz,1H),8.16(d,J＝7.8Hz,1H),8.10(d,J＝6.6Hz,1H),7.74(dd,J＝13.3,5.4Hz,2H),5.10(d,J＝5.2Hz,2H),4.37(s,2H),4.18(d,J＝21.1Hz,2H),3.89(d,J＝7.7Hz,2H),3.58(s,2H),2.35(t,J＝6.7Hz,2H)；HRMS(ESI)m/z calcd for C 21H 23N 6O 2(M+H)+391.1882,found 391.1881. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.15 (s, 1H), 10.95 (s, 1H), 9.09 (d, J = 2.7 Hz, 1H), 8.71 (s, 2H), 8.56 (d, J = 8.3 Hz, 1H), 8.16 (d, J = 7.8 Hz, 1H), 8.10 (d, J = 6.6 Hz, 1H), 7.74 (dd, J = 13.3, 5.4 Hz, 2H), 5.10 (d, J = 5.2Hz, 2H), 4.37 (s, 2H), 4.18 (d, J = 21.1 Hz, 2H), 3.89 (d, J = 7.7 Hz, 2H), 3.58 (s, 2H), 2.35 (t, J＝6.7Hz, 2H); HRMS(ESI)m/z calcd for C 21 H 23 N 6 O 2 (M+H)+391.1882,found 391.1881.
实施例67Example 67
2-(2-(异喹啉-8-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-24)的制备2-(2-(Isoquinolin-8-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (Compound I-24 ) Preparation
Figure PCTCN2019077356-appb-000076
Figure PCTCN2019077356-appb-000076
将实施例4中的I-1d替换成I-24d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-24。Replace I-1d in Example 4 with I-24d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-24 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.92(s,1H),11.15(s,1H),10.29(s,1H),8.79(d,J＝6.3Hz,1H),8.76–8.69(m,2H),8.54(d,J＝6.3Hz,1H),8.41(d,J＝8.2Hz,1H),8.30(d,J＝6.8Hz,1H),8.26–8.20(m,1H),5.21(d,J＝5.5Hz,2H),4.44–4.36(m,2H),4.07(dd,J＝14.5,7.8Hz,3H),3.91(d,J＝8.0Hz,2H),3.59(t,J＝6.9Hz,2H),2.41–2.32(m,2H)；HRMS(ESI)m/z calcd for C 21H 23N 6O 2(M+H)+391.1882,found 391.1883. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.92 (s, 1H), 11.15 (s, 1H), 10.29 (s, 1H), 8.79 (d, J = 6.3 Hz, 1H), 8.76-8.69 ( m, 2H), 8.54 (d, J = 6.3 Hz, 1H), 8.41 (d, J = 8.2 Hz, 1H), 8.30 (d, J = 6.8 Hz, 1H), 8.26-8.20 (m, 1H), 5.21(d,J=5.5Hz,2H),4.44-4.36(m,2H),4.07(dd,J=14.5,7.8Hz,3H),3.91(d,J=8.0Hz,2H),3.59(t ,J=6.9Hz,2H),2.41–2.32(m,2H); HRMS(ESI)m/z calcd for C 21 H 23 N 6 O 2 (M+H)+391.1882,found 391.1883.
实施例68Example 68
2-(2-(异喹啉-5-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-25)的制备2-(2-(Isoquinolin-5-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (Compound I-25 ) Preparation
Figure PCTCN2019077356-appb-000077
Figure PCTCN2019077356-appb-000077
将实施例4中的I-1d替换成I-25d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-25。Replace I-1d in Example 4 with I-25d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-25 is obtained through a three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.97(s,1H),11.18(s,1H),9.95(s,1H),8.88(s,2H),8.73(d,J＝12.4Hz,2H),8.62(d,J＝8.1Hz,1H),8.51(d,J＝5.5Hz,1H),8.08(t,J＝7.7Hz,1H),5.13(s,2H),4.36(dd,J＝16.2,9.9Hz,2H),4.18–4.05(m,2H),3.91(d,J＝25.6Hz,2H),3.59(t,J＝5.7Hz,2H),2.38(dt,J＝14.2,6.9Hz,2H)；HRMS(ESI)m/z calcd for C 21H 23N 6O 2(M+H)+391.1882,found 391.1883. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.97 (s, 1H), 11.18 (s, 1H), 9.95 (s, 1H), 8.88 (s, 2H), 8.73 (d, J = 12.4 Hz, 2H), 8.62 (d, J = 8.1 Hz, 1H), 8.51 (d, J = 5.5 Hz, 1H), 8.08 (t, J = 7.7 Hz, 1H), 5.13 (s, 2H), 4.36 (dd, J = 16.2, 9.9 Hz, 2H), 4.18–4.05 (m, 2H), 3.91 (d, J = 25.6 Hz, 2H), 3.59 (t, J = 5.7 Hz, 2H), 2.38 (dt, J = 14.2 ,6.9Hz,2H); HRMS(ESI)m/z calcd for C 21 H 23 N 6 O 2 (M+H)+391.1882,found 391.1883.
实施例69Example 69
2-(2-(喹啉-4-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-26)的制备2-(2-(quinolin-4-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (compound I-26) Preparation
Figure PCTCN2019077356-appb-000078
Figure PCTCN2019077356-appb-000078
将实施例4中的I-1d替换成I-26d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-26。Replace I-1d in Example 4 with I-26d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-26 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.81(s,1H),11.14(s,1H),9.13(d,J＝4.6Hz,1H),8.72(s,2H),8.42(d,J＝8.0Hz,1H),8.24(d,J＝8.3Hz,1H),8.02–7.96(m,1H),7.90(dd,J＝16.9,8.8Hz,2H),5.22(s,2H),4.36(s,2H),4.29(s,2H),3.98(s,1H),3.88(s,1H),3.65–3.57(m,2H),2.43(s,1H),2.36(d,J＝6.7Hz,1H)；HRMS(ESI)m/z calcd for C 21H 23N 6O 2(M+H)+391.1882,found 391.1883. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.81 (s, 1H), 11.14 (s, 1H), 9.13 (d, J = 4.6 Hz, 1H), 8.72 (s, 2H), 8.42 (d, J = 8.0Hz, 1H), 8.24 (d, J = 8.3Hz, 1H), 8.02–7.96 (m, 1H), 7.90 (dd, J = 16.9, 8.8 Hz, 2H), 5.22 (s, 2H), 4.36(s,2H), 4.29(s,2H), 3.98(s,1H), 3.88(s,1H), 3.65–3.57(m,2H),2.43(s,1H), 2.36(d,J= 6.7Hz,1H); HRMS(ESI)m/z calcd for C 21 H 23 N 6 O 2 (M+H)+391.1882,found 391.1883.
实施例70Example 70
2-(2-(异喹啉-4-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-27)的制备2-(2-(Isoquinolin-4-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (Compound I-27 ) Preparation
Figure PCTCN2019077356-appb-000079
Figure PCTCN2019077356-appb-000079
将实施例4中的I-1d替换成I-27d，其余所需原料，试剂及制备方法同实施例4-6， 经三步反应得I-27。Replace I-1d in Example 4 with I-27d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-27 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ12.03(d,J＝34.1Hz,1H),11.17(s,1H),9.98(s,1H),9.06(s,1H),8.72(s,2H),8.61(d,J＝8.2Hz,1H),8.31(t,J＝7.5Hz,1H),8.10(t,J＝7.5Hz,1H),5.19(s,2H),4.43(s,2H),4.14(s,2H),3.92(d,J＝24.0Hz,2H),3.64–3.58(m,2H),2.45–2.34(m,2H)；HRMS(ESI)m/z calcd for C 21H 23N 6O 2(M+H)+391.1882,found 391.1883. 1 H NMR(400MHz,DMSO-d 6 )δ12.03(d,J=34.1Hz,1H), 11.17(s,1H), 9.98(s,1H), 9.06(s,1H), 8.72(s, 2H), 8.61 (d, J = 8.2 Hz, 1H), 8.31 (t, J = 7.5 Hz, 1H), 8.10 (t, J = 7.5 Hz, 1H), 5.19 (s, 2H), 4.43 (s, 2H),4.14(s,2H),3.92(d,J=24.0Hz,2H),3.64-3.58(m,2H),2.45-2.34(m,2H); HRMS(ESI)m/z calcd for C 21 H 23 N 6 O 2 (M+H)+391.1882,found 391.1883.
实施例71Example 71
2-(2-(咪唑并[1,2-]吡啶-3-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-28)的制备2-(2-(imidazo[1,2-]pyridin-3-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamic acid Preparation of salt (compound I-28)
Figure PCTCN2019077356-appb-000080
Figure PCTCN2019077356-appb-000080
将实施例4中的I-1d替换成I-28d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-28。Replace I-1d in Example 4 with I-28d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-28 is obtained through a three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ12.24(s,1H),11.20(s,1H),9.31(d,J＝6.8Hz,1H),8.72(s,2H),8.56(s,1H),8.08(d,J＝3.9Hz,2H),7.65(dt,J＝8.0,4.1Hz,1H),5.09(s,2H),4.40(s,2H),4.10(s,2H),3.86(s,2H),3.60(s,2H),2.37(d,J＝6.3Hz,2H)；HRMS(ESI)m/z calcd for C 19H 22N 7O 2(M+H)+380.1835,found 380.1836. 1 H NMR (400MHz, DMSO-d 6 ) δ 12.24 (s, 1H), 11.20 (s, 1H), 9.31 (d, J = 6.8 Hz, 1H), 8.72 (s, 2H), 8.56 (s, 1H), 8.08 (d, J = 3.9 Hz, 2H), 7.65 (dt, J = 8.0, 4.1 Hz, 1H), 5.09 (s, 2H), 4.40 (s, 2H), 4.10 (s, 2H), 3.86(s,2H),3.60(s,2H),2.37(d,J=6.3Hz,2H); HRMS(ESI)m/z calcd for C 19 H 22 N 7 O 2 (M+H)+380.1835 ,found 380.1836.
实施例72Example 72
2-(2-((1-甲基-1H-苯并[d]咪唑-2-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-29)的制备2-(2-((1-methyl-1H-benzo[d]imidazol-2-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5- Preparation of Hydroxamate Hydrochloride (Compound I-29)
Figure PCTCN2019077356-appb-000081
Figure PCTCN2019077356-appb-000081
将实施例4中的I-1d替换成I-29d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-29。Replace I-1d in Example 4 with I-29d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-29 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.93–11.59(m,1H),11.15(s,1H),8.74(s,2H),7.73(s,2H),7.39(dd,J＝15.3,7.6Hz,2H),5.00(s,2H),4.40(s,4H),3.96–3.88(m,3H),3.73(dd,J＝19.3,12.8Hz,2H),3.64(d,J＝7.0Hz,2H),2.39(s,2H)；HRMS(ESI)m/z calcd for C 20H 24N 7O 2(M+H)+394.1991,found 394.1990. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.93-11.59 (m, 1H), 11.15 (s, 1H), 8.74 (s, 2H), 7.73 (s, 2H), 7.39 (dd, J = 15.3) ,7.6Hz,2H),5.00(s,2H),4.40(s,4H),3.96-3.88(m,3H),3.73(dd,J＝19.3,12.8Hz,2H),3.64(d,J＝ 7.0Hz,2H),2.39(s,2H); HRMS(ESI)m/z calcd for C 20 H 24 N 7 O 2 (M+H)+394.1991,found 394.1990.
实施例73Example 73
2-(2-((1-甲基-1H-吡咯并[2,3-b]吡啶-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-30)的制备2-(2-((1-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl) Preparation of pyrimidine-5-hydroxamate hydrochloride (compound I-30)
Figure PCTCN2019077356-appb-000082
Figure PCTCN2019077356-appb-000082
将实施例4中的I-1d替换成I-30d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-30。Replace I-1d in Example 4 with I-30d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-30 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.37(d,J＝27.4Hz,2H),8.73(d,J＝9.7Hz,2H),8.45(d,J＝7.9Hz,1H),8.39(d,J＝4.7Hz,1H),7.88(d,J＝5.7Hz,1H),7.31–7.24(m,1H),4.59(d,J＝4.5Hz,2H),4.19(dd,J＝16.1,6.5Hz,2H),4.05–3.95(m,2H),3.85(d,J＝33.7Hz,5H),3.58(dd,J＝14.1,7.0Hz,2H),2.31(dt,J＝29.1,6.9Hz,3H)；HRMS(ESI)m/z calcd for C 20H 24N 7O 2(M+H)+394.1991,found 394.1992. 1 H NMR(400MHz,DMSO-d 6 )δ11.37(d,J=27.4Hz,2H), 8.73(d,J=9.7Hz,2H), 8.45(d,J=7.9Hz,1H), 8.39 (d,J=4.7Hz,1H), 7.88(d,J=5.7Hz,1H), 7.31–7.24(m,1H), 4.59(d,J=4.5Hz,2H), 4.19(dd,J= 16.1, 6.5 Hz, 2H), 4.05-3.95 (m, 2H), 3.85 (d, J = 33.7 Hz, 5H), 3.58 (dd, J = 14.1, 7.0 Hz, 2H), 2.31 (dt, J = 29.1 ,6.9Hz,3H); HRMS(ESI)m/z calcd for C 20 H 24 N 7 O 2 (M+H)+394.1991,found 394.1992.
实施例74Example 74
2-(2-((1-甲基-1H-吡咯并[2,3-c]吡啶-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-31)的制备2-(2-((1-methyl-1H-pyrrolo[2,3-c]pyridin-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl) Preparation of pyrimidine-5-hydroxamate hydrochloride (Compound I-31)
Figure PCTCN2019077356-appb-000083
Figure PCTCN2019077356-appb-000083
将实施例4中的I-1d替换成I-31d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-31。Replace I-1d in Example 4 with I-31d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-31 is obtained through a three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ12.18–11.90(m,1H),11.18(s,1H),9.48(s,1H),8.77–8.68(m,2H),8.55(t,J＝5.7Hz,2H),8.49(d,J＝6.4Hz,1H),4.75(t,J＝5.8Hz,2H),4.25(d,J＝6.5Hz,2H),4.12(s,3H),3.97(d,J＝6.8Hz,2H),3.91(s,1H),3.83(s,1H),3.58(dd,J＝13.1,6.6Hz,2H),2.34(dt,J＝26.4,6.9Hz,2H)；HRMS(ESI)m/z calcd for C 20H 24N 7O 2(M+H)+394.1991,found 394.1992. 1 H NMR (400MHz, DMSO-d 6 ) δ 12.18-11.90 (m, 1H), 11.18 (s, 1H), 9.48 (s, 1H), 8.77-8.68 (m, 2H), 8.55 (t, J ＝5.7Hz,2H),8.49(d,J＝6.4Hz,1H),4.75(t,J＝5.8Hz,2H), 4.25(d,J＝6.5Hz,2H), 4.12(s,3H), 3.97(d,J＝6.8Hz,2H),3.91(s,1H),3.83(s,1H),3.58(dd,J＝13.1,6.6Hz,2H),2.34(dt,J＝26.4,6.9Hz ,2H); HRMS(ESI)m/z calcd for C 20 H 24 N 7 O 2 (M+H)+394.1991,found 394.1992.
实施例75Example 75
2-(2-((1-甲基-1H-吡咯并[3,2-b]吡啶-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-32)的制备2-(2-((1-methyl-1H-pyrrolo[3,2-b]pyridin-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl) Preparation of pyrimidine-5-hydroxamate hydrochloride (compound I-32)
Figure PCTCN2019077356-appb-000084
Figure PCTCN2019077356-appb-000084
将实施例4中的I-1d替换成I-32d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-32。Replace I-1d in Example 4 with I-32d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-32 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.73(s,1H),11.18(s,1H),8.86–8.80(m,2H),8.72(s,2H),8.46(s,1H),7.81(dd,J＝8.2,5.8Hz,1H),4.93(s,2H),4.39(s,2H),4.07(s,3H),4.00(d,J＝9.5Hz,2H),3.84(s,2H),3.59(s,2H),2.36(d,J＝6.7Hz,2H)；HRMS(ESI)m/z calcd for C 20H 24N 7O 2(M+H)+394.1991,found 394.1989. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.73 (s, 1H), 11.18 (s, 1H), 8.86-8.80 (m, 2H), 8.72 (s, 2H), 8.46 (s, 1H), 7.81(dd,J=8.2,5.8Hz,1H), 4.93(s, 2H), 4.39(s, 2H), 4.07(s, 3H), 4.00(d, J=9.5Hz, 2H), 3.84(s ,2H),3.59(s,2H),2.36(d,J=6.7Hz,2H); HRMS(ESI)m/z calcd for C 20 H 24 N 7 O 2 (M+H)+394.1991,found 394.1989 .
实施例76Example 76
2-(2-((1-甲基-1H-吲唑-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-33)的制备2-(2-((1-methyl-1H-indazol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamic acid Preparation of Hydrochloride (Compound I-33)
Figure PCTCN2019077356-appb-000085
Figure PCTCN2019077356-appb-000085
将实施例4中的I-1d替换成I-33d，其余所需原料，试剂及制备方法同实施例4-6， 经三步反应得I-33。Replace I-1d in Example 4 with I-33d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-33 is obtained through a three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.53(s,1H),11.24(s,1H),8.73(s,2H),8.06(d,J＝8.1Hz,1H),7.73(d,J＝8.5Hz,1H),7.55–7.46(m,1H),7.27(t,J＝7.5Hz,1H),4.85(d,J＝5.3Hz,2H),4.37–4.24(m,2H),4.19–4.06(m,5H),3.86(d,J＝47.4Hz,2H),3.61(s,1H),3.59–3.54(m,1H),2.32(dt,J＝37.4,6.9Hz,2H)；HRMS(ESI)m/z calcd for C 20H 24N 7O 2(M+H)+394.1991,found 394.1990. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.53 (s, 1H), 11.24 (s, 1H), 8.73 (s, 2H), 8.06 (d, J = 8.1 Hz, 1H), 7.73 (d, J=8.5Hz,1H), 7.55–7.46(m,1H), 7.27(t,J=7.5Hz,1H), 4.85(d,J=5.3Hz,2H), 4.37–4.24(m,2H), 4.19–4.06(m,5H), 3.86(d,J=47.4Hz,2H),3.61(s,1H),3.59–3.54(m,1H),2.32(dt,J=37.4,6.9Hz,2H) ；HRMS(ESI)m/z calcd for C 20 H 24 N 7 O 2 (M+H)+394.1991,found 394.1990.
实施例77Example 77
2-(2-((1-甲基-1H-吡咯并[3,2-c]吡啶-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-34)的制备2-(2-((1-methyl-1H-pyrrolo[3,2-c]pyridin-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl) Preparation of pyrimidine-5-hydroxamate hydrochloride (Compound I-34)
Figure PCTCN2019077356-appb-000086
Figure PCTCN2019077356-appb-000086
将实施例4中的I-1d替换成I-34d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-34。Replace I-1d in Example 4 with I-34d, and the remaining raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-34 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ12.01(s,1H),11.19(s,1H),9.73(s,1H),8.77–8.70(m,2H),8.60(d,J＝6.4Hz,1H),8.22(t,J＝7.7Hz,2H),4.78(t,J＝5.5Hz,2H),4.27(d,J＝6.3Hz,2H),4.04(s,3H),4.00–3.94(m,2H),3.88(d,J＝25.1Hz,2H),3.59(s,2H),2.34(dt,J＝19.2,6.8Hz,2H)；HRMS(ESI)m/z calcd for C 20H 24N 7O 2(M+H)+394.1991,found 394.1992. 1 H NMR (400MHz, DMSO-d 6 ) δ 12.01 (s, 1H), 11.19 (s, 1H), 9.73 (s, 1H), 8.77-8.70 (m, 2H), 8.60 (d, J = 6.4 Hz, 1H), 8.22 (t, J = 7.7 Hz, 2H), 4.78 (t, J = 5.5 Hz, 2H), 4.27 (d, J = 6.3 Hz, 2H), 4.04 (s, 3H), 4.00– 3.94(m,2H),3.88(d,J=25.1Hz,2H),3.59(s,2H),2.34(dt,J=19.2,6.8Hz,2H); HRMS(ESI)m/z calcd for C 20 H 24 N 7 O 2 (M+H)+394.1991,found 394.1992.
实施例78Example 78
2-(2-(蒽-9-基甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-35)的制备2-(2-(Anthracene-9-ylmethyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-hydroxamate hydrochloride (compound I-35) preparation
Figure PCTCN2019077356-appb-000087
Figure PCTCN2019077356-appb-000087
将实施例4中的I-1d替换成I-35d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-35。Replace I-1d in Example 4 with I-35d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-35 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.15(s,1H),10.54(d,J＝26.4Hz,1H),8.86(s,1H),8.78–8.65(m,4H),8.24(d,J＝8.4Hz,2H),7.80–7.74(m,2H),7.66(t,J＝7.5Hz,2H),5.65(d,J＝4.4Hz,2H),4.49–4.43(m,2H),4.08(dd,J＝18.4,12.7Hz,2H),3.88(d,J＝17.8Hz,2H),3.55(dd,J＝11.5,6.7Hz,2H),2.33(dt,J＝31.0,6.9Hz,2H)；HRMS(ESI)m/z calcd for C 26H 26N 5O 2(M+H)+440.2087,found 440.2088. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.15 (s, 1H), 10.54 (d, J = 26.4 Hz, 1H), 8.86 (s, 1H), 8.78-8.65 (m, 4H), 8.24 ( d,J=8.4Hz,2H),7.80–7.74(m,2H),7.66(t,J=7.5Hz,2H), 5.65(d,J=4.4Hz,2H), 4.49–4.43(m,2H ), 4.08 (dd, J = 18.4, 12.7 Hz, 2H), 3.88 (d, J = 17.8 Hz, 2H), 3.55 (dd, J = 11.5, 6.7 Hz, 2H), 2.33 (dt, J = 31.0, 6.9Hz, 2H); HRMS(ESI)m/z calcd for C 26 H 26 N 5 O 2 (M+H)+440.2087, found 440.2088.
实施例79Example 79
2-(2-((6-甲氧基-1-甲基-1H-吲哚-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-36)的制备2-(2-((6-Methoxy-1-methyl-1H-indol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine- Preparation of 5-Hydroxamate Hydrochloride (Compound I-36)
Figure PCTCN2019077356-appb-000088
Figure PCTCN2019077356-appb-000088
将实施例4中的I-1d替换成I-36d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-36。Replace I-1d in Example 4 with I-36d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-36 is obtained through a three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.09(d,J＝74.7Hz,2H),8.71(s,2H),7.76(dd,J＝8.6,4.4Hz,1H),7.50(d,J＝4.6Hz,1H),7.04(s,1H),6.81(dd,J＝8.7,2.2Hz,1H),4.52(d,J＝5.0Hz,2H),4.22–4.12(m,2H),4.00–3.94(m,2H),3.88(s,2H),3.85(s,3H),3.79(s,3H),3.60–3.56(m,2H),2.29(dt,J＝34.0,6.9Hz,2H)；HRMS(ESI)m/z calcd for C 22H 27N 6O 3(M+H)+423.2145,found 423.2144. 1 H NMR(400MHz,DMSO-d 6 )δ11.09(d,J=74.7Hz,2H),8.71(s,2H),7.76(dd,J=8.6,4.4Hz,1H),7.50(d, J = 4.6Hz, 1H), 7.04 (s, 1H), 6.81 (dd, J = 8.7, 2.2Hz, 1H), 4.52 (d, J = 5.0Hz, 2H), 4.22-4.12 (m, 2H), 4.00–3.94(m,2H), 3.88(s,2H), 3.85(s,3H), 3.79(s,3H), 3.60–3.56(m,2H), 2.29(dt,J=34.0,6.9Hz, 2H); HRMS(ESI)m/z calcd for C 22 H 27 N 6 O 3 (M+H)+423.2145,found 423.2144.
实施例80Example 80
2-(2-((1,6-二甲基-1H-吲哚-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-37)的制备2-(2-((1,6-Dimethyl-1H-indol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5-iso Preparation of Hydroxamate Hydrochloride (Compound I-37)
Figure PCTCN2019077356-appb-000089
Figure PCTCN2019077356-appb-000089
将实施例4中的I-1d替换成I-37d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-37。Replace I-1d in Example 4 with I-37d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-37 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.17(s,1H),10.87(d,J＝29.2Hz,1H),8.72(d,J＝8.5Hz,2H),7.76(dd,J＝8.1,4.4Hz,1H),7.55(d,J＝4.7Hz,1H),7.31(s,1H),7.00(d,J＝8.2Hz,1H),4.53(d,J＝5.0Hz,2H),4.20–4.15(m,2H),3.96(d,J＝10.5Hz,2H),3.88(s,1H),3.80(s,3H),3.78(s,1H),3.62–3.52(m,2H),2.47(s,3H),2.28(dt,J＝31.1,6.9Hz,2H)；HRMS(ESI)m/z calcd for C 22H 27N 6O 2(M+H)+407.2195,found 407.2196. 1 H NMR (400MHz, DMSO-d 6 ) δ 11.17 (s, 1H), 10.87 (d, J = 29.2 Hz, 1H), 8.72 (d, J = 8.5 Hz, 2H), 7.76 (dd, J = 8.1,4.4Hz,1H),7.55(d,J=4.7Hz,1H),7.31(s,1H),7.00(d,J=8.2Hz,1H),4.53(d,J=5.0Hz,2H) ,4.20–4.15(m,2H),3.96(d,J=10.5Hz,2H), 3.88(s,1H), 3.80(s,3H), 3.78(s,1H), 3.62–3.52(m,2H) ),2.47(s,3H),2.28(dt,J=31.1,6.9Hz,2H); HRMS(ESI)m/z calcd for C 22 H 27 N 6 O 2 (M+H)+407.2195,found 407.2196 .
实施例81Example 81
2-(2-((6-氰基-1-甲基-1H-吲哚-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-38)的制备2-(2-((6-cyano-1-methyl-1H-indol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5 -Preparation of Hydroxamate Hydrochloride (Compound I-38)
Figure PCTCN2019077356-appb-000090
Figure PCTCN2019077356-appb-000090
将实施例4中的I-1d替换成I-38d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-38。Replace I-1d in Example 4 with I-38d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-38 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.34(d,J＝30.0Hz,2H),8.71(s,2H),8.18(s,1H),8.11(dd,J＝8.2,4.0Hz,1H),7.96(d,J＝4.8Hz,1H),7.52(d,J＝8.3Hz,1H),4.61(d,J＝5.2Hz,2H),4.20(dd,J＝19.4,13.3Hz,2H),4.05–3.94(m,2H),3.92(s,3H),3.85(d,J＝34.8Hz,2H),3.57(dd,J＝14.8,7.8Hz,2H),2.31(dt,J＝28.4,6.8Hz,2H)；HRMS(ESI)m/z calcd for C 22H 24N 7O 2(M+H)+418.1991,found 418.1992. 1 H NMR(400MHz,DMSO-d 6 )δ11.34(d,J=30.0Hz,2H), 8.71(s,2H), 8.18(s,1H), 8.11(dd,J=8.2,4.0Hz, 1H), 7.96 (d, J = 4.8 Hz, 1H), 7.52 (d, J = 8.3 Hz, 1H), 4.61 (d, J = 5.2 Hz, 2H), 4.20 (dd, J = 19.4, 13.3 Hz, 2H),4.05–3.94(m,2H),3.92(s,3H), 3.85(d,J＝34.8Hz,2H),3.57(dd,J＝14.8,7.8Hz,2H),2.31(dt,J ＝28.4,6.8Hz,2H); HRMS(ESI)m/z calcd for C 22 H 24 N 7 O 2 (M+H)+418.1991,found 418.1992.
实施例82Example 82
2-(2-((6-溴-1-甲基-1H-吲哚-3-基)甲基)-2,6-二氮杂螺[3.4]辛-6-基)嘧啶-5-异羟肟酸盐酸盐(化合物I-39)的制备2-(2-((6-Bromo-1-methyl-1H-indol-3-yl)methyl)-2,6-diazaspiro[3.4]oct-6-yl)pyrimidine-5- Preparation of Hydroxamate Hydrochloride (Compound I-39)
Figure PCTCN2019077356-appb-000091
Figure PCTCN2019077356-appb-000091
将实施例4中的I-1d替换成I-39d，其余所需原料，试剂及制备方法同实施例4-6，经三步反应得I-39。Replace I-1d in Example 4 with I-39d, and the remaining required raw materials, reagents and preparation methods are the same as those in Example 4-6, and I-39 is obtained through three-step reaction.
1H NMR(400MHz,DMSO-d 6)δ11.03(d,J＝30.8Hz,2H),8.72(d,J＝9.0Hz,2H),7.87(dd,J＝8.5,4.4Hz,1H),7.81(s,1H),7.68(d,J＝4.8Hz,1H),7.34–7.28(m,1H),4.57(t,J＝4.8Hz,2H),4.17(d,J＝7.9Hz,2H),3.99–3.94(m,2H),3.88(s,1H),3.84(s,3H),3.80(s,1H),3.57(dd,J＝16.2,7.2Hz,2H),2.29(dt,J＝27.9,6.8Hz,2H)；HRMS(ESI)m/z calcd for C 21H 24N 6O 2Br(M+H)+471.1144,found 471.1143. 1 H NMR(400MHz,DMSO-d 6 )δ11.03(d,J=30.8Hz,2H), 8.72(d,J=9.0Hz,2H), 7.87(dd,J=8.5,4.4Hz,1H) ,7.81(s,1H),7.68(d,J=4.8Hz,1H),7.34–7.28(m,1H),4.57(t,J=4.8Hz,2H),4.17(d,J=7.9Hz, 2H),3.99–3.94(m,2H), 3.88(s,1H), 3.84(s,3H), 3.80(s,1H), 3.57(dd,J=16.2,7.2Hz,2H), 2.29(dt ,J=27.9,6.8Hz,2H); HRMS(ESI)m/z calcd for C 21 H 24 N 6 O 2 Br(M+H)+471.1144,found 471.1143.
实施例83Example 83
化合物对疟原虫3D7和Dd2的体外生长半数有效抑制浓度(IC 50)的测定 Determination of the in vitro growth half effective inhibitory concentration (IC 50 ) of the compound against Plasmodium 3D7 and Dd2
(1)疟原虫培养：疟原虫培养使用PRMI(含NaHCO 3、HEPES、Albumax I、Hypoxanthine、Genaotamicin)完全培养基(complete medium)，在37℃培养箱(5％CO 2、5％O 2)中培养。 (1) Plasmodium culture: Use PRMI (containing NaHCO 3 , HEPES, Albumax I, Hypoxanthine, Genaotamicin) complete medium (complete medium) for plasmodium culture, in a 37°C incubator (5% CO 2 , 5% O 2 ) In the cultivation.
(2)化合物对疟原虫的体外生长半数有效抑制浓度(IC 50)的测定：100μL完全培养基加入到96孔板中，在第一孔加入适量的200μM的化合物并用完全培养基定容至200μL，使化合物终浓度为1000nM，然后按1/2的比例进行梯度稀释(11个浓度梯度)，二氢青蒿素(DHA)作为阳性药，不加任何化合物作为阴性组，不加疟原虫和化合物作为空白组。之后每孔加入100μL的疟原虫培养物(红细胞压积4％，原虫率1％)使最终红细胞压积2％，原虫率1％。加样完成，96孔板置于37℃培养箱(5％CO 2、5％O 2)中培养72h。培养完成，每孔除去100μL上清液，加入100μL裂解液(10×SYBR Green I、0.5％v/v Triton X-100，0.5mg/mL saponin、0.75％EDTA/Tris-Cl缓冲液)，混合均匀，室温下避光孵育2h。用孔板荧光读数机读取每孔数值(最大激发光/最大接受光：485nm/535nm)。根据荧光值计算每孔抑制率，抑制率％＝(阴性组-实验组)/(阴性组-空白组)×100％，根据浓度-抑制率，在Graphpad Prism中绘制生长抑制曲线并计算IC 50值。 (2) Determination of the half effective inhibitory concentration (IC 50 ) of the compound against the growth of Plasmodium in vitro: 100 μL of complete medium was added to a 96-well plate, and an appropriate amount of 200 μM compound was added to the first well and the volume was adjusted to 200 μL with complete medium , Make the final concentration of the compound 1000nM, and then perform a gradient dilution of 1/2 (11 concentration gradients), dihydroartemisinin (DHA) as the positive drug, no compound as the negative group, no Plasmodium and The compound is used as a blank group. After that, 100 μL of the malaria parasite culture (4% hematocrit and 1% protozoan rate) was added to each well to make the final hematocrit 2% and the protozoan rate 1%. After the sample is added, the 96-well plate is placed in a 37°C incubator (5% CO 2 , 5% O 2 ) for 72 hours. After culture is completed, remove 100μL of supernatant from each well, add 100μL of lysis buffer (10×SYBR Green I, 0.5% v/v Triton X-100, 0.5 mg/mL saponin, 0.75% EDTA/Tris-Cl buffer), and mix Evenly, incubate for 2h in the dark at room temperature. Use a plate fluorescence reader to read the value of each well (maximum excitation light/maximum acceptance light: 485nm/535nm). Calculate the inhibition rate of each well based on the fluorescence value, the inhibition rate %=(negative group-experimental group)/(negative group-blank group)×100%, according to the concentration-inhibition rate, draw the growth inhibition curve in Graphpad Prism and calculate the IC 50 value.
(3)化合物活性测试结果(3) Compound activity test results
表1 化合物对3D7和Dd2的IC 50值 Table 1 The IC 50 values of the compounds for 3D7 and Dd2
Figure PCTCN2019077356-appb-000092
Figure PCTCN2019077356-appb-000092
Figure PCTCN2019077356-appb-000093
Figure PCTCN2019077356-appb-000093
3D7为野生型虫株，对药物无明显抗性；Dd2对氯喹、奎宁、磺胺多辛、乙胺嘧啶、阿莫地喹具有抗性。表1显示化合物具有较强的体外杀虫活性，部分化合物对3D7和Dd2的IC 50值与DHA相当。 3D7 is a wild-type strain and has no obvious resistance to drugs; Dd2 is resistant to chloroquine, quinine, sulfadoxine, pyrimethamine, and amodiaquine. Table 1 shows that the compounds have strong in vitro insecticidal activity, and the IC 50 values of some compounds against 3D7 and Dd2 are comparable to DHA.
实施例84Example 84
化合物对两株正常人细胞HepG-2和293-T体外生长半数有效抑制浓度(EC 50)的测定 Determination of the half effective inhibitory concentration (EC 50 ) of the compound on the growth of two normal human cells HepG-2 and 293-T in vitro
(1)化合物对两株正常人细胞体外生长抑制活性的测定(1) Determination of the compound's in vitro growth inhibitory activity on two normal human cells
准备HepG-2和293-T细胞，10cm dish中于37℃，5％CO 2细胞培养箱内培养 Prepare HepG-2 and 293-T cells, culture them in a 10cm dish at 37℃, 5% CO 2 cell incubator
第一天first day
胰酶消化重悬细胞并计数，按100μL/孔的体系，7000/孔细胞的量将细胞转接至96孔板中。37℃，5％CO 2细胞培养箱内培养24h； Trypsin digest and resuspend the cells and count them, transfer the cells to a 96-well plate according to a 100μL/well system and 7000/well cells. Cultivate 24h in 37℃, 5% CO 2 cell incubator;
第二天the next day
1、准备化合物梯度浓度体系，2倍稀释，体系为100μL/孔。1. Prepare a compound gradient concentration system, 2 times dilution, the system is 100μL/well.
2、去掉第一天中96孔板细胞培养体系中的上清，并将新配置好的药物浓度体系对应加入到培养细胞的培养板孔内(设置双复孔)。37℃，5％CO 2细胞培养箱内培养72h。 2. Remove the supernatant from the 96-well plate cell culture system on the first day, and add the newly configured drug concentration system to the culture plate wells of the cultured cells (set up double multiple wells). Cultivate for 72h in 37℃, 5% CO 2 cell incubator.
第五天Fifth day
1、细胞培养结束之后，去掉96孔板细胞培养体系中的上清，每孔加入100μL检测溶液(含10％CCK-8的培养基)，37℃，5％CO 2细胞培养箱内孵育1h，然后取出用酶标仪测定450nm下的吸光度。 1. After the cell culture is finished, remove the supernatant from the cell culture system of the 96-well plate, add 100 μL of detection solution (medium containing 10% CCK-8) to each well, and incubate in a cell incubator at 37°C and 5% CO 2 for 1 hour , And then take it out and measure the absorbance at 450nm with a microplate reader.
2、进行数据处理，计算不同浓度下化合物对细胞生长的抑制率，将抑制率输入 GraphPad Prism，按照非线性回归方法，计算每个药物的EC 50。抑制率计算公式如下： 2. Perform data processing, calculate the inhibition rate of the compound on cell growth at different concentrations, input the inhibition rate into GraphPad Prism, and calculate the EC 50 of each drug according to the nonlinear regression method. The calculation formula of the inhibition rate is as follows:
抑制率(％)＝[A(0)-A(加药)]/[A(0)-A(空白)]×100％Inhibition rate (%)=[A(0)-A(dosing)]/[A(0)-A(blank)]×100%
A(加药)：具有细胞、CCK-8溶液和药物溶液的孔的吸光度A (Dosing): Absorbance of wells with cells, CCK-8 solution and drug solution
A(空白)：具有培养基和CCK-8溶液而没有细胞的孔的吸光度A (blank): absorbance of wells with medium and CCK-8 solution but no cells
A(0)：具有细胞、CCK-8溶液而没有药物溶液的孔的吸光度A(0): Absorbance of wells with cells and CCK-8 solution but no drug solution
(2)选择性指数SI的计算(2) Calculation of selectivity index SI
SI＝EC 50(细胞)/IC 50(3D7) SI=EC 50 (cell)/IC 50 (3D7)
(3)化合物测试结果(3) Compound test results
表2 化合物对HepG-2和293-T的EC 50及SI值 Table 2 EC 50 and SI values of the compounds against HepG-2 and 293-T
Figure PCTCN2019077356-appb-000094
Figure PCTCN2019077356-appb-000094
Figure PCTCN2019077356-appb-000095
Figure PCTCN2019077356-appb-000095
结果汇总于上表2，结果显示，化合物对正常细胞的生长抑制较弱，选择性可以达到数千。The results are summarized in Table 2 above. The results show that the compound has weak growth inhibition on normal cells, and the selectivity can reach thousands.
实施例85Example 85
部分化合物对小鼠肝微粒体代谢稳定性实验Metabolic stability test of some compounds on mouse liver microsomes
(1)化合物对小鼠肝微粒体代谢稳定性测试(1) Metabolic stability test of the compound on mouse liver microsomes
实验选用小鼠肝微粒体(0.5mg/mL)，采购自Corning公司。阳性对照选用酮舍林(ketanserin)，待测化合物首先配成10mM的DMSO溶液，用乙腈稀释至0.5mM；取上述0.5mM溶液加入含肝微粒体的缓冲液中，使化合物浓度为1.5μM；取上述1.5μM化合物/肝微粒体混合液30μL，加入15μL 6mM的NADPH溶液，使化合物最终浓度为1.5μM，NADPH最终浓度为2mM。化合物/肝微粒体测试液置于测试板上，在37℃水浴孵育，在每个时间点(0、5、15、30、45min)各加入135μL乙腈淬灭。待全部样品淬灭完毕，用振荡器(IKA,MTS 2/4)振荡样品10min(600rpm/min)，然后以5594g离心15min(Thermo Multifuge×3R)。取上层清液，按1:1加入蒸馏水稀释，用LC-MS进行分析。将化合物在5、15、30、45min的峰面积响应比率(PARR)与时间0的PARR进行比较，以确定在每个时间点保留的测试化合物的百分比。使用Excel软件拟合单相指数衰减方程计算半衰期。The experiment uses mouse liver microsomes (0.5mg/mL), purchased from Corning. The positive control is ketanserin. The test compound is first prepared into a 10 mM DMSO solution and diluted to 0.5 mM with acetonitrile; the above 0.5 mM solution is added to the buffer containing liver microsomes to make the compound concentration 1.5 μM; Take 30 μL of the 1.5 μM compound/liver microsome mixture and add 15 μL of 6 mM NADPH solution to make the final concentration of the compound 1.5 μM and the final concentration of NADPH 2 mM. The compound/liver microsome test solution was placed on the test plate, incubated in a 37°C water bath, and quenched by adding 135 μL of acetonitrile at each time point (0, 5, 15, 30, 45 min). After all the samples are quenched, shake the samples with a shaker (IKA, MTS 2/4) for 10 minutes (600 rpm/min), and then centrifuge at 5594g for 15 minutes (Thermo Multifuge×3R). Take the supernatant, dilute it with distilled water 1:1, and analyze by LC-MS. The peak area response ratio (PARR) of the compound at 5, 15, 30, and 45 min was compared with the PARR at time 0 to determine the percentage of test compound retained at each time point. Use Excel software to fit the single-phase exponential decay equation to calculate the half-life.
(2)化合物测试结果(2) Compound test results
表3 化合物对小鼠肝微粒体代谢稳定性Table 3 Metabolic stability of compounds to mouse liver microsomes
序号Serial number 编号Numbering T 1/2(min) T 1/2 (min) CL int(mL/min/kg) CL int (mL/min/kg)
11 I-20I-20 134.00134.00 40.7340.73
22 I-29I-29 179.79179.79 30.3530.35
33 I-32I-32 398.53398.53 13.6913.69
44 I-37I-37 83.9883.98 64.9864.98
55 I-38I-38 629.70629.70 8.538.53
66 I-39I-39 80.1780.17 68.0768.07
77 ketanserinketanserin 23.2523.25 234.74234.74
实施例86Example 86
优选化合物对五株临床耐药株的半数有效抑制浓度(IC 50)的测定 Determination of the half effective inhibitory concentration (IC 50 ) of the preferred compound against five clinically resistant strains
选取杀虫活性、细胞毒性及体外肝微粒体代谢稳定性较优的化合物，按照实施例83的方法，测试它们对五株具有不同药物抗性的临床虫株的IC 50值，结果如下： The compounds with better insecticidal activity, cytotoxicity, and in vitro liver microsomal metabolic stability were selected, and their IC 50 values were tested against five clinical strains with different drug resistance according to the method of Example 83. The results are as follows:
表4 化合物对五株临床虫株的IC 50值 Table 4 IC 50 values of compounds against five clinical strains
Figure PCTCN2019077356-appb-000096
Figure PCTCN2019077356-appb-000096
GB4对氯喹具有抗性；C2A对奎宁具有抗性；CP286对磺胺多辛、乙胺嘧啶、甲氟喹具有抗性；6218与6320对青蒿素类药物具有时间依赖的抗性，只在环期同步化后6h以内显示。综合表1和表4，化合物的72h IC 50值均与DHA相当，显示化合物具有治疗对目前抗疟一线和二线药物耐受的疟疾，应对疟疾耐药性的潜力。 GB4 is resistant to chloroquine; C2A is resistant to quinine; CP286 is resistant to sulfadoxine, pyrimethamine, and mefloquine; 6218 and 6320 have time-dependent resistance to artemisinin drugs, only in It will be displayed within 6h after the ring phase is synchronized. Combining Table 1 and Table 4, the 72h IC 50 values of the compounds are equivalent to DHA, indicating that the compounds have the potential to treat malaria that is resistant to current first- and second-line antimalarial drugs and cope with malaria resistance.
实施例87Example 87
优选化合物抑制疟原虫去乙酰化酶活性验证实验Validation experiment of preferred compounds to inhibit the activity of Plasmodium deacetylase
疟原虫培养Plasmodium culture
疟原虫培养使用RPMI(含NaHCO3,HEPES,Albumax I,Hypoxanthine Genaotamicin)完全培养基(Complete Medium)，在37℃培养箱中培养Plasmodium culture uses RPMI (containing NaHCO3, HEPES, Albumax I, Hypoxanthine Genaotamicin) complete medium (Complete Medium), cultivated in a 37℃ incubator
药物配制Drug formulation
根据药物前期测出的IC50值，将药物溶于DMSO中，配制200*20*IC50的初浓度。According to the IC50 value measured in the early stage of the drug, the drug was dissolved in DMSO to prepare an initial concentration of 200*20*IC50.
药物处理(在生物安全柜中进行)Drug handling (in a biological safety cabinet)
取44h左右的恶性疟原虫同完全培养基与红细胞在50mL管中配制混合物(红细胞含量为2％，原虫率为8％-10％)，在6孔板的每孔中加入6mL混合物，再在每孔中加入30μL药物(工作浓度为20*IC50)，加好之后混匀板子，并放入三气培养箱孵育4h。Take about 44 hours of Plasmodium falciparum, complete medium and red blood cells to prepare a mixture (red blood cell content of 2%, protozoa rate 8%-10%) in a 50 mL tube, add 6 mL of the mixture to each well of a 6-well plate, and then Add 30μL of drug (working concentration is 20*IC50) to each well, mix the plate after adding, and incubate it in a three-gas incubator for 4h.
收取蛋白样Collect protein samples
取出6孔板，弃去4mL上清液，余下的2mL虫血混合物转入2mLEP管，离心(4000r/2min，常温)，去除上清，再将各孔用1mL PBS重悬残液并转移至原EP管以减少损失，再次离心去上清；Take out the 6-well plate, discard 4mL supernatant, transfer the remaining 2mL insect blood mixture into a 2mL EP tube, centrifuge (4000r/2min, room temperature), remove the supernatant, and then resuspend the remaining liquid in each well with 1mL PBS and transfer to Original EP tube to reduce loss, centrifuge again to remove the supernatant;
在各EP管中加入2mL裂解液，涡旋仪振荡混匀，冰上裂解10min，离心(12000r/1min，4℃)，弃去上清液；Add 2mL lysate to each EP tube, vortex and mix well, lyse on ice for 10min, centrifuge (12000r/1min, 4℃), discard the supernatant;
各EP管加入1mL PBS，涡旋仪振荡混匀，离心(12000r/1min，4℃)，弃去上清液，再重复此步骤两次；Add 1mL PBS to each EP tube, vortex and mix well, centrifuge (12000r/1min, 4℃), discard the supernatant, and repeat this step twice;
加入90μL 1×PBS重悬并转移至1.5mL超声管，再加入10μL 10％SDS，混匀后超声5min(30s on/30s off)，离心(12000r/10min，4℃)，取上清液，加入loading，振荡，100℃加热10min，样品可保存于-20℃。Add 90μL 1×PBS to resuspend and transfer to a 1.5mL ultrasound tube, then add 10μL 10% SDS, mix and sonicate for 5min (30s on/30s off), centrifuge (12000r/10min, 4℃), take the supernatant, Add loading, shake, and heat at 100°C for 10 minutes. The sample can be stored at -20°C.
Western BlotWestern Blot
SDS-PAGE凝胶电泳：预制胶各孔上样10μL，电压80V跑30min后将电压调至120V再接着跑至loading接近分离胶下边缘。SDS-PAGE gel electrophoresis: Load 10μL in each well of the precast gel, run at 80V for 30 minutes, adjust the voltage to 120V, and then run until the loading is close to the bottom edge of the separation gel.
转膜：切取相应覆盖面积的PVDF膜，采用湿转法，快速转膜buffer，400mA恒流转35min，结束后取出PVDF膜。Transfer membrane: Cut out the PVDF membrane with corresponding coverage area, adopt wet transfer method, fast transfer membrane buffer, 400mA constant current transfer for 35 minutes, and take out the PVDF membrane after finishing.
封闭：将膜放于封闭液(在TBST中加入5％脱脂奶粉)中，摇床摇晃封闭2h。Sealing: Put the membrane in the sealing solution (add 5% skimmed milk powder in TBST), shake and seal for 2h on a shaker.
孵一抗：采用组蛋白histone H3抗体和H3K9乙酰化抗体，与5％脱脂奶粉按照1：2000稀释，于摇床孵育PVDF膜2h后弃去孵育液，加入TBST洗膜三次，每次10min。Incubate the primary antibody: use histone histone H3 antibody and H3K9 acetylated antibody, dilute with 5% skimmed milk powder at a ratio of 1:2000, incubate the PVDF membrane on a shaker for 2 hours, discard the incubation solution, add TBST to wash the membrane three times, each time for 10 minutes.
孵二抗：二抗按照1：5000稀释，于摇床孵育PVDF膜1h后弃去孵育液。Incubate the secondary antibody: Dilute the secondary antibody at 1:5000, incubate the PVDF membrane on a shaker for 1 hour and discard the incubation solution.
显色曝光：临时配制显色液并均匀的铺在PVDF膜上，可根据条带亮度调节曝光时间。Color development and exposure: Temporarily prepare color development solution and spread it evenly on the PVDF film. The exposure time can be adjusted according to the brightness of the strip.
(5)化合物Western Blot实验结果(5) Western Blot experiment results of the compound
实验结果如图1中所示，JL01为阳性对照化合物，对比化合物处理和DMSO处理4h后的疟原虫组蛋白H3乙酰化条带，可以看出化合物上调了乙酰化水平，即抑制了去乙酰化酶的活性，间接证明化合物是泛pfHDAC抑制剂。The experimental results are shown in Figure 1. JL01 is the positive control compound. Comparing the acetylation bands of Plasmodium histone H3 after treatment with compound and DMSO for 4 hours, it can be seen that the compound up-regulated the level of acetylation, that is, inhibited deacetylation. The activity of the enzyme indirectly proves that the compound is a pan pfHDAC inhibitor.
实施例88Example 88
优选化合物小鼠体内药效实验In vivo drug efficacy experiment of the preferred compound in mice
(1)小鼠体内药效实验方法(1) Experimental methods of drug efficacy in mice
体内药效实验选用balb-c小鼠感染P.yoelii模型进行。小鼠选用6-8周龄的雌性，每个剂量组设置5只。阳性药为磷酸哌喹(PPQ)，给药方式为腹腔注射，给药前每组测量平均体重，按15μL/g注射相应体积的药液。化合物注射液的配置方法：先溶于5％v/v的二甲基亚砜，剧烈振荡，使固体部分部分或完全溶解，再加入95％v/v的20％wtβ-羟丙基环糊精水溶液，混匀即可。P.yoelii从-78℃解冻后经转接两只小鼠恢复毒力，取血，加入PBS稀释。每只小鼠接种10 5只疟原虫，感染后24h开始给药，一共给药5次，每次间隔24h。从感染后24h开始定期从小鼠尾静脉取血涂片观察，计算原虫率： In vivo pharmacodynamic experiments were performed using balb-c mice infected with P. yoelii model. The mice were females aged 6-8 weeks, and 5 mice were set for each dose group. The positive drug is piperaquine phosphate (PPQ), the administration method is intraperitoneal injection, the average body weight of each group is measured before administration, and the corresponding volume of liquid medicine is injected at 15 μL/g. Compound injection preparation method: first dissolve in 5% v/v dimethyl sulfoxide, shake vigorously to partially or completely dissolve the solid, then add 95% v/v 20% wt β-hydroxypropyl cyclopaste Essence water solution, just mix well. After P.yoelii was thawed from -78°C, the virulence was restored by transferring two mice, and the blood was taken and diluted with PBS. Inoculated with 10 5 per mouse Plasmodium, 24h after infection start of administration, a total of 5 doses at intervals of 24h. From 24 hours after infection, blood smears were taken regularly from the tail vein of mice to observe and calculate the protozoan rate:
原虫率＝被疟原虫感染的红细胞数/红细胞总数×100％Protozoa rate = number of red blood cells infected by plasmodium/total number of red blood cells×100%
感染后观察30天血涂片，计算相应的原虫率。Observe the blood smear for 30 days after infection and calculate the corresponding protozoan rate.
(2)小鼠体内药效实验结果(2) Results of in vivo drug efficacy experiments in mice
结果如图2和3中所示。原虫率曲线图(图2)显示与空白组相比，化合物I-39在60mg/kg下具有较好的杀虫活性，且存活的小鼠在第30天时原虫率均为0，显示体内疟原虫已被清除。原虫率曲线和生存曲线(图3)综合显示I-39在药效和毒性上具有较好的平衡。The results are shown in Figures 2 and 3. The protozoan rate curve (Figure 2) shows that compared with the blank group, compound I-39 has better insecticidal activity at 60 mg/kg, and the protozoan rate of the surviving mice is 0 on the 30th day. The protozoan has been eliminated. The protozoan rate curve and survival curve (Figure 3) comprehensively show that I-39 has a good balance of efficacy and toxicity.
实施例89Example 89
优选化合物对人源HDAC半数有效抑制浓度(IC 50)的测定 Determination of the half effective inhibitory concentration (IC 50 ) of the preferred compound against human HDAC
(1)hHDAC1-3,6测试方法：通过Echo将250nL DMSO或化合物溶液加入到 OptiPlate TM-384F黑色测定板中，依次将15μL酶溶液、10μL GL-8底物溶液加入测定板中。在25℃下孵育60min，使用Ex350-360/Em450-465(敏感60)的设置读取值。计算抑制率，用GraphPad Prism算出IC 50值。 (1) hHDAC1-3,6 test method: add 250nL DMSO or compound solution to the OptiPlate TM-384F black assay plate via Echo, and add 15 μL enzyme solution and 10 μL GL-8 substrate solution to the assay plate in turn. Incubate at 25°C for 60 minutes and read the value using the setting of Ex350-360/Em450-465 (sensitive 60). The inhibition rate was calculated, and the IC 50 value was calculated with GraphPad Prism.
(2)hHDAC8测试方法：通过Echo将250nL DMSO或化合物溶液加入到OptiPlate TM-384F黑色测定板中，依次加入15μL酶溶液，10μL对应底物溶液，在25℃反应4h。添加10μL终止液终止反应，使用Ex350-360/Em450-465的设置读取值。计算抑制率，用GraphPad Prism算出IC 50值。 (2) hHDAC8 test method: add 250nL DMSO or compound solution to the OptiPlate TM-384F black assay plate via Echo, add 15 μL enzyme solution, 10 μL corresponding substrate solution, and react at 25°C for 4 hours. Add 10μL stop solution to stop the reaction, and use the setting of Ex350-360/Em450-465 to read the value. The inhibition rate was calculated, and the IC 50 value was calculated with GraphPad Prism.
(3)hSirt2测试方法：通过Echo将800nL DMSO或化合物溶液加入到OptiPlate TM-384F黑色测定板中，依次加入10μL酶溶液、10μL对应底物溶液，在25℃反应4h。添加20μL终止液终止反应，使用Ex350-360/Em450-465的设置读取值。计算抑制率，用GraphPad Prism算出IC 50值。 (3) hSirt2 test method: add 800nL DMSO or compound solution to the OptiPlate TM-384F black assay plate via Echo, add 10μL enzyme solution and 10μL corresponding substrate solution in sequence, and react at 25°C for 4h. Add 20μL stop solution to stop the reaction, and use the setting of Ex350-360/Em450-465 to read the value. The inhibition rate was calculated, and the IC 50 value was calculated with GraphPad Prism.
(4)化合物测试结果(4) Compound test results
表5 化合物对hHDACs的IC 50值 Table 5 IC 50 value of the compounds on hHDACs
Figure PCTCN2019077356-appb-000097
Figure PCTCN2019077356-appb-000097
SAHA与suramin为阳性对照。实验结果表明，化合物I-38对hHDACs有一定抑制，其中对hHDAC1-3有较强抑制，对hSirt2几乎无抑制。SAHA and suramin are positive controls. The experimental results show that compound I-38 has a certain inhibitory effect on hHDACs, among which it has strong inhibition on hHDAC1-3 and almost no inhibition on hSirt2.
本发明的2,6-二氮杂螺[3.4]辛烷类嘧啶-异羟肟酸化合物分子结构较为简单，制备工艺简洁，在对与疟原虫生存繁殖有密切关系的HDAC酶抑制实验和体内外杀虫药效实验中均显示出较强的抑制活性，且细胞毒性较弱，选择性指数可到数千。因此，该类化合物不但有望开发成新型的单药给药的抗疟疾药物，而且还可以开发成与现有抗疟疾药物进行组合给药的抗疟疾药物。The 2,6-diazaspiro[3.4]octane pyrimidine-hydroxamic acid compound of the present invention has a relatively simple molecular structure and a simple preparation process. It is useful in HDAC enzyme inhibition experiments and experiments that are closely related to the survival and reproduction of plasmodium. Both internal and external insecticidal efficacy experiments have shown strong inhibitory activity, and the cytotoxicity is weak, and the selectivity index can reach thousands. Therefore, such compounds are not only expected to be developed into a new type of single-drug antimalarial drugs, but also can be developed into antimalarial drugs for combined administration with existing antimalarial drugs.
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。All documents mentioned in the present invention are cited as references in this application, as if each document was individually cited as a reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims (10)

  1. 一种如下式I所示的2,6-二氮杂螺[3.4]辛烷类嘧啶-异羟肟酸化合物，或其药学上可接受的盐或光学异构体，A 2,6-diazaspiro[3.4]octane pyrimidine-hydroxamic acid compound represented by the following formula I, or a pharmaceutically acceptable salt or optical isomer thereof,
    Figure PCTCN2019077356-appb-100001
    Figure PCTCN2019077356-appb-100001
    其中，among them,
    R 1选自下组：取代或未取代的C1～C6烷基、取代或未取代的C3～C8环烷基、取代或未取代的C6-C15单环、二环或三环芳基、取代或未取代的5-15元单环、二环或三环杂环基(包括饱和、部分不饱和或芳香性杂环基)；其中，所述的杂芳基包括一个或多个选自下组的杂原子作为环骨架：N、O或S； R 1 is selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C15 monocyclic, bicyclic or tricyclic aryl, substituted Or unsubstituted 5-15 membered monocyclic, bicyclic or tricyclic heterocyclic group (including saturated, partially unsaturated or aromatic heterocyclic group); wherein, the heteroaryl group includes one or more selected from Group of heteroatoms as the ring skeleton: N, O or S;
    R 2选自下组：NHOH，或OR 3； R 2 is selected from the group consisting of NHOH, or OR 3 ;
    R 3选自下组：氢，取代或未取代的C1～C3烷基，取代或未取代的C3～C8环烷基、取代或未取代的5-15元杂环基； R 3 is selected from the following group: hydrogen, substituted or unsubstituted C1-C3 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 5-15 membered heterocyclic group;
    其中，所述的取代指基团上的氢原子被一个或者多个(例如2个、3个、4个等)选自下组的取代基所取代：卤素、氘代、C1-C6烷氧基、卤代的C1-C6烷氧基、卤代的C3-C8环烷基、甲基砜基、-S(＝O) 2NH 2、氧代(＝O)、-CN、羟基、-NH 2、羧基、或取代或未取代的选自下组的基团：C1-C6烷基、C3-C8环烷基、C1-C6胺基、C6-C10芳基、具有1-3个选自N、S和O的杂原子的5-10元杂芳基、具有1-3个选自N、S和O的杂原子的5-12元杂环基，且所述的取代基选自下组：卤素、C1-C6烷基、C1-C6烷氧基、氧代、-CN、-NH 2、-OH、C6-C10芳基、C1-C6胺基、C1-C6酰胺基、具有1-3个选自N、S和O的杂原子的5-10元杂芳基。 Wherein, the substitution means that the hydrogen atom on the group is replaced by one or more (for example, 2, 3, 4, etc.) substituents selected from the following group: halogen, deuterated, C1-C6 alkoxy Group, halogenated C1-C6 alkoxy, halogenated C3-C8 cycloalkyl, methyl sulfone, -S(=O) 2 NH 2 , oxo(=O), -CN, hydroxyl,- NH 2 , carboxyl group, or substituted or unsubstituted group selected from the following group: C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 amino, C6-C10 aryl, with 1-3 selected A 5-10 membered heteroaryl group from heteroatoms of N, S and O, a 5-12 membered heterocyclic group with 1-3 heteroatoms selected from N, S and O, and the substituents are selected from The following group: halogen, C1-C6 alkyl, C1-C6 alkoxy, oxo, -CN, -NH 2 , -OH, C6-C10 aryl, C1-C6 amine, C1-C6 amide, with A 5-10 membered heteroaryl group with 1-3 heteroatoms selected from N, S and O.
  2. 如权利要求1所述的化合物，其特征在于，所述的R 2选自下组：H、取代或未取代的C1～C6烷基，取代或未取代的C3～C8环烷基，或取代或未取代的选自下组的基团： The compound of claim 1, wherein said R 2 is selected from the group consisting of H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, or substituted Or unsubstituted group selected from the following group:
    Figure PCTCN2019077356-appb-100002
    Figure PCTCN2019077356-appb-100002
    上述各式中，X选自N、O、S。In the above formulas, X is selected from N, O, and S.
  3. 如权利要求1所述的化合物，其特征在于，所述的R 1选自下组：取代或未取代的C1～C6烷基、取代或未取代的C3～C8环烷基，取代或未取代的C6-C15芳基，取代或未取代的5-15元杂芳基；其中，所述的C3～C8环烷基选自下组：环戊基，环己基； The compound of claim 1, wherein said R 1 is selected from the group consisting of substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C15 aryl group, substituted or unsubstituted 5-15 membered heteroaryl group; wherein, the C3-C8 cycloalkyl group is selected from the following group: cyclopentyl, cyclohexyl;
    所述的芳基选自下组：苯基、萘基、菲基；The aryl group is selected from the following group: phenyl, naphthyl, phenanthryl;
    所述的5-15元杂芳基选自下组：
    Figure PCTCN2019077356-appb-100003
    Figure PCTCN2019077356-appb-100004
    The 5-15 membered heteroaryl group is selected from the following group:
    Figure PCTCN2019077356-appb-100003
    Figure PCTCN2019077356-appb-100004
  4. 如权利要求1所述的化合物，其特征在于，所述的取代基选自下组：卤素、OH、NH 2、CN、C1-C6烷基、C3～C8环烷基、C6-C10芳基、C1-C6烷氧基、C1-C6烷氨基。 The compound of claim 1, wherein the substituent is selected from the group consisting of halogen, OH, NH 2 , CN, C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl , C1-C6 alkoxy, C1-C6 alkylamino.
  5. 如权利要求1所述的化合物，其特征在于，所述的式I化合物可选自下组：The compound of claim 1, wherein the compound of formula I can be selected from the following group:
    Figure PCTCN2019077356-appb-100005
    Figure PCTCN2019077356-appb-100005
    Figure PCTCN2019077356-appb-100006
    Figure PCTCN2019077356-appb-100006
  6. 一种药物组合物，其特征在于，所述的药物组合物包括(a)治疗有效量的如权利要求1中所述的化合物、或其药学上可接受的盐、水合物或溶剂化物；和(b)药学上可接受的载体。A pharmaceutical composition, characterized in that, the pharmaceutical composition comprises (a) a therapeutically effective amount of the compound as claimed in claim 1, or a pharmaceutically acceptable salt, hydrate or solvate thereof; and (b) A pharmaceutically acceptable carrier.
  7. 如权利要求6所述的药物组合物，其特征在于，所述的药物组合物用于：The pharmaceutical composition according to claim 6, wherein the pharmaceutical composition is used for:
    (a)治疗或预防疟原虫导致的疾病或病症；或(a) Treatment or prevention of diseases or conditions caused by malaria parasites; or
    (b)治疗或预防与HDAC活性或表达量相关的疾病或病症。(b) Treating or preventing diseases or disorders related to the activity or expression of HDAC.
  8. 如权利要求1所述的式I化合物的用途，其特征在于，用于制备治疗或预防疟原虫导致的疾病或病症的药物组合物。The use of the compound of formula I according to claim 1, characterized in that it is used to prepare a pharmaceutical composition for treating or preventing diseases or disorders caused by malaria parasites.
  9. 如权利要求8所述的用途，其特征在于，所述的疾病或病症为疟疾。The use according to claim 8, wherein the disease or condition is malaria.
  10. 如权利要求1所述的式I化合物的用途，其特征在于，用于制备治疗或预防HDAC活性或表达量相关的疾病或病症的药物组合物。The use of the compound of formula I according to claim 1, characterized in that it is used to prepare a pharmaceutical composition for treating or preventing diseases or disorders related to the activity or expression of HDAC.
PCT/CN2019/077356 2019-03-07 2019-03-07 2,6-diazaspiro[3.4]octane pyrimidine-hydroxamic acid compound and use thereof WO2020177128A1 (en)

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WO2014026467A1 (en) * 2012-08-14 2014-02-20 山东轩竹医药科技有限公司 Bicyclic substituted pyrimidine compounds

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014026467A1 (en) * 2012-08-14 2014-02-20 山东轩竹医药科技有限公司 Bicyclic substituted pyrimidine compounds

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