CN112062713B - ALA-HPO hybrid derivative with iron chelation and PDT activity and preparation method and application thereof - Google Patents
ALA-HPO hybrid derivative with iron chelation and PDT activity and preparation method and application thereof Download PDFInfo
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Abstract
The invention provides an ALA-HPO hybrid derivative shown in a formula (I) or a formula (II), a preparation method thereof and application thereof in preparing photodynamic therapy medicaments;
Description
Technical Field
The invention relates to a Hydroxypyridone (HPO) and 5-aminolevulinic acid (ALA) coupled derivative with iron ion chelating capacity, a preparation method thereof and application in the aspect of photodynamic.
Background
Traditional tumor treatment methods mainly comprise surgery, chemotherapy, radiotherapy and the like, but the treatment methods have some serious defects. In recent years, with the development of medical technology, photodynamic therapy (PDT) has gradually entered the visual field of people, PDT is a method of activating an exogenously applied or endogenously formed photosensitive substance by visible light in the presence of molecular oxygen, and photosensitizers after photoactivation cause the formation of singlet oxygen, which is a strong oxidant and can cause the damage and cell death of various subcellular substrates. Through research on the action mechanism of the photosensitizer, the photosensitizer is found to be a key factor in the PDT treatment process, and the performance of the photosensitizer determines the treatment effect of PDT. Therefore, it is important to develop an ideal photosensitizer which has no dark toxicity, low skin photosensitivity and strong targeting property.
ALA is a second generation photosensitizer that has attracted considerable interest to researchers in the fields of PDT and photobiology. ALA is an early intermediate in the heme biosynthetic pathway and is not itself a photosensitizer. It can generate protoporphyrin PpIX with photoactivity in vivo through a series of reactions. This process occurs more efficiently in rapidly dividing cancer cells and thus ALA is less toxic to normal tissue cells. The disadvantage of ALA arises mainly from ALA itself, which is a zwitterion at physiological pH conditions, and its hydrophilicity severely hampers its ability to pass through the cell membrane, limiting its uptake by the cell.
In order to overcome the problem of low bioavailability associated with ALA, the development of ALA prodrugs has been the subject of most. Based on the mechanism of action of ALA in the body, it was found that PpIX production by ALA was associated with iron chelatase, which forms the basis of therapeutic approaches combining iron chelators with ALA-mediated PDT. In the final step of heme synthesis, the iron chelator chelates Fe with the action of ferrochelatase 2+ Leading to a massive accumulation of PpIX, suggesting that iron is critical for conversion of PpIX to hemoglobin in rapidly proliferating tumor cells.
Therefore, iron chelation strategies in modern anticancer chemotherapy have attracted considerable interest, but cellular iron is always chelated by various molecules, which does not render it non-toxic, since the chelated iron is still capable of generating reactive oxygen species. Deferoxamine was found to be selective in increasing PpIX accumulation after its use as a heme synthesis inhibitor, but the pharmacokinetic profile was poor, which resulted in a longer duration of its distribution, and secondly, higher DFO concentrations may have a detrimental effect on PpIX formation due to the strong antioxidant activity of deferoxamine under these conditions. A group of hydroxypyridone-based chelators, HPO, has recently been reported, which have a lower molecular weight and a higher lipophilicity than deferoxamine, and thus are particularly effective in chelating intracellular iron, thereby enhancing PpIX formation.
Disclosure of Invention
The invention designs and synthesizes novel active compounds with iron chelation and PDT activity based on the principles of reasonable drug design, drug-like property and the like. In particular to a 5-aminolevulinic acid-hydroxypyridone (ALA-HPO) conjugate capable of remarkably improving the drug effect of 5-aminolevulinic acid, and a preparation method and application thereof.
The technical scheme of the invention is as follows:
an ALA-HPO hybrid derivative of formula (I) or formula (II):
in the formula (I) or the formula (II),
R 1 is C1-C6 alkyl, C1-C3 alkoxyC 1-C3 alkyl or C6-C10 arylC 1-C3 alkyl, preferably R 1 Is methyl, ethyl, propyl, butyl, pentyl, hexyl, methoxypropyl, methoxyethyl, ethoxypropyl or phenethyl;
R 2 is C1-C10 alkyl or C1-C3 alkoxyC 1-C3 alkyl, preferably R 2 Is n-butyl, n-pentyl, isopentyl, n-hexyl, n-heptyl, n-octyl, n-decyl, methoxyethyl, methoxypropyl or ethoxypropyl.
The invention also provides a preparation method of the ALA-HPO hybrid derivative shown in the formula (I) or the formula (II).
The preparation method of the ALA-HPO hybrid derivative shown in the formula (I) comprises the following steps:
(1) Dissolving the compound 1, benzyl bromide and an alkaline substance in an organic solvent, reacting for 2-12 h (preferably 3-8 h) at 25-90 ℃ (preferably 25-65 ℃), and then carrying out post-treatment on the reaction solution to obtain a compound 2;
the mass ratio of the compound 1, the benzyl bromide and the alkaline substance is 1:1 to 6:1 to 5, preferably 1:2 to 5:2 to 4;
the alkaline substance is one or a mixture of more than two of potassium carbonate, potassium hydroxide, sodium carbonate, sodium bicarbonate and triethylamine in any proportion, preferably potassium carbonate or potassium hydroxide;
the organic solvent 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 in any proportion, and preferably acetone or N, N-dimethylformamide;
the volume usage of the organic solvent is 1-5 mL/mmol based on the substance of the compound 1;
the post-treatment method comprises the following steps: after the reaction is finished, concentrating the reaction solution under reduced pressure, adding water, extracting with dichloromethane, combining organic layers, washing with saturated saline solution, drying with anhydrous sodium sulfate, and finally evaporating to remove the solvent and drying to obtain a compound 2;
(2) Mixing the compounds 2 and R 1 NH 2 Dissolving an alkaline substance in an organic solvent, reacting for 1-12 h (preferably 8-12 h) at 40-100 ℃ (preferably 60-85 ℃), and then carrying out post-treatment on the reaction solution to obtain a compound 3a;
the compound 2, R 1 NH 2 The ratio of the amount of the alkaline substance is 1:1 to 5:0.5 to 2, preferably 1: 1.5-2: 0.8 to 1.5;
the alkaline substance is one or a mixture of more than two of potassium carbonate, potassium hydroxide, sodium carbonate, sodium bicarbonate and triethylamine in any proportion, preferably sodium hydroxide or potassium hydroxide;
the organic solvent 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 in any proportion, preferably ethanol or a mixed solvent of ethanol and water;
the volume usage of the organic solvent is 2-4 mL/mmol based on the substance of the compound 2;
the post-treatment method comprises the following steps: after completion of the reaction, the reaction mixture was concentrated under reduced pressure and subjected to silica gel column chromatography, and the reaction mixture was purified by dichloromethane: methanol: ammonia (25 to 28 wt%) =10:1:0.1 Separating the mixture as eluent to obtain compound 3a;
(3) Reacting compound 4 with Boc 2 Dissolving O and an alkaline substance in an organic solvent, reacting for 2-24 h (preferably 8-20 h) at 0-50 ℃ (preferably 10-40 ℃), and then carrying out post-treatment on the reaction solution to obtain a compound 5;
the compound 4, boc 2 The ratio of the amount of O to the amount of the basic substance is 1:1 to 2:2 to 10, preferably 1:1:6;
the alkaline substance is one or a mixture of more than two of potassium carbonate, potassium hydroxide, sodium carbonate, sodium bicarbonate, sodium acetate, sodium propionate and triethylamine in any proportion, preferably sodium bicarbonate;
the organic solvent is one or a mixed solvent of more than two of acetone, ethanol, methanol, acetic acid, acetic anhydride, dichloromethane, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide in any proportion, preferably methanol;
the volume usage of the organic solvent is 1-2 mL/mmol based on the substance of the compound 4;
the post-treatment method comprises the following steps: after the reaction, the reaction solution was filtered (to remove solids in the reaction system), the filtrate was distilled under reduced pressure to remove the solvent, the residue was dissolved in water, acidified to pH =2 with 10wt% potassium hydrogen sulfate solution, extracted with ethyl acetate, the organic layer was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent and dried to give compound 5;
(4) Dissolving the compound 5 in an organic solvent, sequentially adding dicyclohexylcarbodiimide and 4-dimethylaminopyridine, reacting for 2-12 h (preferably 4-8 h) at 10-50 ℃ (preferably 25-35 ℃), filtering the reaction solution, and collecting the filtrate; dissolving the compound 3a in an organic solvent, dripping the solution into the collected filtrate at the temperature of between 10 and 50 ℃ (preferably between 25 and 35 ℃), maintaining the temperature for reacting for 6 to 24 hours (preferably between 4 and 8 hours) after dripping, and then carrying out post-treatment on the reaction solution to obtain a compound 6;
the mass ratio of the compound 5, dicyclohexylcarbodiimide, 4-dimethylaminopyridine and the compound 3a is 10:10:1:10;
the organic solvent used for dissolving the compound 5 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 in any proportion, preferably a mixed solvent of dichloromethane and N, N-dimethylacetamide; the definition of the organic solvent used for dissolving the compound 3a is the same as that;
when the compound 5 is dissolved, the volume usage of the organic solvent is 5-15 mL/mmol based on the substance of the compound 5; when the compound 3a is dissolved, the volume usage of the organic solvent is 5-15 mL/mmol based on the substance of the compound 3a;
the post-treatment method comprises the following steps: after the reaction, the reaction mixture was concentrated under reduced pressure and subjected to silica gel column chromatography, and the reaction mixture was purified by dichloromethane: methanol =100 to 20:1 as eluent, and separating to obtain a compound 6;
(5) Dissolving a compound 6 in dichloromethane, dropwise adding an anhydrous dichloromethane solution of 1.0mol/L trifluoroacetic acid under the protection of nitrogen at-10-20 ℃, transferring to room temperature (20-30 ℃) after dropwise adding, continuously stirring and reacting for 12-24 h, and then distilling the reaction solution under reduced pressure to remove the solvent to obtain a compound 7;
the mass ratio of the compound 6 to trifluoroacetic acid is 1:1;
the concentration of the solution obtained by dissolving the compound 6 in dichloromethane is 0.02-0.06 mmol/mL;
(6) Dissolving the compound 7 in an organic solvent, adding palladium carbon, stirring and reacting for 2-24 h under the atmosphere of hydrogen at-5-50 ℃ (preferably 10-30 ℃), and then carrying out post-treatment on reaction liquid to obtain a compound shown in a formula (I);
the mass ratio of the compound 7 to palladium on carbon (calculated as palladium) is 5:1 to 3, preferably 5: 1-2;
the organic solvent 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 in any proportion, and ethanol is preferred;
the concentration of the solution obtained by dissolving the compound 7 in the organic solvent is 0.02-0.06 mmol/mL;
the post-treatment method comprises the following steps: after the reaction is finished, filtering the reaction solution, distilling the filtrate under reduced pressure to remove the solvent, and recrystallizing with ethanol/diethyl ether to obtain the compound shown in the formula (I);
the R is 1 NH 2 In the compound 3a, the compound 6 and the compound 7, R 1 Is as defined in formula (I);
the preparation method of the ALA-HPO hybrid derivative shown in the formula (II) comprises the following steps:
(1) The preparation method is the same as the step (1) of the ALA-HPO hybrid derivative shown in the formula (I);
(2) In the step (2) of the preparation method of the ALA-HPO hybrid derivative shown as the formula (I), R 1 NH 2 Is replaced by R 2 NH 2 Reacting to obtain a compound 3b;
(3) Dissolving the compound 8 in an organic solvent, dropwise adding an anhydrous dichloromethane solution of the compound 3b and triethylamine at-20 ℃ (preferably-10-0 ℃), transferring to room temperature after dropwise addition, continuously stirring and reacting for 4-24 h (preferably 8-12 h), and then carrying out post-treatment on a reaction solution to obtain a compound 9;
the mass ratio of the compound 3b to the compound 8 to the triethylamine is 1:1 to 5:0.5 to 2, preferably 1: 1.5-2: 0.8 to 1.5;
the organic solvent 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 in any proportion, preferably dichloromethane;
the volume usage of the organic solvent is 1-2 mL/mmol based on the substance of the compound 8;
the post-treatment method comprises the following steps: after completion of the reaction, the reaction mixture was concentrated under reduced pressure and subjected to silica gel column chromatography using a dichloromethane: methanol =80:1 (volume ratio) is used as eluent, and a compound 9 is obtained by separation;
(4) Dissolving the compound 11 in an organic solvent, sequentially adding triethylamine and 4-dimethylaminopyridine, and reacting at-10-20 ℃ for 4-24 hours to obtain a reaction mixed solution; dissolving a compound 9 in an organic solvent, dropwise adding the obtained reaction mixed solution at-10-20 ℃, continuously reacting for 2-24 h (preferably 4-8 h) at-10-50 ℃ (preferably-5-10 ℃) after dropwise adding, and then carrying out post-treatment on the reaction solution to obtain a compound 12;
the mass ratio of the compound 9, triethylamine, 4-dimethylaminopyridine and the compound 11 is 1:2:0.1:1.1;
the organic solvent used for dissolving the compound 9 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 in any proportion, preferably dichloromethane; the definition of the organic solvent used for dissolving the compound 11 is the same as that;
when the compound 9 is dissolved, the volume usage of the organic solvent is 5-15 mL/mmol based on the substance of the compound 9; when the compound 11 is dissolved, the volume usage of the organic solvent is 5-15 mL/mmol based on the substance of the compound 11;
the post-treatment method comprises the following steps: after completion of the reaction, the reaction mixture was concentrated under reduced pressure and subjected to silica gel column chromatography using a dichloromethane: methanol =100 to 20:1 (volume ratio) as eluent to carry out gradient elution, and separating to obtain a compound 12;
(5) Replacing a compound 7 with a compound 12 in the step (6) of the preparation method of the ALA-HPO hybrid derivative shown in the formula (I) to obtain a compound shown in a formula (II) through reaction;
the R is 2 NH 2 Compound 3b, compound 9, compound 12, R 2 Is as defined in formula (II).
The ALA-HPO hybrid derivative shown in the formula (I) or the formula (II) can be used for preparing photodynamic therapeutic medicaments, and particularly can be used for preparing medicaments for treating squamous cell carcinoma, skin carcinoma or basal cell carcinoma.
Compared with the prior art, the invention has the beneficial effects that: the invention synthesizes a series of novel compounds with PDT activity.
Drawings
FIG. 1 viability of HeLa cells in example 21 at varying light intensities and compound concentrations of 50. Mu.M.
FIG. 2 viability of HeLa cells in example 21 at varying light intensities, compound concentration 100. Mu.M.
FIG. 3 viability of HeLa cells in example 21 at concentrations of compound 50. Mu.M or 100. Mu.M without light.
FIG. 4 viability of HaCaT cell line at varying light intensity, compound concentration 50. Mu.M in example 22.
FIG. 5 viability of HaCaT cell line at varying light intensities, compound concentration 100. Mu.M in example 22.
FIG. 6 viability of HaCaT cell lines in example 22 in the absence of light, at a concentration of compound 50. Mu.M or 100. Mu.M, cultured for 4h.
FIG. 7 viability of HaCaT cell lines in example 22 in the absence of light, at a concentration of compound 50. Mu.M or 100. Mu.M, cultured for 24 h.
Detailed Description
The present invention is further illustrated by the following specific examples, but the scope of the invention is not limited thereto.
Example 1
A100 mL round bottom flask was charged with ALA hydrochloride (5.00g, 29.8 mmol), naHCO 3 (15.04g, 179mmol) and Boc 2 O (6.51g, 29.8 mmol) and 40mL of anhydrous methanol as a solvent were added, the mixture was stirred at room temperature, and the progress of the reaction was checked by TLC during the reaction. After 12h, the reaction was stopped, at which time the reaction solution was light yellow, the solid was filtered off, the solvent was distilled off under reduced pressure, the residual solid was dissolved in water and acidified to pH =2 with 10% potassium hydrogen sulfate solution, extracted with ethyl acetate, the organic layer was washed with saturated brine (15 mL × 3) a few times, dried over anhydrous sodium sulfate, filtered, and the organic solvent was distilled off under reduced pressure to give N-Boc-ALA (6.24g, 90%) as a white solid.
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Benzyl bromide (10.26g, 60mmol) was placed in a constant pressure funnel, slowly added dropwise to the reaction solution with stirring, and after the dropwise addition was completed, the reflux reaction was continued for 2 hours, during which time the progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was concentrated in vacuo to precipitate a large amount of yellow solid, which was washed with water (50 mL. Times.3) and then filtered to give pyrone (13.5 g) as a pale yellow solid in 91% yield.
Pyrone (2.30g, 10mmol) is added into a 100mL round-bottom flask respectively, dissolved in ethanol and water (1:1) by 30mL, stirred, and after the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is more than 12, methylamine (0.372g, 12mmol) is added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC during the reaction. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and the mixture was purified by a silica gel column, and gradient elution was performed using dichloromethane: methanol (100). The product 1- (2-aminomethyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one was isolated as a yellow oil (1.5 g) in 50% yield.
DCC (2 mmol), DMAP (0.2 mmol) and N-Boc-ALA (2.2 mmol) were dissolved in dichloromethane (20 mL) and DMF (5 mL) under nitrogen and stirred at room temperature for 50min. 1-methyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (2 mmol) was dissolved in dichloromethane (10 mL) and placed in a constant pressure dropping funnel and added dropwise over 30 min. The reaction was stirred at room temperature overnight. The precipitate was filtered off, the solvent was distilled off under reduced pressure, the residue was dissolved in dichloromethane, washed with water and then with saturated sodium bicarbonate solution, the organic layer was concentrated under reduced pressure, and the residue was purified by flash chromatography on silica gel using a gradient of dichloromethane: methanol = 80.
A20 mL round bottom flask was charged with the above yellow oily compound (0.5 mmol), dichloromethane as a solvent, trifluoroacetic acid (2.1g, 3.1mL) diluted with dichloromethane (2.5 mL) in an isopiestic dropping funnel, slowly added dropwise, stirred at 0-5 ℃ for 1h, monitored by TLC for the progress of the reaction, and after completion of the reaction, dichloromethane and trifluoroacetic acid were distilled off under reduced pressure to give the intermediate compound as a yellow oily compound (478 mg) in 91% yield.
0.5mmol of the above compound was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi hydrogen pressure, catalytically reacted for 2 hours, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/ether to give ALA-HPO complex 10a.
ESI-HRMS:m/z calcd for C 12 H 17 N 2 O 5 [M+H] + :269.1132；found:269.1130.
1 H NMR(400MHz,DMSO-d 6 )δ8.20(s,1H),8.16(s,3H),7.21(s,1H),5.26(s,2H),4.11–3.86(m,5H),2.86(t,J＝5.7,2H),2.71(t,J＝5.2Hz,2H). 13 C NMR(100MHz,DMSO-d 6 )δ203.09,171.89,162.61,159.09(q,J＝33.0Hz,COO),145.65,143.85,132.50,118.51,115.57,113.76,65.35,61.31,47.09,42.91,34.58.
Example 2
A100 mL round bottom flask was charged with ALA hydrochloride (5.00g, 29.8 mmol), naHCO 3 (15.04g, 179mmol) and Boc 2 O (6.51g, 29.8 mmol) and 40mL of anhydrous methanol as a solvent were added, the mixture was stirred at room temperature, and the progress of the reaction was checked by TLC during the reaction. After 12h, the reaction was stopped, at which time the reaction solution was light yellow, the solid was filtered off, the solvent was distilled off under reduced pressure, the residual solid was dissolved in water and acidified to pH =2 with 10% potassium hydrogen sulfate solution, extracted with ethyl acetate, the organic layer was washed with saturated brine (15 mL × 3) a few times, dried over anhydrous sodium sulfate, filtered, and the organic solvent was distilled off under reduced pressure to give N-Boc-ALA (6.24g, 90%) as a white solid.
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Benzyl bromide (10.26g, 60mmol) was placed in a constant pressure funnel, slowly added dropwise to the reaction solution with stirring, and after the dropwise addition was completed, the reflux reaction was continued for 2 hours, during which time the progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was concentrated in vacuo to precipitate a large amount of yellow solid, which was washed with water (50 mL. Times.3) and then filtered to give pyrone (13.5 g) as a pale yellow solid with a yield of 91%.
Pyrone (2.30g, 10mmol) is respectively added into a 100mL round-bottom flask, dissolved in ethanol and water (1:1) by 30mL, stirred, and after the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is more than 12, ethylamine (0.54g, 12mmol) is added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC during the reaction. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and purified by a silica gel column, using a gradient elution of dichloromethane: methanol (100). The product, 1- (2-aminoethyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one, was isolated as a yellow oil (1.8 g) in 53% yield.
DCC (2 mmol), DMAP (0.2 mmol) and N-Boc-ALA (2.2 mmol) were dissolved in dichloromethane (20 mL) and DMF (5 mL) under nitrogen and stirred at room temperature for 50min. 1-Ethyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (0.518mg, 2mmol) was dissolved in dichloromethane (10 mL), placed in a constant pressure dropping funnel and added dropwise over 30 min. The reaction was stirred at room temperature overnight. The precipitate was filtered off, the solvent was distilled off under reduced pressure, the residue was dissolved in dichloromethane, washed with water and then with saturated sodium bicarbonate solution, the organic layer was concentrated under reduced pressure, and the residue was purified by flash chromatography on silica gel using a gradient of dichloromethane: methanol = 80.
The benzyl and Boc protected hydroxypyridones (0.236 g,0.5 mmol) were added to a 20mL round bottom flask in the presence of dichloromethane as solvent, trifluoroacetic acid (2.1g, 3.1mL) was diluted with dichloromethane (2.5 mL) in a constant pressure dropping funnel, slowly added dropwise, stirred at 0-5 ℃ for 1h, the progress of the reaction was monitored by TLC, and after completion of the reaction, dichloromethane and trifluoroacetic acid were distilled off under reduced pressure to give benzyl protected hydroxypyridones as yellow oil (0.245 mg) in a yield of 89%.
0.5mmol of the above benzyl-protected hydroxypyridone was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi of hydrogen pressure, catalytically reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to give ALA-HPO complex 10b.
HRMS:m/z calcd for C 13 H 19 N 2 O 5 [M+H] + :283.1288；found:283.1289.
1 H NMR(400MHz,DMSO-d 6 )δ8.24(s,1H),8.17(s,3H),7.19(s,1H),5.27(s,2H),4.26(q,J＝7.2Hz,2H),3.99(s,2H),2.85(t,J＝6.5Hz,2H),2.68(t,J＝6.5Hz,2H),1.41(t,J＝7.2Hz,3H). 13 C NMR(100MHz,DMSO-d 6 )δ203.02,171.90,163.34,159.12(q,J＝33.1Hz,COO),143.38,143.03,130.45,118.52,115.57,114.36,65.35,61.24,50.35,47.10,34.57,27.33.
Example 3
A100 mL round bottom flask was charged with ALA hydrochloride (5.00g, 29.8 mmol), naHCO 3 (15.04g, 179mmol) and Boc 2 O (6.51g, 29.8 mmol), and 40mL of anhydrous methanol as a solvent were added, and the mixture was stirred at room temperature, during which the progress of the reaction was checked by TLC. After 12h, the reaction was stopped, at which point the reaction solution was light yellow, the solid was filtered off, the solvent was distilled off under reduced pressure, the residual solid was dissolved in water and acidified to pH =2 with 10% potassium hydrogen sulfate solution, extracted with ethyl acetate, and the organic layer was diluted with saturated brine (15 mL × 3) a few timesWashing, drying over anhydrous sodium sulfate, filtering, and removing the organic solvent by distillation under reduced pressure gave N-Boc-ALA (6.24g, 90%) as a white solid.
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Benzyl bromide (10.26g, 60mmol) was placed in a constant pressure funnel, slowly added dropwise to the reaction solution with stirring, and after the dropwise addition was completed, the reflux reaction was continued for 2 hours, during which time the progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was concentrated in vacuo to precipitate a large amount of yellow solid, which was washed with water (50 mL. Times.3) and then filtered to give pyrone (13.5 g) as a pale yellow solid with a yield of 91%.
Pyrone (2.30g, 10mmol) is respectively added into a 100mL round-bottom flask, dissolved in ethanol and water (1:1) by 30mL, stirred, when the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is more than 12, propylamine (0.708g, 12mmol) is continuously added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC during the reaction. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and the mixture was purified by a silica gel column, and gradient elution was performed using dichloromethane: methanol (100). The product, 1- (2-aminopropyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one, was isolated as a yellow oil (1.81 g) in 54% yield.
DCC (2 mmol), DMAP (0.2 mmol) and N-Boc-ALA (2.2 mmol) were dissolved in dichloromethane (20 mL) and DMF (5 mL) under nitrogen and stirred at room temperature for 50min. 1-propyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (0.546g, 2mmol) was dissolved in dichloromethane (10 mL) and placed in a constant pressure dropping funnel and added dropwise over 30 min. The reaction was stirred at room temperature overnight. The precipitate was filtered off, the solvent was distilled off under reduced pressure, the residue was dissolved in dichloromethane, washed with water then saturated sodium bicarbonate solution, the organic layer was concentrated under reduced pressure, and the residue was purified by flash chromatography on silica gel using a gradient of dichloromethane: methanol = 80.
A20 mL round bottom flask was charged with benzyl and Boc protected hydroxypyridinones (0.243g, 0.5mmol) in dichloromethane as solvent, trifluoroacetic acid (2.1g, 3.1mL) was diluted with dichloromethane (2.5 mL) in a constant pressure dropping funnel, slowly added dropwise, stirred at 0-5 ℃ for 1h, the progress of the reaction was monitored by TLC, and after completion of the reaction, dichloromethane and trifluoroacetic acid were distilled off under reduced pressure to give benzyl protected hydroxypyridinone (468 mg) in 87% yield.
0.5mmol of benzyl-protected hydroxypyridone was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi of hydrogen pressure, catalytically reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to give ALA-HPO complex 10C.
ESI-HRMS:m/z calcd for C 14 H 21 N 2 O 5 [M+H] + :297.1445；found:297.1446.
1 H NMR(400MHz,DMSO-d 6 )δ8.16(d,J＝9.8Hz,3H),7.10(d,J＝12.4Hz,1H),5.25(s,2H),4.15(t,J＝7.6Hz,2H),3.98(s,2H),2.85(t,J＝6.5Hz,2H),2.67(t,J＝6.4Hz,2H),1.80(h,J＝7.5Hz,2H),0.91(t,J＝7.3Hz,3H). 13 C NMR(150MHz,DMSO-d 6 )δ202.99,171.91,170.11,158.58(q,J＝30.0Hz,COO),147.87,141.78,124.74,116.72,114.85,61.90,52.79,47.14,34.61,31.27,27.40,14.32.
Example 4
A100 mL round bottom flask was charged with ALA hydrochloride (5.00g, 29.8mmol), naHCO 3 (15.04g, 179mmol) and Boc 2 O (6.51g, 29.8 mmol) and 40mL of anhydrous methanol as a solvent were added, the mixture was stirred at room temperature, and the progress of the reaction was checked by TLC during the reaction. After 12h, the reaction was stopped, at which point the reaction was pale yellow, the solid was filtered off,the solvent was distilled off under reduced pressure, and the residual solid was dissolved in water and acidified to pH =2 with 10% potassium hydrogen sulfate solution, extracted with ethyl acetate, and the organic layer was washed with saturated brine (15 mL × 3) a little by little, dried over anhydrous sodium sulfate, filtered, and the organic solvent was distilled off under reduced pressure to give N-Boc-ALA (6.24g, 90%) as a white solid.
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Benzyl bromide (10.26g, 60mmol) was placed in a constant pressure funnel, slowly added dropwise to the reaction solution with stirring, and after the dropwise addition was completed, the reflux reaction was continued for 2 hours, during which time the progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was concentrated in vacuo to precipitate a large amount of yellow solid, which was washed with water (50 mL. Times.3) and then filtered to give pyrone (13.5 g) as a pale yellow solid in 91% yield.
Pyrone (2.30g, 10mmol) reacted above is respectively added into a 100mL round-bottom flask, dissolved in ethanol and water (1:1) for 30mL, stirred, when the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is more than 12, butylamine (0.876g, 12mmol) is continuously added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC during the reaction. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to half the original volume, diluted hydrochloric acid was added to the residue to adjust the pH to 1 to 2, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), the pH of the aqueous layer was adjusted to neutral again, and extraction was performed with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and purified by a silica gel column, using a gradient elution of dichloromethane: methanol (100). The product, 1- (2-aminobutyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.8 g), was isolated as a yellow oil in 55% yield.
DCC (2 mmol), DMAP (0.2 mmol) and N-Boc-ALA (2.2 mmol) were dissolved in dichloromethane (20 mL) and DMF (5 mL) under nitrogen and stirred at room temperature for 50min. 1-butyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (0.574g, 2mmol) was dissolved in dichloromethane (10 mL) and placed in a constant pressure dropping funnel and added dropwise over 30 min. The reaction was stirred at room temperature overnight. The precipitate was filtered off, the solvent was distilled off under reduced pressure, the residue was dissolved in dichloromethane, washed with water and then with saturated sodium bicarbonate solution, the organic layer was concentrated under reduced pressure, and the residue was purified by flash chromatography on silica gel using a gradient of dichloromethane: methanol = 80.
A20 mL round bottom flask was charged with benzyl and Boc protected hydroxypyridinones (0.250g, 0.5 mmol) and dichloromethane as a solvent, trifluoroacetic acid (2.1g, 3.1mL) was diluted with dichloromethane (2.5 mL) in a constant pressure dropping funnel, slowly added dropwise, stirred at 0-5 ℃ for 1h, followed by TLC to monitor the progress of the reaction, and after completion of the reaction, dichloromethane and trifluoroacetic acid were distilled off under reduced pressure to obtain benzyl protected hydroxypyridinones (0.359 mg) with a yield of 79%.
0.5mmol of benzyl-protected hydroxypyridone was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi of hydrogen pressure, catalytically reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to give ALA-HPO complex 10d.
m.p.:51～53℃.
ESI-HRMS:m/z calcd for C 15 H 23 N 2 O 5 [M+H] + :311.1601；found:311.1588.
1 H NMR(400MHz,DMSO-d 6 )δ8.22(s,1H),8.15(s,3H),7.18(s,1H),5.27(s,2H),4.21(t,J＝7.8Hz,2H),3.99(s,2H),2.85(t,J＝6.5Hz,2H),2.67(t,J＝6.5Hz,2H),1.76(p,J＝7.8Hz,2H),1.42–1.27(m,2H),0.91(t,J＝7.4Hz,3H). 13 C NMR(100MHz,DMSO-d 6 )δ202.97,171.88,162.89,159.13(q,J＝34.1Hz,COO),146.06,143.30,131.29,115.40,113.28,61.14,54.88,47.11,34.55,32.75,27.31,19.47,13.89.
Example 5
A100 mL round bottom flask was charged with ALA hydrochloride (5.00)g,29.8mmol)、NaHCO 3 (15.04g, 179mmol) and Boc 2 O (6.51g, 29.8 mmol) and 40mL of anhydrous methanol as a solvent were added, the mixture was stirred at room temperature, and the progress of the reaction was checked by TLC during the reaction. After 12h, the reaction was stopped, at which time the reaction solution was light yellow, the solid was filtered off, the solvent was distilled off under reduced pressure, the residual solid was dissolved in water and acidified to pH =2 with 10% potassium hydrogen sulfate solution, extracted with ethyl acetate, the organic layer was washed with saturated brine (15 mL × 3) a few times, dried over anhydrous sodium sulfate, filtered, and the organic solvent was distilled off under reduced pressure to give N-Boc-ALA (6.24g, 90%) as a white solid.
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Benzyl bromide (10.26g, 60mmol) was placed in a constant pressure funnel, slowly added dropwise to the reaction solution with stirring, and after the dropwise addition was completed, the reflux reaction was continued for 2 hours, during which time the progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was concentrated in vacuo to precipitate a large amount of yellow solid, which was washed with water (50 mL. Times.3) and then filtered to give pyrone (13.5 g) as a pale yellow solid in 91% yield.
Pyrone (2.30g, 10mmol) reacted in the above reaction is respectively added into a 100mL round-bottom flask, dissolved in ethanol and water (1:1) for 30mL, stirred until the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is more than 12, pentylamine (1.044g, 12mmol) is continuously added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC in the period. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and the mixture was purified by a silica gel column, and gradient elution was performed using dichloromethane: methanol (100). The product, 1- (2-aminopentyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.7 g), was isolated as a yellow oil in 52% yield.
DCC (2 mmol), DMAP (0.2 mmol) and N-Boc-ALA (2.2 mmol) were dissolved in dichloromethane (20 mL) and DMF (5 mL) under nitrogen and stirred at room temperature for 50min. 1-pentyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (0.602g, 2mmol) was dissolved in dichloromethane (10 mL) and placed in a constant pressure dropping funnel and added dropwise over 30 min. The reaction was stirred at room temperature overnight. The precipitate was filtered off, the solvent was distilled off under reduced pressure, the residue was dissolved in dichloromethane, washed with water and then with saturated sodium bicarbonate solution, the organic layer was concentrated under reduced pressure, and the residue was purified by flash chromatography on silica gel using a gradient of dichloromethane: methanol = 80.
A20 mL round bottom flask was charged with benzyl and Boc protected hydroxypyridinones (0.250g, 0.5 mmol) and dichloromethane as a solvent, trifluoroacetic acid (2.1g, 3.1mL) was diluted with dichloromethane (2.5 mL) in a constant pressure dropping funnel, slowly added dropwise, stirred at 0-5 ℃ for 1h, followed by TLC to monitor the progress of the reaction, and after completion of the reaction, dichloromethane and trifluoroacetic acid were distilled off under reduced pressure to obtain benzyl protected hydroxypyridinone (0.347 mg) with a yield of 84%.
0.5mmol of benzyl-protected hydroxypyridone was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi of hydrogen pressure, catalytically reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to give ALA-HPO complex 10e.
m.p.:69.6～72.0℃.
ESI-HRMS:m/z calcd for C 16 H 25 N 2 O 5 [M+H] + :325.1758；found:325.1752.
1 H NMR(400MHz,DMSO-d 6 )δ8.14(s,3H),7.40(s,1H),6.30(s,1H),5.25(s,2H),4.02(s,2H),3.68(t,J＝7.6Hz,2H),2.80(t,J＝6.4Hz,2H),2.62(t,J＝6.4Hz,2H),1.56(q,J＝7.4Hz,2H),1.50–1.26(m,4H),0.82(t,J＝7.4Hz,3H). 13 C NMR(100MHz,DMSO-d 6 )δ203.19,170.81,168.30,157.55(q,J＝31.Hz,COO),145.77,142.04,124.21,112.78,61.90,52.79,47.14,34.61,31.27,30.91,27.40,22.42,14.32.
Example 6
A100 mL round bottom flask was charged with ALA hydrochloride (5.00g, 29.8 mmol), naHCO 3 (15.04g, 179mmol) and Boc 2 O (6.51g, 29.8 mmol), and 40mL of anhydrous methanol as a solvent were added, and the mixture was stirred at room temperature, during which the progress of the reaction was checked by TLC. After 12h, the reaction was stopped, at which time the reaction solution was light yellow, the solid was filtered off, the solvent was distilled off under reduced pressure, the residual solid was dissolved in water and acidified to pH =2 with 10% potassium hydrogen sulfate solution, extracted with ethyl acetate, the organic layer was washed with saturated brine (15 mL × 3) a few times, dried over anhydrous sodium sulfate, filtered, and the organic solvent was distilled off under reduced pressure to give N-Boc-ALA (6.24g, 90%) as a white solid.
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Benzyl bromide (10.26g, 60mmol) was placed in a constant pressure funnel, slowly added dropwise to the reaction solution with stirring, and after the dropwise addition was completed, the reflux reaction was continued for 2 hours, during which time the progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was concentrated in vacuo to precipitate a large amount of yellow solid, which was washed with water (50 mL. Times.3) and then filtered to give pyrone (13.5 g) as a pale yellow solid with a yield of 91%.
Pyrone (2.30g, 10mmol) reacted above is respectively added into a 100mL round-bottom flask, dissolved in ethanol and water (1:1) for 30mL, stirred until the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is more than 12, hexylamine (0.101g, 12mmol) is continuously added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC during the reaction. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to half the original volume, diluted hydrochloric acid was added to the residue to adjust the pH to 1 to 2, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), the pH of the aqueous layer was adjusted to neutral again, and extraction was performed with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and the mixture was purified by a silica gel column, and gradient elution was performed using dichloromethane: methanol (100). The product, 1- (2-aminohexyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.2 g), was isolated as a yellow oil in 42% yield.
DCC (2 mmol), DMAP (0.2 mmol) and N-Boc-ALA (2.2 mmol) were dissolved in dichloromethane (20 mL) and DMF (5 mL) under nitrogen and stirred at room temperature for 50min. 1-hexyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (0.630g, 2mmol) was dissolved in dichloromethane (10 mL), and placed in a constant pressure dropping funnel, and added dropwise over 30 min. The reaction was stirred at room temperature overnight. The precipitate was filtered off, the solvent was distilled off under reduced pressure, the residue was dissolved in dichloromethane, washed with water and then with saturated sodium bicarbonate solution, the organic layer was concentrated under reduced pressure, and the residue was purified by flash chromatography on silica gel using a gradient of dichloromethane: methanol = 80.
A20 mL round bottom flask was charged with benzyl and Boc protected hydroxypyridones (0.264 g,0.5 mmol) in dichloromethane as solvent, trifluoroacetic acid (2.1g, 3.1mL) was diluted with dichloromethane (2.5 mL) in a constant pressure dropping funnel, slowly added dropwise, stirred at 0-5 ℃ for 1h, the progress of the reaction was monitored by TLC, and after completion of the reaction, dichloromethane and trifluoroacetic acid were distilled off under reduced pressure to give benzyl protected hydroxypyridones (0.289 mg) in 82% yield.
0.5mmol of benzyl-protected hydroxypyridone was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi of hydrogen pressure, catalytically reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to give ALA-HPO complex 10f.
m.p.:79～82℃.
ESI-HRMS:m/z calcd for C 17 H 27 N 2 O 5 [M+H] + :339.1914；found:339.1901.
1 H NMR(400MHz,DMSO-d 6 )δ8.16(s,3H),7.58(s,1H),6.33(s,1H),5.06(s,2H),3.99(s,2H),3.89(t,J＝7.7Hz,2H),2.84(t,J＝6.4Hz,2H),2.65(t,J＝6.4Hz,2H),1.68(q,J＝7.4Hz,2H),1.50–1.26(m,6H),0.94–0.73(m,3H). 13 C NMR(100MHz,DMSO-d 6 )δ203.10,171.93,169.37,158.75(q,31.0Hz,COO),147.69,141.93,125.45,119.15,114.83,61.83,53.04,47.14,34.60,31.28,30.91,27.38,25.92,22.43,14.33.
Example 7
A100 mL round bottom flask was charged with ALA hydrochloride (5.00g, 29.8 mmol), naHCO 3 (15.04g, 179mmol) and Boc 2 O (6.51g, 29.8 mmol) and 40mL of anhydrous methanol as a solvent were added, the mixture was stirred at room temperature, and the progress of the reaction was checked by TLC during the reaction. After 12h, the reaction was stopped, at which time the reaction solution was light yellow, the solid was filtered off, the solvent was distilled off under reduced pressure, the residual solid was dissolved in water and acidified to pH =2 with 10% potassium hydrogen sulfate solution, extracted with ethyl acetate, the organic layer was washed with saturated brine (15 mL × 3) a few times, dried over anhydrous sodium sulfate, filtered, and the organic solvent was distilled off under reduced pressure to give N-Boc-ALA (6.24g, 90%) as a white solid.
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Benzyl bromide (10.26g, 60mmol) was placed in a constant pressure funnel, slowly added dropwise to the reaction solution with stirring, and after completion of dropwise addition, the reflux reaction was continued for 2h, during which time TLC monitored the progress of the reaction. After completion of the reaction, the solvent was concentrated in vacuo to precipitate a large amount of yellow solid, which was washed with water (50 mL. Times.3) and then filtered to give pyrone (13.5 g) as a pale yellow solid with a yield of 91%.
Adding pyrone (2.30g, 10mmol) reacted in the step (b) into 100mL round-bottomed flasks, dissolving the pyrone in ethanol and water (1:1) for 30mL, stirring, adding sodium hydroxide solution (2.0 mmol/L) dropwise until the pH value of a reaction system is more than 12 after the pyrone is completely dissolved, continuously adding methoxyethylamine (0.90g, 12mmol) dropwise, heating to reflux, reacting for 3 hours, and monitoring the reaction condition by TLC. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and the mixture was purified by a silica gel column, and gradient elution was performed using dichloromethane: methanol (100). The product, 1- (2-aminomethoxyethyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.42 g), was isolated as a yellow oil in 45% yield.
DCC (2 mmol), DMAP (0.2 mmol) and N-Boc-ALA (2.2 mmol) were dissolved in dichloromethane (20 mL) and DMF (5 mL) under nitrogen and stirred at room temperature for 50min. 1-methoxyethyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (0.578g, 2mmol) was dissolved in dichloromethane (10 mL), placed in a constant pressure dropping funnel and added dropwise over 30 min. The reaction was stirred at room temperature overnight. The precipitate was filtered off, the solvent was distilled off under reduced pressure, the residue was dissolved in dichloromethane, washed with water and then with saturated sodium bicarbonate solution, the organic layer was concentrated under reduced pressure, and the residue was purified by flash chromatography on silica gel using a gradient of dichloromethane: methanol = 80.
A20 mL round bottom flask was charged with benzyl and Boc protected hydroxypyridones (0.251g, 0.5 mmol) and dichloromethane as solvent, trifluoroacetic acid (2.1g, 3.1mL) was diluted with dichloromethane (2.5 mL) in a constant pressure dropping funnel, slowly added dropwise, stirred at 0-5 ℃ for 1h, the progress of the reaction was monitored by TLC, and after completion of the reaction, dichloromethane and trifluoroacetic acid were distilled off under reduced pressure to give benzyl protected hydroxypyridones (0.409 mg) in 82% yield.
0.5mmol of benzyl-protected hydroxypyridone was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi of hydrogen pressure, catalytically reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to give 10g of ALA-HPO complex.
ESI-HRMS:m/z calcd for C 14 H 21 N 2 O 6 [M+H] + :313.1394；found:313.1393.
1 H NMR(600MHz,DMSO-d 6 )δ8.13(s,3H),8.12(s,1H),6.16(s,1H),5.25(s,2H),4.43(s,2H),3.99(t,J＝7.5Hz,2H),3.70(t,J＝4.9Hz,2H),3.25(s,3H),2.84(t,J＝6.5Hz,2H),2.65(t,J＝6.5Hz,2H). 13 C NMR(150MHz,DMSO-d6)δ203.03,171.85,163.26,158.78(q,34.5Hz,COO),145.67,143.93,131.65,117.76,115.81,113.97,70.99,61.30,58.90,54.56,47.10,34.57,27.34.
Example 8
A100 mL round bottom flask was charged with ALA hydrochloride (5.00g, 29.8 mmol), naHCO 3 (15.04g, 179mmol) and Boc 2 O (6.51g, 29.8 mmol) and 40mL of anhydrous methanol as a solvent were added, the mixture was stirred at room temperature, and the progress of the reaction was checked by TLC during the reaction. After 12h, the reaction was stopped, at which time the reaction solution was light yellow, the solid was filtered off, the solvent was distilled off under reduced pressure, the residual solid was dissolved in water and acidified to pH =2 with 10% potassium hydrogen sulfate solution, extracted with ethyl acetate, the organic layer was washed with saturated brine (15 mL × 3) a few times, dried over anhydrous sodium sulfate, filtered, and the organic solvent was distilled off under reduced pressure to give N-Boc-ALA (6.24g, 90%) as a white solid.
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Benzyl bromide (10.26g, 60mmol) was placed in a constant pressure funnel, slowly added dropwise to the reaction solution with stirring, and after completion of dropwise addition, the reflux reaction was continued for 2h, during which time TLC monitored the progress of the reaction. After completion of the reaction, the solvent was concentrated in vacuo to precipitate a large amount of yellow solid, which was washed with water (50 mL. Times.3) and then filtered to give pyrone (13.5 g) as a pale yellow solid with a yield of 91%.
Adding pyrone (2.30g, 10mmol) reacted in the step (b) into 100mL round-bottomed flasks, dissolving the pyrone in ethanol and water (1:1) for 30mL, stirring, adding sodium hydroxide solution (2.0 mmol/L) dropwise until the pH value of a reaction system is more than 12 after the pyrone is completely dissolved, continuously adding methoxypropylamine (0.90g, 12mmol) dropwise, heating to reflux, reacting for 3 hours, and monitoring the reaction condition by TLC. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and the mixture was purified by a silica gel column, and gradient elution was performed using dichloromethane: methanol (100). The product, 1- (2-aminomethoxypropyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.50 g) was isolated as a yellow oil in 47% yield.
DCC (2 mmol), DMAP (0.2 mmol) and N-Boc-ALA (2.2 mmol) were dissolved in dichloromethane (20 mL) and DMF (5 mL) under nitrogen and stirred at room temperature for 50min. 1-Methoxypropyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (0.606g, 2mmol) was dissolved in dichloromethane (10 mL) and placed in a constant pressure dropping funnel and added dropwise over 30 min. The reaction was stirred at room temperature overnight. The precipitate was filtered off, the solvent was distilled off under reduced pressure, the residue was dissolved in dichloromethane, washed with water and then with saturated sodium bicarbonate solution, the organic layer was concentrated under reduced pressure, and the residue was purified by flash chromatography on silica gel using a gradient of dichloromethane: methanol = 80.
A20 mL round bottom flask was charged with benzyl and Boc protected hydroxypyridinones (0.258g, 0.5 mmol) and dichloromethane as a solvent, trifluoroacetic acid (2.1g, 3.1mL) was diluted with dichloromethane (2.5 mL) in a constant pressure dropping funnel, slowly added dropwise, stirred at 0-5 ℃ for 1h, the progress of the reaction was monitored by TLC, and after completion of the reaction, dichloromethane and trifluoroacetic acid were distilled off under reduced pressure to give benzyl protected hydroxypyridinone (0.425 mg) in 86% yield.
0.5mmol of benzyl-protected hydroxypyridone was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi of hydrogen pressure, catalytically reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to give ALA-HPO complex for 10h.
m.p.:97.2～100.3℃.
ESI-HRMS:m/z calcd for C 15 H 23 N 2 O 6 [M+H] + :327.1551；found:327.1529.
1 H NMR(400MHz,DMSO-d 6 )δ8.22(s,3H),7.57(s,1H),6.38(s,1H),5.09(s,2H),4.12–3.89(m,4H),3.35(t,J＝5.9Hz,2H),3.26(s,3H),2.87(t,J＝6.4Hz,2H),2.68(t,J＝6.4Hz,2H),2.07–1.88(m,2H). 13 C NMR(100MHz,DMSO-d 6 )δ203.12,171.94,170.11,147.90,142.03,124.80,114.80,68.74,61.84,58.40,50.04,47.13,34.60,30.71,27.39.
Example 9
A100 mL round bottom flask was charged with ALA hydrochloride (5.00g, 29.8 mmol), naHCO 3 (15.04g, 179mmol) and Boc 2 O (6.51g, 29.8 mmol) and 40mL of anhydrous methanol as a solvent were added, the mixture was stirred at room temperature, and the progress of the reaction was checked by TLC during the reaction. After 12h, the reaction was stopped, at which time the reaction solution was light yellow, the solid was filtered off, the solvent was distilled off under reduced pressure, the residual solid was dissolved in water and acidified to pH =2 with 10% potassium hydrogen sulfate solution, extracted with ethyl acetate, the organic layer was washed with saturated brine (15 mL × 3) a few times, dried over anhydrous sodium sulfate, filtered, and the organic solvent was distilled off under reduced pressure to give N-Boc-ALA (6.24g, 90%) as a white solid.
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Benzyl bromide (10.26g, 60mmol) was placed in a constant pressure funnel, slowly added dropwise to the reaction solution with stirring, and after the dropwise addition was completed, the reflux reaction was continued for 2 hours, during which time the progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was concentrated in vacuo to precipitate a large amount of yellow solid, which was washed with water (50 mL. Times.3) and then filtered to give pyrone (13.5 g) as a pale yellow solid with a yield of 91%.
Pyrone (2.30g, 10mmol) reacted above is respectively added into a 100mL round-bottom flask, dissolved in ethanol and water (1:1) for 30mL, stirred until the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is more than 12, ethoxypropylamine (0.372g, 12mmol) is continuously added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC during the reaction. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and purified by a silica gel column, using a gradient elution of dichloromethane: methanol (100). The product, 1- (2-aminoethoxypropyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.5 g) was isolated as a yellow oil in 50% yield.
DCC (2 mmol), DMAP (0.2 mmol) and N-Boc-ALA (2.2 mmol) were dissolved in dichloromethane (20 mL) and DMF (5 mL) under nitrogen and stirred at room temperature for 50min. 1-ethoxypropyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (0.632g, 2mmol) was dissolved in dichloromethane (10 mL) and placed in a dropping funnel at constant pressure and added dropwise over 30 min. The reaction was stirred at room temperature overnight. The precipitate was filtered off, the solvent was distilled off under reduced pressure, the residue was dissolved in dichloromethane, washed with water and then with saturated sodium bicarbonate solution, the organic layer was concentrated under reduced pressure, and the residue was purified by flash chromatography on silica gel using a gradient of dichloromethane: methanol = 80.
A20 mL round-bottomed flask was charged with benzyl and Boc protected hydroxypyridones (0.265g, 0.5 mmol) and dichloromethane as a solvent, trifluoroacetic acid (2.1g, 3.1mL) was diluted with dichloromethane (2.5 mL) in a constant pressure dropping funnel, slowly added dropwise, stirred at 0 to 5 ℃ for 1 hour, the progress of the reaction was monitored by TLC, and after completion of the reaction, dichloromethane and trifluoroacetic acid were distilled off under reduced pressure to give benzyl protected hydroxypyridones (0.329 mg) in a yield of 83%.
0.5mmol of benzyl-protected hydroxypyridone was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi of hydrogen pressure, catalytically reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to give ALA-HPO complex 10i.
m.p.:98～101℃.
ESI-HRMS:m/z calcd for C 16 H 25 N 2 O 6 [M+H] + :341.1707；found:341.1709.
1 H NMR(600MHz,DMSO-d 6 )δ8.21(s,3H),7.64(s,1H),6.47(s,1H),5.10(s,2H),4.03(d,J＝6.7Hz,2H),4.00(s,2H),3.43(ddt,J＝23.4,11.9,6.3Hz,4H),2.85(t,J＝6.4Hz,2H),2.66(t,J＝6.4Hz,2H),1.95(q,J＝6.4Hz,2H),1.11(t,J＝6.3,5.7Hz,3H). 13 C NMR(150MHz,DMSO-d 6 )δ203.05,171.90,169.15,158.95,158.64(q,J＝46.9Hz,COO),147.62,142.26,119.07,116.10,66.46,65.84,61.78,50.43,47.13,34.60,30.75,27.38,15.43.
Example 10
A100 mL round bottom flask was charged with ALA hydrochloride (5.00g, 29.8 mmol), naHCO 3 (15.04g, 179mmol) and Boc 2 O (6.51g, 29.8 mmol) and 40mL of anhydrous methanol as a solvent were added, the mixture was stirred at room temperature, and the progress of the reaction was checked by TLC during the reaction. After 12h, the reaction was stopped, at which time the reaction solution was light yellow, the solid was filtered off, the solvent was distilled off under reduced pressure, the residual solid was dissolved in water and acidified to pH =2 with 10% potassium hydrogen sulfate solution, extracted with ethyl acetate, the organic layer was washed with saturated brine (15 mL × 3) a few times, dried over anhydrous sodium sulfate, filtered, and the organic solvent was distilled off under reduced pressure to give N-Boc-ALA (6.24g, 90%) as a white solid.
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added in 100mL as solvents, respectively, and the reaction system was heated to reflux. Benzyl bromide (10.26g, 60mmol) was placed in a constant pressure funnel, slowly added dropwise to the reaction solution with stirring, and after the dropwise addition was completed, the reflux reaction was continued for 2 hours, during which time the progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was concentrated in vacuo to precipitate a large amount of yellow solid, which was washed with water (50 mL. Times.3) and then filtered to give pyrone (13.5 g) as a pale yellow solid with a yield of 91%.
The pyrone (2.30g, 10mmol) reacted in the reaction is added into a 100mL round-bottom flask respectively, dissolved in ethanol and water (1:1) for 30mL, stirred until the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is more than 12, phenethylamine (1.452g, 12mmol) is added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC in the process. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and the mixture was purified by a silica gel column, and gradient elution was performed using dichloromethane: methanol (100). The product, 1- (2-aminoethoxypropyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (2.1 g), was isolated as a yellow oil in 67% yield.
DCC (2 mmol), DMAP (0.2 mmol) and N-Boc-ALA (2.2 mmol) were dissolved in dichloromethane (20 mL) and DMF (5 mL) under nitrogen and stirred at room temperature for 50min. 1-phenethyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (0.670g, 2mmol) was dissolved in dichloromethane (10 mL), placed in a constant pressure dropping funnel, and added dropwise over 30 min. The reaction was stirred at room temperature overnight. The precipitate was filtered off, the solvent was distilled off under reduced pressure, the residue was dissolved in dichloromethane, washed with water then saturated sodium bicarbonate solution, the organic layer was concentrated under reduced pressure, and the residue was purified by flash chromatography on silica gel using a gradient of dichloromethane: methanol = 80.
A20 mL round bottom flask was charged with benzyl and Boc protected hydroxypyridinones (0.274g, 0.5mmol) in dichloromethane as solvent, trifluoroacetic acid (2.1g, 3.1mL) was diluted with dichloromethane (2.5 mL) in a constant pressure dropping funnel, slowly added dropwise, stirred at 0-5 ℃ for 1h, the progress of the reaction was monitored by TLC, and after completion of the reaction, dichloromethane and trifluoroacetic acid were distilled off under reduced pressure to give benzyl protected hydroxypyridinones (0.290 mg) in 85% yield.
0.5mmol of benzyl-protected hydroxypyridone was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi of hydrogen pressure, catalytically reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to give ALA-HPO complex 10j.
m.p.:115～117℃.
ESI-HRMS:m/z calcd for C 19 H 23 N 2 O 5 [M+H] + :359.1601；found:359.1603.
1 H NMR(600MHz,DMSO-d 6 )δ8.18(s,3H),7.64(s,1H),7.32(t,J＝7.4Hz,2H),7.29–7.22(m,3H),6.39(s,1H),4.98(s,2H),4.17(t,J＝7.7Hz,2H),4.00(s,2H),3.05(t,J＝7.7Hz,2H),2.85(t,J＝6.5Hz,2H),2.67(t,J＝6.5Hz,2H). 13 C NMR(150MHz,DMSO-d 6 )δ203.07,171.91,169.64,158.76(q,31.5Hz,COO),147.71,142.00,137.75,129.40,129.01,127.25,125.35,118.63,114.68,61.82,54.07,47.13,36.80,34.61,27.41.
Example 11
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Placing benzyl bromide (10.26g, 60mmol) in a constant-pressure funnel, slowly dropwise adding the benzyl bromide into the reaction solution while stirring, continuously refluxing for reaction for 2h, monitoring the reaction process by TLC (thin layer chromatography), stopping the reaction after the raw materials are completely converted, concentrating the solvent in vacuum, precipitating a large amount of yellow solid, washing the solid with water (50 mL multiplied by 3), and filtering to obtain a light yellow solid compound pyrone (13.5 g), wherein the yield is 98%.
Pyrone (2.30g, 10mmol) reacted above is respectively added into a 100mL round-bottom flask, dissolved in ethanol and water (1:1) for 30mL, stirred until the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is more than 12, ethoxypropylamine (0.372g, 12mmol) is continuously added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC during the reaction. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and the mixture was purified by a silica gel column, and gradient elution was performed using dichloromethane: methanol (100). The product, 1- (2-aminoethoxypropyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.5 g) was isolated as a yellow oil in 50% yield.
Anhydrous dichloromethane (10 mL) and p-nitrophenyl chloroformate (242mg, 1.2mmol) are sequentially added into a 50mL single-neck bottle, the round-bottom flask is moved into an ice maker to be cooled to-5 ℃ (internal temperature), after 30min, a solution of 1-ethoxypropyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (2458 mmol) in dichloromethane and triethylamine (152mg, 1.5 mmol) is slowly added dropwise into the round-bottom flask, the solution is stirred for 30min, then the solution is moved to room temperature and stirred for 6-8 h, the reaction solution is dried, and an intermediate (151 mg) is obtained by column chromatography (dichloromethane: methanol =100: 1-50).
1mmol of methyl 5-aminolevulinate hydrochloride was dissolved in dichloromethane (6 mL), triethylamine (202mg, 2mmol) and DMAP (12mg, 0.1eq.) were added in this order, and stirred at room temperature for 5 hours, and then the intermediate carbonate (393mg, 1.1eq.) obtained above was dissolved in dichloromethane, and the reaction mixture obtained above was added dropwise at-10 to 20 ℃ and the solution rapidly turned yellow due to the formation of p-nitrophenol. The reaction was complete until the carbonate blue spot observed on TLC became very weak. The reaction mixture was washed once with saturated sodium bicarbonate solution, twice with water, the aqueous layer was back-extracted once with dichloromethane, the combined organic phases were dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by means of a silica gel column to give benzyl-protected hydroxypyridone (231 mg) in 60% yield.
0.5mmol of the benzyl-protected hydroxypyridone obtained in the above step was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi of hydrogen pressure, reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to obtain ALA-HPO complex 13a.
m.p.130～132℃；
ESI-HRMS:m/z calcd for C 18 H 27 N 2 O 8 [M+H] + :399.1762；found:399.1746；
H NMR(400MHz,DMSO-d6)δ7.72(t,J＝5.7Hz,1H),7.73(s,1H),6.26(s,1H),5.00(s,2H),3.92(d,J＝5.6Hz,4H),3.57(s,3H),3.40(q,J＝7.0Hz,2H),3.33(t,J＝5.8Hz,2H),2.70(t,J＝6.5Hz,2H),2.49(d,J＝6.4Hz,2H),1.99–1.83(m,2H),1.11(t,J＝7.0Hz,3H).13C NMR(100MHz,DMSO-d6)δ205.21,172.56,170.43,155.54,147.43,142.41,123.31,113.62,66.03,65.32,61.19,51.37,49.68,49.18,33.74,30.21,27.09,14.97.
Example 12
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Benzyl bromide (10.26g, 60mmol) is placed in a constant-pressure funnel, slowly dropwise added into the reaction liquid while stirring, the reflux reaction is continued for 2h, the reaction process is monitored by TLC (thin layer chromatography), after the raw materials are completely converted, the reaction is stopped, the solvent is concentrated in vacuum, a large amount of yellow solid is separated out, and after the solid is washed by water (50 mL multiplied by 3), the solid is filtered to obtain a light yellow solid compound pyrone (13.5 g), wherein the yield is 97%.
Pyrone (2.30g, 10mmol) reacted above is respectively added into a 100mL round-bottom flask, dissolved in ethanol and water (1:1) for 30mL, stirred until the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is more than 12, isoamylamine (1.044g, 12mmol) is continuously added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC during the reaction. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and the mixture was purified by a silica gel column, and gradient elution was performed using dichloromethane: methanol (100). The product, 1- (2-aminoisoamyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.2 g) was isolated as a yellow oil in 48% yield.
Anhydrous dichloromethane (10 mL) and p-nitrophenyl chloroformate (242mg, 1.2mmol) are sequentially added into a 50mL single-neck bottle, the round-bottom flask is moved to an ice maker to be cooled to-5 ℃ (internal temperature), after 30min, a solution of 1-isoamyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (301mg, 1 mmol) in dichloromethane and triethylamine (152mg, 1.5mmol) is slowly added dropwise into the round-bottom flask, the solution is stirred for 30min, then the solution is moved to room temperature and stirred for 6-8 h, the reaction solution is dried, and an intermediate (151 mg) is obtained by column chromatography (dichloromethane: methanol =100: 1-50).
1mmol of methyl 5-aminolevulinate hydrochloride was dissolved in dichloromethane (6 mL), triethylamine (202mg, 2mmol) and DMAP (12mg, 0.1eq.) were sequentially added thereto, and the mixture was stirred at room temperature for 5 hours, and then the intermediate carbonate (376mg, 1.1eq.) obtained above was dissolved in dichloromethane and the reaction mixture obtained above was added dropwise at-10 to 20 ℃ to rapidly turn yellow due to the formation of p-nitrophenol. The reaction was complete until the carbonate blue spot observed on TLC became very weak. The reaction mixture was washed once with saturated sodium bicarbonate solution, twice with water, the aqueous layer was back-extracted once with dichloromethane, the combined organic phases were dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by means of a silica gel column to give benzyl-protected hydroxypyridone (223 mg) in 60% yield.
0.5mmol of the benzyl-protected hydroxypyridone obtained in the above step was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi hydrogen pressure, reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to obtain ALA-HPO complex 13b.
m.p.145.8～147.1℃.
ESI-HRMS:m/z calcd for C 18 H 27 N 2 O 7 [M+H] + :383.1813；found:383.1808；
1 H NMR(400MHz,DMSO-d 6 )δ7.76(d,J＝5.8Hz,1H),7.50(s,1H),6.27(s,1H),4.99(s,2H),3.92(d,J＝5.9Hz,2H),3.86(d,J＝8.0Hz,2H),3.57(s,3H),2.70(t,J＝6.5Hz,2H),2.53(t,J＝6.4Hz,2H),1.59(dd,J＝9.4,5.6Hz,3H),0.91(d,J＝5.8Hz,6H). 13 C NMR(100MHz,DMSO-d 6 )δ205.71,173.08,170.85,156.03,148.01,142.48,124.05,114.50,61.77,51.19,51.88,50.15,39.66,34.23,27.56,25.98,22.63.
Example 13
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added in 100mL as solvents, respectively, and the reaction system was heated to reflux. Placing benzyl bromide (10.26g, 60mmol) in a constant-pressure funnel, slowly dropwise adding the benzyl bromide into the reaction solution while stirring, continuously refluxing for reaction for 2h, monitoring the reaction process by TLC (thin layer chromatography), stopping the reaction after the raw materials are completely converted, concentrating the solvent in vacuum, precipitating a large amount of yellow solid, washing the solid with water (50 mL multiplied by 3), and filtering to obtain a light yellow solid compound pyrone (13.5 g), wherein the yield is 98%.
The pyrone (2.30g, 10mmol) reacted in the above reaction is respectively added into a 100mL round-bottom flask, dissolved in ethanol and water (1:1) for 30mL, stirred until the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of the reaction system is more than 12, the decylamine (1.884g, 12mmol) is continuously added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the condition of the reaction is monitored by TLC in the period. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and purified by a silica gel column, using a gradient elution of dichloromethane: methanol (100). The product, 1- (2-aminodecyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.5 g) was isolated as a yellow oil in 50% yield.
Anhydrous dichloromethane (10 mL) and p-nitrophenyl chloroformate (242mg, 1.2mmol) are sequentially added into a 50mL single-neck bottle, the round-bottom flask is moved to an ice maker to be cooled to-5 ℃ (internal temperature), after 30min, a solution of 1-decyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (371mg, 1mmol) in dichloromethane and triethylamine (152mg, 1.5mmol) is slowly added dropwise into the round-bottom flask, the solution is stirred for 30min, then the solution is moved to room temperature and stirred for 6-8 h, the reaction solution is dried, and an intermediate (352 mg) is obtained by column chromatography (dichloromethane: methanol =100: 1-50).
1mmol of methyl 5-aminolevulinate hydrochloride was dissolved in dichloromethane (6 mL), triethylamine (202mg, 2mmol) and DMAP (12mg, 0.1eq.) were added in this order, and stirred at room temperature for 5 hours, and then the intermediate carbonate (536mg, 1.1eq.) obtained above was dissolved in dichloromethane, and the reaction mixture obtained above was added dropwise at-10 to 20 ℃ to rapidly turn yellow due to the formation of p-nitrophenol. The reaction was complete until the carbonate blue spot observed on TLC became very weak. The reaction mixture was washed once with saturated sodium bicarbonate solution, twice with water, the aqueous layer was back-extracted once with dichloromethane, the combined organic phases were dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by means of a silica gel column to give benzyl-protected hydroxypyridone (324 mg) in 68% yield.
0.5mmol of the benzyl-protected hydroxypyridone obtained in the above step was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi of hydrogen pressure, reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to obtain ALA-HPO complex 13C.
m.p.:164～167℃.
ESI-HRMS:m/z calcd for C 23 H 37 N 2 O 7 [M+H] + :453.2595；found:453.2582.
1 H NMR(400MHz,DMSO-d 6 )δ7.71(t,J＝5.7Hz,1H),7.47(s,1H),6.25(s,1H),4.98(s,2H),3.91(d,J＝5.8Hz,2H),3.87–3.74(m,2H),3.57(s,3H),2.70(t,J＝6.4Hz,2H),2.48(t,J＝6.4Hz,2H),1.67(s,2H),1.24(m,16H),0.85(t,J＝6.6Hz,3H). 13 C NMR(100MHz,DMSO-d 6 )δ205.14,172.54,170.35,155.54,147.40,142.11,123.37,113.71,61.22,51.97,51.36,49.67,33.72,31.26,30.39,28.94,28.90,28.64,28.55,27.07,25.88,22.06,13.92.
Example 14
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added in 100mL as solvents, respectively, and the reaction system was heated to reflux. Benzyl bromide (10.26g, 60mmol) is placed in a constant-pressure funnel, slowly dropwise added into the reaction liquid while stirring, the reflux reaction is continued for 2h, the reaction process is monitored by TLC (thin layer chromatography), after the raw materials are completely converted, the reaction is stopped, the solvent is concentrated in vacuum, a large amount of yellow solid is separated out, and after the solid is washed by water (50 mL multiplied by 3), the solid is filtered to obtain a light yellow solid compound pyrone (13.5 g), wherein the yield is 98%.
The pyrone (2.30g, 10mmol) reacted in the reaction is added into a 100mL round-bottom flask respectively, dissolved in ethanol and water (1:1) for 30mL, stirred until the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is larger than 12, the heptamine (1.380g, 12mmol) is added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC in the process. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and the mixture was purified by a silica gel column, and gradient elution was performed using dichloromethane: methanol (100). The product, 1- (2-aminoheptyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.8 g), was isolated as a yellow oil in 47% yield.
Anhydrous dichloromethane (10 mL) and p-nitrophenyl chloroformate (242mg, 1.2 mmol) are sequentially added into a 50mL single-neck bottle, the round-bottom flask is moved into an ice maker to be cooled to-5 ℃ (internal temperature), after 30min, a solution of 1-heptyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (329mg, 1mmol) in dichloromethane and triethylamine (152mg, 1.5mmol) is slowly added dropwise into the round-bottom flask, the solution is stirred for 30min, the stirring is further carried out at room temperature for 6-8 h, the reaction solution is dried, and an intermediate (302 mg) is obtained by column chromatography (dichloromethane: methanol =100: 1-50).
1mmol of methyl 5-aminolevulinate hydrochloride was dissolved in dichloromethane (6 mL), triethylamine (202mg, 2mmol) and DMAP (12mg, 0.1eq.) were added in this order, and stirred at room temperature for 5 hours, and then the intermediate carbonate (543mg, 1.1eq.) obtained above was dissolved in dichloromethane, and the reaction mixture obtained above was added dropwise at-10 to 20 ℃ and the solution rapidly turned yellow due to the formation of p-nitrophenol. The reaction was complete until the carbonate blue spot observed on TLC became very weak. The reaction mixture was washed once with saturated sodium bicarbonate solution, twice with water, the aqueous layer was back-extracted once with dichloromethane, the combined organic phases were dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by means of a silica gel column to give benzyl-protected hydroxypyridone (338 mg) in 59% yield.
0.5mmol of the benzyl-protected hydroxypyridone obtained in the above step was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi of hydrogen pressure, reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to obtain ALA-HPO complex 13d.
m.p.:169～171℃.
ESI-HRMS:m/z calcd for C 20 H 31 N 2 O 7 [M+H] + :425.2126；found:411.2113.
1 H NMR(400MHz,DMSO-d 6 )δ7.72(t,J＝5.8Hz,1H),7.48(s,1H),6.26(s,1H),4.99(s,2H),3.92(d,J＝5.9Hz,2H),3.83(t,J＝8.0Hz,2H),3.58(s,3H),2.70(t,J＝6.5Hz,2H),2.49(t,J＝6.4Hz,2H),1.68(s,2H),1.37–1.17(m,8H),0.86(t,J＝6.6Hz,3H). 13 C NMR(100MHz,DMSO-d 6 )δ205.16,172.55,170.36,155.55,147.42,142.12,123.41,113.75,61.23,51.98,51.36,49.68,33.73,31.15,30.40,28.20,27.07,25.83,21.98,13.88.
Example 15
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Placing benzyl bromide (10.26g, 60mmol) in a constant-pressure funnel, slowly dropwise adding the benzyl bromide into the reaction solution while stirring, continuously refluxing for reaction for 2h, monitoring the reaction process by TLC (thin layer chromatography), stopping the reaction after the raw materials are completely converted, concentrating the solvent in vacuum, precipitating a large amount of yellow solid, washing the solid with water (50 mL multiplied by 3), and filtering to obtain a light yellow solid compound pyrone (13.5 g), wherein the yield is 98%.
The pyrone (2.30g, 10mmol) reacted above is respectively added into a 100mL round-bottom flask, dissolved in ethanol and water (1:1) for 30mL, stirred until the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is more than 12, methoxyethylamine (0.90g, 12mmol) is continuously added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC during the reaction. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to half the original volume, diluted hydrochloric acid was added to the residue to adjust the pH to 1 to 2, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), the pH of the aqueous layer was adjusted to neutral again, and extraction was performed with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and the mixture was purified by a silica gel column, and gradient elution was performed using dichloromethane: methanol (100). The product, 1- (2-aminomethoxyethyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.42 g) was isolated as a yellow oil in 45% yield.
Anhydrous dichloromethane (10 mL) and p-nitrophenyl chloroformate (242mg, 1.2mmol) are sequentially added into a 50mL single-neck bottle, the round-bottom flask is moved into an ice maker to be cooled to-5 ℃ (internal temperature), after 30min, a solution of 1-methoxyethyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (289mg, 1mmol) in dichloromethane and triethylamine (152mg, 1.5mmol) is slowly added dropwise into the round-bottom flask, the solution is stirred for 30min, then the solution is moved to room temperature and stirred for 6-8 h, the reaction solution is dried, and an intermediate (351 mg) is obtained through column chromatography (dichloromethane: methanol =100: 1-50).
1mmol of methyl 5-aminolevulinate hydrochloride was dissolved in dichloromethane (6 mL), triethylamine (202mg, 2mmol) and DMAP (12mg, 0.1eq.) were added in this order, and stirred at room temperature for 5 hours, and then the intermediate carbonate (499mg, 1.1eq.) obtained above was dissolved in dichloromethane, and the reaction mixture obtained above was added dropwise at-10 to 20 ℃ and the solution rapidly turned yellow due to the formation of p-nitrophenol. The reaction was complete until the carbonate blue spot observed on TLC became very weak. The reaction mixture was washed once with saturated sodium bicarbonate solution, twice with water, the aqueous layer was back-extracted once with dichloromethane, the combined organic phases were dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by column on silica gel to give benzyl protected hydroxypyridone (252 mg) in 58% yield.
Weighing 0.5mmol of the benzyl protected hydroxypyridone obtained in the above step, dissolving in 3mL of methanol in a 25mL round bottom flask, catalytically hydrogenating with 5% Pd/C, maintaining 30psi of hydrogen pressure, reacting for 2h, filtering, distilling off the solvent under reduced pressure, and recrystallizing with ethanol/diethyl ether to obtain ALA-HPO complex 13e.
m.p.:127～129℃.
ESI-HRMS:m/z calcd for C 16 H 23 N 2 O 8 [M+H] + :371.1449；found:371.1441.
1 H NMR(400MHz,DMSO-d 6 )δ7.72(t,J＝5.7Hz,1H),7.43(s,1H),6.26(s,1H),5.01(s,2H),4.06(t,J＝4.9Hz,2H),3.58(s,3H),3.24(s,3H),3.24(s,3H),2.70(t,J＝6.5Hz,2H),2.50(t,J＝6.4Hz,2H). 13 C NMR(100MHz,DMSO-d6)δ205.73,173.07,171.04,156.01,147.60,143.21,124.41,114.13,71.63,61.94,58.88,51.97,51.87,50.17,34.24,27.58.
Example 16
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Placing benzyl bromide (10.26g, 60mmol) in a constant-pressure funnel, slowly dropwise adding the benzyl bromide into the reaction solution while stirring, continuously refluxing for reaction for 2h, monitoring the reaction process by TLC (thin layer chromatography), stopping the reaction after the raw materials are completely converted, concentrating the solvent in vacuum, precipitating a large amount of yellow solid, washing the solid with water (50 mL multiplied by 3), and filtering to obtain a light yellow solid compound pyrone (13.5 g), wherein the yield is 98%.
The pyrone (2.30g, 10mmol) reacted in the reaction is added into a 100mL round-bottom flask respectively, dissolved in ethanol and water (1:1) for 30mL, stirred until the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is larger than 12, hexylamine (0.101g, 12mmol) is added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC in the process. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and the mixture was purified by a silica gel column, and gradient elution was performed using dichloromethane: methanol (100). The product, 1- (2-aminohexyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.2 g), was isolated as a yellow oil in 42% yield.
Anhydrous dichloromethane (10 mL) and p-nitrophenyl chloroformate (242mg, 1.2mmol) are sequentially added into a 50mL single-neck bottle, a round-bottom flask is moved into an ice maker to be cooled to-5 ℃ (internal temperature), after 30min, a solution of 1-hexyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (315mg, 1mmol) in dichloromethane and triethylamine (152mg, 1.5mmol) is slowly added dropwise into the round-bottom flask, the mixture is stirred for 30min, then moved to room temperature and stirred for 6-8 h, the reaction solution is dried by spinning, and an intermediate (252 mg) is obtained by column chromatography (dichloromethane: methanol = 100.
1mmol of methyl 5-aminolevulinate hydrochloride was dissolved in dichloromethane (6 mL), triethylamine (202mg, 2mmol) and DMAP (12mg, 0.1eq.) were added in this order, and the mixture was stirred at room temperature for 5 hours, then the intermediate carbonate (528mg, 1.1eq.) obtained above was dissolved in dichloromethane, and the reaction mixture obtained above was added dropwise at-10 to 20 ℃ to rapidly turn yellow due to the formation of p-nitrophenol. The reaction was complete until the carbonate blue spot observed on TLC became very weak. The reaction mixture was washed once with saturated sodium bicarbonate solution, twice with water, the aqueous layer was back-extracted once with dichloromethane, the combined organic phases were dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by means of a silica gel column to give benzyl-protected hydroxypyridone (323 mg) in 69% yield.
0.5mmol of the benzyl-protected hydroxypyridone obtained in the above step was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi hydrogen pressure, reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to obtain ALA-HPO complex 13f.
m.p.:147～150℃.
ESI-HRMS:m/z calcd for C 19 H 29 N 2 O 7 [M+H] + :397.1969；found:397.1960.
1 H NMR(400MHz,DMSO-d 6 )δ7.71(t,J＝5.7Hz,1H),7.48(s,1H),6.25(s,1H),4.98(s,2H),3.91(d,J＝5.9Hz,2H),3.88–3.75(m,2H),3.57(s,3H),2.70(t,J＝6.5Hz,2H),2.49(d,J＝6.5Hz,2H),1.67(s,2H),1.27(s,6H),0.87(d,J＝6.0Hz,3H). 13 C NMR(100MHz,DMSO-d 6 )δ205.16,172.78,170.33,155.54,147.40,142.12,123.42,113.74,61.22,51.98,51.36,49.67,33.73,30.72,30.35,27.07,25.52,21.98,13.83.
Example 17
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Placing benzyl bromide (10.26g, 60mmol) in a constant-pressure funnel, slowly dropwise adding the benzyl bromide into the reaction solution while stirring, continuously refluxing for reaction for 2h, monitoring the reaction process by TLC (thin layer chromatography), stopping the reaction after the raw materials are completely converted, concentrating the solvent in vacuum, precipitating a large amount of yellow solid, washing the solid with water (50 mL multiplied by 3), and filtering to obtain a light yellow solid compound pyrone (13.5 g), wherein the yield is 98%.
Pyrones (2.30g, 10mmol) reacted above were respectively added into a 100mL round-bottomed flask, dissolved in ethanol and water (1:1) for 30mL, stirred until the pyrones are completely dissolved, a sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH of the reaction system is more than 12, pentylamine (1.044g, 12mmol) is continuously added dropwise, the temperature is increased to reflux, and the reaction is carried out for 3 hours, wherein the reaction condition is monitored by TLC. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and purified by a silica gel column, using a gradient elution of dichloromethane: methanol (100). The product, 1- (2-aminopentyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.7 g), was isolated as a yellow oil in 52% yield.
Anhydrous dichloromethane (10 mL) and p-nitrophenyl chloroformate (242mg, 1.2mmol) are sequentially added into a 50mL single-neck bottle, the round-bottom flask is moved to an ice maker to be cooled to-5 ℃ (internal temperature), after 30min, a solution of 1-pentyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (330mg, 1mmol) in dichloromethane and triethylamine (152mg, 1.5mmol) is slowly added dropwise into the round-bottom flask, stirring is continued for 30min, the mixture is moved to room temperature and stirred for 6-8 h, the reaction solution is dried by spinning, and an intermediate (251 mg) is obtained by column chromatography (dichloromethane: methanol =100: 1-50).
1mmol of methyl 5-aminolevulinate hydrochloride was dissolved in dichloromethane (6 mL), triethylamine (202mg, 2mmol) and DMAP (12mg, 0.1eq.) were added in this order, and stirred at room temperature for 5 hours, and then the intermediate carbonate (512mg, 1.1eq.) obtained above was dissolved in dichloromethane, and the reaction mixture obtained above was added dropwise at-10 to 20 ℃ to rapidly turn yellow due to the formation of p-nitrophenol. The reaction was complete until the carbonate blue spot observed on TLC became very weak. The reaction mixture was washed once with saturated sodium bicarbonate solution, twice with water, the aqueous layer was back-extracted once with dichloromethane, the combined organic phases were dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column to give benzyl protected hydroxypyridone (335 mg) in 63% yield.
0.5mmol of the benzyl-protected hydroxypyridone obtained in the above step was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi of hydrogen pressure, reacted for 2 hours, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to obtain 13g of ALA-HPO complex.
m.p.:145.3～147.2℃.
ESI-HRMS:m/z calcd for C 18 H 27 N 2 O 7 [M+H] + :383.1813；found:383.1808.
1 H NMR(400MHz,DMSO-d 6 )δ7.76(t,J＝5.9Hz,1H),7.52(s,1H),6.29(s,1H),5.02(s,2H),3.95(d,J＝5.8Hz,2H),3.78(s,2H),3.60(s,3H),2.73(t,J＝6.5Hz,2H),2.53(t,J＝6.4Hz,2H),1.71(p,J＝7.6Hz,2H),1.36–1.25(m,4H),0.89(t,J＝6.9Hz,3H). 13 C NMR(100MHz,DMSO-d 6 )δ205.67,173.05,170.74,156.03,147.88,142.63,124.02,114.26,61.70,52.48,51.86,50.15,34.22,30.60,28.52,27.56,22.20,14.33.
Example 18
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added in 100mL as solvents, respectively, and the reaction system was heated to reflux. Placing benzyl bromide (10.26g, 60mmol) in a constant-pressure funnel, slowly dropwise adding the benzyl bromide into the reaction solution while stirring, continuously refluxing for reaction for 2h, monitoring the reaction process by TLC (thin layer chromatography), stopping the reaction after the raw materials are completely converted, concentrating the solvent in vacuum, precipitating a large amount of yellow solid, washing the solid with water (50 mL multiplied by 3), and filtering to obtain a light yellow solid compound pyrone (13.5 g), wherein the yield is 98%.
The pyrone (2.30g, 10mmol) reacted in the reaction is added into a 100mL round-bottom flask respectively, dissolved in ethanol and water (1:1) for 30mL, stirred until the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is more than 12, octylamine (1.548g, 12mmol) is added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC in the period. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and the mixture was purified by a silica gel column, and gradient elution was performed using dichloromethane: methanol (100). The product, 1- (2-octyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.7 g), was isolated as a yellow oil in 48% yield.
Anhydrous dichloromethane (10 mL) and p-nitrophenylchloroformate (242mg, 1.2mmol) are sequentially added into a 50mL single-neck bottle, the round-bottom flask is moved to an ice maker to be cooled to-5 ℃ (internal temperature), after 30min, a solution of 1-octyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (343mg, 1mmol) in dichloromethane and triethylamine (152mg, 1.5mmol) is slowly added dropwise into the round-bottom flask, the solution is stirred for 30min, then the solution is moved to room temperature and stirred for 6-8 h, the reaction solution is dried, and an intermediate (312 mg) is obtained by column chromatography (dichloromethane: methanol =100: 1-50).
1mmol of methyl 5-aminolevulinate hydrochloride was dissolved in dichloromethane (6 mL), triethylamine (202mg, 2mmol) and DMAP (12mg, 0.1eq.) were sequentially added, stirred at room temperature for 5h, the intermediate carbonate (559mg, 1.1eq.) obtained above was dissolved in dichloromethane, and the reaction mixture obtained above was added dropwise at-10-20 ℃ to rapidly turn yellow due to the formation of p-nitrophenol. The reaction was complete until the carbonate blue spot observed on TLC became very weak. The reaction mixture was washed once with saturated sodium bicarbonate solution, twice with water, the aqueous layer was back-extracted once with dichloromethane, the combined organic phases were dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by column on silica gel to give benzyl protected hydroxypyridone (343 mg) in 61% yield.
0.5mmol of the benzyl protected hydroxypyridone obtained in the above step was weighed out and dissolved in 3mL of methanol in a 25mL round bottom flask, catalytically hydrogenated with 5% Pd/C, maintained at 30psi hydrogen pressure, reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/ether to obtain ALA-HPO complex for 13h.
m.p.:167～169℃.
ESI-HRMS:m/z calcd for C 21 H 33 N 2 O 7 [M+H] + :425.2288；found:425.2262.
1 H NMR(400MHz,DMSO-d 6 )δ7.76(t,J＝5.9Hz,1H),7.51(s,1H),6.28(s,1H),5.01(s,2H),3.94(d,J＝5.8Hz,2H),3.86(t,J＝7.8Hz,2H),3.60(s,3H),2.73(t,J＝6.5Hz,2H),2.60(t,J＝6.5Hz,2H),1.70(t,J＝7.6Hz,2H),1.28(q,J＝4.8Hz,10H),0.93–0.83(t,J＝6.12,3H). 13 C NMR(100MHz,DMSO-d 6 )δ206.15,173.64,171.15,156.57,148.42,122.34,112.61,103.67,71.12,61.22,52.30,52.91,51.36,49.96,34.52,33.72,32.11,30.37,28.58,21.07,14.12.
Example 19
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Placing benzyl bromide (10.26g, 60mmol) in a constant-pressure funnel, slowly dropwise adding the benzyl bromide into the reaction solution while stirring, continuously refluxing for reaction for 2h, monitoring the reaction process by TLC (thin layer chromatography), stopping the reaction after the raw materials are completely converted, concentrating the solvent in vacuum, precipitating a large amount of yellow solid, washing the solid with water (50 mL multiplied by 3), and filtering to obtain a light yellow solid compound pyrone (13.5 g), wherein the yield is 98%.
Pyrone (2.30g, 10mmol) reacted above is respectively added into a 100mL round-bottom flask, dissolved in ethanol and water (1:1) for 30mL, stirred, when the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is more than 12, butylamine (0.876g, 12mmol) is continuously added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC during the reaction. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and purified by a silica gel column, using a gradient elution of dichloromethane: methanol (100). The product, 1- (2-aminobutyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.8 g), was isolated as a yellow oil in 55% yield.
Anhydrous dichloromethane (10 mL) and p-nitrophenyl chloroformate (242mg, 1.2mmol) are sequentially added into a 50mL single-neck bottle, the round-bottom flask is moved to an ice maker to be cooled to-5 ℃ (internal temperature), after 30min, a solution of 1-butyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (287 mg,1 mmol) in dichloromethane and triethylamine (152mg, 1.5mmol) is slowly added dropwise into the round-bottom flask, the solution is stirred for 30min, the mixture is moved to room temperature and stirred for 6-8 h, the reaction solution is dried, and an intermediate (231 mg) is obtained by column chromatography (dichloromethane: methanol =100: 1-50).
1mmol of methyl 5-aminolevulinate hydrochloride was dissolved in dichloromethane (6 mL), triethylamine (202mg, 2mmol) and DMAP (12mg, 0.1eq.) were sequentially added thereto, and the mixture was stirred at room temperature for 5 hours, and then the intermediate carbonate (497mg, 1.1eq.) obtained above was dissolved in dichloromethane and the reaction mixture obtained above was added dropwise at-10 to 20 ℃ to rapidly turn yellow due to the formation of p-nitrophenol. The reaction was complete until the carbonate blue spot observed on TLC became very weak. The reaction mixture was washed once with saturated sodium bicarbonate solution, twice with water, the aqueous layer was back-extracted once with dichloromethane, the combined organic phases were dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by means of a silica gel column to give benzyl-protected hydroxypyridone (385 mg) in 65% yield.
Weighing 0.5mmol of the benzyl protected hydroxypyridone obtained in the above step, dissolving in 3mL of methanol in a 25mL round bottom flask, catalytically hydrogenating with 5% Pd/C, maintaining 30psi of hydrogen pressure, reacting for 2h, filtering, distilling off the solvent under reduced pressure, and recrystallizing with ethanol/diethyl ether to obtain ALA-HPO complex 13i.
m.p.:141～143℃.
ESI-HRMS:m/z calcd for C 17 H 25 N 2 O 7 [M+H] + :369.1656；found:369.1640.
1 H NMR(400MHz,Chloroform-d)δ7.25(s,1H),6.54(s,1H),6.04(s,1H),5.02(s,2H),4.19(s,2H),3.87(t,J＝5.8Hz,2H),3.67(s,3H),2.78(t,J＝6.5,2H),2.66(t,J＝6.4Hz,2H),1.74(q,J＝7.6Hz,2H),1.37(h,J＝7.3Hz,2H),0.96(t,J＝7.3Hz,3H). 13 C NMR(100MHz,DMSO-d 6 )δ205.25,172.62,170.41,155.60,147.47,142.19,123.57,113.85,61.27,51.85,51.42,49.72,33.79,32.44,27.13,19.19,13.49.
Example 20
In a 250mL round-bottom flask, starting materials, kojic acid (4.10 g, 50mmol), potassium carbonate (8.28g, 60mmol), ethanol and water (1:1) were added to 100mL of each as a solvent, and the reaction system was heated to reflux. Placing benzyl bromide (10.26g, 60mmol) in a constant-pressure funnel, slowly dropwise adding the benzyl bromide into the reaction solution while stirring, continuously refluxing for reaction for 2h, monitoring the reaction process by TLC (thin layer chromatography), stopping the reaction after the raw materials are completely converted, concentrating the solvent in vacuum, precipitating a large amount of yellow solid, washing the solid with water (50 mL multiplied by 3), and filtering to obtain a light yellow solid compound pyrone (13.5 g), wherein the yield is 98%.
Pyrone (2.30g, 10mmol) reacted above is respectively added into a 100mL round-bottom flask, dissolved in ethanol and water (1:1) for 30mL, stirred until the pyrone is completely dissolved, sodium hydroxide solution (2.0 mmol/L) is added dropwise until the pH value of a reaction system is more than 12, methoxyethylamine (1.068g, 12mmol) is added dropwise, the temperature is increased to reflux, the reaction is carried out for 3 hours, and the reaction condition is monitored by TLC during the reaction. After the reaction, the solvent was concentrated under reduced pressure, and when the reaction liquid was concentrated to one-half of the original volume, the residue was adjusted to pH 1 to 2 by adding dilute hydrochloric acid, the aqueous layer was washed with ethyl acetate (50 mL. Times.2), adjusted to pH neutral again, and extracted with dichloromethane (20 mL. Times.5). The organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, and purified by a silica gel column, using a gradient elution of dichloromethane: methanol (100). The product, 1- (2-aminomethoxypropyl) -2-hydroxymethyl-5-benzyloxypyridin-4-one (1.39 g), was isolated as a yellow oil in 43% yield.
Anhydrous dichloromethane (10 mL) and p-nitrophenyl chloroformate (242mg, 1.2mmol) are sequentially added into a 50mL single-neck bottle, the round-bottom flask is moved into an ice maker to be cooled to-5 ℃ (internal temperature), after 30min, a solution of 1-methoxypropyl-2-hydroxymethyl-5- (benzyloxy) -pyridin-4-one (303mg, 1mmol) in dichloromethane and triethylamine (152mg, 1.5mmol) is slowly added dropwise into the round-bottom flask, the solution is stirred for 30min, then the solution is moved to room temperature and stirred for 6-8 h, the reaction solution is dried, and an intermediate (362 mg) is obtained by column chromatography (dichloromethane: methanol =100: 1-50).
1mmol of methyl 5-aminolevulinate hydrochloride was dissolved in dichloromethane (6 mL), triethylamine (202mg, 2mmol) and DMAP (12mg, 0.1eq.) were added in this order, and stirred at room temperature for 5 hours, and then the intermediate carbonate (514mg, 1.1eq.) obtained above was dissolved in dichloromethane, and the reaction mixture obtained above was added dropwise at-10 to 20 ℃ to rapidly turn yellow due to the formation of p-nitrophenol. The reaction was complete until the carbonate blue spot observed on TLC became very weak. The reaction mixture was washed once with saturated sodium bicarbonate solution, twice with water, the aqueous layer was back-extracted once with dichloromethane, the combined organic phases were dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by column on silica gel to give benzyl protected hydroxypyridone (328 mg) in 59% yield.
0.5mmol of the benzyl-protected hydroxypyridone obtained in the above step was weighed out and dissolved in 3mL of methanol in a 25mL round-bottomed flask, catalytically hydrogenated with 5% Pd/C, maintained under 30psi hydrogen pressure, reacted for 2h, filtered, distilled under reduced pressure to remove the solvent, and recrystallized with ethanol/diethyl ether to obtain ALA-HPO complex 13j.
m.p.:128～130℃.
HRMS:m/z calcd for C 17 H 25 N 2 O 8 [M+H] + :385.1605；found:385.1596.
1 H NMR(400MHz,DMSO-d 6 )δ7.72(t,J＝5.7Hz,1H),7.43(s,1H),6.26(s,1H),4.99(s,2H),3.91(t,J＝6.9Hz,4H),3.58(s,3H),3.31(t,J＝5.8Hz,2H),3.24(s,3H),2.70(t,J＝6.5Hz,2H),2.51(t,J＝6.4Hz 2H),1.94(p,J＝6.1Hz,2H). 13 C NMR(100MHz,DMSO-d 6 )δ205.71,172.06,170.92,156.02,147.94,142.89,123.79,114.10,68.82,61.63,58.35,51.86,50.17,49.69,34.23,30.60,27.58.
Example 21
The following are the pharmacological test data for some of the compounds of the invention:
1. phototoxicity test
The experimental method comprises the following steps: the cells were incubated at approximately 2.5X 10 5 Density of/well plated in 24-well plates, after 48h of incubation, cells were washed with phosphate buffer. 1mL of compound solutions containing different concentrations (50-10. Mu.M) were added to the designated wells and incubated for 4h. The blank (without compound) and different concentrations of compound were each subjected to 4 parallel experiments, irradiated with blue light for 5min, immediately replaced with fresh medium after irradiation, and the cells were incubated for a further 18h. Cytotoxicity was determined using MTT. The MTT detection method comprises the following steps: all media was removed from the well plate, 1mL MTT-containing media was added per well, capped and placed in an incubator for 4h. Is removed againAll media in the plate, 1mL of dimethyl sulfoxide per well, shaken for 5min, absorbance at 520nm was measured, the average cell viability at each concentration of each prodrug was calculated and expressed as a percentage of control,
the values for cell viability and concentration are tabulated and presented in bar graph form.
Mean cell viability = absorbance (mean of each prodrug)/absorbance (mean of control) x 100%
To determine the "dark" toxicity of the compounds, the previous procedure was repeated, but without irradiation with light.
From the experimental results (fig. 1 and 2) the following conclusions can be drawn:
after 4h incubation of the compound with cells (fig. 1): at a concentration of 50. Mu.M, the illumination intensity was 5J/cm 2 In the cases, the survival percentages of ALA, ALA-DFP and 10 a-10J-treated HeLa cells were 99.8%, 93.4%, 91.3%, 93.5%, 96.8%, 92.24%, 95.58%, 98.26%, 94.24%, 96.49%, 98.41%, 97.19%, respectively, and the light intensity was 10J/cm 2 The survival percentages of ALA, ALA-DFP and 10 a-10 j-treated HeLa cells were 65.34%, 56.73%, 99.81%, 98.41%, 94.87%, 81.64%, 42.00%, 75.64%, 87.47%, 75.51%, 60.33% and 66.57%, respectively. Comparing the viability of the cells at the two light intensities, it is clear that there is a certain dependence between the activity of the compound and the light intensity.
Continuing to increase the concentration of compound to 100 μ M, from the experimental results (fig. 2): the dependence between compound activity and light intensity is more pronounced, with compound 10e having the best cellular activity. These results indicate that ALA-HPO can enter cells, and that cytosolic esterase can disrupt the bond between the PpIX prodrug and HPO molecule to liberate ALA and metabolically produce the photoactive active substance PpIX, so that the synthesized target compound can effectively function in cells.
2. Dark toxicity test
After 4h incubation with compound: as shown in fig. 3, compounds 10 a-10 j inhibit HeLa cells at 50 μ M or 100 μ M concentrations in a histogram with 1% FBS-DMEM cell culture as the blank, and the data indicates: after 4h incubation of the compounds with cells, all compounds showed no dark toxicity at the relevant concentrations of 50 or 100 μ M compared to untreated controls and the positive drugs ALA and ALA-DFP.
Example 22
1. Phototoxicity test
From FIG. 1, it can be seen that: from the experimental results: haCaT cell line was incubated with 50. Mu.M compound and then irradiated with light (5 KJ/M) 2 ) After irradiation, the compounds were found to have a cell killing effect that was substantially equivalent to that of ALA, higher than that of DFP alone, but lower than that of the ALA-DFP combination. At the same concentration, when UV (10 KJ/m) is used 2 ) After irradiation, the activity of 13I and 13j is slightly higher than when ALA is used alone and, in contrast to the structure of the compounds of series I, we have found that the activity is better when the number of carbon atoms in the R substituent is 4.
Similarly, the HaCaT cell line was incubated with 100. Mu.M compound and then incubated with 5KJ/M 2 The percent survival of the HaCaT cell lines treated with ALA, compounds 13a,13 b, 13e,13 i, 13j upon intense UV irradiation was 95.2%, 97.5%, 89.4%, 90.2%, 84.3%, 83.7%, respectively. It is evident that compounds 13b, 13e,13 i, 13j have an increased overall cytotoxicity compared to ALA. At the same concentration, when UV (10 KJ/m) is used 2 ) After irradiation, the activity of the remaining compounds including ALA was not significantly changed, except that the activity of ALA-DFP was significantly enhanced.
In addition, a comparison of the cytotoxicity of the series of compounds at 50. Mu.M and 100. Mu.M with ALA-DFP was also investigated, and the combination of ALA and DFP was found to be more effective in cell killing than the series of compounds, indicating that ALA-DFP combination therapy is the most effective therapeutic approach. Nevertheless, the above results indicate that ALA-HPO can enter cells, and that it is speculated that cytosolic esterases can break the bond between the PpIX prodrug and the HPO molecule, free ALA, and metabolically produce the photoactive active substance PpIX, so that the synthesized series I target compounds can effectively function in cells.
2. Dark toxicity test
After 4h incubation with compound: only compounds 13c and 13f showed dark toxicity at the relevant concentrations; after 24h incubation with compound: compounds 13a,13e,13g and 13j showed no dark toxicity at the relevant concentrations of 50 or 100 μ M, compounds 13c, 13d, 13f, 13h at both concentrations, and compounds 13b and 13i only at a concentration of 100 μ M, compared to the untreated control.
Finally, it is to be noted that the above-listed are 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 the person skilled in the art from the present disclosure are to be considered within the scope of the present invention.
Claims (8)
1. An ALA-HPO hybrid derivative of formula (I):
in the formula (I), the compound is shown in the specification,
R 1 is a pentyl group.
2. A process for the preparation of ALA-HPO hybrid derivatives of formula (I) as claimed in claim 1, wherein:
the preparation method of the ALA-HPO hybrid derivative shown in the formula (I) comprises the following steps:
(1) Dissolving the compound 1, benzyl bromide and an alkaline substance in an organic solvent, reacting for 2-12 h at 25-90 ℃, and then carrying out post-treatment on the reaction solution to obtain a compound 2;
the mass ratio of the compound 1, the benzyl bromide and the alkaline substance is 1:1 to 6:1 to 5;
the alkaline substance is one or a mixture of more than two of potassium carbonate, potassium hydroxide, sodium carbonate, sodium bicarbonate and triethylamine in any proportion;
(2) Mixing the compounds 2 and R 1 NH 2 The alkaline substance is dissolved in the organic solventReacting in the agent for 1-12 h at 40-100 ℃, and then carrying out post-treatment on reaction liquid to obtain a compound 3a;
the compound 2, R 1 NH 2 The ratio of the amount of the alkaline substance is 1:1 to 5:0.5 to 2;
the alkaline substance is one or a mixture of more than two of potassium carbonate, potassium hydroxide, sodium carbonate, sodium bicarbonate and triethylamine in any proportion;
(3) Reacting compound 4 with Boc 2 Dissolving O and an alkaline substance in an organic solvent, reacting for 2-24 h at 0-50 ℃, and then carrying out post-treatment on the reaction liquid to obtain a compound 5;
the compound 4, boc 2 The ratio of the amount of O to the amount of the basic substance is 1:1 to 2:2 to 10;
the alkaline substance is one or a mixture of more than two of potassium carbonate, potassium hydroxide, sodium carbonate, sodium bicarbonate, sodium acetate, sodium propionate and triethylamine in any proportion;
(4) Dissolving the compound 5 in an organic solvent, sequentially adding dicyclohexylcarbodiimide and 4-dimethylaminopyridine, reacting at 10-50 ℃ for 2-12 h, filtering the reaction solution, and collecting the filtrate; dissolving the compound 3a in an organic solvent, dripping the solution into the collected filtrate at the temperature of between 10 and 50 ℃, maintaining the temperature for reacting for 6 to 24 hours after dripping, and then carrying out post-treatment on the reaction solution to obtain a compound 6;
the mass ratio of the compound 5, dicyclohexylcarbodiimide, 4-dimethylaminopyridine and the compound 3a is 10:10:1:10;
(5) Dissolving a compound 6 in dichloromethane, dropwise adding an anhydrous dichloromethane solution of 1.0mol/L trifluoroacetic acid under the protection of nitrogen at-10-20 ℃, transferring to room temperature after dropwise adding, continuously stirring and reacting for 12-24 h, and then distilling the reaction solution under reduced pressure to remove the solvent to obtain a compound 7;
the mass ratio of the compound 6 to trifluoroacetic acid is 1:1;
(6) Dissolving the compound 7 in an organic solvent, adding palladium carbon, stirring and reacting for 2-24 h at-5-50 ℃ under a hydrogen atmosphere, and then carrying out post-treatment on a reaction solution to obtain a compound shown in a formula (I);
the ratio of the compound 7 to palladium carbon in terms of palladium is 5:1 to 3;
the R is 1 NH 2 In the compound 3a, the compound 6 and the compound 7, R 1 The definition of (A) is the same as in formula (I).
3. The process according to claim 2, wherein the organic solvent used in the step (1) of the process for producing the ALA-HPO hybrid derivative represented by formula (I) is one or a mixture of two or more of acetone, ethanol, methanol, dichloromethane, chloroform, carbon tetrachloride, acetonitrile, toluene, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide.
4. The process according to claim 2, wherein the organic solvent used in the step (2) of the process for producing the ALA-HPO hybrid derivative represented by formula (I) is one or a mixture of two or more of water, acetone, ethanol, methanol, dichloromethane, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide.
5. The process according to claim 2, wherein the organic solvent used in the step (3) of the process for producing the ALA-HPO hybrid derivative represented by formula (I) is one or a mixture of two or more selected from the group consisting of acetone, ethanol, methanol, acetic acid, acetic anhydride, methylene chloride, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide.
6. The process according to claim 2, wherein the organic solvent for dissolving compound 5 in the process step (4) for preparing the ALA-HPO hybrid derivative represented by formula (I) is one or a mixture of two or more of acetone, ethanol, methanol, dichloromethane, chloroform, carbon tetrachloride, toluene, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide and N, N-dimethylacetamide in any ratio; the definition of the organic solvent used for dissolving the compound 3a is the same as that.
7. The process according to claim 2, wherein in the step (6), the organic solvent is one or a mixture of two or more selected from acetone, ethanol, methanol, dichloromethane, chloroform, carbon tetrachloride, toluene, acetonitrile, dimethyl sulfoxide, dioxane, N-dimethylformamide, and N, N-dimethylacetamide.
8. Use of ALA-HPO hybrid derivatives of formula (I) as defined in claim 1 for the preparation of a medicament for photodynamic therapy.
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