CN113121423A - ALA hybrid 3-hydroxypyridone derivative and preparation method and application thereof - Google Patents

ALA hybrid 3-hydroxypyridone derivative and preparation method and application thereof Download PDF

Info

Publication number
CN113121423A
CN113121423A CN202110261792.8A CN202110261792A CN113121423A CN 113121423 A CN113121423 A CN 113121423A CN 202110261792 A CN202110261792 A CN 202110261792A CN 113121423 A CN113121423 A CN 113121423A
Authority
CN
China
Prior art keywords
compound
reaction
dichloromethane
solvent
ala
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN202110261792.8A
Other languages
Chinese (zh)
Other versions
CN113121423B (en
Inventor
谢媛媛
袁圣利
高变变
林玉燕
张长俊
张雨佳
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Zhejiang University of Technology ZJUT
Original Assignee
Zhejiang University of Technology ZJUT
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Zhejiang University of Technology ZJUT filed Critical Zhejiang University of Technology ZJUT
Priority to CN202110261792.8A priority Critical patent/CN113121423B/en
Priority to CN202210473051.0A priority patent/CN114736155B/en
Publication of CN113121423A publication Critical patent/CN113121423A/en
Application granted granted Critical
Publication of CN113121423B publication Critical patent/CN113121423B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/62—Oxygen or sulfur atoms
    • C07D213/69—Two or more oxygen atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Epidemiology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention designs and synthesizes the novel anti-tumor active compound with iron chelation and photosensitive activity based on the principles of reasonable drug design, drug-like property and the like. The invention aims to provide a series of novel active compounds of ALA hybrid 3-Hydroxypyridone (HPO) derivatives with iron ion chelating capacity and PDT activity, a preparation method and application in the aspect of tumor resistance. A series of novel compounds with PDT activity synthesized by the invention have significant advantages in the aspect of anti-tumor activity.

Description

ALA hybrid 3-hydroxypyridone derivative and preparation method and application thereof
Technical Field
The invention relates to the field of organic synthesis and pharmaceutical chemistry, in particular to an ALA hybrid 3-hydroxypyridone derivative which has iron ion chelating capacity and photosensitive activity and is obtained by hybrid 3-Hydroxypyridone (HPO) and 5-aminolevulinic acid (ALA), a preparation method thereof and application thereof in Photodynamic therapy (PDT).
Background
Since the twentieth century, with the acceleration of industrialization, the pollution to the human living environment has increased, people have been in frequent contact with carcinogenic factors, the incidence of cancer has increased year by year, and in high income countries, cancer has become the first enemy of human health over cardiovascular diseases. The treatment modes for the tumor mainly comprise three common means of operation treatment, chemotherapy and radiotherapy, and although the three common treatment modes have good curative effect on the treatment of the tumor, the three common treatment modes can also cause irreversible damage to a normal body. In recent years, with the application of photodynamic therapy in antitumor research, PDT has become an emerging method of tumor therapy into the human visual field and has achieved significant efficacy in the treatment of certain cancers.
The clinical application mechanism of PDT is very different compared to traditional cancer treatment methods, and its therapeutic effect depends on the photosensitized oxidation of target cells. First, Photosensitizers (PSs) are orally administered or injected into the body and accumulate in the target tissue, and then the photodynamic effect is activated by irradiation with light of appropriate wavelength, and the singlet oxygen or free radicals produced in the presence of molecular oxygen can cause damage to the relevant biomolecules. By understanding the mechanism of action of PDT, we have recognized that PS plays an important role in the study and development of PDT, and its performance determines the therapeutic efficacy of PDT. At the same time, it is important to develop an ideal photosensitizer which has little toxic side effect, strong phototoxicity and relatively selective retention in target tissues.
In recent years, ALA has been shown to be a prodrug of PS protoporphyrin ix (ppix), and its mediated PDT has become one of the most promising areas of PDT research. Compared with other PS, the light-sensitive effect generated by PpIX generated by metabolism of ALA through a heme biosynthetic pathway has a shorter half-life period, and the light-sensitive duration time to normal tissues and the residence time of the medicine in vivo are reduced, so that the toxic and side effects of the medicine to the normal body tissues are reduced. However, ALA has high hydrophilicity, so the cell absorbs ALA poorly, and the bioavailability of ALA is greatly reduced.
In order to improve the bioavailability of ALA and to increase PpIX levels, scientists have conducted extensive research, mainly focusing on the development of ALA prodrugs with better lipid solubility. By analyzing the action mechanism of ALA in vivo, iron chelate enzyme can catalyze PpIX to be converted into heme without photosensitive activity, so that the treatment effect is reduced, the activity of the iron chelate enzyme is shown to be a key factor influencing the accumulation of the PpIX in cells, and the iron chelator can inhibit the activity of the iron chelate enzyme by clearing unstable iron pools in the cells.
It was found that in ALA-PDT the levels of ALA-induced PpIX production could be increased due to the presence of the iron chelators ethylenediaminetetraacetic acid (EDTA) and deferoxamine, but at the same time also showed some adverse effects on the already produced PpIX and poor pharmacokinetic properties. The iron chelator HPO with lower molecular weight and higher lipophilicity can further improve the bioavailability of ALA in vivo by being hybridized with ALA, and meanwhile, iron ions in a chelate body inhibit the activity of iron chelate synthase and improve the PpIX level.
Disclosure of Invention
In order to solve the problems in the prior art, the invention designs and synthesizes a novel anti-tumor active compound with iron chelation and photosensitive activity based on the principles of reasonable drug design, drug-like property and the like. The invention aims to provide a series of novel active compounds of ALA hybrid 3-Hydroxypyridone (HPO) derivatives with iron ion chelating capacity and PDT activity, a preparation method and application in the aspect of tumor resistance.
In order to solve the technical problems, the invention provides the following technical scheme:
in a first aspect, the present invention provides an ALA hybrid 3-hydroxypyridone derivative of formula (I) or formula (II):
Figure BDA0002970338500000021
in the formula (I), the compound is shown in the specification,
R1is C1-C10 alkyl, C3-C6 alkoxy, C6-C10 aryl, preferably R1N-butyl, pentyl, n-hexyl, octyl, methoxypropyl, ethoxypropyl and phenethyl;
r in the formula (II)2is-H or-OH, wherein n1＝1-5。
Further preferably, the ALA hybrid 3-hydroxypyridone derivative is one of the following compounds:
Figure BDA0002970338500000022
particularly preferably, the ALA hybrid 3-hydroxypyridone derivative is compound a2, a3, a4, a7 or b 5.
In a second aspect, the present invention also provides a process for the preparation of ALA hybrid 3-hydroxypyridone derivatives of formula (I) above, said process comprising:
(5) dissolving a compound 10, benzyl bromide and an alkaline substance E1 in a solvent E1, carrying out reflux reaction for 0.5-2 h (preferably 0.5-1 h, particularly preferably 1h), and carrying out aftertreatment on the obtained reaction liquid E1 to obtain a compound 11, wherein the compound is E1; the mass ratio of the compound 10, the benzyl bromide and the alkaline substance E1 is 1: 1-4: 1-4, preferably 1: 1-2: 1 to 2, particularly preferably 1:1.1: 1.1; the alkaline substance is one or a mixture of more than two of potassium carbonate, potassium hydroxide, sodium carbonate and sodium bicarbonate, preferably potassium carbonate or sodium hydroxide, and particularly preferably potassium carbonate;
Figure BDA0002970338500000031
(6) dissolving the compound 11, 3, 4-dihydro-2H-pyran and p-toluenesulfonic acid in the step (5) in dichloromethane, reacting for 4-12H at 20-60 ℃ (preferably room temperature 25 ℃), and performing aftertreatment on the obtained reaction liquid F1 to obtain F1 to obtain a compound 12; the mass ratio of the compound 11, the 3, 4-dihydro-2H-pyran to the p-toluenesulfonic acid is 1: 2-6: 0.1 to 0.6, preferably 1: 2-3: 0.1 to 0.3, particularly preferably 1:2: 0.2;
Figure BDA0002970338500000032
(7) reacting the compound 12, R in the step (6)1NH2Dissolving in a solvent G1, carrying out reflux reaction for 12-36 h (preferably 14-30 h, particularly preferably 18h), and concentrating the obtained reaction solution under reduced pressure to obtain a crude product containing the compound 13; dissolving the crude product containing the compound 13 in ethanol, adjusting the pH to 1-2 (preferably 1) by using concentrated hydrochloric acid, continuously performing reflux reaction for 2-8 h (preferably 2-5 h), and performing aftertreatment on the obtained reaction liquid G1 to obtain G1 to obtain a compound shown in a formula 2; the compound 12 and R1NH2The ratio of the amounts of substances (1): 4 to 10, preferably 1:4 to 8, particularly preferably 1: 6;
Figure BDA0002970338500000033
(8) dissolving the compound 4, Dicyclohexylcarbodiimide (DCC) and 4-Dimethylaminopyridine (DMAP) in the step (1) in an organic solvent H1 under a protective atmosphere (preferably argon or nitrogen, more preferably argon), stirring for 0.5-1H (preferably 45 minutes) at 10-60 ℃ (preferably room temperature 25 ℃), slowly dropwise adding the compound 2 dissolved in the organic solvent Y1 in the step (7), continuously reacting for 6-24H (preferably 8-12H, particularly preferably 12H) after dropwise adding, and performing aftertreatment on the obtained reaction liquid H1 to obtain a compound 5, wherein the compound is H1; the amount ratio of the compound 4, dicyclohexylcarbodiimide, 4-dimethylaminopyridine to the compound 2 is 1: 1-2: 0.1-0.8: 0.6-1.2 (preferably 1:1:0.17: 0.83);
Figure BDA0002970338500000034
(9) dissolving the compound 5 in the step (8) in ethyl acetate J1, dropwise adding an anhydrous ethyl acetate solution of saturated hydrogen chloride at-10-0 ℃ (preferably-10 ℃), reacting for 1h after dropwise adding, transferring to room temperature (25 ℃) to continue reacting for 2-16 h (preferably 4h), and carrying out aftertreatment J1 on the obtained reaction liquid J1 to obtain a compound 15; the volume of the anhydrous ethyl acetate solution of saturated hydrogen chloride is 1-5 mL/mmol based on the substance of the compound 5;
Figure BDA0002970338500000041
(10) dissolving the compound 15 in the step (9) in an organic solvent K1, adding a palladium-carbon catalyst (mass fraction is 5%), stirring and reacting for 1-16 h (preferably 1-8 h, particularly preferably 2h) at 10-60 ℃ (preferably room temperature 25 ℃) under a hydrogen atmosphere, and carrying out aftertreatment on the obtained reaction liquid K1 with K1 to obtain an ALA hybrid 3-hydroxypyridone derivative shown in the formula (I); the mass ratio of the compound 15 to the palladium carbon catalyst is 1:0.1 to 0.4, preferably 1:0.1 to 0.3;
Figure BDA0002970338500000042
wherein, the compound 2, 5, 13, 15, the formula (I) or R1NH2In, R1Is C1-C10 alkyl, C3-C6 alkoxy, C6-C10 aryl, preferably R1N-butyl, pentyl, n-hexyl, octyl, methoxypropyl, ethoxypropyl and phenethyl.
Preferably, the ALA hybrid 3-hydroxypyridone derivative of formula (I) is one of the following compounds:
Figure BDA0002970338500000043
particularly preferably, the ALA hybrid 3-hydroxypyridone derivative shown in the formula (I) is a compound a2, a3, a4 or a 7.
Further, in the step (5), the solvent E1 is one or a mixed solvent of two or more of water, acetone, ethanol, methanol, dichloromethane, chloroform, carbon tetrachloride, acetonitrile, benzene, toluene, dimethyl sulfoxide, dioxane and N, N-dimethylformamide, and preferably a mixed solvent of water and ethanol in a volume ratio of 1:1.
The volume of the solvent E1 in the step (5) is 1-6 mL/mmol based on the substance of the compound 10.
The post-treatment E1 in the step (5) is as follows: the reaction solution E1 was cooled to room temperature, concentrated, dissolved in dichloromethane, washed with water, dried over anhydrous sodium sulfate, filtered, evaporated to remove the solvent and dried to give compound 11.
The volume of the dichloromethane in the step (6) is 1.5-3 mL/mmol based on the substance of the compound 11.
The post-treatment F1 in the step (6) is as follows: and (3) washing the reaction solution F1 with a sodium carbonate solution with the mass fraction of 5%, then washing with water, drying with anhydrous sodium sulfate, filtering, distilling under reduced pressure to remove the solvent, and drying in vacuum to obtain the compound 12.
In the step (7), the solvent G1 is one or a mixed solvent of more than two of water, acetone, ethanol, methanol, chloroform, carbon tetrachloride, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide, preferably ethanol or a mixed solvent of ethanol and water with a volume ratio of 1: 1;
the volume of the ethanol in the step (7) is 1-5 mL/mmol based on the substance of the compound 13;
the post-treatment G1 in the step (7) is as follows: cooling the reaction solution G1 to room temperature, concentrating under reduced pressure, dissolving in water, washing with diethyl ether, adjusting the pH to 8-10 (10M sodium hydroxide solution), extracting with dichloromethane, combining organic layers, drying with anhydrous sodium sulfate, filtering, concentrating, and recrystallizing with a mixed solution of diethyl ether and methanol at a volume ratio of 12:1 to obtain a compound 2.
In the step (8), the organic solvent H1 is one or more of acetone, ethanol, methanol, dichloromethane, chloroform, carbon tetrachloride, toluene, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide, and is preferably a mixed solvent of dichloromethane and N, N-dimethylformamide in a volume ratio of 2:1. The volume of the organic solvent H1 is 1-10 mL/mmol, preferably 3.75mL/mmol, based on the substance of the compound 4.
In the step (8), the organic solvent Y1 is dichloromethane. The volume usage amount of the organic solvent Y1 is 1-8 mL/mmol, preferably 3mL/mmol, based on the substance amount of the compound 2.
The post-treatment H1 in the step (8) is as follows: filtering the reaction liquid H1, concentrating the filtrate under reduced pressure, adding dichloromethane for dissolving, washing with a saturated sodium bicarbonate solution and a saturated sodium chloride solution in sequence, drying with anhydrous sodium sulfate, filtering, concentrating under reduced pressure, performing silica gel column chromatography, performing gradient elution with a dichloromethane-methanol mixed solution with a volume ratio of 100-20: 1, collecting an eluent containing a target product, concentrating, and drying to obtain a compound 5.
Further, the amount of the substance of the compound 5 in the step (9) is 0.1-2 mL/mmol, preferably 0.17mL/mmol, based on the volume of the ethyl acetate J1.
The post-treatment J1 in the step (9) is as follows: the reaction solution J1 was subjected to distillation under reduced pressure to remove the solvent, and dried to obtain compound 15.
In the step (10), the organic solvent K1 is one or a mixed solvent of more than two of acetone, ethanol, methanol, dichloromethane, chloroform, carbon tetrachloride, toluene, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide, preferably methanol.
The amount of the compound 15 in the step (10) is 0.1 to 0.2mmol/mL based on the volume of the organic solvent G.
The post-treatment K1 in the step (10) is as follows: filtering the reaction solution K1, distilling the filtrate under reduced pressure to remove the solvent, and recrystallizing with a mixed solvent of methanol and diethyl ether (the volume ratio of methanol to diethyl ether is 10:1) to obtain the ALA hybrid 3-hydroxypyridone derivative shown in formula (I).
In a third aspect, the present invention also provides a process for the preparation of ALA hybrid 3-hydroxypyridone derivatives of formula (II) above, said process comprising:
1) dissolving a compound 1 or a compound 9, benzyl bromide and an alkaline substance A2 in an organic solvent A2, carrying out reflux reaction for 4-18 h (preferably 4-12h), and carrying out aftertreatment A2 on the obtained reaction liquid A2 to obtain a compound 16; the amount ratio of the compound 1 or the compound 9 to the benzyl bromide and the alkaline substance A2 is 1: 1-4: 1-4, preferably 1: 1-3: 1-3; the alkaline substance A2 is one or a mixture of more than two of potassium carbonate, potassium hydroxide, sodium carbonate, sodium bicarbonate and triethylamine, preferably potassium carbonate or sodium hydroxide;
Figure BDA0002970338500000051
2) subjecting the compound 16, NH in the step 1)2(CH2)n1COOH and an alkaline substance B2 are dissolved in a solvent B2, reflux reaction is carried out for 4-24 h (preferably 8-12 h), and the obtained reaction liquid B2 is subjected to post-treatment B2 to obtain a compound 3; the compound 16, NH2(CH2)n1The ratio of the amounts of COOH and of the basic substance B2 was 1: 1-4: 2-8, preferably 1: 1-2: 2 to 4, particularly preferably 1:1.2: 2.4; the alkaline substance B2 is one or a mixture of more than two of potassium carbonate, potassium hydroxide, sodium carbonate, sodium bicarbonate and triethylamine, preferably sodium hydroxide or potassium hydroxide;
Figure BDA0002970338500000061
3) dissolving the compound 3 in the step 2) in an organic solvent C2, sequentially adding a compound 6 and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethylurea Hexafluorophosphate (HATU), slowly dropwise adding N-methylmorpholine in an ice water bath, stirring for reacting for 45 minutes, completely dissolving solids in the system, then transferring to 20-50 ℃ (preferably room temperature 25 ℃) for continuously reacting for 2-20 hours (preferably 8-16 hours), and carrying out aftertreatment on the obtained reaction liquid C2 to obtain a compound 7, wherein the reaction liquid C2 is subjected to aftertreatment; the mass ratio of the compound of formula 3, 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate to the compound 6 to N-methylmorpholine is 1: 1-6: 1-6: 2-8, preferably 1: 1-4: 1-4: 2-6 (particularly preferably 1:2:1.5: 3);
Figure BDA0002970338500000062
4) dissolving the compound 7 in the step 3) in an organic solvent D2, adding a palladium-carbon catalyst (the mass fraction is 5%), stirring and reacting for 4-18 h at 10-50 ℃ (preferably at room temperature of 25 ℃) under a hydrogen atmosphere, and carrying out aftertreatment on the obtained reaction liquid D2 to obtain an ALA hybrid 3-hydroxypyridone derivative shown in a formula (II); the mass ratio of the compound of the formula 3 to the palladium carbon is 1:0.1 to 0.4; preferably 1:0.1 to 0.2, particularly preferably 1: 0.2.
Figure BDA0002970338500000063
In the compounds 3, 7 and 16, R3Is H-or PhCHO-; NH (NH)2(CH2)n1COOH or R in the formula (II)2is-H or-OH, wherein n1＝1-5。
Further preferably, the ALA hybrid 3-hydroxypyridone derivative represented by formula (II) is one of the following compounds:
Figure BDA0002970338500000064
Figure BDA0002970338500000071
particularly preferably, the ALA hybrid 3-hydroxypyridone derivative shown in the formula (II) is a compound b 5.
Further, the organic solvent a2 in step 1) is one or a mixed solvent of two or more of acetone, ethanol, methanol, dichloromethane, chloroform, carbon tetrachloride, acetonitrile, toluene, dimethyl sulfoxide, dioxane, N-dimethylformamide, and N, N-dimethylacetamide, preferably acetone or carbon tetrachloride, and particularly preferably acetone.
The volume of the organic solvent A2 in the step 1) is 1-4 mL/mmol based on the substance of the compound 9.
The post-treatment A2 in the step 1) is as follows: cooling the reaction solution A2 to room temperature, concentrating under reduced pressure, dissolving with dichloromethane, washing with water, drying with anhydrous sodium sulfate, filtering, concentrating under reduced pressure, performing silica gel column chromatography, performing gradient elution with a mixed solution of n-hexane and ethyl acetate at a volume ratio of 8-3: 1, collecting an eluent containing a target product, concentrating, and drying to obtain the compound 16.
Further, in the step 2), the solvent B2 is one or a mixed solvent of two or more of water, acetone, ethanol, methanol, dichloromethane, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide, and preferably ethanol or a mixed solvent of ethanol and water in a volume ratio of 1:1.
The volume of the solvent B2 in the step 2) is 4-10 mL/mmol based on the substance of the compound 16;
the post-treatment B2 in the step 2) is as follows: cooling the reaction solution B2 to room temperature, concentrating under reduced pressure, adding water for dissolving, washing with dichloromethane, adjusting the pH to 1-2 by using 2mol/L hydrochloric acid, extracting with dichloromethane, combining organic layers, drying with anhydrous sodium sulfate, filtering, evaporating the filtrate under reduced pressure to remove the solvent, drying in vacuum, and recrystallizing the obtained solid with a mixed solvent of methanol and ether in a volume ratio of 1:10 to obtain the compound 3.
The organic solvent C2 in the step 3) is one or a mixed solvent of more than two of acetone, ethanol, methanol, acetic acid, dichloromethane, acetonitrile, dimethyl sulfoxide, dioxane, piperidine, N-dimethylformamide and N, N-dimethylacetamide, preferably N, N-dimethylformamide.
The post-treatment C2 in the step 3) is as follows: adding water into the reaction liquid C2, extracting with dichloromethane, combining organic layers, washing with 0.1M hydrochloric acid and saturated sodium bicarbonate in sequence, drying with anhydrous sodium sulfate, filtering, concentrating, performing silica gel column chromatography, and performing silica gel column chromatography according to the volume ratio of 80: -20: the mixed solution of dichloromethane and methanol of 1 is used as eluent for gradient elution, the eluent containing the target product is collected, concentrated and dried to obtain the compound 7.
The amount of the compound of formula 3 in the step 4) is 0.01-0.12 mmol/mL based on the volume of the organic solvent D2.
The organic solvent D2 in the step 4) is one or a mixed solvent of more than two of acetone, ethanol, methanol, dichloromethane, chloroform, carbon tetrachloride, toluene, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide, and preferably methanol or ethanol.
The post-treatment D2 in the step 4) is as follows: after the reaction is finished, filtering the reaction solution, concentrating the filtrate under reduced pressure, and recrystallizing with methanol/diethyl ether to obtain the compound shown in the formula (II); filtering the reaction liquid D2, taking the filtrate, distilling the filtrate under reduced pressure to remove the solvent, and recrystallizing with a mixed solvent of methanol and diethyl ether (the volume ratio of the methanol to the diethyl ether is 1:10) to obtain the ALA hybrid 3-hydroxypyridone derivative shown in the formula (II).
The invention also recommends the preparation of the above compounds 9, 10 according to the following method:
(1) compound 14, di-tert-butyl dicarbonate (Boc)2O) and an alkaline substance A1 are dissolved in an organic solvent A1, the mixture is reacted for 18 to 36 hours (preferably 20 to 25 hours) at the temperature of 20 to 60 ℃ (preferably 25 to 50 ℃, particularly preferably room temperature), and the obtained reaction liquid A1 is subjected to post-treatment A1 to obtain a compound 4 (ALA protected by Boc); the mass ratio of the compound 14, the di-tert-butyl dicarbonate to the basic substance A1 is 1: 1-1.5: 4-12, preferably 1:1.1: 6; the alkaline substance A1 is one or a mixture of more than two of potassium carbonate, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium bicarbonate or triethylamine (preferably sodium bicarbonate or potassium bicarbonate);
Figure BDA0002970338500000081
further, in the step (1), the organic solvent a1 is one or a mixed solvent of two or more of acetone, ethanol, methanol, dichloromethane, chloroform, carbon tetrachloride, toluene, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide, and N, N-dimethylacetamide, preferably methanol.
Preferably, the volume of the organic solvent A1 in the step (1) is 2-4 mL/mmol based on the substance of the compound 14.
Further, the post-treatment a1 in step (1) is: and filtering the reaction solution A1, distilling the filtrate under reduced pressure to remove the solvent, dissolving the solvent in water, acidifying the solution to pH 1-2 with a 10% potassium bisulfate solution by mass fraction, extracting the solution with ethyl acetate, combining organic layers, washing the organic layers with saturated saline solution, drying the organic layers with anhydrous sodium sulfate, filtering, distilling the filtrate under reduced pressure to remove the solvent, and drying to obtain the compound 4.
(2) Reacting the compound 1 with thionyl chloride at room temperature for 3-9 h (preferably 4-6 h), and carrying out aftertreatment on the obtained reaction liquid B1 to obtain a compound 8, wherein B1 is the first-mentioned chemical substance; the volume of thionyl chloride is 0.5 to 1mL/mmol (preferably 0.6mL/mmol) based on the amount of the substance of Compound 1; in this case thionyl chloride acts as both solvent and reactant.
Figure BDA0002970338500000082
The post-treatment B1 in the step (2) is as follows: adding water into the reaction liquid B1 for quenching, filtering to obtain a crude product (a light yellow substance) containing the compound 8, washing the crude product containing the compound 8 by using n-hexane, and drying to obtain the compound 8.
(3) Dissolving the compound 8 in the step (2) in water C1, stirring for 5 minutes at 40-60 ℃ (preferably 40-55 ℃), then adding zinc powder with the amount equivalent of 1-6 substances (preferably 2-3 eq), and then slowly dropwise adding concentrated hydrochloric acid, wherein the temperature is controlled at 70-80 ℃. After the dropwise addition of the concentrated hydrochloric acid is finished, maintaining the temperature at 75 ℃, continuing to react for 6-8 hours, and performing aftertreatment on the obtained reaction liquid C1 to obtain a compound 9 through C1; the mass ratio of the compound 8 to the HCl in the concentrated hydrochloric acid is 1: 0.8-2, preferably 1: 0.8 to 1.5, particularly preferably 1: 1.17;
Figure BDA0002970338500000083
the volume of the water C1 in the step (3) is 0.5-1mL/mmol based on the substance of the compound 8;
the post-treatment C1 in the step (3) is as follows: filtering the reaction solution C1 while the reaction solution is hot, extracting the filtrate with dichloromethane, combining organic layers, drying with anhydrous sodium sulfate, finally removing the solvent by reduced pressure distillation, recrystallizing isopropanol, and drying to obtain a compound 9;
(4) dissolving the compound 9 in the step (3) by using 1.1mol/L sodium hydroxide aqueous solution, stirring for 40 minutes at room temperature, slowly dropwise adding 37% by mass of formaldehyde aqueous solution, reacting for 18-24 hours, and performing aftertreatment on the obtained reaction liquid D1 to obtain D1 to obtain a compound 10; the ratio of the amount of the compound 9 to the amount of formaldehyde in the aqueous formaldehyde solution is 1: 1-2, preferably 1: 1.1;
Figure BDA0002970338500000091
further, the volume of the 1.1mol/L aqueous solution of sodium hydroxide in the step (4) is 1 to 3mL/mmol based on the substance of the compound 8.
The post-treatment D1 in the step (4) is as follows: the reaction solution D1 was adjusted to pH 1 with concentrated hydrochloric acid, cooled in an ice bath to precipitate crystals, filtered, and the filter cake was dried to obtain compound 10.
In a fourth aspect, the invention also provides application of the ALA hybrid 3-hydroxypyridone derivative shown in the formula (I) or the formula (II) in preparing a photodynamic therapy medicament for preventing or treating tumors.
Further, the tumor is squamous cell carcinoma, skin cancer, basal cell carcinoma, or skin cancer.
Preferably, the tumor cells are HeLa cells, MCF-7 cells or A375 cells.
Further preferably, the ALA hybrid 3-hydroxypyridone derivative is compound a2, a3, a4, a7 or b 5.
Further, the light is blue light.
Compared with the prior art, the invention has the beneficial effects that: the invention synthesizes a series of novel compounds with PDT activity, and has obvious advantages in the aspect of anti-tumor activity.
Drawings
FIG. 1: the phototoxicity of ALA, ALA + CP20 and ALA-HPO hybrid a1-a7 (concentration 100-400 mu M) in a HeLa cell line is evaluated by an MTT method, blue light irradiation (2.5J cm < -2 >), and the compound and cells are incubated for 4h.
FIG. 2: the phototoxicity of ALA, ALA + CP20 and ALA-HPO hybrid a1-a7 (concentration is 100-400 mu M) in an MCF-7 cell line is evaluated by an MTT method, blue light irradiation (2.5J. cm < -2 >), and the compound and cells are incubated for 4h.
FIG. 3: the phototoxicity of ALA, ALA + CP20 and ALA-HPO hybrid a1-a7 (concentration 100-400 μ M) in an A375 cell line was evaluated by MTT method, blue light irradiation (2.5J. cm-2) was performed, and the compound was incubated with the cells for 4h.
FIG. 4: dark toxicity of ALA, ALA + CP20 and ALA-HPO hybrid a1-a7 (concentration 100 μ M) in the a375 cell line was evaluated by MTT method, (a) compound was incubated with HeLa cells for 4h, (B) compound was incubated with MCF-7 cells for 4h, and (C) compound was incubated with a375 cells for 4h.
FIG. 5: the phototoxicity of ALA, ALA + CP20 and ALA-HPO hybrid B1-B10 (the concentration is 100-400 mu M) in a HeLa cell line is evaluated by an MTT method, blue light is irradiated (5J cm < -2 >), the compound (A) and the cell are incubated for 4h, and the compound (B) and the cell are incubated for 24h.
FIG. 6: the phototoxicity of ALA, ALA + CP20 and ALA-HPO hybrid B1-B10 (concentration is 100-400 mu M) in an MCF-7 cell line is evaluated by an MTT method, blue light is irradiated (5J cm < -2 >), (A) the compound is incubated with cells for 4h, and (B) the compound is incubated with the cells for 24h.
FIG. 7: the phototoxicity of ALA, ALA + CP20 and ALA-HPO hybrid B1-B10 (concentration is 100-400 mu M) in an A375 cell line is evaluated by an MTT method, blue light irradiation is carried out (5J cm < -2 >), the compound (A) and the cell are incubated for 4h, and the compound (B) and the cell are incubated for 24h.
FIG. 8: evaluating the dark toxicity of ALA, ALA + CP20 and ALA-HPO hybrid B1-B10 (the concentration is 100-400 mu M) in a HeLa cell line by using an MTT method, wherein the compound is incubated for 4h with the cells, and the compound is incubated for 24h with the cells.
FIG. 9: evaluating the dark toxicity of ALA, ALA + CP20 and ALA-HPO hybrid B1-B10 (the concentration is 100-400 mu M) in an MCF-7 cell line by using an MTT method, wherein the compound is incubated for 4h with the cells, and the compound is incubated for 24h with the cells.
FIG. 10: evaluating the dark toxicity of ALA, ALA + CP20 and ALA-HPO hybrid B1-B10 (the concentration is 100-400 mu M) in an A375 cell line by using an MTT method, wherein the compound A and the cell are incubated for 4h, and the compound B and the cell are incubated for 24h.
Detailed Description
The present invention will be further described with reference to specific examples, but the present invention is not limited to these examples.
The mass fraction of concentrated hydrochloric acid used below was 37%.
The formula for calculating the yield (irrespective of purity) in the following examples is:
Y＝(myield of the product/MProduct of)/NRaw materials
mYield of the productTo preserve the quality of the product of impurities, MProduct ofIs the relative molecular mass of the target product, NRaw materialsThe amount of material that is the reactant for which the amount of material is smaller.
Example 1
(1) In a 250mL round bottom flask was added ALA hydrochloride (4.20g,25.0mmol), NaHCO3(12.60g,150mmol) and Boc2O (6.00g,27.5mmol) and 75mL of absolute methanol as a solvent are added, the mixture is stirred at room temperature under the protection of argon, the progress of the reaction is detected by TLC during the reaction, and the color is developed in an iodine cylinder. After 24h, the reaction was stopped, at which point the reaction solution appeared beige, and unreacted NaHCO was added3The solid was removed by filtration using a buchner funnel and washed with methanol, the filtrate was distilled under reduced pressure and dissolved in 30mL of water, then acidified with 10% by mass potassium hydrogensulfate solution to pH 1-2, extracted 3 times with ethyl acetate, the organic layers were combined and washed with saturated brine (3 × 25mL), dried over anhydrous sodium sulfate for 12h, filtered to remove the solid, distilled under reduced pressure to remove the solvent, and dried under vacuum to give N-Boc-ALA (5.05g, 87%) as a pale yellow solid with HPLC assay content of not less than 98%.
(2) Kojic acid (23.86g,168mmol) and 101mL of thionyl chloride are added into a 500mL round-bottom flask, the mixture reacts for 5 hours at room temperature, water is added for quenching, sodium hydroxide solution is used for absorbing tail gas to obtain yellow solid suspended matter, a Buchner funnel is used for filtering, a filter cake is washed by n-hexane and dried in vacuum, 22.80g of 2-chloromethyl-5-hydroxypyran-4-one white solid compound is obtained, the yield is 95%, and the HPLC analysis content is not less than 96%.
(3) 2-chloromethyl-5-hydroxypyran-4-one (25.69g, 160.0mmol) and 112mL of water were charged into a 500mL round-bottomed flask, stirred well in an oil bath at 50 ℃ and then zinc powder (20.92g,320.0mmol) was added thereto, stirring was continued for 5 minutes, 96mL of concentrated hydrochloric acid was slowly dropped into the reaction system, maintaining the temperature of the system at 70-80 ℃, maintaining the temperature at 75 ℃ after dropwise adding concentrated hydrochloric acid, detecting the reaction condition by TLC, after 8h of reaction, the mixture is filtered while the mixture is hot, dichloromethane is used for extraction (6X 60mL), organic phases are combined, dried by anhydrous sodium sulfate, the organic solvent is removed by reduced pressure distillation to obtain brown solid, the brown solid is recrystallized by using isopropanol and then naturally dried by air to obtain 13.10g of a white solid compound of 2-methyl-5-hydroxypyran-4-one, the yield is 65%, and the HPLC analysis content is more than or equal to 97%.
(4) 2-methyl-5-hydroxy-pyran-4-one (13.10g, 104.0mmol) and 112mL of 1.1mol/L sodium hydroxide solution are added into a 500mL round bottom flask, stirred for 40 minutes at room temperature, aqueous formaldehyde solution (3.43g, 114.4mmol) with the mass fraction of 37% is slowly dropped into the flask, the reaction is monitored for 24 hours, the pH value is adjusted to 1 by concentrated hydrochloric acid after the reaction is finished, solid is precipitated after ice bath cooling, filtered and dried in vacuum, and 12.17g of 3-hydroxy-2-hydroxymethyl-6-methyl-pyran-4-one white solid compound is obtained, the yield is 75%, and the HPLC analysis content is more than or equal to 97%.
(5) 3-hydroxy-2-hydroxymethyl-6-methyl pyran-4-ketone (12.17g, 78mmol), potassium carbonate (12.92g,93.6mmol), ethanol and water (1:1)140mL, benzyl bromide (16g,93.6mmol) are sequentially added into a 250mL round bottom flask, reflux reaction is carried out for 1h, the reaction progress is detected by using TLC during the reaction, after the reaction is finished, the mixture is cooled to room temperature, the solvent is concentrated in vacuum, dichloromethane is dissolved, washed for 3 times by using water, dried over anhydrous sodium sulfate overnight, filtered, the solvent is removed by reduced pressure distillation, and dried in vacuum to obtain light yellow solid 3-benzyloxy-2-hydroxymethyl-6-methyl pyran-4-ketone 18.23g, the yield is 95%, and the HPLC analysis content is more than or equal to 96%.
(6) 3-benzyloxy-2-hydroxymethyl-6-methyl pyran-4-one (18.23g, 74.1mmol), p-toluenesulfonic acid (2.55g, 14.8mmol), 120mL dichloromethane and 3, 4-dihydro-2H-pyran (12.47g, 148.2mmol) are sequentially added into a 250mL round bottom flask, the mixture is reacted for 6H at room temperature, the reaction progress is monitored by TLC, after the reaction is finished, the mixture is washed by 5% sodium carbonate solution and water for 3 times respectively, dried over anhydrous sodium sulfate overnight, filtered, the solvent is removed by reduced pressure distillation, and after vacuum drying, 23.71g of yellow oily product is obtained, the yield is 97%, and the HPLC analysis content is more than or equal to 95%.
(7) The yellow oily substance (3.30g, 10.0mmol) obtained above and n-butylamine (4.39g, 60.0mmol) were added to a 100mL round bottom flask, 20mL of ethanol and water (V: V ═ 1:1) were used as solvents, the reaction was carried out under reflux for 18 hours while monitoring the progress of the reaction by TLC, after the completion of the reaction, the solvent was distilled off under reduced pressure to obtain a brown oily substance, the solvent was dissolved in ethanol and adjusted to pH 1 with concentrated hydrochloric acid, the reflux was continued for 4 hours, the progress of the reaction was monitored by TLC, after the completion of the reaction, the reaction was cooled to room temperature, the solvent was distilled off under reduced pressure, the residue was dissolved in water and washed twice with ether, then adjusted to pH 9 with 10mol/L sodium hydroxide solution, dichloromethane was extracted (3 × 20mL), the organic layers were combined, dried over anhydrous sodium sulfate, filtered, the solvent was distilled off under reduced pressure to obtain a pale yellow solid, and the ether/methanol (V: V ═ 12:1) was recrystallized to obtain 3-benzyloxy-1-butyl-2- 0.96g of white solid compound of 6-methylpyridine-4-ketone, the yield is 32 percent, and the HPLC analysis content is more than or equal to 96 percent.
(8) DCC (0.50g, 2.4mmol), DMAP (0.049g, 0.4mmol) and N-Boc-ALA (0.555g, 2.4mmol) were dissolved in dichloromethane (6mL) and DMF (3mL) under argon and stirred at room temperature for 45 min. Placing 3-benzyloxy-1-butyl-2-hydroxymethyl-6-methylpyridin-4-one (0.602g and 2mmol) dissolved in dichloromethane (6mL) in a constant-pressure dropping funnel, dropwise adding the mixture within 40 minutes, reacting the reaction solution at room temperature for 12 hours, monitoring the reaction progress by TLC, filtering out precipitates after the reaction is finished, distilling the filtrate under reduced pressure to remove the solvent, dissolving the residue in dichloromethane, washing the residue with saturated sodium bicarbonate solution and saturated sodium chloride solution in sequence, drying the residue with anhydrous sodium sulfate, filtering, concentrating the organic layer under reduced pressure, purifying the organic layer by silica gel column chromatography (dichloromethane: methanol is 100-20: 1 volume ratio, gradient elution), collecting the eluent when the dichloromethane: methanol is 40: 1-20: 1, concentrating and drying to obtain 0.504g of a yellow oily product under reduced pressure, the yield is 49%, and the HPLC analysis content is more than or equal to 98%.
(9) The yellow oily compound (0.257g, 0.5mmol) is added into a 25mL round bottom flask, dissolved in 3mL ethyl acetate solution, saturated ethyl acetate solution of hydrogen chloride (3mL) is placed in a constant pressure dropping funnel, slowly dropped at-10 ℃, continuously reacted for 1h, then moved to room temperature for continuous reaction for 4h, the reaction progress is monitored by TLC, after the reaction is finished, the organic solvent is removed by reduced pressure distillation, vacuum drying is carried out, and the obtained intermediate compound is 0.205g of yellow oily matter, the yield is 91%, and the HPLC analysis content is more than or equal to 97%.
(10) 0.205g of the above compound was weighed out and dissolved in 4mL of methanol and placed in a 25mL round-bottomed flask, and the reaction was carried out with 0.041g of 5% Pd/C (Saien chemical technology, Inc., E060062-25g) by catalytic hydrogenation under 30psi of hydrogen gas pressure and stirred at room temperature for 2 hours while monitoring the progress of the reaction by TLC, after the completion of the reaction, Pd/C was removed by filtration, the filtrate was distilled under reduced pressure to remove the solvent, and recrystallized from ether/methanol (V: 10:1) and air-dried to obtain a white solid compound a 10.079g, with a yield of 48% and an HPLC analytical content of 99% or more.
m.p.＝149.1-151.9℃.1H NMR(400MHz,DMSO-d6)δ8.32(s,3H),7.27(s,1H),5.37(s,2H),4.23(t,J＝5.2Hz,2H),3.96(m,2H),2.85(t,J＝4.4Hz,2H),2.63(s,3H),2.61(t,J＝4.4Hz,2H),1.73(m,2H),1.24(m,2H),0.94(t,J＝4.8Hz,3H),13C NMR(400MHz,DMSO-d6)δ203.0,172.0,160.9,148.9,144.7,135.5,114.3,56.6,51.4,46.9,34.7,31.5,27.4,20.4,19.8,13.9；ESI-HRMS:m/z calcd for C16H25N2O5[M+H]+:325.1758；found:325.1762.
Example 2
(1) To a 100mL round bottom flask were added the yellow oil (3.30g, 10.0mmol) obtained in example 1(6) and pentylamine (5.23g, 60.0mmol), ethanol and 20mL of water (V: V ═ 1:1) as solvents, and the reaction was carried out under reflux for 18 hours while monitoring the progress of the reaction by TLC, after the completion of the reaction, the solvent was distilled off under reduced pressure to obtain a brown oil, the solvent was dissolved in ethanol and adjusted to pH 1 with concentrated hydrochloric acid, followed by refluxing for 4 hours, the progress of the reaction was monitored by TLC, after the completion of the reaction, the reaction was cooled to room temperature, the solvent was distilled off under reduced pressure to obtain a residue, the residue was dissolved in water and washed twice with diethyl ether, then adjusted to pH 9 with 10mol/L sodium hydroxide solution, dichloromethane was extracted (3 × 20mL), the organic layers were combined, dried over anhydrous sodium sulfate, filtered, the solvent was distilled off under reduced pressure to obtain a pale yellow solid, and the pale yellow solid was recrystallized from diethyl ether/methanol (V: V ═ 12:1) to obtain 3-benzyloxy-1 0.946g of white solid compound of 2-hydroxymethyl-6-methylpyridine-4-ketone, 30 percent of yield and more than or equal to 96 percent of HPLC analysis content.
(2) DCC (0.50g, 2.4mmol), DMAP (0.049g, 0.4mmol) and N-Boc-ALA (0.555g, 2.4mmol) prepared in example 1(1) were dissolved in dichloromethane (6mL) and DMF (3mL) under an argon atmosphere and stirred at room temperature for 45 min. Placing 3-benzyloxy-1-pentyl-2-hydroxymethyl-6-methylpyridin-4-one (0.63g, 2mmol) dissolved in dichloromethane (6mL) in a constant pressure dropping funnel, dropwise adding in 40 minutes, reacting the reaction solution at room temperature for 12h, monitoring the reaction progress by TLC, filtering out the precipitate after the reaction is finished, distilling the filtrate under reduced pressure to remove the solvent, dissolving the residue in dichloromethane, washing the residue with saturated sodium bicarbonate solution and sodium chloride solution in sequence, drying with anhydrous sodium sulfate, filtering, concentrating the organic layer under reduced pressure, purifying by silica gel column chromatography (gradient elution of dichloromethane: methanol 100: 1-20: 1), collecting the eluent when dichloromethane: methanol is 40: 1-20: 1, concentrating under reduced pressure, drying to obtain 0.539g of yellow oily product, the yield is 51 percent, and the HPLC analysis content is more than or equal to 95 percent.
(3) The yellow oily compound (0.264g, 0.5mmol) is added into a 25mL round-bottom flask, dissolved in 3mL ethyl acetate solution, saturated ethyl acetate solution (3mL) of hydrogen chloride is placed in a constant-pressure dropping funnel, slowly dropped at-10 ℃, continuously reacted for 1h, then moved to room temperature for continuous reaction for 4h, the reaction progress is monitored by TLC, after the reaction is finished, the organic solvent is removed by reduced pressure distillation, vacuum drying is carried out, the obtained intermediate compound is 0.214g of yellow oily matter, the yield is 92%, and the HPLC analysis content is more than or equal to 96%.
(4) 0.214g of the above compound was weighed out and dissolved in 4mL of methanol and placed in a 25mL round bottom flask, and then catalyzed and hydrogenated with 0.043g of 5% Pd/C (Saien chemical technology, Inc., E060062-25g) while maintaining 30psi of hydrogen pressure, stirred and reacted at room temperature for 2 hours, while monitoring the progress of the reaction by TLC, after the reaction was completed, palladium/carbon was removed by filtration, the filtrate was distilled under reduced pressure to remove the solvent, and recrystallized with methanol/ether (V: V ═ 10:1) to obtain 20.079g of a white solid compound a, the yield was 46%, and the HPLC analysis content was 99% or more.
m.p.＝149.2-151.4℃.1H NMR(400MHz,DMSO-d6)δ8.40(s,3H),7.37(s,1H),5.37(s,2H),4.24(t,J＝7.6Hz,2H),3.94(m,2H),2.85(t,J＝6.0Hz,2H),2.64(s,3H),2.61(t,J＝6.0Hz,2H),1.74(m,2H),1.35(m,4H),0.90(t,J＝6.4Hz,3H),13C NMR(400MHz,DMSO-d6)δ203.0,172.0,161.1,148.9,144.8,135.5,114.3,56.6,51.6,47.0,34.7,29.4,28.5,27.4,22.1,20.4,14.3；ESI-HRMS:m/z calcd.for C17H27N2O5[M+H]+:339.1914；found:339.1909.
Example 3
(1) The yellow oily substance (3.30g, 10.0mmol) obtained in example 1(6) and n-hexylamine (6.07g, 60.0mmol) were added to a 100mL round bottom flask, 20mL of ethanol and water (V: V ═ 1:1) were used as solvents, and the reaction was carried out under reflux for 18 hours while monitoring the progress of the reaction by TLC, after the completion of the reaction, the solvent was distilled off under reduced pressure to obtain a brown oily substance, the solvent was dissolved in ethanol and adjusted to pH 1 with concentrated hydrochloric acid, the reflux was continued for 4 hours, the progress of the reaction was monitored by TLC, after the completion of the reaction, the reaction was cooled to room temperature, the solvent was distilled off under reduced pressure to obtain a residue, the residue was dissolved in water and washed twice with diethyl ether, then adjusted to pH 9 with 10mol/L sodium hydroxide solution, dichloromethane was extracted (3 × 20mL), the organic layers were combined, dried over anhydrous sodium sulfate, filtered, the solvent was distilled off under reduced pressure, and dried under vacuum to obtain a pale yellow solid, which was recrystallized to obtain 3-benzyloxy-1 (V: 12:1) and 1.22g of hexyl-2-hydroxymethyl-6-methylpyridin-4-one white solid compound, the yield is 37 percent, and the HPLC analysis content is more than or equal to 98 percent.
(2) DCC (0.50g, 2.4mmol), DMAP (0.049g, 0.4mmol) and N-Boc-ALA (0.555g, 2.4mmol) prepared in example 1(1) were dissolved in dichloromethane (6mL) and DMF (3mL) under an argon atmosphere and stirred at room temperature for 45 min. Placing 3-benzyloxy-1-hexyl-2-hydroxymethyl-6-methylpyridin-4-one (0.66g and 2mmol) dissolved in dichloromethane (6mL) in a constant-pressure dropping funnel, dropwise adding the mixture within 40 minutes, reacting the reaction solution at room temperature for 12 hours, monitoring the reaction process by TLC, filtering out precipitates after the reaction is finished, distilling the filtrate under reduced pressure to remove the solvent, dissolving the residue in dichloromethane, washing the residue with saturated sodium bicarbonate solution and sodium chloride solution in sequence, drying the residue with anhydrous sodium sulfate, filtering, concentrating the organic layer under reduced pressure, purifying the organic layer by silica gel column chromatography (gradient elution of dichloromethane: methanol is 100: 1-20: 1), collecting the eluent when the dichloromethane: methanol is 40: 1-20: 1, concentrating under reduced pressure, drying to obtain 0.575g of yellow oily product, the yield is 53 percent, and the content of HPLC analysis is more than or equal to 97 percent.
(3) The yellow oily compound (0.271g, 0.5mmol) is added into a 25mL round-bottom flask, dissolved in 3mL ethyl acetate solution, saturated ethyl acetate solution of hydrogen chloride (3mL) is placed in a constant-pressure dropping funnel, slowly dropped at-10 ℃, continuously reacted for 1h, then moved to room temperature for continuous reaction for 4h, the reaction progress is monitored by TLC, after the reaction is finished, the organic solvent is removed by reduced pressure distillation, vacuum drying is carried out, the obtained intermediate compound is 0.213g of yellow oily matter, the yield is 89%, and the HPLC analysis content is more than or equal to 98%.
(4) 0.213g of the above compound was weighed out and dissolved in 4mL of methanol and placed in a 25mL round bottom flask, and then catalyzed and hydrogenated with 0.043g of 5% Pd/C (Saien chemical technology, Shanghai, Ltd., E060062-25g), and the reaction was stirred at room temperature for 2 hours while monitoring the progress of the reaction by TLC, after the completion of the reaction, palladium/carbon was removed by filtration, the solvent was removed from the filtrate by distillation under reduced pressure, and the white solid compound a30.08g was obtained by recrystallization from methanol/ether (V: V ═ 10:1), the yield was 50%, and the HPLC analysis content was 99% or more.
m.p.＝139.3-141.0℃.1H NMR(400MHz,DMSO-d6)δ8.40(s,3H),7.35(s,1H),5.36(s,2H),4.24(t,J＝8.0Hz,2H),3.94(m,2H),2.85(t,J＝6.4Hz,2H),2.64(s,3H),2.60(t,J＝6.4Hz,2H),1.72(m,2H),1.38(m,2H),1.30(m,4H),0.88(t,J＝6.8Hz,3H),13C NMR(400MHz,DMSO-d6)δ202.9,172.0,161.1,148.9,144.8,135.4,114.3,56.6,51.6,47.0,34.7,31.1,29.6,27.4,26.1,22.5,20.4,14.4；ESI-HRMS:m/z calcd.for C18H29N2O5[M+H]+:353.2071；found:353.2074.
Example 4
(1) The yellow oily substance (3.30g, 10.0mmol) obtained in example 1(6) and octylamine (7.75g, 60.0mmol) were added to a 100mL round bottom flask, 20mL of ethanol and water (V: V ═ 1:1) were used as solvents, and the reaction was carried out under reflux for 18 hours while monitoring the progress of the reaction by TLC, after the completion of the reaction, the solvent was distilled off under reduced pressure to obtain a brown oily substance, the solvent was dissolved in ethanol and adjusted to pH 1 with concentrated hydrochloric acid, the reflux was continued for 4 hours, the progress of the reaction was monitored by TLC, after the completion of the reaction, the reaction was cooled to room temperature, the solvent was distilled off under reduced pressure to obtain a residue, the residue was dissolved in water and washed twice with diethyl ether, then adjusted to pH 9 with 10mol/L sodium hydroxide solution, dichloromethane was extracted (3 × 20mL), the organic layers were combined, dried over anhydrous sodium sulfate, filtered, the solvent was distilled off under reduced pressure, and dried under vacuum to obtain a pale yellow solid, which was recrystallized to obtain 3-benzyloxy-1-octylamine, and further recrystallized to obtain 1.25g of white solid compound of 2-hydroxymethyl-6-methylpyridin-4-one, the yield is 35%, and the HPLC analysis content is more than or equal to 97%.
(2) DCC (0.50g, 2.4mmol), DMAP (0.049g, 0.4mmol) and N-Boc-ALA (0.555g, 2.4mmol) prepared in example 1(1) were dissolved in dichloromethane (6mL) and DMF (3mL) under an argon atmosphere and stirred at room temperature for 45 min. Placing 3-benzyloxy-1-octyl-2-hydroxymethyl-6-methylpyridin-4-one (0.71g and 2mmol) dissolved in dichloromethane (6mL) in a constant-pressure dropping funnel, dropwise adding the mixture within 40 minutes, reacting the reaction solution at room temperature for 12 hours, monitoring the reaction progress by TLC, filtering out precipitates after the reaction is finished, distilling the filtrate under reduced pressure to remove the solvent, dissolving the residue in dichloromethane, washing the residue with saturated sodium bicarbonate solution and sodium chloride solution in sequence, drying the residue with anhydrous sodium sulfate, filtering, concentrating the organic layer under reduced pressure, purifying the organic layer by silica gel column chromatography (gradient elution of dichloromethane: methanol is 100: 1-20: 1), collecting the eluent when the dichloromethane: methanol is 40: 1-20: 1, concentrating under reduced pressure, drying to obtain 0.548g of yellow oily product, the yield is 48%, and the HPLC analysis content is more than or equal to 96%.
(3) The yellow oily compound (0.285g, 0.5mmol) is added into a 25mL round-bottom flask, dissolved in 3mL ethyl acetate solution, saturated ethyl acetate solution of hydrogen chloride (3mL) is placed in a constant-pressure dropping funnel, slowly dropped at-10 ℃, continuously reacted for 1h, then moved to room temperature for continuous reaction for 4h, the reaction progress is monitored by TLC, after the reaction is finished, the organic solvent is removed by reduced pressure distillation, vacuum drying is carried out, the obtained intermediate compound is 0.215g of yellow oily matter, the yield is 85%, and the HPLC analysis content is more than or equal to 95%.
(4) 0.215g of the above compound was weighed out and dissolved in 4mL of methanol and placed in a 25mL round bottom flask, and then catalyzed and hydrogenated with 0.043g of 5% Pd/C (Saien chemical technology, Shanghai, Ltd., E060062-25g), maintaining 30psi of hydrogen pressure, stirred and reacted at room temperature for 2 hours, while monitoring the progress of the reaction by TLC, after the reaction was completed, palladium/carbon was removed by filtration, the filtrate was distilled under reduced pressure to remove the solvent, and recrystallized with methanol/ether (V: V ═ 10:1) to obtain 40.081g of white solid compound a, yield 46%, HPLC assay content of 99% or more.
m.p.＝80.2-82.1℃.1H NMR(400MHz,DMSO-d6)δ8.35(s,3H),7.35(s,1H),5.36(s,2H),4.23(t,J＝8.0Hz,2H),3.94(m,2H),2.84(t,J＝6.4Hz,3H),2.64(s,3H),2.61(t,J＝6.4Hz,2H),1.72(m,2H),1.29(m,10H),0.85(t,J＝6.8Hz,3H),13C NMR(400MHz,DMSO-d6)δ202.9,172.0,161.1,148.8,144.8,135.4,114.3,56.6,51.5,47.0,34.7,31.7,29.7,29.0,28.9,27.4,26.4,22.5,20.4,14.4；ESI-HRMS:m/z calcd.for C20H33N2O5[M+H]+:381.2384；found:381.2384.
Example 5
(1) A yellow oily substance (3.30g, 10.0mmol) obtained in example 1(6) and 3-methoxypropylamine (5.35g, 60.0mmol) were added to a 100mL round-bottom flask, and 20mL of ethanol and water (V: V ═ 1:1) were used as solvents, and the reaction was carried out under reflux for 18 hours while monitoring the progress of the reaction by TLC, and after the completion of the reaction, the solvent was distilled off under reduced pressure to obtain a brown oily substance, the reaction was dissolved in ethanol and adjusted to pH 1 with concentrated hydrochloric acid, followed by refluxing for 4 hours, and the progress of the reaction was monitored by TLC, and after the completion of the reaction, the reaction was cooled to room temperature, the solvent was distilled off under reduced pressure, the residue was dissolved in water and washed twice with diethyl ether, then adjusted to pH 9 with 10mol/L sodium hydroxide solution, dichloromethane was extracted (3 × 20mL), the organic layers were combined, dried over anhydrous sodium sulfate, filtered, the solvent was distilled off under reduced pressure, and dried in vacuo, recrystallization from ether/methanol (V: V ═ 12:1) gave 1.33g of 3-benzyloxy-2-hydroxymethyl-1- (3-methoxypropyl) -6-methylpyridin-4-one as a white solid, 42% yield, with HPLC assay content ≥ 96%.
(2) DCC (0.50g, 2.4mmol), DMAP (0.049g, 0.4mmol) and N-Boc-ALA (0.555g, 2.4mmol) prepared in example 1(1) were dissolved in dichloromethane (6mL) and DMF (3mL) under an argon atmosphere and stirred at room temperature for 45 min. Placing 3-benzyloxy-2-hydroxymethyl-1- (3-methoxypropyl) -6-methylpyridin-4-one (0.63g, 2mmol) dissolved in dichloromethane (6mL) in a constant-pressure dropping funnel, adding dropwise in 40 minutes, reacting the reaction solution at room temperature for 12 hours, monitoring the reaction progress by TLC, filtering out the precipitate after the reaction is finished, distilling the filtrate under reduced pressure to remove the solvent, dissolving the residue in dichloromethane, washing the residue with saturated sodium bicarbonate solution and sodium chloride solution in sequence, drying with anhydrous sodium sulfate, filtering, concentrating the organic layer under reduced pressure, purifying by silica gel column chromatography (gradient elution of dichloromethane: methanol 100: 1-20: 1), collecting the eluent of dichloromethane: methanol 40: 1-20: 1, concentrating under reduced pressure, drying, 0.594g of the product is obtained as yellow oil, the yield is 56%, and the content is more than or equal to 97% by HPLC analysis.
(3) The yellow oily compound (0.265g, 0.5mmol) is added into a 25mL round-bottom flask, dissolved in 3mL ethyl acetate solution, saturated ethyl acetate solution of hydrogen chloride (3mL) is placed in a constant pressure dropping funnel, slowly dropped at-10 ℃, continuously reacted for 1h, then moved to room temperature for continuous reaction for 4h, the reaction progress is monitored by TLC, after the reaction is finished, the organic solvent is removed by reduced pressure distillation, vacuum drying is carried out, and the obtained intermediate compound is 0.219g of yellow oily matter, the yield is 94%, and the HPLC analysis content is more than or equal to 96%.
(4) Weighing 0.219g of the compound, dissolving in 4mL of methanol, placing in a 25mL round bottom flask, carrying out catalytic hydrogenation with 0.044g of 5% Pd/C (Saien chemical technology, Shanghai, Ltd., E060062-25g), maintaining 30psi hydrogen pressure, stirring at room temperature for 2h, monitoring the reaction progress by using TLC, filtering to remove palladium carbon after the reaction is finished, removing the solvent from the filtrate by reduced pressure distillation, recrystallizing with methanol/diethyl ether (V: V ═ 10:1) to obtain 50.095 g of a white solid compound, wherein the yield is 54%, and the HPLC analysis content is more than or equal to 99%.
m.p.＝138.1-140.3℃.1H NMR(400MHz,DMSO-d6)δ8.42(s,3H),7.37(s,1H),5.37(s,2H),4.34(t,J＝7.8Hz,2H),3.94(m,2H),3.42(t,J＝5.6Hz,2H),3.26(s,3H),2.84(t,J＝6.4Hz,3H),2.64(s,3H),2.61(t,J＝6.4Hz,2H),2.00(m,2H),13C NMR(400MHz,DMSO-d6)δ202.9,171.9,161.2,149.1,144.7,135.6,114.3,68.9,58.6,56.6,49.2,47.0,34.7,29.7,27.4,20.4；ESI-HRMS:m/z calcd.for C16H25N2O6[M+H]+:341.1707；found:341.1713.
Example 6
(1) A yellow oily substance (3.30g, 10.0mmol) obtained in example 1(6) and 3-ethoxypropylamine (6.19g, 60.0mmol) were added to a 100mL round bottom flask, and 20mL of ethanol and water (V: V ═ 1:1) were used as solvents, and the reaction was carried out under reflux for 18 hours while monitoring the progress of the reaction by TLC, and after the completion of the reaction, the solvent was distilled off under reduced pressure to obtain a brown oily substance, the reaction was dissolved in ethanol and adjusted to pH 1 with concentrated hydrochloric acid, followed by refluxing for 4 hours, the progress of the reaction was monitored by TLC, and after the completion of the reaction, the reaction was cooled to room temperature, the solvent was distilled off under reduced pressure, the residue was dissolved in water and washed twice with diethyl ether, then adjusted to pH 9 with 10mol/L sodium hydroxide solution, dichloromethane was extracted (3 × 20mL), the organic layers were combined, dried over anhydrous sodium sulfate, filtered, the solvent was distilled off under reduced pressure, and dried in vacuo to obtain, recrystallization from ether/methanol (V: V ═ 12:1) gave 1.56g of 3-benzyloxy-2-hydroxymethyl-1- (3-ethoxypropyl) -6-methylpyridin-4-one as a white solid compound in 47% yield and at a HPLC assay content of 98% or more.
(2) DCC (0.50g, 2.4mmol), DMAP (0.049g, 0.4mmol) and N-Boc-ALA (0.555g, 2.4mmol) prepared in example 1(1) were dissolved in dichloromethane (6mL) and DMF (3mL) under an argon atmosphere and stirred at room temperature for 45 min. Placing 3-benzyloxy-2-hydroxymethyl-1- (3-ethoxypropyl) -6-methylpyridin-4-one (0.66g and 2mmol) dissolved in dichloromethane (6mL) in a constant-pressure dropping funnel, dropwise adding the mixture within 40 minutes, reacting the reaction solution at room temperature for 12 hours, monitoring the reaction progress by TLC, filtering out precipitates after the reaction is finished, distilling the filtrate under reduced pressure to remove the solvent, dissolving the residue in dichloromethane, washing the residue with saturated sodium bicarbonate solution and sodium chloride solution in sequence, drying the residue with anhydrous sodium sulfate, filtering, concentrating the organic layer under reduced pressure, purifying the organic layer by silica gel column chromatography (gradient elution of dichloromethane: methanol is 100: 1-20: 1), collecting the eluent of dichloromethane: methanol is 40: 1-20: 1, concentrating the eluent under reduced pressure, drying, 0.62g of yellow oily product is obtained, the yield is 57%, and the content of HPLC analysis is more than or equal to 97%.
(3) The yellow oily compound (0.272g, 0.5mmol) is added into a 25mL round-bottom flask, dissolved in 3mL ethyl acetate solution, saturated ethyl acetate solution of hydrogen chloride (3mL) is placed in a constant pressure dropping funnel, slowly dropped at-10 ℃, continuously reacted for 1h, then moved to room temperature for continuous reaction for 4h, the reaction progress is monitored by TLC, after the reaction is finished, the organic solvent is removed by reduced pressure distillation, vacuum drying is carried out, and the obtained intermediate compound is 0.209g of yellow oily matter, the yield is 87%, and the HPLC analysis content is more than or equal to 97%.
(4) 0.209g of the above compound was weighed out and dissolved in 4mL of methanol and placed in a 25mL round bottom flask, and then catalyzed and hydrogenated with 0.042g of 5% Pd/C (Saien chemical technology, Shanghai, Ltd., E060062-25g), maintaining 30psi of hydrogen pressure, stirred and reacted at room temperature for 2 hours, while monitoring the progress of the reaction by TLC, after the reaction was completed, palladium/carbon was removed by filtration, the filtrate was distilled under reduced pressure to remove the solvent, and recrystallized with methanol/ether (V: V ═ 12:1) to obtain 60.083g of a white solid compound a, yield 49%, HPLC assay content of 99% or more.
m.p.＝140.1-142.3℃.1H NMR(400MHz,DMSO-d6)δ8.39(s,3H),7.37(s,1H),5.37(s,2H),4.35(t,J＝7.2Hz,2H),3.95(m,2H),3.44(m,4H),2.84(t,J＝6.4Hz,3H),2.64(s,3H),2.60(t,J＝6.4Hz,2H),2.00(m,2H),1.10(t,J＝6.8Hz,3H),13C NMR(400MHz,DMSO-d6)δ202.9,171.9,161.2,149.1,144.7,135.6,114.3,66.7,66.0,56.7,49.2,47.0,34.7,29.8,27.4,20.4,15.5；ESI-HRMS:m/z calcd.for C17H27N2O6[M+H]+:355.1864；found:355.1863.
Example 7
(1) A yellow oily substance (3.30g, 10.0mmol) obtained in example 1(6) and 3-ethoxypropylamine (6.19g, 60.0mmol) were added to a 100mL round bottom flask, and 20mL of ethanol and water (V: V ═ 1:1) were used as solvents, and the reaction was carried out under reflux for 18 hours while monitoring the progress of the reaction by TLC, and after the completion of the reaction, the solvent was distilled off under reduced pressure to obtain a brown oily substance, the reaction was dissolved in ethanol and adjusted to pH 1 with concentrated hydrochloric acid, followed by refluxing for 4 hours, the progress of the reaction was monitored by TLC, and after the completion of the reaction, the reaction was cooled to room temperature, the solvent was distilled off under reduced pressure, the residue was dissolved in water and washed twice with diethyl ether, then adjusted to pH 9 with 10mol/L sodium hydroxide solution, dichloromethane was extracted (3 × 20mL), the organic layers were combined, dried over anhydrous sodium sulfate, filtered, the solvent was distilled off under reduced pressure, and dried in vacuo to obtain, recrystallization from ether/methanol (V: V ═ 12:1) gave 2.06g of 3-benzyloxy-2-hydroxymethyl-6-methyl-1-phenethylpyridin-4-one as a white solid, 59% yield and HPLC assay content ≥ 96%.
(2) DCC (0.50g, 2.4mmol), DMAP (0.049g, 0.4mmol) and N-Boc-ALA (0.555g, 2.4mmol) were dissolved in dichloromethane (6mL) and DMF (3mL) under argon and stirred at room temperature for 45 min. Placing 3-benzyloxy-2-hydroxymethyl-6-methyl-1-phenethylpyridin-4-one (0.70g, 2mmol) dissolved in dichloromethane (6mL) in a constant pressure dropping funnel, adding dropwise over 40 minutes, reacting the reaction solution at room temperature for 12h, monitoring the reaction progress by TLC, filtering off the precipitate after the reaction is finished, distilling the filtrate under reduced pressure to remove the solvent, dissolving the residue in dichloromethane, washing the residue with saturated sodium bicarbonate solution and sodium chloride solution in sequence, drying with anhydrous sodium sulfate, filtering, concentrating the organic layer under reduced pressure, purifying by silica gel column chromatography (gradient elution with dichloromethane: methanol: 100: 1-20: 1), collecting the eluent with dichloromethane: methanol: 40: 1-20: 1, concentrating under reduced pressure, drying to obtain 0.58g of yellow oily product, the yield is 52%, and the HPLC analysis content is more than or equal to 97%.
(3) The yellow oily compound (0.281g, 0.5mmol) is added into a 25mL round-bottom flask, dissolved in 3mL ethyl acetate solution, saturated ethyl acetate solution of hydrogen chloride (3mL) is placed in a constant pressure dropping funnel, slowly dropped at-10 ℃, continuously reacted for 1h, then moved to room temperature for continuous reaction for 4h, the reaction progress is monitored by TLC, after the reaction is finished, the organic solvent is removed by reduced pressure distillation, vacuum drying is carried out, and the obtained intermediate compound is 0.222g of yellow oily matter, the yield is 89%, and the HPLC analysis content is more than or equal to 98%.
(4) 0.222g of the above compound was weighed out and dissolved in 4mL of methanol and placed in a 25mL round bottom flask, and then catalyzed and hydrogenated with 0.044g of 5% Pd/C (Saien chemical technology, Shanghai, Ltd., E060062-25g), and the reaction was stirred at room temperature for 2 hours while monitoring the progress of the reaction by TLC, after the completion of the reaction, palladium/carbon was removed by filtration, the solvent was removed from the filtrate by distillation under reduced pressure, and the white solid compound a 70.112g was obtained by recrystallization from methanol/ether (V: V ═ 10:1), the yield was 62%, and the HPLC analysis content was 99% or more.
m.p.＝147.2-148.9℃.1H NMR(400MHz,DMSO-d6)δ8.40(t,J＝5.2Hz,3H),7.25-7.37(m,6H),5.37(s,2H),4.51(t,J＝7.6Hz,2H),3.94(m,2H),3.12(t,J＝7.6Hz,2H),2.84(t,J＝6.4Hz,2H),2.61(t,J＝6.4Hz,2H),2.59(s,3H),13C NMR(400MHz,DMSO-d6)δ203.0,172.0,161.2,149.3,144.8,137.2,135.7,129.4,129.2,127.6,114.3,56.6,52.4,46.9,35.3,34.7,27.4,20.6；ESI-HRMS:m/z calcd.for C20H25N2O5[M+H]+:373.1758；found:373.1759.
Example 8
(1) Kojic acid (34.08g,240mmol) and 144mL of thionyl chloride are added into a 500mL round-bottom flask, the mixture reacts for 5 hours at room temperature, water is added for quenching, sodium hydroxide solution is used for absorbing tail gas to obtain yellow solid suspended matter, a Buchner funnel is used for filtering, a filter cake is washed by n-hexane and dried in vacuum, and 36.71g of 2-chloromethyl-5-hydroxypyran-4-one white solid compound is obtained, the yield is 95%, and the HPLC analysis content is not less than 96%.
(2) Adding 2-chloromethyl-5-hydroxypyran-4-one (36.71g, 228mmol) and 160mL of water into a 500mL round-bottom flask, fully stirring at 50 ℃ in an oil bath, adding zinc powder (29.82g,456.0mmol), continuously stirring for 5 minutes, slowly dropwise adding 137mL of concentrated hydrochloric acid into a reaction system, maintaining the temperature of the system at 70-80 ℃, maintaining the temperature at 75 ℃ after dropwise adding of concentrated hydrochloric acid, detecting the reaction condition by TLC, filtering while hot after reacting for 8 hours, extracting dichloromethane (6 multiplied by 60mL), combining organic phases, drying anhydrous sodium sulfate, distilling under reduced pressure to remove an organic solvent to obtain brown solid, recrystallizing by using isopropanol to obtain 18.67g of a white solid compound of 2-methyl-5-hydroxypyran-4-one, wherein the yield is 65%, and the HPLC analysis content is not less than 97%.
(3) A500 mL round-bottom flask was charged with the starting material 2-methyl-5-hydroxypyran-4-one (18.67g,148.2mmol), anhydrous K2CO3(24.54g,177.8mmol), benzyl bromide (30.4g,177.8mmol) and acetone (288mL), the reaction was heated to reflux, TLC monitored the progress of the reaction, after completion of the reaction, the mixture was cooled to room temperature, the solvent was removed by distillation under reduced pressure, the residue was dissolved in dichloromethane (120mL), washed with water (3 × 100mL), dried over anhydrous sodium sulfate, filtered, the organic layer was concentrated under reduced pressure, purified by silica gel column chromatography (gradient elution of n-hexane: ethyl acetate ═ 8:1 to 3: 1), and n-hexane: ethyl acetate ═ 5: concentrating the eluent at the ratio of 1-3: 1 under reduced pressure, and drying to obtain 29.77g of a yellow oily product, wherein the yield is 93%, and the HPLC analysis content is more than or equal to 95%.
(4) Glycine (0.90g,12.0mmol), NaOH (0.96g,24.0mmol) were dissolved in H2To 60mL of a mixed solution of EtOH (V: V ═ 1:1), after the solid was completely dissolved, the above-mentioned pyrone (2.16g,10.0mmol) as a yellow oil was added, and the mixture was refluxed for 9 hours, during which the progress of the reaction was checked by TLC. After the reaction, the reaction mixture was cooled to room temperature, ethanol was removed by distillation under reduced pressure, the residue was dissolved in 30mL of water, washed with dichloromethane (3 × 40mL), adjusted to pH 2 with dilute hydrochloric acid, extracted with dichloromethane (5 × 40mL), the combined organic layers were dried over anhydrous sodium sulfate, filtered, the solvent was removed by distillation under reduced pressure from the filtrate, dried under vacuum to give a pale yellow solid, and recrystallized with methanol/diethyl ether (V: V ═ 1:10) to give 1.48g of a white solid, the yield was 54%, and the HPLC assay content was 99% or more.
(5) The white solid intermediate (0.41g,1.5mmol) was added to a 25mL round bottom flask and dissolved in N, N-dimethylformamide (6mL), HATU (1.14g,3mmol) and ALA methyl ester hydrochloride (0.33g,1.8mmol) were added, N-methylmorpholine (0.45g,4.5mmol) was slowly added dropwise in an ice water bath, the reaction was stirred for 45 minutes, and after the solid in the system had completely dissolved, the reaction was transferred to room temperature and continued for 12h, during which the progress of the reaction was monitored by TLC. After the reaction is finished, adding water, extracting with dichloromethane (3 × 20mL), combining organic layers, washing the organic layer with 0.1mol/L dilute hydrochloric acid and saturated sodium bicarbonate solution in sequence, drying with anhydrous sodium sulfate, filtering, concentrating the solvent in vacuum, concentrating the organic layer under reduced pressure, purifying by silica gel column chromatography (gradient elution is carried out on dichloromethane: methanol 80: 1-20: 1), collecting eluent when dichloromethane: methanol is 40: 1-20: 1, concentrating under reduced pressure, and drying to obtain 0.36g of yellow oily product, wherein the yield is 60%, and the content of HPLC analysis is more than or equal to 98%.
(6) The yellow oily intermediate (0.2g, 0.5mmol), 5% Pd/C (Saien chemical technology, Ltd., E060062-25g) (0.04g) and 6mL of methanol were charged into a 25mL round-bottomed flask, and the mixture was stirred under 30psi of hydrogen gas for reaction for 4 hours, followed by TLC to detect the progress of the reaction, after completion of the reaction, insoluble impurities were removed by filtration, the filtrate was concentrated by distillation under reduced pressure, and recrystallized from methanol/diethyl ether (V: 1:10) to obtain b10.14g as a white solid, the yield was 88%, and the HPLC analysis content was 99% or more.
m.p.＝183.5-184.9℃.1H NMR(400MHz,DMSO-d6)δ8.48(t,J＝4.8Hz,1H),7.34(s,1H),6.08(s,1H),4.65(s,2H),4.05(d,J＝5.2Hz,2H),3.57(s,3H),2.72(t,J＝6.4Hz,2H),2.52(t,J＝6.8Hz,2H),2.17(s,3H)；13C NMR(400MHz,DMSO-d6)δ205.2,173.1,171.4,167.4,146.6,145.9,124.7,113.9,55.0,51.9,48.8,34.5,27.6,18.8；ESI-HRMS:m/z calcd for C14H19N2O6[M+H]+:311.1243；found:311.1224.
Example 9
(1) Alanine (1.07g,12.0mmol), NaOH (0.96g,24.0mmol) were dissolved in H2To 60mL of a mixed solution of EtOH (V: V ═ 1:1), after the solid was completely dissolved, pyrone (2.16g,10.0mmol) as a yellow oil prepared in example 8(3) was added, and the mixture was refluxed for 9 hours, during which the progress of the reaction was checked by TLC. After the reaction, the reaction mixture was cooled to room temperature, ethanol was removed by distillation under the reduced pressure, the residue was dissolved in water, washed with dichloromethane (3 × 40mL), adjusted to pH 2 with dilute hydrochloric acid, extracted with dichloromethane (5 × 40mL), the combined organic layers were dried over anhydrous sodium sulfate, filtered, the solvent was removed by distillation under the reduced pressure from the filtrate, dried under vacuum to give a pale yellow solid, and the product was quenched with methanol/diethyl ether (V: V ═ 1:10)1.35g of white solid is obtained by crystallization, the yield is 47 percent, and the HPLC analysis content is more than or equal to 96 percent.
(2) The white solid intermediate (0.43g,1.5mmol) was added to a 25mL round bottom flask and dissolved in N, N-dimethylformamide (6mL), HATU (1.14g,3mmol) and ALA methyl ester hydrochloride (0.33g,1.8mmol) were added, N-methylmorpholine (0.45g,4.5mmol) was slowly added dropwise to the flask in an ice water bath, the reaction was stirred for 45 minutes, and after the solid in the system had completely dissolved, the flask was allowed to shift to room temperature and the reaction was continued for 12h while the progress of the reaction was monitored by TLC. After the reaction is finished, adding water, extracting with dichloromethane (3 × 20mL), combining organic layers, washing the organic layer with 0.1mol/L dilute hydrochloric acid and saturated sodium bicarbonate solution in sequence, drying with anhydrous sodium sulfate, filtering, concentrating the solvent in vacuum, concentrating the organic layer under reduced pressure, purifying by silica gel column chromatography (gradient elution of dichloromethane: methanol is 80: 1-20: 1), collecting eluent when dichloromethane: methanol is 40: 1-20: 1, concentrating under reduced pressure, and drying to obtain 0.36g of yellow oily product, wherein the yield is 63%, and the content of HPLC analysis is more than or equal to 97%.
(3) The yellow oily intermediate (0.21g, 0.5mmol), 5% Pd/C (Sahn chemical technology, E060062-25g) (0.041g) and 6mL of methanol were added to a 25mL round-bottomed flask, and stirred under 30psi of hydrogen gas for 4 hours, the progress of the reaction was checked by TLC, after the completion of the reaction, insoluble impurities were removed by filtration, the filtrate was concentrated by distillation under reduced pressure, and recrystallized from methanol/diethyl ether (V: 1:10) to give b20.13g as a white solid compound with a yield of 83% and an HPLC analysis content of 99% or more.
m.p.＝192.0-194.2℃.
1H NMR(400MHz,DMSO-d6)δ8.30(t,J＝4.8Hz,1H),7.39(s,1H),6.20(s,1H),4.41(s,2H),4.11(t,J＝6.8Hz,2H),3.96(d,J＝5.2Hz,2H),2.64(m,4H),2.48(t,J＝6.0Hz,2H),2.27(s,3H)；13C NMR(400MHz,DMSO-d6)δ205.6,173.1,171.0,169.9,147.1,145.1,122.9,114.0,51.9,51.9,48.8,36.1,34.3,27.6,18.8；ESI-HRMS:m/z calcd for C15H21N2O6[M+H]+:325.1400；found 325.1390.
Example 10
(1) 4-aminobutyric acid (1.24g,12.0mmol), NaOH (0.96g,24.0mmol) were dissolved in H2To 60mL of a mixed solution of EtOH (V: V ═ 1:1), after the solid was completely dissolved, pyrone (2.16g,10.0mmol) as a yellow oil prepared in example 8(3) was added, and the mixture was refluxed for 9 hours, during which the progress of the reaction was checked by TLC. After the reaction, the reaction mixture was cooled to room temperature, ethanol was removed by distillation under reduced pressure, the residue was dissolved in water (30mL), washed with dichloromethane (3 × 40mL), adjusted to pH 2 with dilute hydrochloric acid, extracted with dichloromethane (5 × 40mL), the combined organic layers were dried over anhydrous sodium sulfate, filtered, the solvent was removed by distillation under reduced pressure from the filtrate, and dried under vacuum to give a pale yellow solid, which was recrystallized from methanol/ether (V: V ═ 1:10) to give 1.36g of a white solid with a yield of 45% and an HPLC assay content of 97% or more.
(2) The white solid intermediate (0.45g,1.5mmol) was added to a 25mL round bottom flask and dissolved in N, N-dimethylformamide (6mL), HATU (1.14g,3mmol) and ALA methyl ester hydrochloride (0.33g,1.8mmol) were added, N-methylmorpholine (0.45g,4.5mmol) was slowly added dropwise in an ice water bath, the reaction was stirred for 45 minutes, and after the solid in the system had completely dissolved, the reaction was transferred to room temperature and continued for 12h, during which the progress of the reaction was monitored by TLC. After the reaction is finished, adding water, extracting with dichloromethane (3 × 20mL), combining organic layers, washing the organic layer with 0.1mol/L dilute hydrochloric acid and saturated sodium bicarbonate solution in sequence, drying with anhydrous sodium sulfate, filtering, concentrating the solvent in vacuum, concentrating the organic layer under reduced pressure, purifying by silica gel column chromatography (gradient elution is carried out on dichloromethane: methanol 80: 1-20: 1), collecting eluent when dichloromethane: methanol is 40: 1-20: 1, concentrating under reduced pressure, and drying to obtain 0.36g of yellow oily product, wherein the yield is 57%, and the content of HPLC analysis is more than or equal to 96%.
(3) The yellow oily intermediate (0.21g, 0.5mmol), 5% Pd/C (sahn chemical technology (shanghai) ltd., E060062-25g) (0.041g) and 6mL of methanol were added to a 25mL round-bottomed flask, stirred and reacted for 4 hours under 30psi of hydrogen gas, the progress of the reaction was checked by TLC, after the completion of the reaction, insoluble impurities were removed by filtration, the filtrate was concentrated by distillation under reduced pressure, and recrystallized from methanol/ether (V: V ═ 1:10) to give b30.15g of a white solid compound, the yield was 88%, and the HPLC assay content was 99% or more.
m.p.＝117.0-117.9℃.1H NMR(400MHz,DMSO-d6)δ8.23(t,J＝5.6Hz,1H),7.37(s,1H),6.06(s,1H),3.96(d,J＝5.6Hz,2H),3.83(t,J＝7.6Hz,2H),3.57(s,3H),2.70(t,J＝6.4Hz,2H),2.49(t,J＝6.4Hz,2H),2.25(s,3H),2.21(t,J＝7.2Hz,2H),1.86(m,2H)；13C NMR(400MHz,DMSO-d6)δ205.9,173.1,172.0,170.9,147.2,145.0,122.9,114.0,51.9,51.8,48.8,34.4,31.8,27.6,26.4,18.7；ESI-HRMS:m/z calcd for C16H23N2O6[M+H]+:339.1556；found 339.1542.
Example 11
(1) 5-amino valeric acid (1.41g,12.0mmol), NaOH (0.96g,24.0mmol) were dissolved in H2To 60mL of a mixed solution of EtOH (V: V ═ 1:1), after the solid was completely dissolved, pyrone (2.16g,10.0mmol) as a yellow oil prepared in example 8(3) was added, and the mixture was refluxed for 9 hours, during which the progress of the reaction was checked by TLC. After the reaction, the reaction mixture was cooled to room temperature, ethanol was removed by distillation under reduced pressure, the residue was dissolved in 30mL of water, washed with dichloromethane (3 × 40mL), adjusted to pH 2 with dilute hydrochloric acid, extracted with dichloromethane (5 × 40mL), the combined organic layers were dried over anhydrous sodium sulfate, filtered, the solvent was removed by distillation under reduced pressure from the filtrate, dried under vacuum to give a pale yellow solid, and recrystallized from methanol/ether (V: V ═ 1:10) to give 1.36g of a white solid, yield 43%, HPLC assay content ≥ 96%.
(2) The white solid intermediate (0.47g,1.5mmol) was added to a 25mL round bottom flask and dissolved in N, N-dimethylformamide (6mL), HATU (1.14g,3mmol) and ALA methyl ester hydrochloride (0.33g,1.8mmol) were added, N-methylmorpholine (0.45g,4.5mmol) was slowly added dropwise in an ice water bath, the reaction was stirred for 45 minutes, and after the solid in the system had completely dissolved, the reaction was transferred to room temperature and continued for 12h, during which the progress of the reaction was monitored by TLC. After the reaction is finished, adding water, extracting with dichloromethane (3 × 20mL), combining organic layers, washing the organic layer with 0.1mol/L dilute hydrochloric acid and saturated sodium bicarbonate solution in sequence, drying with anhydrous sodium sulfate, filtering, concentrating the solvent in vacuum, concentrating the organic layer under reduced pressure, purifying by silica gel column chromatography (gradient elution is carried out on dichloromethane: methanol 80: 1-20: 1), collecting eluent when dichloromethane: methanol is 40: 1-20: 1, concentrating under reduced pressure, and drying to obtain 0.36g of yellow oily product, wherein the yield is 51%, and the content of HPLC analysis is more than or equal to 97%.
(3) The yellow oily intermediate (0.22g, 0.5mmol), 5% Pd/C (sahn chemical technology (shanghai) ltd., E060062-25g) (0.044g) and 6mL of methanol were charged into a 25mL round-bottomed flask, stirred and reacted for 4 hours under 30psi of hydrogen gas, the progress of the reaction was checked by TLC, after the completion of the reaction, insoluble impurities were removed by filtration, the filtrate was concentrated by distillation under reduced pressure, and recrystallized from methanol/ether (V: V ═ 1:10) to give b40.15g of a white solid compound, the yield was 86%, and the HPLC assay content was not less than 99%.
m.p.＝84.2-85.2℃.1H NMR(400MHz,DMSO-d6)δ8.17(t,J＝4.4Hz,1H),7.39(s,1H),6.06(s,1H),3.94(d,J＝5.2Hz,2H),3.83(t,J＝7.2Hz,2H),3.57(s,3H),2.69(t,J＝6.4Hz,2H),2.48(t,J＝6.4Hz,2H),2.25(s,3H),2.18(t,J＝6.8Hz,2H),1.62(m,2H),1.52(m,2H)；13C NMR(400MHz,DMSO-d6)δ205.9,173.1,172.6,170.9,147.1,144.8,123.2,114.1,52.4,51.9,48.7,34.9,34.4,29.9,27.6,22.5,18.7；ESI-HRMS:m/z calcd for C17H25N2O6[M+H]+:353.1713；found 353.1708.
Example 12
(1) 6-aminocaproic acid (1.57g,12.0mmol), NaOH (0.96g,24.0mmol) were dissolved in H2To 60mL of a mixed solution of EtOH (V: V ═ 1:1), after the solid was completely dissolved, pyrone (2.16g,10.0mmol) as a yellow oil prepared in example 8(3) was added, and the mixture was refluxed for 9 hours, during which the progress of the reaction was checked by TLC. After the reaction, the reaction mixture was cooled to room temperature, ethanol was removed by distillation under reduced pressure, the residue was dissolved in 30mL of water, washed with dichloromethane (3 × 40mL), adjusted to pH 2 with dilute hydrochloric acid, extracted with dichloromethane (5 × 40mL), the combined organic layers were dried over anhydrous sodium sulfate, filtered, the solvent was removed by distillation under reduced pressure from the filtrate, dried under vacuum to give a pale yellow solid, and recrystallized from methanol/ether (V: V ═ 1:10) to give 1.88g of a white solid, yield 57%, HPLC assay content ≥ 96%.
(2) The white solid intermediate (0.49g,1.5mmol) was added to a 25mL round bottom flask and dissolved in N, N-dimethylformamide (6mL), HATU (1.14g,3mmol) and ALA methyl ester hydrochloride (0.33g,1.8mmol) were added, N-methylmorpholine (0.45g,4.5mmol) was slowly added dropwise to the flask in an ice water bath, the reaction was stirred for 45 minutes, and after the solid in the system had completely dissolved, the flask was allowed to shift to room temperature and the reaction was continued for 12h while the progress of the reaction was monitored by TLC. After the reaction is finished, adding water, extracting with dichloromethane (3 × 20mL), combining organic layers, washing the organic layer with 0.1mol/L dilute hydrochloric acid and saturated sodium bicarbonate solution in sequence, drying with anhydrous sodium sulfate, filtering, concentrating the solvent in vacuum, concentrating the organic layer under reduced pressure, purifying by silica gel column chromatography (gradient elution is carried out on dichloromethane: methanol 80: 1-20: 1), collecting eluent when dichloromethane: methanol is 40: 1-20: 1, concentrating under reduced pressure, and drying to obtain 0.36g of yellow oily product, wherein the yield is 48%, and the content of HPLC analysis is more than or equal to 97%.
(3) The yellow oily intermediate (0.23g, 0.5mmol), 5% Pd/C (sahn chemical technology, shanghai, ltd., E060062-25g) (0.046g) and 6mL of methanol were charged into a 25mL round-bottomed flask, stirred and reacted for 4 hours under 30psi of hydrogen gas, the progress of the reaction was checked by TLC, insoluble impurities were removed by filtration after the completion of the reaction, the filtrate was concentrated by distillation under reduced pressure, and recrystallized from methanol/ether (V: V ═ 1:10) to give b50.16g as a white solid compound with a yield of 86% and an HPLC assay content of 99% or more.
m.p.＝65.2-67.2℃.1H NMR(400MHz,DMSO-d6)δ8.13(t,J＝5.6Hz,1H),7.40(s,1H),6.07(s,1H),3.92(d,J＝5.6Hz,2H),3.81(t,J＝7.6Hz,2H),3.57(s,3H),2.68(t,J＝6.4Hz,2H),2.48(t,J＝6.4Hz,2H),2.26(s,3H),2.15(t,J＝7.2Hz,2H),1.63(m,2H),1.53(m,2H),1.27(m,2H)；13C NMR(400MHz,DMSO-d6)δ206.0,173.2,172.8,170.7,147.1,144.8,123.3,114.0,52.5,51.9,48.7,35.2,34.3,30.1,27.6,25.8,25.2,18.8；ESI-HRMS:m/z calcd for C18H27N2O6[M+H]+:367.1869；found 367.1864.
Example 13
(1) The method comprises the steps of adding raw materials of kojic acid (14.2g,100mmol), sodium hydroxide (8.4g,210mmol), benzyl bromide (35.9g,210mmol) and acetone (200mL) into a 500mL round-bottom flask, heating a reaction system to reflux, monitoring the progress of the reaction by TLC, cooling a mixture to room temperature after the reaction is completed, distilling under reduced pressure to remove a solvent, dissolving a residue with dichloromethane (160mL), washing with water (3X 100mL), drying with anhydrous sodium sulfate, filtering, concentrating under reduced pressure, purifying by silica gel column chromatography (n-hexane: ethyl methylacetate is subjected to gradient elution, and collecting an eluent when n-hexane: ethyl acetate is 5: 1-3: 1, concentrating under reduced pressure, and drying to obtain 28.04g of a yellow oily product, wherein the yield is 87%, and the HPLC analysis content is not less than 98%.
(2) Glycine (0.90g,12.0mmol), NaOH (0.96g,24.0mmol) were dissolved in H2To 60mL of a mixed solution of EtOH (V: V ═ 1:1), after the solid was completely dissolved, the above-mentioned pyrone (3.22g,10.0mmol) as a yellow oil was added, and the mixture was refluxed for 9 hours, during which the progress of the reaction was checked by TLC. After the reaction, the reaction mixture was cooled to room temperature, ethanol was removed by distillation under reduced pressure, the residue was dissolved in water (30mL), washed with dichloromethane (3 × 40mL), adjusted to pH 2 with dilute hydrochloric acid, extracted with dichloromethane (5 × 40mL), the combined organic layers were dried over anhydrous sodium sulfate, filtered, the solvent was removed by distillation under reduced pressure from the filtrate, and dried under vacuum to give a pale yellow solid, which was recrystallized from methanol/ether (V: V ═ 1:10) to give 1.86g of a white solid with a yield of 49% and a HPLC assay content of 96% or more.
(3) The white solid intermediate (0.57g,1.5mmol) was added to a 25mL round bottom flask and dissolved in N, N-dimethylformamide (6mL), HATU (1.14g,3mmol) and ALA methyl ester hydrochloride (0.33g,1.8mmol) were added, N-methylmorpholine (0.45g,4.5mmol) was slowly added dropwise to the flask in an ice water bath, the reaction was stirred for 45 minutes, and after the solid in the system had completely dissolved, the flask was allowed to shift to room temperature and the reaction was continued for 12h while the progress of the reaction was monitored by TLC. After the reaction is finished, adding water, extracting with dichloromethane (3 × 20mL), combining organic layers, washing the organic layer with 0.1mol/L dilute hydrochloric acid and saturated sodium bicarbonate solution in sequence, drying with anhydrous sodium sulfate, filtering, concentrating the solvent in vacuum, concentrating the organic layer under reduced pressure, purifying by silica gel column chromatography (gradient elution is carried out on dichloromethane: methanol 80: 1-20: 1), collecting eluent when dichloromethane: methanol is 40: 1-20: 1, concentrating under reduced pressure, and drying to obtain 0.36g of yellow oily product, wherein the yield is 51%, and the content of HPLC analysis is more than or equal to 97%.
(4) The yellow oily intermediate (0.2g, 0.5mmol), 5% Pd/C (Saien chemical technology, Ltd., E060062-25g) (0.051g) and 6mL of methanol were added to a 25mL round-bottomed flask, stirred and reacted for 16 hours under 30psi of hydrogen, the progress of the reaction was checked by TLC, after the completion of the reaction, insoluble impurities were removed by filtration, the filtrate was concentrated by distillation under reduced pressure, and recrystallized from methanol/diethyl ether (V: 1:10) to give a white solid compound b60.11g, the yield was 67%, and the HPLC analysis content was 99% or more.
m.p.＝186.2-188.4℃.1H NMR(400MHz,DMSO-d6)δ8.48(t,J＝6.0Hz,1H),7.33(s,1H),6.22(s,1H),4.73(s,2H),4.27(s,2H),4.04(d,J＝5.2Hz,2H),3.57(s,3H),2.71(t,J＝6.4Hz,2H),2.50(t,J＝6.8Hz,2H)；13C NMR(400MHz,DMSO-d6)δ205.2,173.1,171.6,167.6,148.2,146.8,125.3,112.5,59.8,53.9,51.9,48.9,34.5,27.6；ESI-HRMS:m/z calcd for C14H18N2NaO7[M+Na]+:349.1012；found 349.1006.
Example 14
(1) Alanine (1.07g,12.0mmol), NaOH (0.96g,24.0mmol) were dissolved in H2To 60mL of a mixed solution of EtOH (V: V ═ 1:1), after the solid was completely dissolved, pyrone (3.22g,10.0mmol) as a yellow oil prepared in example 13(1) was added, and the mixture was refluxed for 9 hours, during which the progress of the reaction was checked by TLC. After the reaction, the reaction mixture was cooled to room temperature, ethanol was removed by distillation under reduced pressure, the residue was dissolved in water (30mL), washed with dichloromethane (3 × 40mL), adjusted to pH 2 with dilute hydrochloric acid, extracted with dichloromethane (5 × 40mL), the combined organic layers were dried over anhydrous sodium sulfate, filtered, the solvent was removed by distillation under reduced pressure from the filtrate, and dried under vacuum to give a pale yellow solid, which was recrystallized from methanol/ether (V: V ═ 1:10) to give 1.69g of a white solid with a yield of 43% and a content of not less than 96% by HPLC analysis.
(2) The white solid intermediate (0.59g,1.5mmol) was added to a 25mL round bottom flask and dissolved in N, N-dimethylformamide (6mL), HATU (1.14g,3mmol) and ALA methyl ester hydrochloride (0.33g,1.8mmol) were added, N-methylmorpholine (0.45g,4.5mmol) was slowly added dropwise in an ice water bath, the reaction was stirred for 45 minutes, and after the solid in the system had completely dissolved, the reaction was transferred to room temperature and continued for 12h, during which the progress of the reaction was monitored by TLC. After the reaction is finished, adding water, extracting with dichloromethane (3 × 20mL), combining organic layers, washing the organic layer with 0.1mol/L dilute hydrochloric acid and saturated sodium bicarbonate solution in sequence, drying with anhydrous sodium sulfate, filtering, concentrating the solvent in vacuum, concentrating the organic layer under reduced pressure, purifying by silica gel column chromatography (gradient elution is carried out on dichloromethane: methanol 80: 1-20: 1), collecting eluent when dichloromethane: methanol is 40: 1-20: 1, concentrating under reduced pressure, and drying to obtain 0.36g of yellow oily product, wherein the yield is 49%, and the content of HPLC analysis is more than or equal to 96%.
(3) The yellow oily intermediate (0.26g, 0.5mmol), 5% Pd/C (Saien chemical technology, Ltd., E060062-25g) (0.052g) and 6mL of methanol were charged into a 25mL round-bottomed flask, stirred and reacted for 16 hours under 30psi of hydrogen gas, the progress of the reaction was checked by TLC, after the completion of the reaction, insoluble impurities were removed by filtration, the filtrate was concentrated by distillation under reduced pressure, and recrystallized from methanol/diethyl ether (V: 1:10) to give a white solid compound, b70.092g, in 54% yield and 99% or more by HPLC analysis.
m.p.＝176.7-177.4℃.1H NMR(400MHz,DMSO-d6)δ8.30(t,J＝4.8Hz,1H),7.39(s,1H),6.20(s,1H),4.41(s,2H),4.11(t,J＝6.8Hz,2H),3.96(d,J＝5.2Hz,2H),3.57(s,3H),2.66(m,4H),2.48(t,J＝8.0Hz,2H)；13C NMR(400MHz,DMSO-d6)δ205.7,173.1,171.2,170.1,147.7,147.4,123.4,113.0,60.0,51.9,48.9,48.1,36.5,34.4,27.7；ESI-HRMS:m/z calcd for C15H20N2NaO7[M+Na]+:363.1168；found 363.1180.
Example 15
(1) 4-aminobutyric acid (1.24g,12.0mmol), NaOH (0.96g,24.0mmol) were dissolved in H2To 60mL of a mixed solution of EtOH (V: V ═ 1:1), after the solid was completely dissolved, pyrone (3.22g,10.0mmol) as a yellow oil prepared in example 13(1) was added, and the mixture was refluxed for 9 hours, during which the progress of the reaction was checked by TLC. After completion of the reaction, the reaction mixture was cooled to room temperature, ethanol was distilled off under reduced pressure, the residue was dissolved in water (30mL), the mixture was washed with dichloromethane (3 × 40mL), the pH was adjusted to 2 with dilute hydrochloric acid, dichloromethane was extracted (5 × 40mL), and the combined organic layers were extracted with anhydrous sodium sulfateDrying, filtering, distilling the filtrate under reduced pressure to remove the solvent, drying in vacuum to obtain a light yellow solid, and recrystallizing with methanol/diethyl ether (V: V ═ 1:10) to obtain 1.71g of a white solid, wherein the yield is 42%, and the HPLC analysis content is more than or equal to 97%.
(2) The white solid intermediate (0.61g,1.5mmol) was added to a 25mL round bottom flask and dissolved in N, N-dimethylformamide (6mL), HATU (1.14g,3mmol) and ALA methyl ester hydrochloride (0.33g,1.8mmol) were added, N-methylmorpholine (0.45g,4.5mmol) was slowly added dropwise in an ice water bath, the reaction was stirred for 45 minutes, and after the solid in the system had completely dissolved, the reaction was transferred to room temperature and continued for 12h, during which the progress of the reaction was monitored by TLC. After the reaction is finished, adding water, extracting with dichloromethane (3 × 20mL), combining organic layers, washing the organic layer with 0.1mol/L dilute hydrochloric acid and saturated sodium bicarbonate solution in sequence, drying with anhydrous sodium sulfate, filtering, concentrating the solvent in vacuum, concentrating the organic layer under reduced pressure, purifying by silica gel column chromatography (gradient elution is carried out on dichloromethane: methanol 80: 1-20: 1), collecting eluent when dichloromethane: methanol is 40: 1-20: 1, concentrating under reduced pressure, and drying to obtain 0.36g of yellow oily product, wherein the yield is 51%, and the content of HPLC analysis is more than or equal to 98%.
(3) The yellow oily intermediate (0.27g, 0.5mmol), 5% Pd/C (Saien chemical technology, Ltd., E060062-25g) (0.053g) and 6mL of methanol were added to a 25mL round-bottomed flask, stirred and reacted for 16 hours under 30psi of hydrogen gas, the progress of the reaction was checked by TLC, after the completion of the reaction, insoluble impurities were removed by filtration, the filtrate was concentrated by distillation under reduced pressure, and recrystallized from methanol/diethyl ether (V: 1:10) to give b80.105g of a white solid compound, the yield was 59%, and the HPLC analysis content was 99% or more.
m.p.＝157.6-158.2℃.1H NMR(400MHz,DMSO-d6)δ8.21(t,J＝5.2Hz,1H),7.40(s,1H),6.23(s,1H),4.39(s,2H),3.96(d,J＝5.6Hz,2H),3.87(t,J＝7.2Hz,2H),3.58(s,3H),2.70(t,J＝6.4Hz,2H),2.50(t,J＝7.6Hz,2H),2.21(t,J＝7.2Hz,2H),1.90(m,2H)；13C NMR(400MHz,DMSO-d6)δ205.9,173.1,172.1,171.1,147.8,147.4,123.4,112.7,59.7,51.9,51.2,48.8,34.4,32.0,27.6,26.9；ESI-HRMS:m/z calcd for C16H22N2NaO7[M+Na]+:377.1325；found 377.1332.
Example 16
(1) 5-amino valeric acid (1.41g,12.0mmol), NaOH (0.96g,24.0mmol) were dissolved in H2To 60mL of a mixed solution of EtOH (V: V ═ 1:1), after the solid was completely dissolved, pyrone (3.22g,10.0mmol) as a yellow oil prepared in example 13(1) was added, and the mixture was refluxed for 9 hours, during which the progress of the reaction was checked by TLC. After the reaction, the reaction mixture was cooled to room temperature, ethanol was removed by distillation under reduced pressure, the residue was dissolved in water (30mL), washed with dichloromethane (3 × 40mL), adjusted to pH 2 with dilute hydrochloric acid, extracted with dichloromethane (5 × 40mL), the combined organic layers were dried over anhydrous sodium sulfate, filtered, the solvent was removed by distillation under reduced pressure from the filtrate, and dried under vacuum to give a pale yellow solid, which was recrystallized from methanol/ether (V: V ═ 1:10) to give 2.19g of a white solid with a yield of 52% and a content of not less than 96% by HPLC analysis.
(2) The white solid intermediate (0.63g,1.5mmol) was added to a 25mL round bottom flask and dissolved in N, N-dimethylformamide (6mL), HATU (1.14g,3mmol) and ALA methyl ester hydrochloride (0.33g,1.8mmol) were added, N-methylmorpholine (0.45g,4.5mmol) was slowly added dropwise in an ice water bath, the reaction was stirred for 45 minutes, and after the solid in the system had completely dissolved, the reaction was transferred to room temperature and continued for 12h, during which the progress of the reaction was monitored by TLC. After the reaction is finished, adding water, extracting with dichloromethane (3 × 20mL), combining organic layers, washing the organic layer with 0.1mol/L dilute hydrochloric acid and saturated sodium bicarbonate solution in sequence, drying with anhydrous sodium sulfate, filtering, concentrating the solvent in vacuum, concentrating the organic layer under reduced pressure, purifying by silica gel column chromatography (gradient elution is carried out on dichloromethane: methanol 80: 1-20: 1), collecting eluent when dichloromethane: methanol is 40: 1-20: 1, concentrating under reduced pressure, and drying to obtain 0.36g of yellow oily product, wherein the yield is 55%, and the content of HPLC analysis is more than or equal to 96%.
(3) The yellow oily intermediate (0.27g, 0.5mmol), 5% Pd/C (Saien chemical technology, Ltd., E060062-25g) (0.053g) and 6mL of methanol were added to a 25mL round-bottomed flask, stirred and reacted for 16 hours under 30psi of hydrogen gas, the progress of the reaction was checked by TLC, after the completion of the reaction, insoluble impurities were removed by filtration, the filtrate was concentrated by distillation under reduced pressure, and recrystallized from methanol/diethyl ether (V: V ═ 1:10) to give b90.103g of a white solid compound, the yield was 56%, and the HPLC analysis content was 99% or more.
m.p.＝169.0-170.2℃.1H NMR(400MHz,DMSO-d6)δ8.15(t,J＝5.2Hz,1H),7.41(s,1H),6.22(s,1H),4.38(s,2H),3.94(d,J＝5.6Hz,2H),3.87(t,J＝7.2Hz,2H),3.57(s,3H),2.68(t,J＝6.4Hz,2H),2.48(t,J＝6.4Hz,2H),2.18(t,J＝7.2Hz,2H),1.68(m,2H),1.51(m,2H)；13C NMR(400MHz,DMSO-d6)δ205.9,173.1,172.6,171.0,147.6,147.4,123.5,112.7,59.8,51.9,51.6,48.7,34.9,34.4,30.5,27.6,22.6；ESI-HRMS:m/z calcd for C17H25N2O7[M+H]+:369.1662；found 369.1640.
Example 17
(1) 6-aminocaproic acid (1.57g,12.0mmol), NaOH (0.96g,24.0mmol) were dissolved in H2To 60mL of a mixed solution of EtOH (V: V ═ 1:1), after the solid was completely dissolved, pyrone (3.22g,10.0mmol) as a yellow oil prepared in example 13(1) was added, and the mixture was refluxed for 9 hours, during which the progress of the reaction was checked by TLC. After the reaction, the reaction mixture was cooled to room temperature, ethanol was removed by distillation under reduced pressure, the residue was dissolved in water (30mL), washed with dichloromethane (3 × 40mL), adjusted to pH 2 with dilute hydrochloric acid, extracted with dichloromethane (5 × 40mL), the combined organic layers were dried over anhydrous sodium sulfate, filtered, the solvent was removed by distillation under reduced pressure from the filtrate, and dried under vacuum to give a pale yellow solid, which was recrystallized from methanol/ether (V: V ═ 1:10) to give 1.79g of a white solid with a yield of 41% and an HPLC assay content of 97% or more.
(2) The white solid intermediate (0.65g,1.5mmol) was added to a 25mL round bottom flask and dissolved in N, N-dimethylformamide (6mL), HATU (1.14g,3mmol) and ALA methyl ester hydrochloride (0.33g,1.8mmol) were added, N-methylmorpholine (0.45g,4.5mmol) was slowly added dropwise in an ice water bath, the reaction was stirred for 45 minutes, and after the solid in the system had completely dissolved, the reaction was transferred to room temperature and continued for 12h, during which the progress of the reaction was monitored by TLC. After the reaction is finished, adding water, extracting with dichloromethane (3 × 20mL), combining organic layers, washing the organic layer with 0.1mol/L dilute hydrochloric acid and saturated sodium bicarbonate solution in sequence, drying with anhydrous sodium sulfate, filtering, concentrating the solvent in vacuum, concentrating the organic layer under reduced pressure, purifying by silica gel column chromatography (gradient elution is carried out on dichloromethane: methanol 80: 1-20: 1), collecting eluent when dichloromethane: methanol is 40: 1-20: 1, concentrating under reduced pressure, and drying to obtain 0.36g of yellow oily product, wherein the yield is 44%, and the content of HPLC analysis is more than or equal to 98%.
(3) The yellow oily intermediate (0.28g, 0.5mmol), 5% Pd/C (sahn chemical technology (shanghai) ltd., E060062-25g) (0.056g) and 6mL of methanol were charged into a 25mL round-bottomed flask, stirred and reacted for 16 hours under 30psi of hydrogen gas, the progress of the reaction was checked by TLC, insoluble impurities were removed by filtration after the completion of the reaction, the filtrate was concentrated by distillation under reduced pressure, and recrystallized from methanol/diethyl ether (V: V ═ 1:10) to give b100.094g as a white solid compound with a yield of 49% and an HPLC assay content of 99% or more.
m.p.＝151.2-152.4℃.1H NMR(400MHz,DMSO-d6)δ8.11(t,J＝5.2Hz,1H),7.42(s,1H),6.22(s,1H),4.37(s,2H),3.92(d,J＝5.6Hz,2H),3.85(t,J＝7.6Hz,2H),3.57(s,3H),2.69(t,J＝6.4Hz,2H),2.50(t,J＝9.2Hz,2H),2.14(t,J＝7.2Hz,2H),1.67(m,2H),1.53(m,2H),1.28(m,2H)；13C NMR(400MHz,DMSO-d6)δ206.0,173.1,172.8,170.9,147.6,147.4,123.6,112.7,59.9,51.9,51.8,48.7,35.3,34.4,30.6,27.6,25.9,25.2；ESI-HRMS:m/z calcd for C18H26N2NaO7[M+Na]+:405.1638；found 405.1644.
Example 18
The following are the pharmacological experimental data for ALA-HPO hybrid derivatives of formula (I) according to the invention:
1. phototoxicity test
The experimental method comprises the following steps: different tumor cells were treated at a rate of about 1.0X 104Density of/well in 96-well plates, after 48h of incubation, cells were washed with phosphate buffer. Adding 100 μ L of compound solutions containing different concentrations (20-100 μ M) into a specified well, wherein the positive drugs are ALA and mixed administration of ALA and 3-hydroxy-1, 2-dimethylpyridin-4-one (CP20), culturing for 4 hr, and culturing at 2.5J cm-2After 5 minutes of blue light irradiation, the medium containing the drug was aspirated off, and the culture was continued for 18 hours using a medium containing no serum. MiningCytotoxicity was determined using MTT. The MTT detection method comprises the following steps: all media was removed from the well plate, 100. mu.L of media containing 0.5% MTT was added to each well, and the plate was covered and placed in an incubator for 4h. All media in the plate was removed again, 100 μ L of dimethyl sulfoxide was added to each well, shaken for 5 minutes, absorbance at 520nm was measured, and the average cell viability at each concentration of each prodrug was calculated and expressed as a percentage of the control.
The values for cell viability and concentration are tabulated and presented as line graphs.
The average cell survival rate is absorbance (average of each prodrug)/absorbance (average of control group) × 100%.
From Table 1 it can be concluded that the most phototoxic conjugate in the HeLa cell line is a2, the LD of which50The value was 46.11. mu.M; conjugates a2 and a3 both showed strong cytotoxicity in MCF-7 cell line, LD50Values of 52.74. mu.M and 45.42. mu.M, respectively; while the conjugates A3, a4 and a7 showed strong cytotoxicity in A375 cell line, their LD50The values were 33.22. mu.M, 21.32. mu.M and 34.89. mu.M, respectively. Further, the ester bond compound can rapidly enter cells, break the bond between the PpIX prodrug and the HPO molecule under the action of intracellular peptidase, liberate ALA and generate high-concentration photosensitive active substance PpIX in cells through metabolism.
TABLE 1 LD of ALA, ALA-CP 20 mixture and ALA-HPO hybrid derivatives of formula (I) in different tumor cell lines50The value is obtained.
Figure BDA0002970338500000241
2. Dark toxicity test
To determine the "dark" toxicity of a compound, the phototoxicity experimental procedure was repeated, but without irradiation with light.
The values for cell viability and concentration are tabulated and presented in bar graph form.
The average cell survival rate is absorbance (average of each prodrug)/absorbance (average of control group) × 100%.
From fig. 4, the following conclusions can be drawn: the ALA-HPO hybrid derivative shown in the formula (I) is incubated with HeLa, MCF-7 and A375 tumor cell lines for 4h at the concentration of 100 mu M, and the series of compounds are found not to show dark cytotoxicity. The series of compounds are safe to cells under the condition without light.
Example 19
The following are the pharmacological experimental data for ALA-HPO hybrid derivatives of formula (II) according to the present invention:
1. phototoxicity test
The experimental method comprises the following steps: different tumor cells were treated at a rate of about 1.0X 104Density of/well in 96-well plates, after 48h of incubation, cells were washed with phosphate buffer. Adding 100 μ L of compound solutions with different concentrations (100-400 μ M) into a designated well, wherein the positive drugs are ALA, 3-hydroxy-1, 2-dimethylpyridin-4-one (CP20), and ALA and 3-hydroxy-1, 2-dimethylpyridin-4-one (CP20), respectively, mixing, culturing for 4 hr or 24 hr, and culturing with 5J cm-2After 5 minutes of blue light irradiation, the medium containing the drug was aspirated off, and the culture was continued for 18 hours using a medium containing no serum. Cytotoxicity was determined using MTT. The MTT detection method comprises the following steps: all media was removed from the well plate, 100. mu.L of media containing 0.5% MTT was added to each well, and the plate was covered and placed in an incubator for 4h. All media in the plate was removed again, 100 μ L of dimethyl sulfoxide was added to each well, shaken for 5 minutes, absorbance at 520nm was measured, and the average cell viability at each concentration of each prodrug was calculated and expressed as a percentage of the control.
The values for cell viability and concentration are tabulated and presented in bar graph form.
The average cell survival rate is absorbance (average of each prodrug)/absorbance (average of control group) × 100%.
From fig. 5(a), fig. 6(a) and fig. 7(a), the following conclusions can be drawn: incubating ALA-HPO hybrid derivative shown in formula (II) with HeLa, MCF-7 and A375 tumor cell line for 4h, using 5J cm-2After blue light irradiation, the compounds b1, b2, b3, b4, b6, b7, b8, b9, b 3538,The concentration of b10 is 100-400 mu M, no obvious cell killing effect is generated on tumor cells, the compound b5 has obvious cell killing effect on three different tumor cell lines, the cell killing effect is higher than that of ALA or CP20, but is obviously lower than that of ALA and CP20 mixed administration. Wherein the survival percentages of the HeLa cell line treated with ALA and compound b5 at concentrations of 100 μ M, 200 μ M and 400 μ M were 94.5%, 82.1%, 70.5%, 70.4%, 64.9% and 48.6%, respectively; the survival percentages of the MCF-7 cell lines treated with ALA and compound b5 at concentrations of 100. mu.M, 200. mu.M and 400. mu.M were 98.4%, 57.8%, 28.0%, 40.2%, 32.1% and 28.9%, respectively; the percent survival of the A375 cell line treated with ALA, compound b5 at 100. mu.M, 200. mu.M, 400. mu.M concentrations was 73.0%, 52.7%, 43.6%, 47.0%, 36.7%, 28.3%, respectively.
In addition, the cytotoxicity of the series of compounds shown in the formula (II) after 24h of incubation with HeLa, MCF-7 and A375 tumor cell lines is also studied, and the dose of the used blue light is also 5J cm-2. From fig. 5(B), fig. 6(B) and fig. 7(B), it is found that the compounds B1, B2, B3, B4, B6, B7, B8, B9 and B10 do not show obvious cell killing effect on three different tumor cells at the concentration of 100-200 μ M, but show certain cytotoxicity at the concentration of 400 μ M, but the cell killing effect is lower than that of ALA at the same concentration. Wherein the survival percentages of the HeLa cell lines treated by the ALA and the compound b5 under the concentration of 100-400 mu M are 65.5%, 64.1%, 48.6%, 40.0%, 32.8% and 25.0% respectively; the survival percentages of MCF-7 cell lines treated by ALA and compound b5 under the concentration of 100-400 mu M are 92.5%, 61.7%, 13.1%, 32.6%, 24.6% and 18.3%, respectively; the survival percentages of the A375 cell lines treated by ALA and the compound b5 under the concentration of 100-400 μ M are 72.2%, 57.3%, 27.2%, 32.6%, 24.4% and 19.8%, respectively;
the above results show that compound b5 can better enter cells and break the bond between the PpIX prodrug and the HPO molecule under the action of intracellular peptidase, liberate ALA and generate the photosensitive active substance PpIX by metabolism.
2. Dark toxicity test
To determine the "dark" toxicity of a compound, the phototoxicity experimental procedure was repeated, but without irradiation with light.
The values for cell viability and concentration are tabulated and presented in bar graph form.
The average cell survival rate is absorbance (average of each prodrug)/absorbance (average of control group) × 100%.
From fig. 8(a), fig. 9(a) and fig. 10(a), the following conclusions can be drawn: the ALA-HPO hybrid derivative shown in the formula (II) is incubated with HeLa, MCF-7 and A375 tumor cell lines for 4 hours, and the series of compounds are found to have no obvious dark cytotoxicity to tumor cells under the concentration of 100-400 mu M. When the incubation time is prolonged to 24h, the series of compounds do not show obvious dark toxicity to HeLa cells at the concentration of 100-400 mu M from the graph of FIG. 8(B), FIG. 9(B) and FIG. 10 (B); and b2, b3, b5, b6, b7, b8, b9 and b10 in MCF-7 cell line are more obviously cytotoxic to dark cell after being incubated for 24 hours at the concentration of 400 mu M; when this series of compounds was incubated with a375 cells for 24h, only compounds b8 and b10 showed significant dark toxicity at 400 μ M concentration.
Finally, it is to be noted that the above-mentioned list is only a few specific embodiments of the present invention. It is obvious that the invention is not limited to the above embodiments, but that many variations are possible. All modifications which can be derived or suggested by a person skilled in the art from the disclosure of the present invention are to be considered within the scope of the invention.

Claims (10)

1. An ALA hybrid 3-hydroxypyridone derivative of formula (I) or formula (II):
Figure FDA0002970338490000011
in the formula (I), R1Is C1-C10 alkyl, C3-C6 alkoxy and C6-C10 aryl;
r in the formula (II)2is-H or-OH, wherein n1＝1-5。
2. The ALA hybrid 3-hydroxypyridone derivative of formula (I) or formula (II) as claimed in claim 1, wherein said ALA hybrid 3-hydroxypyridone derivative is one of the following compounds:
Figure FDA0002970338490000012
3. a process for the preparation of ALA hybrid 3-hydroxypyridone derivatives of formula (I) as claimed in claim 1, wherein said process comprises:
(5) dissolving a compound 10, benzyl bromide and an alkaline substance E1 in a solvent E1, carrying out reflux reaction for 0.5-2 h, and carrying out aftertreatment on the obtained reaction liquid E1 to obtain an E1 compound 11; the mass ratio of the compound 10, the benzyl bromide and the alkaline substance E1 is 1: 1-4: 1-4; the alkaline substance is one or a mixture of more than two of potassium carbonate, potassium hydroxide, sodium carbonate and sodium bicarbonate;
Figure FDA0002970338490000021
(6) dissolving the compound 11, 3, 4-dihydro-2H-pyran and p-toluenesulfonic acid in the step (5) in dichloromethane, reacting for 4-12H at 20-60 ℃, and performing aftertreatment on the obtained reaction liquid F1 to obtain F1 to obtain a compound 12; the mass ratio of the compound 11, the 3, 4-dihydro-2H-pyran to the p-toluenesulfonic acid is 1: 2-6: 0.1 to 0.6;
Figure FDA0002970338490000022
(7) reacting the compound 12, R in the step (6)1NH2Dissolving in a solvent G1, carrying out reflux reaction for 12-36 h, and carrying out reduced pressure concentration on the obtained reaction liquid to obtain a crude product containing the compound 13; dissolving the crude product containing the compound 13 in ethanol, adjusting the pH value to 1-2 by using concentrated hydrochloric acid,continuously carrying out reflux reaction for 2-8 h, and carrying out aftertreatment on the obtained reaction liquid G1 to G1 to obtain a compound shown in the formula 2; the compound 12 and R1NH2The ratio of the amounts of substances (1): 4-10;
Figure FDA0002970338490000023
(8) dissolving the compound 4, dicyclohexylcarbodiimide and 4-dimethylaminopyridine in the step (1) in an organic solvent H1 under a protective atmosphere, stirring at 10-60 ℃ for 0.5-1H, slowly dropwise adding the compound 2 dissolved in the organic solvent Y1 in the step (7), continuing to react for 6-24H after dropwise adding, and carrying out aftertreatment on the obtained reaction liquid H1 with H1 to obtain a compound 5; the amount ratio of the compound 4, dicyclohexylcarbodiimide, 4-dimethylaminopyridine to the compound 2 is 1: 1-2: 0.1-0.8: 0.6-1.2;
Figure FDA0002970338490000024
(9) dissolving the compound 5 in the step (8) in ethyl acetate J1, dropwise adding an anhydrous ethyl acetate solution of saturated hydrogen chloride at-10-0 ℃, reacting for 1h after dropwise adding, transferring to room temperature, and continuing to react for 2-16 h, wherein the obtained reaction solution J1 is subjected to post-treatment J1 to obtain a compound 15; the volume of the anhydrous ethyl acetate solution of saturated hydrogen chloride is 1-5 mL/mmol based on the substance of the compound 5;
Figure FDA0002970338490000031
(10) dissolving the compound 15 in the step (9) in an organic solvent K1, adding a palladium-carbon catalyst, stirring and reacting for 1-16 h at 10-60 ℃ under a hydrogen atmosphere, and carrying out aftertreatment on the obtained reaction solution K1 with K1 to obtain an ALA hybrid 3-hydroxypyridone derivative shown in the formula (I); the mass ratio of the compound 15 to the palladium carbon catalyst is 1: 01-04;
Figure FDA0002970338490000032
wherein, the compound 2, 5, 13, 15, the formula (I) or R1NH2In, R1Is C1-C10 alkyl, C3-C6 alkoxy and C6-C10 aryl.
4. A process for the preparation of ALA hybrid 3-hydroxypyridone derivatives of formula (I) as claimed in claim 3, wherein: in the step (5), the solvent E1 is one or a mixed solvent of more than two of water, acetone, ethanol, methanol, dichloromethane, chloroform, carbon tetrachloride, acetonitrile, benzene, toluene, dimethyl sulfoxide, dioxane and N, N-dimethylformamide;
the solvent G1 in the step (7) is one or a mixed solvent of more than two of water, acetone, ethanol, methanol, chloroform, carbon tetrachloride, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide;
the organic solvent H1 in the step (8) is one or a mixed solvent of more than two of acetone, ethanol, methanol, dichloromethane, chloroform, carbon tetrachloride, toluene, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide; the organic solvent Y1 is dichloromethane;
in the step (10), the organic solvent K1 is one or a mixed solvent of more than two of acetone, ethanol, methanol, dichloromethane, chloroform, carbon tetrachloride, toluene, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide.
5. A process for the preparation of ALA hybrid 3-hydroxypyridone derivatives of formula (I) as claimed in claim 3, wherein:
the post-treatment E1 in the step (5) is as follows: cooling the reaction solution E1 to room temperature, concentrating, dissolving dichloromethane, washing with water, drying with anhydrous sodium sulfate, filtering, evaporating to remove the solvent, and drying to obtain a compound 11;
the post-treatment F1 in the step (6) is as follows: washing the reaction solution F1 with a sodium carbonate solution with the mass fraction of 5%, then washing with water, drying with anhydrous sodium sulfate, filtering, distilling under reduced pressure to remove the solvent, and drying in vacuum to obtain a compound 12;
the post-treatment G1 in the step (7) is as follows: cooling the reaction solution G1 to room temperature, concentrating under reduced pressure, dissolving in water, washing with diethyl ether, adjusting the pH value to 8-10, extracting with dichloromethane, combining organic layers, drying with anhydrous sodium sulfate, filtering, concentrating, and recrystallizing with a mixed solution of diethyl ether and methanol in a volume ratio of 12:1 to obtain a compound 2;
the post-treatment H1 in the step (8) is as follows: filtering the reaction liquid H1, concentrating the filtrate under reduced pressure, adding dichloromethane for dissolving, washing with a saturated sodium bicarbonate solution and a saturated sodium chloride solution in sequence, drying with anhydrous sodium sulfate, filtering, concentrating under reduced pressure, performing silica gel column chromatography, performing gradient elution with a mixed solution of dichloromethane and methanol in a volume ratio of 100-20: 1, collecting an eluent containing a target product, concentrating, and drying to obtain a compound 5;
the post-treatment J1 in the step (9) is as follows: distilling the reaction solution J1 under reduced pressure to remove the solvent, and drying to obtain a compound 15;
the post-treatment K1 in the step (10) is as follows: filtering the reaction solution K1, distilling the filtrate under reduced pressure to remove the solvent, and recrystallizing with a mixed solvent of methanol and diethyl ether at a volume ratio of 10:1 to obtain the ALA hybrid 3-hydroxypyridone derivative shown in formula (I).
6. A process for the preparation of ALA hybrid 3-hydroxypyridone derivatives of formula (II) as claimed in claim 1, wherein said process comprises:
1) dissolving a compound 1 or a compound 9, benzyl bromide and an alkaline substance A2 in an organic solvent A2, carrying out reflux reaction for 4-18 h, and carrying out aftertreatment on an obtained reaction liquid A2A 2 to obtain a compound 16; the amount ratio of the compound 1 or the compound 9 to the benzyl bromide and the alkaline substance A2 is 1: 1-4: 1-4; the alkaline substance A2 is one or a mixture of more than two of potassium carbonate, potassium hydroxide, sodium carbonate, sodium bicarbonate and triethylamine;
Figure FDA0002970338490000041
2) subjecting the compound 16, NH in the step 1)2(CH2)n1COOH and an alkaline substance B2 are dissolved in a solvent B2, reflux reaction is carried out for 4-24 hours, and the obtained reaction liquid B2 is subjected to post-treatment B2 to obtain a compound 3; the compound 16, NH2(CH2)n1The ratio of the amounts of COOH and of the basic substance B2 was 1: 1-4: 2-8; the alkaline substance B2 is one or a mixture of more than two of potassium carbonate, potassium hydroxide, sodium carbonate, sodium bicarbonate and triethylamine;
Figure FDA0002970338490000042
3) dissolving the compound 3 in the step 2) in an organic solvent C2, sequentially adding a compound 6 and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate, slowly dropwise adding N-methylmorpholine in an ice water bath, stirring for reacting for 45 minutes, completely dissolving the solid, transferring to the temperature of 20-50 ℃ for further reaction for 2-20 hours, and carrying out aftertreatment on the obtained reaction liquid C2 to obtain a compound 7, wherein the compound 3 is C2; the mass ratio of the compound of formula 3, 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate to the compound 6 to N-methylmorpholine is 1: 1-6: 1-6: 2-8;
Figure FDA0002970338490000051
4) dissolving the compound 7 in the step 3) in an organic solvent D2, adding a palladium-carbon catalyst, stirring and reacting for 4-18 h at 10-50 ℃ under a hydrogen atmosphere, and carrying out aftertreatment on the obtained reaction liquid D2 to obtain an ALA hybrid 3-hydroxypyridone derivative shown in a formula (II) after D2; the mass ratio of the compound of the formula 3 to the palladium carbon is 1:0.1 to 0.4;
Figure FDA0002970338490000052
in the compounds 3, 7 and 16, R3Is H-or PhCHO-; NH (NH)2(CH2)n1COOH or R in the formula (II)2is-H or-OH, wherein n1＝1-5。
7. A process for the preparation of ALA hybrid 3-hydroxypyridone derivatives of formula (II) as claimed in claim 6, wherein: the organic solvent A2 in the step 1) is one or a mixed solvent of more than two of acetone, ethanol, methanol, dichloromethane, chloroform, carbon tetrachloride, acetonitrile, toluene, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide;
the solvent B2 in the step 2) is one or a mixed solvent of more than two of water, acetone, ethanol, methanol, dichloromethane, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide;
the organic solvent C2 in the step 3) is one or a mixed solvent of more than two of acetone, ethanol, methanol, acetic acid, dichloromethane, acetonitrile, dimethyl sulfoxide, dioxane, piperidine, N-dimethylformamide and N, N-dimethylacetamide;
the organic solvent D2 in the step 4) is one or a mixed solvent of more than two of acetone, ethanol, methanol, dichloromethane, chloroform, carbon tetrachloride, toluene, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide.
8. A process for the preparation of ALA hybrid 3-hydroxypyridone derivatives of formula (II) as claimed in claim 6, wherein: the post-treatment A2 in the step 1) is as follows: cooling the reaction solution A2 to room temperature, concentrating under reduced pressure, dissolving with dichloromethane, washing with water, drying with anhydrous sodium sulfate, filtering, concentrating under reduced pressure, performing silica gel column chromatography, performing gradient elution with a mixed solution of n-hexane and ethyl acetate at a volume ratio of 8-3: 1, collecting an eluent containing a target product, concentrating, and drying to obtain a compound 16;
the post-treatment B2 in the step 2) is as follows: cooling the reaction solution B2 to room temperature, concentrating under reduced pressure, adding water for dissolution, washing with dichloromethane, adjusting the pH to 1-2 by using 2mol/L hydrochloric acid, extracting with dichloromethane, combining organic layers, drying with anhydrous sodium sulfate, filtering, evaporating the filtrate under reduced pressure to remove the solvent, drying in vacuum, and recrystallizing the obtained solid with a mixed solvent of methanol and ether in a volume ratio of 1:10 to obtain a compound 3;
the post-treatment C2 in the step 3) is as follows: adding water into the reaction liquid C2, extracting with dichloromethane, combining organic layers, washing with 0.1M hydrochloric acid and saturated sodium bicarbonate in sequence, drying with anhydrous sodium sulfate, filtering, concentrating, performing silica gel column chromatography, and performing silica gel column chromatography according to the volume ratio of 80: -20: 1, gradient elution is carried out by taking a mixed solution of dichloromethane and methanol as an eluent, the eluent containing a target product is collected, concentrated and dried to obtain a compound 7;
the post-treatment D2 in the step 4) is as follows: after the reaction is finished, filtering the reaction solution, concentrating the filtrate under reduced pressure, and recrystallizing with methanol/diethyl ether to obtain the compound shown in the formula (II); filtering the reaction liquid D2, taking the filtrate, distilling the filtrate under reduced pressure to remove the solvent, and recrystallizing with a mixed solvent of methanol and diethyl ether with the volume ratio of 1:10 to obtain the ALA hybrid 3-hydroxypyridone derivative shown in the formula (II).
9. Use of an ALA hybrid 3-hydroxypyridone derivative of formula (I) or formula (II) as defined in claim 1 for the preparation of a photodynamic therapy medicament for the prevention or treatment of a tumor.
10. The use of claim 9, wherein: the tumor cells are HeLa cells, MCF-7 cells or A375 cells.
CN202110261792.8A 2021-03-10 2021-03-10 ALA hybrid 3-hydroxypyridone derivative and preparation method and application thereof Active CN113121423B (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CN202110261792.8A CN113121423B (en) 2021-03-10 2021-03-10 ALA hybrid 3-hydroxypyridone derivative and preparation method and application thereof
CN202210473051.0A CN114736155B (en) 2021-03-10 2021-03-10 ALA hybrid 3-hydroxypyridone derivative, and preparation method and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110261792.8A CN113121423B (en) 2021-03-10 2021-03-10 ALA hybrid 3-hydroxypyridone derivative and preparation method and application thereof

Related Child Applications (1)

Application Number Title Priority Date Filing Date
CN202210473051.0A Division CN114736155B (en) 2021-03-10 2021-03-10 ALA hybrid 3-hydroxypyridone derivative, and preparation method and application thereof

Publications (2)

Publication Number Publication Date
CN113121423A true CN113121423A (en) 2021-07-16
CN113121423B CN113121423B (en) 2022-07-22

Family

ID=76773012

Family Applications (2)

Application Number Title Priority Date Filing Date
CN202210473051.0A Active CN114736155B (en) 2021-03-10 2021-03-10 ALA hybrid 3-hydroxypyridone derivative, and preparation method and application thereof
CN202110261792.8A Active CN113121423B (en) 2021-03-10 2021-03-10 ALA hybrid 3-hydroxypyridone derivative and preparation method and application thereof

Family Applications Before (1)

Application Number Title Priority Date Filing Date
CN202210473051.0A Active CN114736155B (en) 2021-03-10 2021-03-10 ALA hybrid 3-hydroxypyridone derivative, and preparation method and application thereof

Country Status (1)

Country Link
CN (2) CN114736155B (en)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998054138A1 (en) * 1997-05-29 1998-12-03 Btg International Limited Novel orally active iron (iii) chelators
CN106478493A (en) * 2016-10-12 2017-03-08 浙江工商大学 5 aminolevulinic acid/3 Hydroxypyridinone conjugates and its preparation method and purposes
CN112023040A (en) * 2020-08-04 2020-12-04 浙江工业大学 Derivative of antineoplastic photosensitizer ALA hybrid 3-hydroxypyridine-4H-ketone and preparation method and application thereof
CN112062713A (en) * 2020-08-04 2020-12-11 浙江工业大学 ALA-HPO hybrid derivative with iron chelation and PDT activity and preparation method and application thereof

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104262241B (en) * 2014-08-26 2016-04-06 浙江工商大学 The conjugate of 5-ALA and 3-pyridone-4-ketone and preparation method thereof and purposes

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998054138A1 (en) * 1997-05-29 1998-12-03 Btg International Limited Novel orally active iron (iii) chelators
CN106478493A (en) * 2016-10-12 2017-03-08 浙江工商大学 5 aminolevulinic acid/3 Hydroxypyridinone conjugates and its preparation method and purposes
CN112023040A (en) * 2020-08-04 2020-12-04 浙江工业大学 Derivative of antineoplastic photosensitizer ALA hybrid 3-hydroxypyridine-4H-ketone and preparation method and application thereof
CN112062713A (en) * 2020-08-04 2020-12-11 浙江工业大学 ALA-HPO hybrid derivative with iron chelation and PDT activity and preparation method and application thereof

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ROLAND TRESSL ET AL.: "Formation of Pyrroles, 2-Pyrrolidones, and Pyridones by Heating of 4-Aminobutyric Acid and Reducing Sugars", 《J.AGRIC.FOOD CHEM.》 *
SARAH FAKIH ET AL.: "Targeting the Lysosome: Fluorescent Iron(III) Chelators To Selectively Monitor Endosomal/Lysosomal Labile Iron Pools", 《J.MED.CHEM.》 *
朱纯风: "新型医用3-羟基吡啶-4-酮衍生物的设计与合成", 《中国优秀硕士学位论文全文数据库（医药卫生科技辑）》 *
董琳琳: "3-羟基-4-吡啶酮类螯合剂在镓放射性药物中的研究进展及展望", 《核化学与放射化学》 *

Also Published As

Publication number Publication date
CN113121423B (en) 2022-07-22
CN114736155B (en) 2023-12-05
CN114736155A (en) 2022-07-12

Similar Documents

Publication Publication Date Title
CN101665484B (en) Method for preparing lenalidomide
CA3132620C (en) Salts and prodrugs of 1-methyl-d-tryptophan
CN113121423B (en) ALA hybrid 3-hydroxypyridone derivative and preparation method and application thereof
CN108947859B (en) Derivatives of kutkin dimer analogue JJA-D0 or pharmaceutically acceptable salts thereof, preparation method and application
CN107698521B (en) Preparation and application of 5-fluorouracil substituted carboxylic acid derivative
CN107522654A (en) New α aminoamide derivatives and its medical usage
CN102126993A (en) Resveratrol derivative and application thereof to preparation of antitumor medicaments
CN110938109B (en) O-quinone compound, preparation method and medical application thereof
CN110804045B (en) Coumarin hybrid pyridone amide derivative with potential anti-AD activity and preparation method and application thereof
CN112023040B (en) Derivative of antineoplastic photosensitizer ALA hybrid 3-hydroxypyridine-4H-ketone and preparation method and application thereof
CN112062713B (en) ALA-HPO hybrid derivative with iron chelation and PDT activity and preparation method and application thereof
CN113717174B (en) Tumor targeting drug resistant O6-thienylmethylguanine-indoloquinone-chloroethylnitrosourea combined molecule and preparation method thereof
CN111205314A (en) O-carborane dibenzonitrile derivative and modified compound thereof, and synthesis methods thereof
CN112552232B (en) Pyridone hexa-alkynylamine modified derivative and preparation method and application thereof
CN110218207B (en) Coumarin hybrid pyridone compound with iron chelation and monoamine oxidase B inhibition activity and preparation and application thereof
CN114751953A (en) Phthalocyanine derivative for targeted photodynamic therapy of postmenopausal breast cancer and preparation method thereof
CN103601673B (en) The fluorinated derivatives of 3-pyridone-4-ketone
CN105985320B (en) Benzyl phthalazine compound and its preparation method and application
CN113173955A (en) Two-photon diagnosis and treatment prodrug with targeted Golgi pH response and preparation thereof
Pessoa-Mahana et al. STUDIES ON QUINONES. PART 36.1 SYNTHESIS AND TRYPANOCIDAL ACTIVITY OF 2-ALKOXYCARBONYLBENZO [b] THIOPHENE-4, 7-QUINONES
CN113968860B (en) Reversible BTK inhibitor and synthesis method and application thereof
CN112679470B (en) (E) -1-phenyl-4-alkenyl-1H-pyrazole compound and medical application thereof
CN103864779A (en) Preparation of 1-(phenyl)-2,3,4,9-tetrahydro-1H-pyridino[3,4-b]indole derivatives and application of derivatives in antitumor drugs
CN116375685A (en) Fasudil derivative and preparation method and application thereof
CN112442091B (en) Replication protein A targeted platinum compound

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant