WO2020098443A1 - Bacterial chlorophyll derivative and preparation method therefor - Google Patents
Bacterial chlorophyll derivative and preparation method therefor Download PDFInfo
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- WO2020098443A1 WO2020098443A1 PCT/CN2019/111601 CN2019111601W WO2020098443A1 WO 2020098443 A1 WO2020098443 A1 WO 2020098443A1 CN 2019111601 W CN2019111601 W CN 2019111601W WO 2020098443 A1 WO2020098443 A1 WO 2020098443A1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
- C07D471/22—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed systems contains four or more hetero rings
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/439—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom the ring forming part of a bridged ring system, e.g. quinuclidine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
- C07D487/22—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains four or more hetero rings
Definitions
- the invention relates to the field of pharmaceuticals, in particular to a chlorophyll derivative of bacteria, a preparation method and application thereof.
- Photodynamic therapy refers to a treatment method that uses visible light to sensitize a photosensitizer located inside a human diseased tissue to kill the diseased tissue.
- a photosensitizer that has an affinity for tumor cells and other biologically active tissues.
- the photosensitizer will selectively enrich in the lesion area, and then irradiate the lesion area with light of a certain wavelength.
- the photosensitizer absorbs light energy It is then converted into a highly reactive active substance, which, with the participation of oxygen, chemically reacts with the biological molecules of the diseased tissue to kill it.
- This is a new type of clinical treatment technology under research and development, which is mainly used in clinical treatment of various tumors.
- chlorophyll (a) derivatives and bacterial chlorophyll derivatives have strong absorption characteristics in the 650-850nm spectral region and are easily degraded after treatment, they are considered as excellent photosensitizers for photodynamic therapy of tumors.
- photosensitizers of chlorophyll (a) derivatives and bacterial chlorophyll derivatives It is the first generation of photosensitizer.
- the photoperiod-free treatment period needs 4-6 weeks, the absorption wavelength is 630nm, and the penetration depth in the tissue is less than 4mm.
- HPPH currently in Phase II clinical stage, is a second-generation photosensitizer with a maximum absorption wavelength of 665 nm and a penetration depth of about 0.5 cm in tissues.
- the photoperiod-free period after photodynamic therapy is one week. Long, in life, patients will inevitably be exposed to light, so HPHP treatment of the skin will cause erythema and other damage.
- WO2004 / 002476 reports a bacterial chlorophyll derivative with an absorption wavelength of 700-850 nm. Since it is a fat-soluble compound, it has a long metabolism time, high content in skin, and high skin phototoxicity. Similarly, the compounds reported in the literature (Bioconjug Chem. 2009 February; 20 (2): 274–282) have not been metabolized in the main tissues or organs after 96 hours of administration, indicating that the compounds are metabolized in vivo Slowness indicates that the compound has a longer light-shielding period.
- the present invention provides a bacterial chlorophyll derivative with fast metabolism and short light-proof period, which can be used as a photosensitizer for photodynamic therapy research, and patients can achieve the same day Treatment, go home on the same day, thereby reducing the financial burden of patients and their families.
- the object of the present invention is to provide a compound of formula (I) or its stereoisomer, or a pharmaceutically acceptable salt thereof:
- R 1 is hydrogen or (CH 2 ) m COOH
- R 2 is selected from hydrogen, hydroxyl, halogen, C 1 -C 6 alkyl or C 1 -C 6 alkoxy;
- R 3 and R 4 are each independently selected from C 1 -C 6 alkyl
- n 1, 2, 3 or 4;
- n 1, 2, 3 or 4.
- a compound of formula (I) or its stereoisomer, or a pharmaceutically acceptable salt thereof is a compound of formula (II) or its stereoisomer, or its Medicinal salt:
- R 2 , R 3 , R 4 , m and n are defined as in formula (I).
- a compound of formula (I) or its stereoisomer, or a pharmaceutically acceptable salt thereof is a compound of formula (III) or its stereoisomer, or its Medicinal salt:
- R 2 , R 3 , R 4 and n are as defined in formula (I).
- a compound of formula (I), formula (II) or formula (III) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof wherein R 2 is selected from hydrogen, halogen , C 1 -C 6 alkyl or C 1 -C 6 alkoxy.
- Representative compounds of the present invention include, but are not limited to:
- the present invention provides a method for preparing a compound of formula (II) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.
- the method includes:
- the compound of formula (IIA) undergoes a hydrolysis reaction under basic conditions, and then acid is added for acid hydrolysis to obtain the compound of formula (II);
- R 2 , R 3 , R 4 , m and n are as defined in formula (I).
- the present invention provides a method for preparing a compound of formula (II) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, the method comprising: the base used in the alkaline condition is selected from alkali metal carbonates Or ammonium carbonate, preferably alkali metal carbonate, more preferably sodium carbonate, potassium carbonate or lithium carbonate;
- the acid is selected from inorganic acid or organic acid, wherein the inorganic acid is selected from hydrochloric acid with a mass fraction of 1-30% or sulfuric acid with a mass fraction of 1-50%, preferably hydrochloric acid with a mass fraction of 1-10% or a mass Sulfuric acid with a fraction of 1-10%, the organic acid is selected from C 1 -C 4 organic acids, preferably formic acid, acetic acid or oxalic acid.
- the present invention provides a method for preparing a compound of formula (III) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.
- the method includes:
- R 2 , R 3 , R 4 and n are as defined in formula (I).
- the present invention provides a method for preparing a compound of formula (III) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, the method comprising: the base used in the alkaline condition is selected from alkali metal carbonates , Ammonium carbonate or organic amine, preferably alkali metal carbonate or C 1 -C 6 primary amine, more preferably sodium carbonate, potassium carbonate, lithium carbonate, n-propylamine or butylamine.
- the base used in the alkaline condition is selected from alkali metal carbonates , Ammonium carbonate or organic amine, preferably alkali metal carbonate or C 1 -C 6 primary amine, more preferably sodium carbonate, potassium carbonate, lithium carbonate, n-propylamine or butylamine.
- the present invention provides a compound represented by formula (IIA) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
- R 2 , R 3 , R 4 , m and n are as defined in formula (I).
- the present invention provides a compound represented by formula (IIIA) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
- R 2 , R 3 , R 4 and n are as defined in formula (I).
- the present invention provides a pharmaceutical composition containing an effective dose of a compound of formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and excipient Or a combination of them.
- the present invention provides a compound of formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for use in the preparation of a medicament for photodynamic therapy of cancer, wherein the cancer is preferably One or more of lung cancer, prostate cancer, esophageal cancer, melanoma, bile duct cancer, head and neck cancer, bladder cancer, stomach cancer, nasopharyngeal cancer, colon cancer.
- the maximum absorption wavelength of the compound of the present invention is around 780nm, and the penetration depth of the tissue is 0.7-1cm, which can effectively kill tumor cells with deep tissues; due to the introduction of hydrophilic groups, carboxyl groups or The phenolic hydroxyl groups have good water solubility, so the metabolic time is short and the dark period is short; at the same time, the compounds of the present invention have good chemical stability.
- Alkyl when taken as a group or part of a group refers to a C 1 -C 6 linear or branched saturated aliphatic hydrocarbon group.
- C 1 -C 6 refers to containing 1 to 6 carbon atoms, for example, may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms .
- alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-di Methylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1 -Ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethyl Butyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl Wait.
- alkoxy refers to the group (alkyl-O-). Among them, alkyl refers to the relevant definitions herein.
- Examples of C 1 -C 6 alkoxy groups include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, t-butoxy, n- Pentoxy, 1,1-dimethylpropoxy, 1,2-dimethylpropoxy, 2,2-dimethylpropoxy, 1-ethylpropoxy, 2-methylbutoxy Oxygen, 3-methylbutoxy, n-hexyloxy, 1-ethyl-2-methylpropoxy, 1,1,2-trimethylpropoxy, 1,1-dimethylbutoxy Group, 1,2-dimethylbutoxy, 2,2-dimethylbutoxy, 1,3-dimethylbutoxy, 2-ethylbutoxy, 2-methylpentoxy , 3-methylpentoxy, 4-methylpentoxy, 2,3-dimethylbutoxy.
- Halogen means fluorine, chlorine, bromine and iodine.
- Amino refers to -NH 2 .
- Niro refers to -NO 2 .
- Benzyl refers to -CH 2 - phenyl.
- Carboxy refers to -C (O) OH.
- Carboxylate group refers to -C (O) O (alkyl), where alkyl is as defined above.
- Boc refers to tert-butoxycarbonyl
- DMSO dimethyl sulfoxide
- Substituted refers to one or more hydrogen atoms in the group, preferably up to 5, more preferably 1-3 hydrogen atoms are independently substituted with a corresponding number of substituents for each other. It goes without saying that the substituents are only at their possible chemical positions, and those skilled in the art can determine (through experiment or theory) possible or impossible substitutions without undue effort. For example, an amino group or hydroxyl group having free hydrogen may be unstable when combined with a carbon atom having an unsaturated (eg, olefinic) bond.
- “Pharmaceutically acceptable salts” refers to certain salts of the above compounds that can retain their original biological activity and are suitable for medical uses.
- the pharmaceutically acceptable salt of the compound represented by formula (I) may be a metal salt or a salt with a suitable acid.
- Stepoisomer means including but not limited to diastereomers, enantiomers, atropisomers and mixtures thereof, such as racemic mixtures, forming part of the present invention.
- Diastereomers can be separated into individual diastereomers by chromatography, crystallization, distillation or sublimation based on their physical and chemical differences.
- Enantiomers can be converted into diastereoisomeric mixtures by separation, by reacting with appropriate optically active compounds (such as chiral auxiliary agents such as chiral alcohols or Mosher's acid chloride) , Separation of diastereomers, and the conversion of individual diastereomers to the corresponding pure enantiomers.
- optically active compounds such as chiral auxiliary agents such as chiral alcohols or Mosher's acid chloride
- the intermediates and compounds of the present invention may also exist in different tautomeric forms, and all such forms are included within the scope of the present invention.
- the prefixes D, L or R, S are used to denote the absolute configuration of the molecular chiral center.
- the prefixes d, l, or (+), (-) are used to name the symbol for rotation of plane polarized light of the compound, (-) or l means the compound is left-handed, and the prefix (+) or d means the compound is right-handed.
- the atoms or groups of these stereoisomers are connected in the same order, but their stereostructures are different.
- the specific stereoisomer may be an enantiomer, and a mixture of isomers is generally called an enantiomeric mixture.
- a 50:50 mixture of enantiomers is called a racemic mixture or a racemate, which may result in no stereoselectivity or stereospecificity during the chemical reaction.
- the terms "racemic mixture” and “racemate” refer to an equimolar mixture of two enantiomers, lacking optical activity.
- Alkali metal carbonate means lithium carbonate, sodium carbonate, potassium carbonate, rubidium carbonate, cesium carbonate, and francium carbonate.
- “Pharmaceutical composition” means a mixture comprising a compound of the present application or a physiologically acceptable salt or prodrug thereof with other chemical components, and other components such as physiologically acceptable carriers and excipients.
- the purpose of the pharmaceutical composition is to promote the administration to the organism, facilitate the absorption of the active ingredient and thus exert the biological activity.
- Figure 1 shows the effect of different doses of compound (I-1) on the volume of NCI-H460 transplanted tumor
- Fig. 2 is a blood concentration-time curve of compound (I-1) (3.0 mg / kg) in different tissues;
- Fig. 3 is the blood concentration-time curve of HPPH (0.6mg / kg) in different tissues.
- Mass spectrometry is measured by LC / MS instrument, and the ionization method can be ESI or APCI.
- UV data was measured with SHIMADZU's UV-2600 ultraviolet spectrometer.
- the thin-layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate.
- the specification of the thin-layer chromatography (TLC) silica gel plate is 0.15mm-0.2mm, and the specification of the thin-layer chromatography separation and purification product is 0.4mm. -0.5mm.
- DMSO Deuterated dimethyl sulfoxide.
- Nitrogen reaction means that the reaction bottle is connected to a nitrogen balloon with a volume of about 1L.
- the ratio of the removal agent is the volume ratio, where the volume ratio of the solvent is different according to the polarity of the compound, and a small amount of acidic or basic reagents can also be added for adjustment, such as acetic acid or triethylamine.
- the compound (IIB-1) in the present invention is synthesized according to the method of the literature (J. Med. Chem., 2016, 59, 9774-9787), compound (IIB-2), compound (IIB-3) and compound (IIB-4 ) Are synthesized according to the preparation method of compound (IIB-1), and the structural characterization data are as follows:
- the compound (IIA-1) was added to a 500 mL three-necked round bottom flask, 100 mL of acetonitrile and 40 mL of deionized water were added, stirred, and then potassium carbonate (3.8 g, 27.5 mmol) was added, and the reaction was stirred at 40 ° C under nitrogen protection. The reaction was monitored by TLC. After the reaction was completed, a 10% aqueous acetic acid solution was added to adjust the pH to 2-3.
- UV Wavelength (Absorbance): 781.5 (0.244), 537.0 (0.214), 416.0 (0.241), 365.5 (0.589), 344.0 (0.301), 291.5 (0.10), 225 (0.156), 201.0 (0.256), 194.5 (0.123) ;
- UV Wavelength (Absorbance): 782.2 (0.18), 537.0 (0.155), 416.0 (0.195), 365.6 (0.438), 202.0 (0.40);
- reaction was quenched with 10% dilute acetic acid aqueous solution, followed by addition of 150mL of dichloromethane and 100mL of water, and the organic phase was separated , Dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a black solid, which was directly subjected to the next reaction;
- UV Wavelength (Absorbance): 781.8 (0.094), 536.6 (0.083), 416.0 (0.096), 365.6 (0.231), 200.4 (0.110);
- UV Wavelength (Absorbance): 782.2 (0.141), 537. (0.122), 415.6 (0.148), 365.8 (0.348), 202.8 (0.235);
- Feeding conditions Animals are housed in the IVC system of the experimental animal house, 5 animals / cage, light period 10h / 14, indoor temperature 20-26 °C, relative humidity 40-70%, all cages, bedding and drinking water are required before use Sterilization, the feed is special sterilized feed, cages, feed and water are replaced once a week, all replacement operations are carried out on the ultra-clean workbench.
- Test Example 1 Tumor inhibition rate of different doses of compound (I-1) on nude mice transplanted with NCI-H460 (human lung cancer)
- NCI-H460 cells were cultured in RPMI-1640 complete culture medium containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 ⁇ g / mL streptomycin in 5% CO 2 , 37 ° C, saturated humidity constant temperature incubator. Cells in the logarithmic growth phase were planted in the axilla of the right forelimb of nude mice, and the number of cells inoculated was 2 ⁇ 10 6 cells.
- NCI-H460 tumor mass (generations 3-10) inoculated in the axillary of nude mice at a rapid proliferation stage, remove the necrotic tissue in the center of the tumor body, and cut the tumor mass into small tumor masses of 1mm ⁇ 1mm ⁇ 1mm under sterile conditions Use a trocar to inoculate subcutaneously in the right forelimb of nude mice.
- the tumor grows to 60-120 mm 3 and then start grouping according to body weight, a total of 5 groups, 8 animals in each group, are tested.
- the optical fiber probe is fixed with a fixed frame, the spot area is one square centimeter, the light intensity is set to 150mw / cm 2 , the energy density is 150J / cm 2 , the illumination time is 20min; 780nm PDT, the probe is about 7.0cm away from the tumor surface, the spot area is one square In centimeters, the light intensity is set to 100mw / cm 2 and the light duration is 15min.
- the animals were irradiated, they were kept in the IVC system, and the volume of the transplanted tumor was measured every 4 days from the 11th day after the light irradiation until the end of the experiment on the 31st day.
- Tumor inhibition rate (%) [(average tumor weight of model control-average tumor weight of experimental group) / average tumor weight of experimental group] ⁇ 100%
- Table 1 The effect of different doses of compound (I-1) on the volume of NCI-H460 transplanted tumor (unit: mm 3 )
- HPPH The structural formula of HPPH is as follows:
- NCI-H460 cells were cultured in RPMI-1640 complete culture medium containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 ⁇ g / mL streptomycin in 5% CO 2 , 37 ° C, saturated humidity constant temperature incubator. Cells in the logarithmic growth phase were planted in the axilla of the right forelimb of nude mice, and the number of cells inoculated was 2 ⁇ 10 6 cells.
- NCI-H460 tumor mass (generations 3-10) inoculated in the axillary of nude mice at a rapid proliferation stage, remove the necrotic tissue in the center of the tumor body, and cut the tumor mass into small tumor masses of 1mm ⁇ 1mm ⁇ 1mm under sterile conditions
- Use a trocar to inoculate subcutaneously in the right forelimb of nude mice.
- After the tumor grows to 80-150 mm 3 combine the body weight to start grouping experiments, so that the average tumor volume of each group of mice tends to be consistent.
- the tumor-bearing nude mice have a tumor volume of 80-150 mm 3 , a male body weight of 21-25 g.
- nude mice After inoculated with tumors, nude mice were divided into 3 groups of 10 groups. Conventional tail vein injection, cardiac puncture and blood collection, dipotassium ethylenediaminetetraacetate (EDTA-K2) as anticoagulant. Killed at 1, 24, 48, and 96 hours after administration of compound (I-1) (3.0 mg / kg) at 1, 2, 4, 8, 12 hours, and after HPPH (0.6 mg / kg) administration, respectively Dead nude mice obtained anticoagulated plasma; after tumor, skin and muscle tissue homogenate and methanol precipitation extraction, the sample was injected into a high-performance liquid system for content determination.
- EDTA-K2 dipotassium ethylenediaminetetraacetate
- the drug concentration-time curve of compound (I-1) in different tissues is shown in the figure As shown in 2, the drug concentration at the corresponding time point is shown in Table 3; the drug concentration-time curve of HPPH in different tissues is shown in Figure 3, and the drug concentration of each HPPH tissue is shown in Table 4.
- BLOQ It means that the drug content is extremely low, which is lower than the detection limit of high performance liquid phase.
- BLOQ It means that the drug content is extremely low, which is lower than the detection limit of high performance liquid phase.
- compound (I-1) has a better tissue distribution selectivity after about 1 hour of administration, which can reduce side effects and weaken the Damage to surrounding tissues; it is rarely detected in the skin and the drug is metabolized faster, which indicates a shorter period of avoiding light after clinical treatment.
- the plasma drug concentration of compound (I-1) decreases from 7217 ng / ml at the first hour to 2600 ng / kg at the second hour, and the drug concentration decreases by about 2/3;
- the 1873ng / ml at 5 hours dropped to 633ng / ml at 24h, and the drug concentration also decreased by about 2/3 after 19h.
- the half-life (t 1/2 ) of HPPH is about 19 times that of compound (I-1). It can be seen that, compared with HPPH, Compound (I-1) significantly shortens the half-life of the drug, and can clinically significantly reduce the time for avoiding light after photodynamic therapy.
- Test Example 1 the compound (I-1) exemplified the dose-effect relationship of the tumor suppressive effect on nude mice xenografts between 0.67 mg / kg and 1.5 mg / kg, and proved its effectiveness; At lower doses, the tumor inhibition rate of the tumor strains in this test case was significantly higher than that of the HPPH group, which had an advantage.
- Test Example 2 after the compound (I-1) was administered at a dose of 3.0 mg / kg in the tail vein, its distribution in the skin and muscle tissue was extremely small, and it had good selectivity. It can be calculated from the data in Tables 3 and 4 that the half-life (t 1/2 ) of HPPH is about 19 times that of compound (I-1), and compound (I-1) is metabolized faster than HPPH In the clinic, it can greatly reduce the time for patients to avoid light and stay in hospital after treatment.
- the compound of the present invention has advantages over HPPH: the compound of the present invention has better tumor suppressing effect and better tumor selectivity; the compound of the present invention has a shorter half-life, which is beneficial to shorten the treatment cycle in clinical application.
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Abstract
The present invention provides a bacterial chlorophyll derivative, a preparation method therefor and application thereof in medicine. Particularly, the present invention relates to a bacterial chlorophyll derivative represented by formula (I), a preparation method therefor and a pharmaceutically acceptable salt thereof, and application thereof in photodynamic therapy of tumors. The definition of each substituent in formula (I) is the same as that in the description. (I)
Description
本发明涉及制药领域,具体涉及一种细菌叶绿素衍生物、其制备方法及其应用。The invention relates to the field of pharmaceuticals, in particular to a chlorophyll derivative of bacteria, a preparation method and application thereof.
光动力疗法(photodynamic therap,简称PDT)是指利用可见光敏化位于人体病变组织内部的光敏剂而杀死病变组织的治疗方法。在临床上,通过静脉注射对肿瘤细胞等生物活性组织有亲和作用的光敏剂,光敏剂会选择性地富集于病灶区,然后用一定波长的光辐照病灶区,光敏剂吸收光能后转变成高反应活性的活性物质,在氧的参与下与病变组织生物分子发生化学反应而将其杀死。这是一种正在研究发展中的新型临床治疗技术,其主要应用于临床上对各种肿瘤的治疗。Photodynamic therapy (PDT) refers to a treatment method that uses visible light to sensitize a photosensitizer located inside a human diseased tissue to kill the diseased tissue. In clinical practice, intravenous injection of a photosensitizer that has an affinity for tumor cells and other biologically active tissues. The photosensitizer will selectively enrich in the lesion area, and then irradiate the lesion area with light of a certain wavelength. The photosensitizer absorbs light energy It is then converted into a highly reactive active substance, which, with the participation of oxygen, chemically reacts with the biological molecules of the diseased tissue to kill it. This is a new type of clinical treatment technology under research and development, which is mainly used in clinical treatment of various tumors.
在所有光敏剂中,卟啉类光敏剂一直是热点研究对象。由于叶绿素(a)衍生物和细菌叶绿素衍生物在650-850nm光谱区域有强吸收特点以及治疗后容易降解,被认为是光动力治疗肿瘤的优良光敏剂。Among all photosensitizers, porphyrin photosensitizers have always been a hot research object. Because chlorophyll (a) derivatives and bacterial chlorophyll derivatives have strong absorption characteristics in the 650-850nm spectral region and are easily degraded after treatment, they are considered as excellent photosensitizers for photodynamic therapy of tumors.
在叶绿素(a)衍生物和细菌叶绿素衍生物光敏剂中,
是第一代光敏剂,光动力治疗后的避光周期需4-6周、吸收波长630nm,在组织中穿透深度小于4毫米。目前处于Ⅱ期临床阶段的HPPH是第二代光敏剂,其最大吸收波长为665nm,在组织中穿透深度约为0.5cm,光动力治疗后的避光周期为一周,由于其避光周期较长,在生活中,患者难免会接触光的照射,因此,HPPH治疗后皮肤会产生红斑等损伤。WO2004/002476中报道了一种细菌叶绿素衍生物,吸收波长为700-850nm,由于是脂溶性化合物,代谢时间长、皮肤中含量高、皮肤光毒性大。同样,文献(Bioconjug Chem.2009 February;20(2):274–282)中报道的化合物在给药96小时后,各主要组织或器官中,药物均没有代谢完,说明该类化合物在体内代谢缓慢,预示着该类化合物的避光周期比较长。
In photosensitizers of chlorophyll (a) derivatives and bacterial chlorophyll derivatives, It is the first generation of photosensitizer. The photoperiod-free treatment period needs 4-6 weeks, the absorption wavelength is 630nm, and the penetration depth in the tissue is less than 4mm. HPPH, currently in Phase II clinical stage, is a second-generation photosensitizer with a maximum absorption wavelength of 665 nm and a penetration depth of about 0.5 cm in tissues. The photoperiod-free period after photodynamic therapy is one week. Long, in life, patients will inevitably be exposed to light, so HPHP treatment of the skin will cause erythema and other damage. WO2004 / 002476 reports a bacterial chlorophyll derivative with an absorption wavelength of 700-850 nm. Since it is a fat-soluble compound, it has a long metabolism time, high content in skin, and high skin phototoxicity. Similarly, the compounds reported in the literature (Bioconjug Chem. 2009 February; 20 (2): 274–282) have not been metabolized in the main tissues or organs after 96 hours of administration, indicating that the compounds are metabolized in vivo Slowness indicates that the compound has a longer light-shielding period.
由此可见,现有的叶绿素(a)或细菌叶绿素衍生物类光敏剂都存在代谢缓慢,光动力治疗后的避光周期长的局限性,限制了光动力治疗在肿瘤治疗中的广泛应用。It can be seen that the existing photosensitizers of chlorophyll (a) or bacterial chlorophyll derivatives have the limitations of slow metabolism and long photoperiod avoidance period after photodynamic therapy, which limits the wide application of photodynamic therapy in tumor therapy.
发明内容Summary of the invention
为了解决现有光敏剂代谢缓慢、避光周期长的问题,本发明提供了一种代谢 快、避光周期短的细菌叶绿素衍生物,其可以作为光敏剂进行光动力治疗研究,病人可以实现当天治疗,当天回家,进而减轻病人和家属的经济负担。In order to solve the problems of slow metabolism of the existing photosensitizer and long light-proof period, the present invention provides a bacterial chlorophyll derivative with fast metabolism and short light-proof period, which can be used as a photosensitizer for photodynamic therapy research, and patients can achieve the same day Treatment, go home on the same day, thereby reducing the financial burden of patients and their families.
本发明目的在于提供一种式(I)化合物或其立体异构体、或其可药用的盐:The object of the present invention is to provide a compound of formula (I) or its stereoisomer, or a pharmaceutically acceptable salt thereof:
其中,among them,
R
1为氢或(CH
2)
mCOOH;
R 1 is hydrogen or (CH 2 ) m COOH;
R
2选自氢、羟基、卤素、C
1-C
6烷基或C
1-C
6烷氧基;
R 2 is selected from hydrogen, hydroxyl, halogen, C 1 -C 6 alkyl or C 1 -C 6 alkoxy;
R
3,R
4各自独立地选自C
1-C
6烷基;
R 3 and R 4 are each independently selected from C 1 -C 6 alkyl;
m为1、2、3或4;且m is 1, 2, 3 or 4; and
n为1、2、3或4。n is 1, 2, 3 or 4.
本发明的优选方案,一种式(I)化合物或其立体异构体、或其可药用的盐,其为一种式(II)所示的化合物或其立体异构体、或其可药用的盐:In a preferred embodiment of the present invention, a compound of formula (I) or its stereoisomer, or a pharmaceutically acceptable salt thereof, is a compound of formula (II) or its stereoisomer, or its Medicinal salt:
其中,R
2,R
3,R
4,m和n的定义如式(I)中所述。
Wherein, R 2 , R 3 , R 4 , m and n are defined as in formula (I).
本发明的优选方案,一种式(I)化合物或其立体异构体、或其可药用的盐, 其为一种式(III)所示的化合物或其立体异构体、或其可药用的盐:In a preferred embodiment of the present invention, a compound of formula (I) or its stereoisomer, or a pharmaceutically acceptable salt thereof, is a compound of formula (III) or its stereoisomer, or its Medicinal salt:
其中,R
2,R
3,R
4和n的定义如式(I)中所述。
Wherein, R 2 , R 3 , R 4 and n are as defined in formula (I).
本发明的优选方案,一种式(I)、式(II)或式(III)所述的化合物或其立体异构体,或其可药用的盐,其中,R
3为甲基、乙基、丙基、丁基或己基。
In a preferred embodiment of the present invention, a compound of formula (I), formula (II), or formula (III) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein R 3 is methyl, ethyl Group, propyl, butyl or hexyl.
本发明的优选方案,一种式(I)、式(II)或式(III)所述的化合物或其立体异构体,或其可药用的盐,其中,R
4为甲基、乙基、丙基、丁基或己基。
In a preferred embodiment of the present invention, a compound of formula (I), formula (II) or formula (III) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein R 4 is methyl, ethyl Group, propyl, butyl or hexyl.
本发明的优选方案,一种式(I)、式(II)或式(III)所述的化合物或其立体异构体、或其可药用的盐,其中,R
2选自氢、卤素、C
1-C
6烷基或C
1-C
6烷氧基。
In a preferred embodiment of the present invention, a compound of formula (I), formula (II) or formula (III) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein R 2 is selected from hydrogen, halogen , C 1 -C 6 alkyl or C 1 -C 6 alkoxy.
本发明的优选方案,一种式(I)、式(II)或式(III)所述的化合物或其立体异构体、或其可药用的盐,其中,R
2选自氢、氟、甲基或甲氧基。
In a preferred embodiment of the present invention, a compound of formula (I), formula (II), or formula (III) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein R 2 is selected from hydrogen, fluorine , Methyl or methoxy.
本发明的代表化合物包括,但不限于:Representative compounds of the present invention include, but are not limited to:
或其立体异构体,或其可药用的盐。Or its stereoisomers, or their pharmaceutically acceptable salts.
本发明提供一种式(II)化合物或其立体异构体、或其可药用的盐的制备方法,该方法包括:The present invention provides a method for preparing a compound of formula (II) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. The method includes:
式(IIA)化合物在碱性条件下进行水解反应,然后加酸进行酸解得到式(II)化合物;The compound of formula (IIA) undergoes a hydrolysis reaction under basic conditions, and then acid is added for acid hydrolysis to obtain the compound of formula (II);
其中:R
2,R
3,R
4,m和n的定义如式(I)中所述。
Wherein: R 2 , R 3 , R 4 , m and n are as defined in formula (I).
进一步地,本发明提供一种式(II)化合物或其立体异构体、或其可药用的盐的制备方法,该方法包括:所述碱性条件所用的碱选自碱金属碳酸盐或碳酸铵,优选碱金属碳酸盐,更优选碳酸钠、碳酸钾或碳酸锂;Further, the present invention provides a method for preparing a compound of formula (II) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, the method comprising: the base used in the alkaline condition is selected from alkali metal carbonates Or ammonium carbonate, preferably alkali metal carbonate, more preferably sodium carbonate, potassium carbonate or lithium carbonate;
所述的酸选自无机酸或有机酸,其中,无机酸选自质量分数为1-30%的盐酸或质量分数为1-50%的硫酸,优选质量分数为1-10%的盐酸或质量分数为1-10%的硫酸,有机酸选自C
1-C
4的有机酸,优选甲酸、乙酸或草酸。
The acid is selected from inorganic acid or organic acid, wherein the inorganic acid is selected from hydrochloric acid with a mass fraction of 1-30% or sulfuric acid with a mass fraction of 1-50%, preferably hydrochloric acid with a mass fraction of 1-10% or a mass Sulfuric acid with a fraction of 1-10%, the organic acid is selected from C 1 -C 4 organic acids, preferably formic acid, acetic acid or oxalic acid.
本发明提供一种式(III)化合物或其立体异构体、或其可药用的盐的制备方法,该方法包括:The present invention provides a method for preparing a compound of formula (III) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. The method includes:
式(IIIA)化合物在碱性条件下水解得到式(III)化合物,The compound of formula (IIIA) is hydrolyzed under basic conditions to obtain the compound of formula (III),
其中:R
2,R
3,R
4和n的定义如式(I)中所述。
Wherein: R 2 , R 3 , R 4 and n are as defined in formula (I).
进一步地,本发明提供一种式(III)化合物或其立体异构体、或其可药用的盐的制备方法,该方法包括:所述碱性条件所用的碱选自碱金属碳酸盐、碳酸铵或有机胺,优选碱金属碳酸盐或C
1-C
6的伯胺,更优选碳酸钠、碳酸钾、碳酸锂、正丙胺或丁胺。
Further, the present invention provides a method for preparing a compound of formula (III) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, the method comprising: the base used in the alkaline condition is selected from alkali metal carbonates , Ammonium carbonate or organic amine, preferably alkali metal carbonate or C 1 -C 6 primary amine, more preferably sodium carbonate, potassium carbonate, lithium carbonate, n-propylamine or butylamine.
本发明提供一种式(IIA)所示的化合物或其立体异构体、或其可药用的盐:The present invention provides a compound represented by formula (IIA) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
其中:R
2,R
3,R
4,m和n的定义如式(I)中所述。
Wherein: R 2 , R 3 , R 4 , m and n are as defined in formula (I).
本发明提供一种式(IIIA)所示的化合物或其立体异构体、或其可药用的盐:The present invention provides a compound represented by formula (IIIA) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
其中:R
2,R
3,R
4和n的定义如式(I)中所述。
Wherein: R 2 , R 3 , R 4 and n are as defined in formula (I).
本发明提供一种药物组合物,所述的药物组合物含有有效剂量的式(I)化合物或其立体异构体、或其可药用的盐,及其可药用的载体、赋形剂或它们的组合。The present invention provides a pharmaceutical composition containing an effective dose of a compound of formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and excipient Or a combination of them.
本发明提供一种式(I)化合物或其立体异构体、或其可药用的盐,或其药物组合物在制备用于光动力治疗癌症的药物中的应用,其中所述癌症优选为肺癌、前列腺癌、食管癌、黑色素瘤、胆管癌、头颈癌、膀胱癌、胃癌、鼻咽癌、结肠癌中的一种或多种。The present invention provides a compound of formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for use in the preparation of a medicament for photodynamic therapy of cancer, wherein the cancer is preferably One or more of lung cancer, prostate cancer, esophageal cancer, melanoma, bile duct cancer, head and neck cancer, bladder cancer, stomach cancer, nasopharyngeal cancer, colon cancer.
本发明的化合物最大吸收波长都在780nm左右,对组织的穿透深度为0.7-1cm,对组织较深的肿瘤细胞可进行有效的杀灭;由于分子结构中引入了亲水基团,羧基或酚羟基,因而都具有较好的水溶性,因而代谢时间短,避光周期短;同时本发明的化合物都具有很好的化学稳定性。The maximum absorption wavelength of the compound of the present invention is around 780nm, and the penetration depth of the tissue is 0.7-1cm, which can effectively kill tumor cells with deep tissues; due to the introduction of hydrophilic groups, carboxyl groups or The phenolic hydroxyl groups have good water solubility, so the metabolic time is short and the dark period is short; at the same time, the compounds of the present invention have good chemical stability.
发明的详细说明Detailed description of the invention
除非有相反陈述,否则本发明在说明书和权利要求书中所使用的部分术语定 义如下:Unless stated to the contrary, some terms used in the specification and claims of the present invention are defined as follows:
“烷基”当作一基团或一基团的一部分时是指包括C
1-C
6直链或者带有支链的饱和脂肪烃基团。例如,C
1-C
6是指包含1至6个碳原子,例如,可包含1个碳原子、2个碳原子、3个碳原子、4个碳原子、5个碳原子、6个碳原子。烷基基团的实施例包括但不限于甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、仲丁基、正戊基、1,1-二甲基丙基、1,2-二甲基丙基、2,2-二甲基丙基、1-乙基丙基、2-甲基丁基、3-甲基丁基、正己基、1-乙基-2-甲基丙基、1,1,2-三甲基丙基、1,1-二甲基丁基、1,2-二甲基丁基、2,2-二甲基丁基、1,3-二甲基丁基、2-乙基丁基、2-甲基戊基、3-甲基戊基、4-甲基戊基、2,3-二甲基丁基等。
"Alkyl" when taken as a group or part of a group refers to a C 1 -C 6 linear or branched saturated aliphatic hydrocarbon group. For example, C 1 -C 6 refers to containing 1 to 6 carbon atoms, for example, may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms . Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-di Methylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1 -Ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethyl Butyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl Wait.
“烷氧基”是指(烷基-O-)的基团。其中,烷基见本文有关定义。C
1-C
6的烷氧基实例包括,但不限于:甲氧基、乙氧基、正丙氧基、异丙氧基、正丁氧基、异丁氧基、叔丁氧基、正戊氧基、1,1-二甲基丙氧基、1,2-二甲基丙氧基、2,2-二甲基丙氧基、1-乙基丙氧基、2-甲基丁氧基、3-甲基丁氧基、正己氧基、1-乙基-2-甲基丙氧基、1,1,2-三甲基丙氧基、1,1-二甲基丁氧基、1,2-二甲基丁氧基、2,2-二甲基丁氧基、1,3-二甲基丁氧基、2-乙基丁氧基、2-甲基戊氧基、3-甲基戊氧基、4-甲基戊氧基、2,3-二甲基丁氧基。
"Alkoxy" refers to the group (alkyl-O-). Among them, alkyl refers to the relevant definitions herein. Examples of C 1 -C 6 alkoxy groups include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, t-butoxy, n- Pentoxy, 1,1-dimethylpropoxy, 1,2-dimethylpropoxy, 2,2-dimethylpropoxy, 1-ethylpropoxy, 2-methylbutoxy Oxygen, 3-methylbutoxy, n-hexyloxy, 1-ethyl-2-methylpropoxy, 1,1,2-trimethylpropoxy, 1,1-dimethylbutoxy Group, 1,2-dimethylbutoxy, 2,2-dimethylbutoxy, 1,3-dimethylbutoxy, 2-ethylbutoxy, 2-methylpentoxy , 3-methylpentoxy, 4-methylpentoxy, 2,3-dimethylbutoxy.
“羟基”指-OH基团。"Hydroxy" refers to the -OH group.
“卤素”是指氟、氯、溴和碘。"Halogen" means fluorine, chlorine, bromine and iodine.
“氨基”指-NH
2。
"Amino" refers to -NH 2 .
“氰基”指-CN。"Cyano" means -CN.
“硝基”指-NO
2。
"Nitro" refers to -NO 2 .
“苄基”指-CH
2-苯基。
"Benzyl" refers to -CH 2 - phenyl.
“羧基”指-C(O)OH。"Carboxy" refers to -C (O) OH.
“羧酸酯基”指-C(O)O(烷基),其中烷基的定义如上所述。"Carboxylate group" refers to -C (O) O (alkyl), where alkyl is as defined above.
“Boc”指叔丁氧基羰基。"Boc" refers to tert-butoxycarbonyl.
“DMSO”指二甲基亚砜。"DMSO" refers to dimethyl sulfoxide.
“取代的”指基团中的一个或多个氢原子,优选为最多5个,更优选为1-3个氢原子彼此独立地被相应数目的取代基取代。不言而喻,取代基仅处在它们的可能的化学位置,本领域技术人员能够在不付出过多努力的情况下确定(通过实验 或理论)可能或不可能的取代。例如,具有游离氢的氨基或羟基与具有不饱和(如烯属)键的碳原子结合时可能是不稳定的。"Substituted" refers to one or more hydrogen atoms in the group, preferably up to 5, more preferably 1-3 hydrogen atoms are independently substituted with a corresponding number of substituents for each other. It goes without saying that the substituents are only at their possible chemical positions, and those skilled in the art can determine (through experiment or theory) possible or impossible substitutions without undue effort. For example, an amino group or hydroxyl group having free hydrogen may be unstable when combined with a carbon atom having an unsaturated (eg, olefinic) bond.
“可药用的盐”是指上述化合物能保持原有生物活性并且适合于医药用途的某些盐类。式(I)所表示的化合物的可药用的盐可以为金属盐、与合适的酸形成的盐。"Pharmaceutically acceptable salts" refers to certain salts of the above compounds that can retain their original biological activity and are suitable for medical uses. The pharmaceutically acceptable salt of the compound represented by formula (I) may be a metal salt or a salt with a suitable acid.
“立体异构体”是指包括但绝不限于非对映体、对映异构体、阻转异构体及其它们的混合物,如外消旋混合物,组成了本发明的一部分。非对映异构体可以以其物理化学差异为基础,通过层析、结晶、蒸馏或升华等方法被分离为个别非对映异构体。对映异构体可以通过分离,使手性异构混合物转化为非对映异构混合物,其方式是与适当光学活性化合物(例如手性辅助剂,譬如手性醇或Mosher氏酰氯)的反应,分离非对映异构体,且使个别非对映异构体转化为相应的纯对映异构体。本发明的中间体与化合物也可以以不同互变异构形式存在,且所有此种形式被包含在本发明的范围内。很多有机化合物都以光学活性形式存在,即它们有能力旋转平面偏振光的平面。在描述光学活性化合物时,前缀D、L或R、S用来表示分子手性中心的绝对构型。前缀d、l或(+)、(-)用来命名化合物平面偏振光旋转的符号,(-)或l是指化合物是左旋的,前缀(+)或d是指化合物是右旋的。这些立体异构体的原子或原子团互相连接次序相同,但是它们的立体结构不一样。特定的立体异构体可以是对映体,异构体的混合物通常称为对映异构体混合物。50:50的对映体混合物被称为外消旋混合物或外消旋体,这可能导致化学反应过程中没有立体选择性或立体定向性。术语“外消旋混合物”和“外消旋体”是指等摩尔的两个对映异构体的混合物,缺乏光学活性。"Stereoisomer" means including but not limited to diastereomers, enantiomers, atropisomers and mixtures thereof, such as racemic mixtures, forming part of the present invention. Diastereomers can be separated into individual diastereomers by chromatography, crystallization, distillation or sublimation based on their physical and chemical differences. Enantiomers can be converted into diastereoisomeric mixtures by separation, by reacting with appropriate optically active compounds (such as chiral auxiliary agents such as chiral alcohols or Mosher's acid chloride) , Separation of diastereomers, and the conversion of individual diastereomers to the corresponding pure enantiomers. The intermediates and compounds of the present invention may also exist in different tautomeric forms, and all such forms are included within the scope of the present invention. Many organic compounds exist in optically active forms, that is, they have the ability to rotate the plane of plane polarized light. When describing optically active compounds, the prefixes D, L or R, S are used to denote the absolute configuration of the molecular chiral center. The prefixes d, l, or (+), (-) are used to name the symbol for rotation of plane polarized light of the compound, (-) or l means the compound is left-handed, and the prefix (+) or d means the compound is right-handed. The atoms or groups of these stereoisomers are connected in the same order, but their stereostructures are different. The specific stereoisomer may be an enantiomer, and a mixture of isomers is generally called an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or a racemate, which may result in no stereoselectivity or stereospecificity during the chemical reaction. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomers, lacking optical activity.
“碱金属碳酸盐”是指碳酸锂、碳酸钠、碳酸钾、碳酸铷、碳酸铯和碳酸钫。"Alkali metal carbonate" means lithium carbonate, sodium carbonate, potassium carbonate, rubidium carbonate, cesium carbonate, and francium carbonate.
“药物组合物”表示包含本申请的化合物或其生理学上可接受的盐或前体药物与其他化学组分的混合物,以及其他组分例如生理学上可接受的载体和赋形剂。药物组合物的目的是促进对生物体的给药,利于活性成分的吸收进而发挥生物活性。"Pharmaceutical composition" means a mixture comprising a compound of the present application or a physiologically acceptable salt or prodrug thereof with other chemical components, and other components such as physiologically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to promote the administration to the organism, facilitate the absorption of the active ingredient and thus exert the biological activity.
图1为不同剂量的化合物(I-1)对NCI-H460移植瘤体积的影响;Figure 1 shows the effect of different doses of compound (I-1) on the volume of NCI-H460 transplanted tumor;
图2为化合物(I-1)(3.0mg/kg)在不同组织的血药浓度-时间曲线图;Fig. 2 is a blood concentration-time curve of compound (I-1) (3.0 mg / kg) in different tissues;
图3为HPPH(0.6mg/kg)在不同组织的血药浓度-时间曲线图。Fig. 3 is the blood concentration-time curve of HPPH (0.6mg / kg) in different tissues.
下文将参照具体实施例详细说明本发明。应该理解,本领域技术人员基于此处公开的内容,可以对本发明进行各种不偏离本发明精神和范围内的各种修改和改进,它们应当都落在本发明申请的权利要求定义的专利保护范围内。此外,应当理解,此处提供的实施例仅用于说明本发明的目的,而不应解释为对本发明的限制。Hereinafter, the present invention will be described in detail with reference to specific embodiments. It should be understood that based on the content disclosed herein, those skilled in the art can make various modifications and improvements to the present invention without departing from the spirit and scope of the present invention, and they should all fall within the patent protection defined by the claims of the present application Within range. In addition, it should be understood that the embodiments provided herein are for illustrative purposes only and should not be construed as limiting the invention.
以下结合实施例用于进一步描述本发明,但这些实施例并非意在限制本发明的范围。The following examples are used to further describe the present invention, but these examples are not intended to limit the scope of the present invention.
实施例Examples
实施例给出了式(I)所表示的代表性化合物的制备及相关结构鉴定数据。需要说明的是,下述实施例是用于说明本发明而不是对本发明的限制。The examples give the preparation of representative compounds represented by formula (I) and related structural identification data. It should be noted that the following examples are for illustrating the present invention rather than limiting the present invention.
NMR图谱是用Bruker仪器(400MHz)测定而得,化学位移用ppm表示。使用四甲基硅烷内标准物(0.00ppm)。
1H NMR的表示方法:s=单峰,d=双重峰,t=三重峰,m=多重峰,br=变宽的,dd=双重峰的双重峰,dt=三重峰的双重峰。若提供偶合常数时,其单位为Hz。
The NMR spectrum is measured with a Bruker instrument (400 MHz), and the chemical shift is expressed in ppm. Use tetramethylsilane internal standard (0.00 ppm). Representation method of 1 H NMR: s = single peak, d = doublet, t = triplet, m = multiplet, br = broadened, dd = doublet of doublet, dt = doublet of triplet. If the coupling constant is provided, the unit is Hz.
质谱是用LC/MS仪测定得到,离子化方式可为ESI或APCI。Mass spectrometry is measured by LC / MS instrument, and the ionization method can be ESI or APCI.
紫外数据是用SHIMADZU的UV-2600紫外光谱仪测定得到。Ultraviolet data was measured with SHIMADZU's UV-2600 ultraviolet spectrometer.
薄层层析硅胶板使用烟台黄海HSGF254或青岛GF254硅胶板,薄层色谱法(TLC)使用的硅胶板采用的规格是0.15mm-0.2mm,薄层层析分离纯化产品采用的规格是0.4mm-0.5mm。The thin-layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The specification of the thin-layer chromatography (TLC) silica gel plate is 0.15mm-0.2mm, and the specification of the thin-layer chromatography separation and purification product is 0.4mm. -0.5mm.
柱层析一般使用烟台黄海硅胶200-300目硅胶为载体。Column chromatography generally uses Yantai Yellow Sea silica gel 200-300 mesh silica gel as a carrier.
在下列实施例中,除非另有指明,所有温度为摄氏温度,除非另有指明,各种起始原料和试剂来自市售或者是根据已知的方法合成,市售原料和试剂均不经进一步纯化直接使用,除非另有指明,市售厂家包括但不限于Aldrich Chemical Company,ABCR GmbH&Co.KG,Acros Organics和国药集团化学试剂有限公司等处购买。In the following examples, unless otherwise specified, all temperatures are in degrees Celsius. Unless otherwise specified, various starting materials and reagents are commercially available or synthesized according to known methods. Purification is used directly, unless otherwise specified, commercially available manufacturers include but are not limited to Aldrich Chemical Company, ABCR GmbH & Co. KG, Acros Organics and Sinopharm Chemical Reagent Co., Ltd. and other places to buy.
CDCl
3:氘代氯仿。
CDCl 3 : deuterated chloroform.
DMSO:氘代二甲基亚砜。DMSO: Deuterated dimethyl sulfoxide.
氮气反应是指反应瓶连接一个约1L容积的氮气气球。Nitrogen reaction means that the reaction bottle is connected to a nitrogen balloon with a volume of about 1L.
实施例中无特殊说明,反应中的溶液是指水溶液。There is no special description in the examples, and the solution in the reaction refers to an aqueous solution.
对化合物进行纯化,采用硅胶柱层析洗脱剂体系和薄层色谱法,其中洗脱剂体系选自:石油醚和乙酸乙酯体系;二氯甲烷和丙酮体系;二氯甲烷和甲醇,洗脱剂比值为体积比,其中溶剂的体积比根据化合物的极性不同而不同,也可以加入少量的酸性或碱性试剂进行调节,如醋酸或三乙胺等。To purify the compound, use silica gel column chromatography eluent system and thin layer chromatography, where the eluent system is selected from: petroleum ether and ethyl acetate system; dichloromethane and acetone system; dichloromethane and methanol, wash The ratio of the removal agent is the volume ratio, where the volume ratio of the solvent is different according to the polarity of the compound, and a small amount of acidic or basic reagents can also be added for adjustment, such as acetic acid or triethylamine.
本发明中的化合物(IIB-1)按照文献(J.Med.Chem.,2016,59,9774-9787)方法合成,化合物(IIB-2),化合物(IIB-3)和化合物(IIB-4)均参照化合物(IIB-1)的制备方法合成,结构表征数据如下:The compound (IIB-1) in the present invention is synthesized according to the method of the literature (J. Med. Chem., 2016, 59, 9774-9787), compound (IIB-2), compound (IIB-3) and compound (IIB-4 ) Are synthesized according to the preparation method of compound (IIB-1), and the structural characterization data are as follows:
实施例1化合物(I-1)的合成Example 1 Synthesis of Compound (I-1)
第一步:2-(4-羟甲基)苯氧乙酸甲酯(IV-1)的合成Step 1: Synthesis of methyl 2- (4-hydroxymethyl) phenoxyacetate (IV-1)
在一个500mL的三口圆底烧瓶中,加入对羟基苯甲醛(10g,82mmol),再加入120mL乙腈,搅拌溶解,再加入溴乙酸甲酯(10mL,105mmol)和碳酸钾(15g,9.2mmol),常温搅拌反应,TLC监控反应,反应结束后,加入150mL乙酸乙酯稀释反应液,过滤,滤液减压浓缩,得到黄色油状物,直接进行下一步反应;In a 500mL three-necked round bottom flask, add p-hydroxybenzaldehyde (10g, 82mmol), then add 120mL acetonitrile, stir to dissolve, then add methyl bromoacetate (10mL, 105mmol) and potassium carbonate (15g, 9.2mmol), The reaction was stirred at room temperature, and the reaction was monitored by TLC. After the reaction was completed, 150 mL of ethyl acetate was added to dilute the reaction solution, filtered, and the filtrate was concentrated under reduced pressure to obtain a yellow oil, which was directly subjected to the next reaction;
将得到的黄色油状物转入到一个500mL的三口圆底烧瓶中,加入100mL二氯甲烷和50mL甲醇,搅拌,冰浴冷却下,缓慢加入硼氢化钠(3.0g,79mmol),加完后,常温搅拌反应,TLC监控反应,反应结束后,用10%的稀盐酸水溶液淬灭反应,接着加入150mL二氯甲烷和100mL水,分出有机相,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积比为:石油醚:乙酸乙酯=4:1,得到8.5g 2-(4-羟甲基)苯氧乙酸甲酯(IV-1),化合物(IV-1)为白色固体;总收率:52.86%。Transfer the obtained yellow oil to a 500mL three-necked round bottom flask, add 100mL of dichloromethane and 50mL of methanol, stir, and slowly add sodium borohydride (3.0g, 79mmol) under ice bath cooling. After the addition, Stir the reaction at room temperature and monitor the reaction by TLC. After the reaction is completed, quench the reaction with 10% dilute hydrochloric acid aqueous solution, then add 150mL of dichloromethane and 100mL of water, separate the organic phase, concentrate to dryness under reduced pressure, and use silica gel column chromatography Purification, eluent and volume ratio: petroleum ether: ethyl acetate = 4: 1, to obtain 8.5g methyl 2- (4-hydroxymethyl) phenoxyacetate (IV-1), compound (IV-1) It is a white solid; total yield: 52.86%.
1H NMR(400MHz,CDCl
3)δ:2.18(s,1H),3.77(s,3H),4.56(s,2H),4.6(s,2H),6.85(d,J=8.36Hz,2H),7.25(d,J=8.28Hz,2H);
1 H NMR (400 MHz, CDCl 3 ) δ: 2.18 (s, 1H), 3.77 (s, 3H), 4.56 (s, 2H), 4.6 (s, 2H), 6.85 (d, J = 8.36 Hz, 2H) , 7.25 (d, J = 8.28 Hz, 2H);
13C NMR(100MHz,CDCl
3)δ:52.30,64.70,65.31,114.63,128.63,134.37,157.25,169.45。
13 C NMR (100 MHz, CDCl 3 ) δ: 52.30, 64.70, 65.31, 114.63, 128.63, 134.37, 157.25, 169.45.
第二步:化合物(IIA-1)的合成Step 2: Synthesis of compound (IIA-1)
在一个500mL三口圆底烧瓶中,称入化合物(IIB-1)(1.4g,2.15mmol),加入100mL二氯甲烷和10mL甲醇,搅拌溶解,冰浴冷却下,缓慢加入硼氢化钠(0.815g,21.5mmol),加完后,常温搅拌反应,TLC监控反应,反应结束后,用10%的稀醋酸水溶液淬灭反应,接着加入150mL二氯甲烷和100mL水,分出有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,得到黑色固体,直接进行下一步反应;In a 500mL three-necked round bottom flask, weigh compound (IIB-1) (1.4g, 2.15mmol), add 100mL of dichloromethane and 10mL of methanol, stir to dissolve, and slowly add sodium borohydride (0.815g) under ice bath cooling , 21.5mmol), after the addition, the reaction was stirred at room temperature, the reaction was monitored by TLC, after the reaction was completed, the reaction was quenched with 10% dilute acetic acid aqueous solution, followed by the addition of 150mL of methylene chloride and 100mL of water, the organic phase was separated, anhydrous sulfuric acid Sodium was dried, filtered, and the filtrate was concentrated under reduced pressure to obtain a black solid, which was directly subjected to the next reaction;
将黑色固体全部转入一个250mL单口圆底烧瓶中,加入150mL二氯甲烷,缓慢搅拌,通入溴化氢气体至饱和,继续搅拌,TLC监控反应,反应结束后,减压浓缩至干,直接进行下一步反应;Transfer all the black solids into a 250mL single-necked round-bottom flask, add 150mL of dichloromethane, slowly stir, pass hydrogen bromide gas to saturation, continue stirring, TLC monitor the reaction, after the reaction is completed, concentrate to dryness under reduced pressure, directly Proceed to the next reaction;
将上述所得的浓缩物转入一个500mL三口圆底烧瓶中,加入化合物2-(4-羟甲基)苯氧乙酸甲酯(IV-1)(1.9g,10mmol),加入100mL二氯甲烷,搅拌,冰浴冷却下,缓慢滴加50mL三乙胺的二氯甲烷溶液(含三乙胺2mL,14.35mmol)加完后,常温反应,TLC监控反应,反应结束后,加入100mL二氯甲烷和100mL 去离子水,分出有机相,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积比为:二氯甲烷:丙酮=100:2,得到1.5g化合物(IIA-1),MS(ESI):[M+H
+]:832.4250。
The concentrate obtained above was transferred to a 500 mL three-necked round bottom flask, compound 2- (4-hydroxymethyl) phenoxyacetic acid methyl ester (IV-1) (1.9 g, 10 mmol) was added, and 100 mL of dichloromethane was added, After stirring and cooling in an ice bath, slowly add 50 mL of triethylamine in dichloromethane (containing triethylamine 2 mL, 14.35 mmol). After the addition, the reaction was performed at room temperature, and the reaction was monitored by TLC. After the reaction was completed, 100 mL of dichloromethane and 100mL of deionized water, the organic phase was separated, concentrated to dryness under reduced pressure, purified by silica gel column chromatography, eluent and volume ratio: dichloromethane: acetone = 100: 2, to obtain 1.5g compound (IIA-1 ), MS (ESI): [M + H + ]: 832.4250.
第三步:化合物(I-1)的合成Step 3: Synthesis of compound (I-1)
将化合物(IIA-1)加入到一个500mL三口圆底烧瓶中,加入100mL乙腈和40mL去离子水,搅拌,再加入碳酸钾(3.8g,27.5mmol),氮气保护下,40℃搅拌反应。TLC监控反应,反应结束后,加入10%的乙酸水溶液,调节pH值至2-3。将反应液转入一个500mL的分液漏斗中,加入200mL乙酸乙酯和100mL去离子水,分出有机相,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积比为:二氯甲烷:甲醇=100:10,得到0.8g化合物(I-1),总收率45.1%。The compound (IIA-1) was added to a 500 mL three-necked round bottom flask, 100 mL of acetonitrile and 40 mL of deionized water were added, stirred, and then potassium carbonate (3.8 g, 27.5 mmol) was added, and the reaction was stirred at 40 ° C under nitrogen protection. The reaction was monitored by TLC. After the reaction was completed, a 10% aqueous acetic acid solution was added to adjust the pH to 2-3. The reaction solution was transferred to a 500mL separatory funnel, 200mL ethyl acetate and 100mL deionized water were added, the organic phase was separated, concentrated to dryness under reduced pressure, purified by silica gel column chromatography, eluent and volume ratio is : Dichloromethane: methanol = 100: 10, 0.8 g of compound (I-1) was obtained, and the total yield was 45.1%.
化合物(I-1)的表征数据如下:The characterization data of compound (I-1) is as follows:
UV Wavelength(Absorbance):781.5(0.244),537.0(0.214),416.0(0.241),365.5(0.589),344.0(0.301),291.5(0.10),225(0.156),201.0(0.256),194.5(0.123);UV Wavelength (Absorbance): 781.5 (0.244), 537.0 (0.214), 416.0 (0.241), 365.5 (0.589), 344.0 (0.301), 291.5 (0.10), 225 (0.156), 201.0 (0.256), 194.5 (0.123) ;
MS(ESI):[M+H
+]:818.4092,[M+Na
+]:840.3926;
MS (ESI): [M + H + ]: 818.4092, [M + Na + ]: 840.3926;
1H NMR(400M Hz,CDCl
3)δ-0.28(s,1H),0.06(s,1H),0.90-0.97(m,6H),1.36-1.46(m,2H),1.58-1.66(m,6H),1.70-1.77(m,3H),1.83-1.93(m,4H),2.17-2.32(m,3H),2.47-2.55(m,1H),3.22(s,3H),3.38(s,3H),3.42(s,3H),3.84-3.86(m,1H),4.02-4.13(m,3H),4.24-4.30(m,1H),4.43-4.54(m,2H),4.64(s,2H),5.06(d,J=7.96Hz,1H),5.81-5.86(m,1H),6.84-6.86(m,2H),7.22(d,J=8.08Hz,2H),8.58-8.61(m,2H),8.78(s,1H),8.86(s,1H),13.01(s,1H);
1 H NMR (400M Hz, CDCl 3 ) δ-0.28 (s, 1H), 0.06 (s, 1H), 0.90-0.97 (m, 6H), 1.36-1.46 (m, 2H), 1.58-1.66 (m, 6H), 1.70-1.77 (m, 3H), 1.83-1.93 (m, 4H), 2.17-2.32 (m, 3H), 2.47-2.55 (m, 1H), 3.22 (s, 3H), 3.38 (s, 3H), 3.42 (s, 3H), 3.84-3.86 (m, 1H), 4.02-4.13 (m, 3H), 4.24-4.30 (m, 1H), 4.43-4.54 (m, 2H), 4.64 (s, 2H), 5.06 (d, J = 7.96 Hz, 1H), 5.81-5.86 (m, 1H), 6.84-6.86 (m, 2H), 7.22 (d, J = 8.08 Hz, 2H), 8.58-8.61 (m , 2H), 8.78 (s, 1H), 8.86 (s, 1H), 13.01 (s, 1H);
13C NMR(100MHz,CDCl
3)δ10.77,11.01,11.08,14.27,20.48,22.43,22.81,23.68,24.13,24.36,29.84,30.89,31.33,32.07,47.79,48.76,51.55,53.55,54.58,64.95,70.24,70.49,71.40,95.73,99.28,101.48,113.13,114.62,128.19,129.79,131.03,131.24,132.54,134.06,138.03,138.41,141.58,157.69,161.25,162.84,167.02,170.62,171.37,171.60,172.68,173.43,175.45。
13 C NMR (100 MHz, CDCl 3 ) δ 10.77, 11.01, 11.08, 14.27, 20.48, 22.43, 22.81,23.68, 24.13, 24.36, 29.84, 30.89, 31.33, 32.07, 47.79, 48.76, 51.55, 53.55, 54.58, 64.95 , 70.24,70.49,71.40,95.73,99.28,101.48,113.13,114.62,128.19,129.79,131.03,131.24,132.54,134.06,138.03,138.41,141.58,157.69,161.25,162.84,167.02,170.62,171.37,171.60,172. , 173.43,175.45.
实施例2化合物(I-2)的合成:Example 2 Synthesis of Compound (I-2):
第一步:5-(3-甲基-4-羟甲基)苯氧戊酸甲酯(IV-2)的合成Step 1: Synthesis of methyl 5- (3-methyl-4-hydroxymethyl) phenoxyvalerate (IV-2)
在一个500mL的三口圆底烧瓶中,加入3-甲基-4-羟基苯甲醛(5.0g,36.7mmol),再加入80mL乙腈,搅拌溶解,再加入5-溴戊酸甲酯(8.0克,41.0mmol)和碳酸钾(7.0g,50.6mmol),60℃下,搅拌反应,TLC监控反应,反应结束后,加入150mL乙酸乙酯稀释反应液,过滤,滤液减压浓缩,得到黄色油状物;In a 500mL three-necked round bottom flask, add 3-methyl-4-hydroxybenzaldehyde (5.0g, 36.7mmol), then add 80mL of acetonitrile, stir to dissolve, and then add 5-bromovaleric acid methyl ester (8.0g, 41.0mmol) and potassium carbonate (7.0g, 50.6mmol), the reaction was stirred at 60 ° C, the reaction was monitored by TLC, after the reaction was completed, 150mL of ethyl acetate was added to dilute the reaction solution, filtered, and the filtrate was concentrated under reduced pressure to obtain a yellow oil;
将得到的黄色油状物转入一个500mL的三口圆底烧瓶中,加入100mL二氯甲烷和50mL甲醇,搅拌,冰浴冷却下,缓慢加入硼氢化钠(0.75g,2.04mmol),加完后,常温搅拌反应,TLC监控反应,反应结束后,用10%的稀盐酸水溶液淬灭反应,接着加入150mL二氯甲烷和100mL水,分出有机相,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积比为:石油醚:乙酸乙酯=4:1,得到7.0g5-(3-甲基-4-羟甲基)苯氧戊酸甲酯(IV-2),浅黄色液体,总收率76.9%。Transfer the resulting yellow oil to a 500mL three-necked round bottom flask, add 100mL of dichloromethane and 50mL of methanol, stir, and slowly add sodium borohydride (0.75g, 2.04mmol) under ice bath cooling. After the addition, Stir the reaction at room temperature and monitor the reaction by TLC. After the reaction is completed, quench the reaction with 10% dilute hydrochloric acid aqueous solution, then add 150mL of dichloromethane and 100mL of water, separate the organic phase, concentrate to dryness under reduced pressure, and use silica gel column chromatography Purification, eluent and volume ratio: petroleum ether: ethyl acetate = 4: 1, to obtain 7.0g 5- (3-methyl-4-hydroxymethyl) phenoxyvaleric acid methyl ester (IV-2), shallow Yellow liquid, total yield 76.9%.
1H NMR(400MHz,CDCl
3)δ1.73(s,1H),1.86(s,4H),2.23(s,3H),2.43(s,2H), 3.69(s,3H),3.99(s,2H),4.59(s,2H),6.78(d,J=8.0Hz,1H),7.13-7.16(m,2H);
1 H NMR (400 MHz, CDCl 3 ) δ 1.73 (s, 1H), 1.86 (s, 4H), 2.23 (s, 3H), 2.43 (s, 2H), 3.69 (s, 3H), 3.99 (s, 2H), 4.59 (s, 2H), 6.78 (d, J = 8.0Hz, 1H), 7.13-7.16 (m, 2H);
13C NMR(100MHz,CDCl
3)δ16.22,21.73,28.76,33.69,51.55,65.10,67.42,110.77,125.77,127.01,129.94,132.70,156.68,173.99.
13 C NMR (100 MHz, CDCl 3 ) δ 16.22, 21.73, 28.76, 33.69, 51.55, 65.10, 67.42, 110.77, 125.77, 127.01, 129.94, 132.70, 156.68, 173.99.
第二步:化合物(IIA-2)的合成Step 2: Synthesis of compound (IIA-2)
在一个500mL三口圆底烧瓶中,称入化合物(IIB-1)(1.9g,2.91mmol),加入100mL二氯甲烷和20mL甲醇,搅拌溶解,冰浴冷却下,缓慢加入硼氢化钠(110mg,2.90mmol),加完后,常温搅拌反应,TLC监控反应,反应结束后,用10%的稀醋酸水溶液淬灭反应,接着加入150mL二氯甲烷和100mL水,分出有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,得到黑色固体,直接进行下一步反应;In a 500 mL three-necked round bottom flask, weigh compound (IIB-1) (1.9 g, 2.91 mmol), add 100 mL of dichloromethane and 20 mL of methanol, stir to dissolve, and slowly add sodium borohydride (110 mg, 2.90mmol), after the completion of the reaction, the reaction was stirred at room temperature, the reaction was monitored by TLC, after the reaction was completed, the reaction was quenched with 10% dilute acetic acid aqueous solution, followed by the addition of 150mL of methylene chloride and 100mL of water, the organic phase was separated, anhydrous sodium sulfate Dry, filter, and concentrate the filtrate under reduced pressure to obtain a black solid, and proceed directly to the next reaction;
将上述所得的黑色固体转入一个250mL单口圆底烧瓶中,加入150mL二氯甲烷,缓慢搅拌,通入溴化氢气体至饱和,继续搅拌,TLC监控反应,反应结束后,减压浓缩至干,直接进行下一步反应;Transfer the black solid obtained above into a 250mL single-necked round-bottom flask, add 150mL of dichloromethane, slowly stir, pass hydrogen bromide gas to saturation, continue stirring, and monitor the reaction by TLC. After the reaction is completed, concentrate to dryness under reduced pressure , Proceed directly to the next reaction;
将上述所得的浓缩物转入一个500mL三口圆底烧瓶中,称入化合物(IV-2)(1.5g,5.95mmol),加入100mL二氯甲烷,搅拌,冰浴冷却下,缓慢滴加50mL三乙胺的二氯甲烷溶液(含三乙胺5mL,35.8mmol)加完后,常温反应,TLC监控反应,反应结束后,加入100mL二氯甲烷和100mL去离子水,分出有机相,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积比为:二氯甲烷:丙酮=100:2,得到化合物(IIA-2),MS(ESI):[M+H
+]:888.55。
Transfer the concentrate obtained above into a 500mL three-necked round-bottom flask, weigh in compound (IV-2) (1.5g, 5.95mmol), add 100mL of dichloromethane, stir, under ice bath cooling, slowly add 50mL After the addition of ethylamine in dichloromethane (containing 5mL of triethylamine, 35.8mmol), the reaction was carried out at room temperature. The reaction was monitored by TLC. After the reaction was completed, 100mL of dichloromethane and 100mL of deionized water were added, the organic phase was separated, and the pressure was reduced. Concentrate to dryness and purify by silica gel column chromatography. The eluent and volume ratio are: dichloromethane: acetone = 100: 2 to obtain compound (IIA-2), MS (ESI): [M + H + ]: 888.55.
第三步:化合物(I-2)的合成Step 3: Synthesis of compound (I-2)
将上述所得的化合物(IIA-2)加入到一个500mL单口圆底烧瓶中,加入100mL乙腈和40mL去离子水,搅拌,再加入碳酸钾(2.1g,15.2mmol),氮气保护下,40℃搅拌反应4-6小时,TLC监控反应,反应结束后,加入10%的乙酸水溶液,调节pH值至2-3。将反应液转入一个500mL的分液漏斗中,加入200mL乙酸乙酯和100mL去离子水,分出有机相,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积比为:二氯甲烷:甲醇=100:10,得到0.1g化合物(I-2),总收率为3.9%。Add the compound (IIA-2) obtained above to a 500mL single-necked round-bottom flask, add 100mL of acetonitrile and 40mL of deionized water, stir, and then add potassium carbonate (2.1g, 15.2mmol), stirring under nitrogen at 40 ° C After 4-6 hours of reaction, the reaction was monitored by TLC. After the reaction was completed, a 10% aqueous acetic acid solution was added to adjust the pH to 2-3. The reaction solution was transferred to a 500mL separatory funnel, 200mL ethyl acetate and 100mL deionized water were added, the organic phase was separated, concentrated to dryness under reduced pressure, purified by silica gel column chromatography, eluent and volume ratio is : Dichloromethane: methanol = 100: 10, 0.1 g of compound (I-2) was obtained, and the total yield was 3.9%.
化合物(I-2)的表征数据如下:The characterization data of compound (I-2) is as follows:
UV Wavelength(Absorbance):782.2(0.18),537.0(0.155),416.0(0.195),365.6(0.438),202.0(0.40);UV Wavelength (Absorbance): 782.2 (0.18), 537.0 (0.155), 416.0 (0.195), 365.6 (0.438), 202.0 (0.40);
MS(ESI):[M
-]:872.56,[M]:873.56,[M+H
+]:874.56;
MS (ESI): [M - ]: 872.56, [M]: 873.56, [M + H +]: 874.56;
1H NMR(400M Hz,DMSO)δ-0.31(s,1H),0.07(s,1H),0.79(t,J=6.34Hz,3H),0.94-0.98(m,6H),1.15-1.19(m,6H),1.39-1.45(m,2H),1.59-1.64(m,6H),1.70-1.77(m,3H)1.82-1.93(m,4H),1.99-2.01(m,8H),2.21-2.28(m,3H),3.20(s,3H),3.42(s,3H),3.77-3.86(m,1H),3.84-3.86(m,1H),4.02-4.05(m,3H),4.13-4.28(m,1H),4.38-4.49(m,2H),5.06(s,1H),5.79(s,1H),6.67-6.69(m,1H),6.96-7.01(m,2H),8.58(s,2H),8.78(d,J=6.34Hz,1H);
1 H NMR (400M Hz, DMSO) δ-0.31 (s, 1H), 0.07 (s, 1H), 0.79 (t, J = 6.34 Hz, 3H), 0.94-0.98 (m, 6H), 1.15-1.19 ( m, 6H), 1.39-1.45 (m, 2H), 1.59-1.64 (m, 6H), 1.70-1.77 (m, 3H) 1.82-1.93 (m, 4H), 1.99-2.01 (m, 8H), 2.21 -2.28 (m, 3H), 3.20 (s, 3H), 3.42 (s, 3H), 3.77-3.86 (m, 1H), 3.84-3.86 (m, 1H), 4.02-4.05 (m, 3H), 4.13 -4.28 (m, 1H), 4.38-4.49 (m, 2H), 5.06 (s, 1H), 5.79 (s, 1H), 6.67-6.69 (m, 1H), 6.96-7.01 (m, 2H), 8.58 (s, 2H), 8.78 (d, J = 6.34 Hz, 1H);
13C NMR(100MHz,DMSO)δ10.77,10.97,11.83,14.24,14.51,16.20,20.46,21.17,21.67,22.43,22.72,23.65,24.15,28.59,29.91,30.90,31.33,32.07,33.72,53.55,60.18,67.55,70.79,99.28,101.45,111.19,113.12,125.86,127.22,128.14,128.23,130.08,130.76,131.02,132.55,134.03,138.07,138.64,138.73,141.62,156.53,161.24,162.87,167.05,170.76,171.32,171.57,172.68,173.42,175.46,176.08。
13 C NMR (100MHz, DMSO) δ 10.77, 10.97, 11.83, 14.24, 14.51, 16.20, 20.46, 21.17, 21.67, 22.43, 22.72, 23.65, 24.15, 28.59, 29.91, 30.90, 31.33, 32.07, 33.72, 53.55, 60.18,67.55,70.79,99.28,101.45,111.19,113.12,125.86,127.22,128.14,128.23,130.08,130.76,131.02,132.55,134.03,138.07,138.64,138.73,141.62,156.53,161.24,162.87,167.05,170 171.32,171.57,172.68,173.42,175.46,176.08.
实施例3化合物(I-3)的合成:Example 3 Synthesis of Compound (I-3):
第一步:2-(3-甲基-(4-羟甲基))苯氧乙酸甲酯(IV-3)的合成Step 1: Synthesis of methyl 2- (3-methyl- (4-hydroxymethyl)) phenoxyacetate (IV-3)
在一个500mL的三口圆底烧瓶中,加入3-甲基-4羟基苯甲醛(5.02g,25.5mmol),再加入80mL乙腈,搅拌溶解,再加入溴乙酸甲酯(2.5mL,2.55mmol)和碳酸钾(5.0g,3.61mmol),常温搅拌反应,TLC监控反应,反应结束后,加入150mL乙酸乙酯稀释反应液,过滤,滤液减压浓缩至干,得到黄色油状物,直接进行下一步反应;In a 500mL three-necked round bottom flask, add 3-methyl-4hydroxybenzaldehyde (5.02g, 25.5mmol), then add 80mL of acetonitrile, stir to dissolve, then add methyl bromoacetate (2.5mL, 2.55mmol) and Potassium carbonate (5.0g, 3.61mmol), stirring the reaction at room temperature, and monitoring the reaction by TLC. After the reaction was completed, 150mL of ethyl acetate was added to dilute the reaction solution, and the filtrate was concentrated to dryness under reduced pressure to obtain a yellow oil. ;
将得到的黄色液体转入一个500mL的三口圆底烧瓶中,加入100mL二氯甲烷和50mL甲醇,搅拌,冰浴冷却下,缓慢加入硼氢化钠(0.7g,1.9mmol),加完后,常温搅拌反应,TLC监控反应,反应结束后,用10%的稀盐酸水溶液淬灭反应,接着加入150mL二氯甲烷和100mL水,分出有机相,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积比为:石油醚:乙酸乙酯=4:1,得到5.8g2-(3-甲基-(4-羟甲基))苯氧乙酸甲酯(IV-3),白色固体,总收率75.3%。Transfer the obtained yellow liquid to a 500mL three-necked round bottom flask, add 100mL of methylene chloride and 50mL of methanol, stir, and slowly add sodium borohydride (0.7g, 1.9mmol) under ice bath cooling. Stir the reaction and monitor the reaction by TLC. After the reaction is completed, quench the reaction with 10% dilute hydrochloric acid aqueous solution, then add 150 mL of dichloromethane and 100 mL of water, separate the organic phase, concentrate to dryness under reduced pressure, and purify by silica gel column chromatography , The eluent and volume ratio are: petroleum ether: ethyl acetate = 4: 1, to obtain 5.8g of 2- (3-methyl- (4-hydroxymethyl)) phenoxyacetic acid methyl ester (IV-3), white Solid, total yield 75.3%.
1H NMR(400MHz,CDCl
3)δ1.77(s,1H),2.31(s,3H),3.81(S,3H),4.59(s,2H),4.67(s,2H),6.69(d,J=8.24Hz,1H),7.13(d,J=8.24Hz,1H),7.19(s,1H);
1 H NMR (400 MHz, CDCl 3 ) δ 1.77 (s, 1H), 2.31 (s, 3H), 3.81 (S, 3H), 4.59 (s, 2H), 4.67 (s, 2H), 6.69 (d, J = 8.24 Hz, 1H), 7.13 (d, J = 8.24 Hz, 1H), 7.19 (s, 1H);
13CNMR(100MHz,CDCl
3)δ16.19,52.2,64.97,65.63,111.15,125.67,127.55,130.27,133.96,155.66,169.60.
13 CNMR (100MHz, CDCl 3 ) δ 16.19, 52.2, 64.97, 65.63, 111.15, 125.67, 127.55, 130.27, 133.96, 155.66, 169.60.
第二步:化合物(IIA-3)的合成Step 2: Synthesis of compound (IIA-3)
在一个500mL三口圆底烧瓶中,称入化合物(IIB-2)(1.7g,2.26mmol),加入 100mL二氯甲烷和20mL甲醇,搅拌溶解,冰浴冷却下,缓慢加入硼氢化钠(45mg,1.13mmol),加完后,常温搅拌反应,TLC监控反应,反应结束后,用10%的稀醋酸水溶液淬灭反应,接着加入150mL二氯甲烷和100mL水,分出有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,得到黑色固体,接进行下一步反应;In a 500mL three-necked round bottom flask, weigh compound (IIB-2) (1.7g, 2.26mmol), add 100mL dichloromethane and 20mL methanol, stir to dissolve, and slowly add sodium borohydride (45mg, 1.13mmol), after the addition, the reaction was stirred at room temperature, and the reaction was monitored by TLC. After the reaction was completed, the reaction was quenched with 10% dilute acetic acid aqueous solution, followed by addition of 150mL of methylene chloride and 100mL of water, and the organic phase was separated, anhydrous sodium sulfate Dry, filter, and concentrate the filtrate under reduced pressure to obtain a black solid, and then proceed to the next reaction;
将上述所得的黑色固体转入一个250mL单口圆底烧瓶中,加入100mL二氯甲烷,缓慢搅拌,通入溴化氢气体至饱和,继续搅拌,TLC监控反应,反应结束后,减压浓缩至干,直接进行下一步反应;Transfer the black solid obtained above to a 250mL single-necked round-bottom flask, add 100mL of dichloromethane, slowly stir, pass hydrogen bromide gas to saturation, continue to stir, and monitor the reaction by TLC. After the reaction is completed, concentrate to dryness under reduced pressure , Proceed directly to the next reaction;
将上述所得的浓缩物转入一个500mL三口圆底烧瓶中,称入化合物(IV-3)(1.5g,7.14mmol),加入100mL二氯甲烷,搅拌,冰浴冷却下,缓慢滴加50mL三乙胺的二氯甲烷溶液(含三乙胺3mL,21.5mmol),加完后,常温反应,TLC监控反应,反应结束后,加入100mL二氯甲烷和100mL去离子水,分出有机相,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积比为:二氯甲烷:丙酮=100:2,得到化合物(ⅡA-3),MS(ESI):[M+H
+]:944.56。
Transfer the concentrate obtained above into a 500mL three-necked round bottom flask, weigh in compound (IV-3) (1.5g, 7.14mmol), add 100mL of dichloromethane, stir, and slowly drop 50mL of trihydrate under ice bath cooling A solution of ethylamine in dichloromethane (containing 3mL of triethylamine, 21.5mmol). After the addition, the reaction was performed at room temperature. The reaction was monitored by TLC. After the reaction, 100mL of dichloromethane and 100mL of deionized water were added to separate the organic phase. Concentrate to dryness by pressure, purify by silica gel column chromatography, eluent and volume ratio: methylene chloride: acetone = 100: 2, to obtain compound (ⅡA-3), MS (ESI): [M + H + ] : 944.56.
第三步:化合物(I-3)的合成Step 3: Synthesis of compound (I-3)
将上述所得的化合物(IIA-3)全部加入到一个500mL单口圆底烧瓶中,加入100mL乙腈和40mL去离子水,搅拌,再加入碳酸钾(2.0g,14.5mmol),氮气保护下,40℃搅拌反应,TLC监控反应,反应结束后,加入10%的乙酸水溶液,调节pH值至2-3。将反应液转入一个500mL的分液漏斗中,加入200mL乙酸乙酯和100mL去离子水,分出有机相,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积为:二氯甲烷:甲醇=100:10,得到0.33g化合物(I-3),总收率为15.7%。Add all the above-obtained compound (IIA-3) to a 500mL single-necked round bottom flask, add 100mL of acetonitrile and 40mL of deionized water, stir, and then add potassium carbonate (2.0g, 14.5mmol), under nitrogen protection, 40 ℃ The reaction was stirred, and the reaction was monitored by TLC. After the reaction was completed, a 10% aqueous acetic acid solution was added to adjust the pH to 2-3. Transfer the reaction solution to a 500 mL separatory funnel, add 200 mL of ethyl acetate and 100 mL of deionized water, separate the organic phase, concentrate to dryness under reduced pressure, and purify by silica gel column chromatography. The eluent and volume are: Dichloromethane: methanol = 100: 10, 0.33 g of compound (I-3) was obtained, and the total yield was 15.7%.
化合物(I-3)的表征数据如下:The characterization data of compound (I-3) is as follows:
UV Wavelength(Absorbance):781.8(0.123),537.2(0.108),415.2(0.135),365.8(0.309),202.0(0.212);UV Wavelength (Absorbance): 781.8 (0.123), 537.2 (0.108), 415.2 (0.135), 365.8 (0.309), 202.0 (0.212);
MS(ESI):[M+H
+]:930.55;
MS (ESI): [M + H + ]: 930.55;
1H NMR(400M Hz,DMSO)δ-0.30(s,1H),0.10(s,1H),0.61(t,J=6.34Hz,3H),0.74-0.84(m,4H),0.88-1.02(m,9H),1.09-1.08(m,2H),1.18-1.37(m,7H),1.60-1.73(m,7H),1.82-1.93(m,4H),2.08-2.28(m,6H),2.38-2.45(m,1H),3.21(s,3H),3.40(s,3H),3.54-3.68(m,3H),3.84-3.91(m,1H),4.02-4.16(m,3H),4.22-4.28(m,1H),4.39-4.50(m,2H),4.62-4.68(m,2H),5.10(d,J=7.48Hz,1H),5.80-5.83(t, J=5.68Hz,1H),6.73(d,J=8.16Hz,1H),6.96-7.12(m,2H),8.60(s,2H),8.88(d,J=19.8Hz,1H);
1 H NMR (400M Hz, DMSO) δ-0.30 (s, 1H), 0.10 (s, 1H), 0.61 (t, J = 6.34Hz, 3H), 0.74-0.84 (m, 4H), 0.88-1.02 ( m, 9H), 1.09-1.08 (m, 2H), 1.18-1.37 (m, 7H), 1.60-1.73 (m, 7H), 1.82-1.93 (m, 4H), 2.08-2.28 (m, 6H), 2.38-2.45 (m, 1H), 3.21 (s, 3H), 3.40 (s, 3H), 3.54-3.68 (m, 3H), 3.84-3.91 (m, 1H), 4.02-4.16 (m, 3H), 4.22-4.28 (m, 1H), 4.39-4.50 (m, 2H), 4.62-4.68 (m, 2H), 5.10 (d, J = 7.48Hz, 1H), 5.80-5.83 (t, J = 5.68Hz, 1H), 6.73 (d, J = 8.16 Hz, 1H), 6.96-7.12 (m, 2H), 8.60 (s, 2H), 8.88 (d, J = 19.8 Hz, 1H);
13C NMR(100MHz,DMSO)δ10.78,11.04,11.84,14.02,14.28,16.35,16.42,22.18,22.46,22.72,22.84,23.66,25.20,26.82,28.11,28.64,31.05,31.49,32.07,53.49,53.53,63.99,65.26,70.67,70.89,99.39,111.41,113.15,126.14,127.06,128.18,128.28,130.84,130.93,131.04,131.08,132.54,134.11,138.05,138.60,138.68,141.58,141.68,155.89,161.25,162.85,167.02,170.67,171.33,172.62,172.85,175.39,175.44,175.47,176.04。
13 C NMR (100MHz, DMSO) δ 10.78, 11.04, 11.84, 14.02, 14.28, 16.35, 16.42, 22.18, 22.46, 22.72, 22.84, 23.66, 25.20, 26.82, 28.11, 28.64, 31.05, 31.49, 32.07, 53.49, 53.53,63.99,65.26,70.67,70.89,99.39,111.41,113.15,126.14,127.06,128.18,128.28,130.84,130.93,131.04,131.08,132.54,134.11,138.05,138.60,138.68,141.58,141.68,155.89,16 162.85,167.02,170.67,171.33,172.62,172.85,175.39,175.44,175.47,176.04.
实施例4化合物(I-4)的合成:Example 4 Synthesis of Compound (I-4):
第一步:2-(4-羟丁基)苯氧乙酸甲酯(IV-4)的合成Step 1: Synthesis of methyl 2- (4-hydroxybutyl) phenoxyacetate (IV-4)
在一个500mL的三口圆底烧瓶中,加入4-羟基苯丁酸(5.0g,27.7mmol),再加入80mL二氯甲烷和50mL甲醇,搅拌溶解,冰水浴冷却下,滴加二氯亚砜(6.5mL,89.5mmol)加完后,常温搅拌反应,TLC监控反应。反应结束后,加入200mL二氯甲烷和100mL水,分出有机相。有机相用饱和碳酸氢钠水溶液洗涤2次后,无水硫酸镁干燥,过滤,滤液减压浓缩至干,得到黄色油状物,直接进行下一步反应;In a 500mL three-necked round-bottom flask, add 4-hydroxyphenylbutyric acid (5.0g, 27.7mmol), then add 80mL of dichloromethane and 50mL of methanol, stir to dissolve, under ice water bath cooling, dropwise add dichlorosulfoxide ( After the addition of 6.5mL, 89.5mmol), the reaction was stirred at room temperature, and the reaction was monitored by TLC. After the reaction was completed, 200 mL of dichloromethane and 100 mL of water were added, and the organic phase was separated. The organic phase was washed twice with saturated aqueous sodium bicarbonate solution, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain a yellow oil, which was directly subjected to the next reaction;
在一个500mL的三口圆底烧瓶中,称入四氢铝锂(3.0g,79.1mmol),加入150mL四氢呋喃,搅拌,冰浴冷却下,滴加上述步骤得到的黄色油状物的四氢呋喃溶液(含30mL四氢呋喃),滴加完毕后,常温搅拌反应,TLC监控反应,反应结束后,加入10%的盐酸水溶液,搅拌30分钟无气泡放出后,加入200mL二氯甲烷,分出有机相,无水硫酸镁干燥,过滤,滤液减压浓缩至干,直接用于下步反应;In a 500mL three-necked round-bottom flask, weigh lithium aluminum hydride (3.0g, 79.1mmol), add 150mL of tetrahydrofuran, stir, and cool in an ice bath, dropwise add the yellow oily tetrahydrofuran solution (containing 30mL) obtained in the above step Tetrahydrofuran), after the dropwise addition, the reaction was stirred at room temperature, and the reaction was monitored by TLC. After the reaction was completed, 10% aqueous hydrochloric acid was added. After stirring for 30 minutes without bubbles, 200 mL of dichloromethane was added to separate the organic phase and anhydrous magnesium sulfate. Dry, filter, and concentrate the filtrate to dryness under reduced pressure, which is directly used in the next reaction;
将上述浓缩物转入一个500mL的单口圆底烧瓶中,再加入150mL乙腈,搅拌,再加入溴乙酸甲酯(6.0mL,61.2mmol)和碳酸钾(14.0g,102mmol),常温下搅拌反应,TLC监控反应,反应结束后,加入150mL乙酸乙酯,过滤,滤液减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积比为:石油醚:乙酸乙酯=4:1,得到5.0g 2-(4-羟丁基)苯氧乙酸甲酯(IV-4),浅黄色液体,总收率为75.7%。Transfer the above concentrate to a 500mL single-necked round bottom flask, add 150mL of acetonitrile, stir, then add methyl bromoacetate (6.0mL, 61.2mmol) and potassium carbonate (14.0g, 102mmol), and stir the reaction at room temperature, The reaction was monitored by TLC. After the reaction was completed, 150 mL of ethyl acetate was added and filtered. The filtrate was concentrated to dryness under reduced pressure and purified by silica gel column chromatography. The eluent and volume ratio were: petroleum ether: ethyl acetate = 4: 1. 5.0 g of methyl 2- (4-hydroxybutyl) phenoxyacetate (IV-4) was obtained as a pale yellow liquid with a total yield of 75.7%.
1H NMR(400MHz,CDCl
3)δ1.58-1.72(m,4H),2.61(t,J=7.38Hz,2H),3.67(t,J=6.36Hz,2H),3.83(s,3H),4.63(s,2H),6.86(d,J=8.64Hz,2H),7.12(d,J=8.64Hz,2H);
1 H NMR (400 MHz, CDCl 3 ) δ 1.58-1.72 (m, 4H), 2.61 (t, J = 7.38 Hz, 2H), 3.67 (t, J = 6.36 Hz, 2H), 3.83 (s, 3H) , 4.63 (s, 2H), 6.86 (d, J = 8.64 Hz, 2H), 7.12 (d, J = 8.64 Hz, 2H);
13CNMR(100MHz,CDCl
3)δ27.67,32.21,34.71,52.22,62.65,65.48,114.52,129.40,135.71,155.94,169.66.
13 CNMR (100MHz, CDCl 3 ) δ 27.67, 32.21, 34.71, 52.22, 62.65, 65.48, 114.52, 129.40, 135.71, 155.94, 169.66.
第二步:化合物(IIA-4)的合成Step 2: Synthesis of compound (IIA-4)
在一个500mL三口圆底烧瓶中,称入化合物(IIB-3)(4.50g,6.24mmol),加入100mL二氯甲烷和20mL甲醇,搅拌溶解,冰浴冷却下,缓慢加入硼氢化钠(0.24g,6.3mmol),加完后,常温搅拌反应1-2小时,TLC监控反应,反应结束后,用10%的稀醋酸水溶液淬灭反应,接着加入150mL二氯甲烷和100mL水,分出有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,得到黑色固体,直接 进行下一步反应;In a 500mL three-necked round bottom flask, weigh compound (IIB-3) (4.50g, 6.24mmol), add 100mL of dichloromethane and 20mL of methanol, stir to dissolve, and slowly add sodium borohydride (0.24g) under ice bath cooling , 6.3mmol), after the addition was completed, the reaction was stirred at room temperature for 1-2 hours, and the reaction was monitored by TLC. After the reaction was completed, the reaction was quenched with 10% dilute acetic acid aqueous solution, followed by addition of 150mL of dichloromethane and 100mL of water, and the organic phase was separated , Dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a black solid, which was directly subjected to the next reaction;
将上述所得的黑色固体转入一个250mL单口圆底烧瓶中,加入100mL二氯甲烷,缓慢搅拌,通入溴化氢气体至饱和,继续搅拌,TLC监控反应,反应结束后,减压浓缩至干,直接进行下一步反应;Transfer the black solid obtained above to a 250mL single-necked round-bottom flask, add 100mL of dichloromethane, slowly stir, pass hydrogen bromide gas to saturation, continue to stir, and monitor the reaction by TLC. After the reaction is completed, concentrate to dryness under reduced pressure , Proceed directly to the next reaction;
将上述所得的浓缩物转入一个500mL三口圆底烧瓶中,称入化合物(IV-4)(1.0g,4.20mmol),加入100mL二氯甲烷,搅拌,冰浴冷却下,缓慢滴加50mL三乙胺的二氯甲烷溶液(含三乙胺3mL,21.5mmol)加完后,常温反应,TLC监控反应,反应结束后,加入100mL二氯甲烷和100mL去离子水,分出有机相,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积比为:二氯甲烷:丙酮=100:2,得到化合物(IIA-4),MS(ESI):[M+H
+]:944.66。
Transfer the concentrate obtained above into a 500mL three-necked round bottom flask, weigh in compound (IV-4) (1.0g, 4.20mmol), add 100mL of dichloromethane, stir, and slowly add 50mL After the addition of ethylamine in dichloromethane (containing 3mL of triethylamine, 21.5mmol), the reaction was carried out at room temperature. The reaction was monitored by TLC. After the reaction was completed, 100mL of dichloromethane and 100mL of deionized water were added, the organic phase was separated, and the pressure was reduced. Concentrate to dryness and purify by silica gel column chromatography. The eluent and volume ratio are: dichloromethane: acetone = 100: 2 to obtain compound (IIA-4), MS (ESI): [M + H + ]: 944.66.
第三步:化合物(I-4)的合成Step 3: Synthesis of compound (I-4)
将上述所得的化合物(IIA-4)加入到一个500mL三口圆底烧瓶中,加入100mL乙腈和40mL去离子水,搅拌,再加入碳酸钾(1.9g,13.7mmol),氮气保护下,40℃搅拌反应,TLC监控反应,反应结束后,加入10%的乙酸水溶液,调节pH值至2-3。将反应液转入一个500mL的分液漏斗中,加入150mL乙酸乙酯和100mL去离子水,分出有机相,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积为:二氯甲烷:甲醇=100:10,得到0.65g化合物(I-4),总收率为11.2%。Add the above-obtained compound (IIA-4) to a 500mL three-necked round bottom flask, add 100mL of acetonitrile and 40mL of deionized water, stir, and then add potassium carbonate (1.9g, 13.7mmol), under nitrogen, stirring at 40 ℃ The reaction was monitored by TLC. After the reaction was completed, a 10% aqueous acetic acid solution was added to adjust the pH to 2-3. Transfer the reaction solution to a 500 mL separatory funnel, add 150 mL of ethyl acetate and 100 mL of deionized water, separate the organic phase, concentrate to dryness under reduced pressure, and purify by silica gel column chromatography. The eluent and volume are: Dichloromethane: methanol = 100: 10, 0.65 g of compound (I-4) was obtained, and the total yield was 11.2%.
化合物(I-4)的表征数据如下:The characterization data of compound (I-4) is as follows:
UV Wavelength(Absorbance):781.8(0.094),536.6(0.083),416.0(0.096),365.6(0.231),200.4(0.110);UV Wavelength (Absorbance): 781.8 (0.094), 536.6 (0.083), 416.0 (0.096), 365.6 (0.231), 200.4 (0.110);
MS(ESI):[M
-]:928.57,[M]:929.63,[M+H
+]:930.64;
MS (ESI): [M - ]: 928.57, [M]: 929.63, [M + H +]: 930.64;
1H NMR(400M Hz,CDCl
3)δ-0.26(s,1H),0.04(s,1H),0.86(t,J=7.16Hz,3H),0.90-0.92(m,1H),0.97(t,J=6.84Hz,3H),1.07-1.17(m,3H),1.20-1.34(m,5H),1.42-1.54(m,6H),1.58-1.84(m,12H),1.92-2.12(m,6H),2.30-2.46(m,3H),2.46-2.56(m,1H),2.58-2.68(m,1H),3.24(s,3H),3.31-3.40(m,1H),3.52-3.73(m,5H),3.94-3.45(m,3H),4.16-4.28(m,2H),4.36-4.48(m,2H),4.58(s,1H),5.28-5.30(m,1H),5.67-5.60(m,1H),6.71-6.75(m,1H),6.95-7.28(m,2H),8.36(s,1H),8.62(s,1H),8.80(d,J=11.24Hz,1H);
1 H NMR (400M Hz, CDCl 3 ) δ-0.26 (s, 1H), 0.04 (s, 1H), 0.86 (t, J = 7.16 Hz, 3H), 0.90-0.92 (m, 1H), 0.97 (t , J = 6.84 Hz, 3H), 1.07-1.17 (m, 3H), 1.20-1.34 (m, 5H), 1.42-1.54 (m, 6H), 1.58-1.84 (m, 12H), 1.92-2.12 (m , 6H), 2.30-2.46 (m, 3H), 2.46-2.56 (m, 1H), 2.58-2.68 (m, 1H), 3.24 (s, 3H), 3.31-3.40 (m, 1H), 3.52-3.73 (m, 5H), 3.94-3.45 (m, 3H), 4.16-4.28 (m, 2H), 4.36-4.48 (m, 2H), 4.58 (s, 1H), 5.28-5.30 (m, 1H), 5.67 -5.60 (m, 1H), 6.71-6.75 (m, 1H), 6.95-7.28 (m, 2H), 8.36 (s, 1H), 8.62 (s, 1H), 8.80 (d, J = 11.24 Hz, 1H );
13C NMR(100MHz,CDCl
3)δ10.77,10.83,11.90,13.60,14.11,19.03,22.65,22.76, 23.57,24.38,27.19,28.21,28.97,29.65,30.25,30.54,31.17,31.80,32.46,34.78,40.27,48.19,49.26,53.91,55.39,62.88,64.27,65.05,69.34,69.41,72.58,94.79,99.80,101.36,113.97,114.61,128.70,129.37,129.49,131.53,132.62,133.31,135.82,137.82,138.06,141.36,155.56,161.2,163.93,167.71,170.38,172.55,173.70,173.80,173.89。
13 C NMR (100 MHz, CDCl 3 ) δ 10.77, 10.83, 11.90, 13.60, 14.11, 19.03, 22.65, 22.76, 23.57, 24.38, 27.19, 28.21, 28.97, 29.65, 30.25, 30.54, 31.17, 31.80, 32.46, 34.78 , 40.27,48.19,49.26,53.91,55.39,62.88,64.27,65.05,69.34,69.41,72.58,94.79,99.80,101.36,113.97,114.61,128.70,129.37,129.49,131.53,132.62,133.31,135.82,137.82,138.06,137.82 , 141.36,155.56,161.2,163.93,167.71,170.38,172.55,173.70,173.80,173.89.
实施例5化合物(I-5)的合成:Example 5 Synthesis of Compound (I-5):
第一步:2-(3-甲氧基-4-羟甲基)苯氧乙酸甲酯(IV-5)的合成Step 1: Synthesis of methyl 2- (3-methoxy-4-hydroxymethyl) phenoxyacetate (IV-5)
在一个500mL的三口圆底烧瓶中,加入3甲氧基-4-羟基苯甲醛(8.1g,53.2mmol),再加入100mL乙腈,搅拌溶解,再加入溴乙酸甲酯(5.2mL,53.08mmol)和碳酸钾(10.0g,72.3mmol),常温搅拌反应,TLC监控反应,反应结束后,加入100mL乙酸乙酯稀释反应液,过滤,滤液减压浓缩,得到黄色油状物,直接进行下一步反应;In a 500mL three-necked round bottom flask, add 3methoxy-4-hydroxybenzaldehyde (8.1g, 53.2mmol), then add 100mL of acetonitrile, stir to dissolve, and then add methyl bromoacetate (5.2mL, 53.08mmol) With potassium carbonate (10.0g, 72.3mmol), the reaction was stirred at room temperature, and the reaction was monitored by TLC. After the reaction was completed, 100mL of ethyl acetate was added to dilute the reaction solution, filtered, and the filtrate was concentrated under reduced pressure to obtain a yellow oil, which was directly subjected to the next reaction;
将得到的黄色油状物转入一个500mL的三口圆底烧瓶中,加入100mL二氯甲烷和30mL甲醇,搅拌,冰浴冷却下,缓慢加入硼氢化钠(0.686g,1.9mmol),加完后,常温搅拌反应,TLC监控反应进程,反应结束后,用10%的稀盐酸水溶液淬灭反应,接着加入150mL二氯甲烷和100mL水,分出有机相,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积比为:石油醚:乙酸乙酯=3:1,得到10.9g 2-(3-甲氧基-4-羟甲基)苯氧乙酸甲酯(IV-5),浅黄色液体,总收率为90.9%。Transfer the resulting yellow oil to a 500mL three-necked round bottom flask, add 100mL of dichloromethane and 30mL of methanol, stir, and slowly add sodium borohydride (0.686g, 1.9mmol) under ice bath cooling. After the addition, The reaction was stirred at room temperature, and the progress of the reaction was monitored by TLC. After the reaction was completed, the reaction was quenched with 10% diluted hydrochloric acid aqueous solution, followed by addition of 150 mL of dichloromethane and 100 mL of water, the organic phase was separated, concentrated to dryness under reduced pressure, and silica gel column chromatography Method purification, eluent and volume ratio: petroleum ether: ethyl acetate = 3: 1, to obtain 10.9g methyl 2- (3-methoxy-4-hydroxymethyl) phenoxyacetate (IV-5) , Light yellow liquid, the total yield is 90.9%.
1H NMR(400MHz,CDCl
3)δ2.98(s,1H),3.70(s,3H),3.77(s,3H),4.48(s,2H),4.59(s,2H),6.66-6.86(m,2H),7.28(s,1H);
1 H NMR (400 MHz, CDCl 3 ) δ 2.98 (s, 1H), 3.70 (s, 3H), 3.77 (s, 3H), 4.48 (s, 2H), 4.59 (s, 2H), 6.66-6.86 ( m, 2H), 7.28 (s, 1H);
13C NMR(100MHz,CDCl
3)δ52.13,55.76,64.58,66.39,110.99,114.14,118.99,135.67,146.38,149.54,169.57.
13 C NMR (100 MHz, CDCl 3 ) δ 52.13, 55.76, 64.58, 66.39, 110.99, 114.14, 118.99, 135.67, 146.38, 149.54, 169.57.
第二步:化合物(IIA-5)的合成Step 2: Synthesis of compound (IIA-5)
在一个500mL三口圆底烧瓶中,称入化合物(IIB-4)(1.2g,1.76mmol),加入100mL二氯甲烷和20mL甲醇,搅拌溶解,冰浴冷却下,缓慢加入硼氢化钠(0.1g,2.6mmol),加完后,常温搅拌反应1-2小时,TLC监控反应进程,反应结束后,用10%的稀醋酸水溶液淬灭反应,接着加入150mL二氯甲烷和100mL水,分出有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,得到黑色固体,直接进行下一步反应;In a 500mL three-necked round bottom flask, weigh compound (IIB-4) (1.2g, 1.76mmol), add 100mL of dichloromethane and 20mL of methanol, stir to dissolve, and slowly add sodium borohydride (0.1g) under ice bath cooling , 2.6mmol). After the addition, the reaction was stirred at room temperature for 1-2 hours. TLC monitored the progress of the reaction. After the reaction was completed, the reaction was quenched with 10% dilute acetic acid aqueous solution, followed by the addition of 150mL of dichloromethane and 100mL of water. Phase, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a black solid, which was directly subjected to the next reaction;
将上述所得的黑色固体转入一个250mL三口圆底烧瓶中,加入100mL二氯甲烷,缓慢搅拌,通入溴化氢气体至饱和,继续搅拌,TLC监控反应,反应结束后,减压浓缩至干,直接进行下一步反应;Transfer the black solid obtained above to a 250mL three-necked round-bottom flask, add 100mL of dichloromethane, slowly stir, pass hydrogen bromide gas to saturation, continue stirring, and monitor the reaction by TLC. After the reaction is completed, concentrate to dryness under reduced pressure , Proceed directly to the next reaction;
将上述所得的浓缩物转入一个500mL三口圆底烧瓶中,称入化合物(IV-5)(2.0g,8.84mmol),加入100mL二氯甲烷,搅拌,冰浴冷却下,缓慢滴加30mL三乙胺的二氯甲烷溶液(含三乙胺3mL,21.5mmol),加完后,常温反应,TLC 监控反应,反应结束后,加入100mL二氯甲烷和100mL去离子水,分出有机相,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积比为:二氯甲烷:丙酮=100:2,得到化合物(IIA-5),MS(ESI):[M+H
+]:889.56;
Transfer the concentrate obtained above into a 500mL three-necked round-bottom flask, weigh in compound (IV-5) (2.0g, 8.84mmol), add 100mL of dichloromethane, stir, under ice bath cooling, slowly add 30mL A solution of ethylamine in dichloromethane (containing 3mL of triethylamine, 21.5mmol). After the addition, the reaction was performed at room temperature. The reaction was monitored by TLC. After the reaction, 100mL of dichloromethane and 100mL of deionized water were added to separate the organic phase. Concentrate to dryness under pressure and purify by silica gel column chromatography. The eluent and volume ratio are: dichloromethane: acetone = 100: 2, to obtain compound (IIA-5), MS (ESI): [M + H + ] : 889.56;
第三步:化合物(I-5)的合成Step 3: Synthesis of compound (I-5)
将上述所得的化合物(IIA-5)加入到一个500mL三口圆底烧瓶中,加入100mL乙腈和40mL去离子水,搅拌,再加入碳酸钾(3.7g,26.7mmol),氮气保护下,40℃搅拌反应,TLC监控反应,反应结束后,加入10%的乙酸水溶液,调节pH值至2-3。将反应液转入一个500mL的分液漏斗中,加入150mL乙酸乙酯和100mL去离子水,分出有机相,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积比为:二氯甲烷:甲醇=100:10,得到0.33g化合物(I-5),总收率为21.3%。Add the above-obtained compound (IIA-5) to a 500mL three-necked round bottom flask, add 100mL of acetonitrile and 40mL of deionized water, stir, and then add potassium carbonate (3.7g, 26.7mmol), stirring under nitrogen at 40 ° C The reaction was monitored by TLC. After the reaction was completed, a 10% aqueous acetic acid solution was added to adjust the pH to 2-3. The reaction solution was transferred to a 500mL separatory funnel, 150mL ethyl acetate and 100mL deionized water were added, the organic phase was separated, concentrated to dryness under reduced pressure, purified by silica gel column chromatography, eluent and volume ratio is : Dichloromethane: methanol = 100: 10, 0.33 g of compound (I-5) was obtained, and the total yield was 21.3%.
化合物(I-5)的表征数据如下:The characterization data of compound (I-5) is as follows:
UV Wavelength(Absorbance):782.2(0.141),537.(0.122),415.6(0.148),365.8(0.348),202.8(0.235);UV Wavelength (Absorbance): 782.2 (0.141), 537. (0.122), 415.6 (0.148), 365.8 (0.348), 202.8 (0.235);
MS(ESI):[M+H
+]:876.55;
MS (ESI): [M + H + ]: 876.55;
1H NMR(400M Hz,CDCl
3)δ-0.28(s,1H),0.04(s,1H),0.98(t,J=6.66Hz,3H),1.16(t,J=6.90Hz,3H),1.42-1.51(m,4H),1.60-1.64(m,3H),1.71-1.84(m,6H),1.96--2.13(m,7H),2.32-2.43(m,3H),2.65-2.71(m,1H),3.27(s,3H),3.61(s,3H),3.68(s,3H),3.84(s,3H),4.05(br,1H),4.20-4.28(m,2H),4.42-4.58(m,3H),4.66(d,J=7.92Hz,1H),4.74(s,2H),5.33(d,J=8.12Hz,1H),5.80(t,J=6.04Hz,1H),6.88-6.92(m,3H),6.98(d,J=13.44Hz,1H),8.42(s,1H),8.68(s,1H),8.31(d,J=23.84Hz,1H);
1 H NMR (400M Hz, CDCl 3 ) δ-0.28 (s, 1H), 0.04 (s, 1H), 0.98 (t, J = 6.66 Hz, 3H), 1.16 (t, J = 6.90 Hz, 3H), 1.42-1.51 (m, 4H), 1.60-1.64 (m, 3H), 1.71-1.84 (m, 6H), 1.96--2.13 (m, 7H), 2.32-2.43 (m, 3H), 2.65-2.71 ( m, 1H), 3.27 (s, 3H), 3.61 (s, 3H), 3.68 (s, 3H), 3.84 (s, 3H), 4.05 (br, 1H), 4.20-4.28 (m, 2H), 4.42 -4.58 (m, 3H), 4.66 (d, J = 7.92 Hz, 1H), 4.74 (s, 2H), 5.33 (d, J = 8.12 Hz, 1H), 5.80 (t, J = 6.04 Hz, 1H) , 6.88-6.92 (m, 3H), 6.98 (d, J = 13.44 Hz, 1H), 8.42 (s, 1H), 8.68 (s, 1H), 8.31 (d, J = 23.84 Hz, 1H);
13C NMR(100MHz,CDCl
3)δ10.77,10.94,14.13,22.75,23.56,,24.19,24.36,27.19,28.98,30.24,31.27,31.81,32.26,40.30,48.19,49.32,51.52,53.84,55.38,55.93,67.38,70.99,71.52,95.01,98.98,99.16,99.78,101.49,112.19,114.15,115.76,120.79,129.91,131.75,132.77,133.44,137.10,137.45,140.79,146.84,146.87,149.84,161.17,163.93,167.74,170.33,171.80,172.53,173.65,174.11。
13 C NMR (100 MHz, CDCl 3 ) δ 10.77, 10.94, 14.13, 22.75, 23.56 ,, 24.19, 24.36, 27.19, 28.98, 30.24, 31.27, 31.81, 32.26, 40.30, 48.19, 49.32, 51.52, 53.84, 55.38, 55.93,67.38,70.99,71.52,95.01,98.98,99.16,99.78,101.49,112.19,114.15,115.76,120.79,129.91,131.75,132.77,133.44,137.10,137.45,140.79,146.84,146.87,149.84,161.17,163.93 167.74,170.33,171.80,172.53,173.65,174.11.
实施例6化合物(I-6)的合成:Example 6 Synthesis of Compound (I-6):
第一步:3-氟-4-乙酰氧基苯甲醇(IV-6)的合成Step 1: Synthesis of 3-fluoro-4-acetoxybenzyl alcohol (IV-6)
在一个500mL三口圆底烧瓶中,称入3-氟-4羟基苯甲醛(5g,35.7mmol),加入250mL二氯甲烷和乙酸酐(5mL,52.9mmol),搅拌溶解,冰浴冷却下,再加入三乙胺(8mL,70.6mmol),常温搅拌反应,TLC监测反应,反应结束后,加入20mL乙醇淬灭反应,减压浓缩,得到黄色油状物,直接进行下一步反应;In a 500mL three-necked round bottom flask, weigh in 3-fluoro-4hydroxybenzaldehyde (5g, 35.7mmol), add 250mL of dichloromethane and acetic anhydride (5mL, 52.9mmol), stir to dissolve, and cool in an ice bath, then Triethylamine (8mL, 70.6mmol) was added, the reaction was stirred at room temperature, and the reaction was monitored by TLC. After the reaction was completed, 20mL of ethanol was added to quench the reaction and concentrated under reduced pressure to obtain a yellow oil, which was directly subjected to the next reaction;
将该黄色油状物转入一个500mL的三口圆底烧瓶中,加入160mL二氯甲烷和40mL甲醇,冰浴冷却下,缓慢加入硼氢化钠(0.76g,20mmol),加完后,常温搅拌反应,TLC监控反应,反应结束后,用10%的稀盐酸水溶液淬灭反应,接着加入150mL二氯甲烷和100mL去离子水,分出有机相,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积比为:石油醚:乙酸乙酯=4:1,得到4.5g 3-氟-4-乙酰氧基苯甲醇(IV-6),白色固体,总收率为68.5%。Transfer the yellow oil to a 500mL three-necked round-bottom flask, add 160mL of dichloromethane and 40mL of methanol, and slowly add sodium borohydride (0.76g, 20mmol) under ice-cooling. After the addition is complete, stir the reaction at room temperature, The reaction was monitored by TLC. After the reaction was completed, the reaction was quenched with 10% dilute hydrochloric acid aqueous solution, followed by addition of 150 mL of dichloromethane and 100 mL of deionized water, the organic phase was separated, concentrated to dryness under reduced pressure, and purified by silica gel column chromatography. The eluent and volume ratio are: petroleum ether: ethyl acetate = 4: 1, to obtain 4.5 g of 3-fluoro-4-acetoxybenzyl alcohol (IV-6), white solid, the total yield is 68.5%.
1H NMR(400MHz,CDCl
3)δ2.30(s,3H),2.99(br,1H),4.53(s,2H),7.03-7.12(m,2H),7.10(d,J=10.8Hz,1H);
1 H NMR (400 MHz, CDCl 3 ) δ 2.30 (s, 3H), 2.99 (br, 1H), 4.53 (s, 2H), 7.03-7.12 (m, 2H), 7.10 (d, J = 10.8 Hz, 1H);
13C NMR(100MHz,CDCl
3)δ20.40,63.69,114.82,122.43,123.49,136.91,140.84,152.67,155.15,168.80.
13 C NMR (100 MHz, CDCl 3 ) δ 20.40, 63.69, 114.82, 122.43, 123.49, 136.91, 140.84, 152.67, 155.15, 168.80.
第二步:化合物(IIIA-1)的合成Step 2: Synthesis of compound (IIIA-1)
在一个500mL三口圆底烧瓶中,称入化合物(IIB-1)(0.65g,0.99mmol),加入100mL二氯甲烷和10mL甲醇,搅拌,冰浴冷却下,缓慢加入硼氢化钠(0.50g,13.2mmol),加完后,常温搅拌反应,TLC监控反应,反应结束后,用10%的稀醋酸水溶液淬灭反应,接着加入150mL二氯甲烷和100mL水,分出有机相,无水硫酸钠干燥,过滤,滤液减压浓缩,得到黑色固体,直接进行下一步反应;In a 500mL three-necked round bottom flask, weigh compound (IIB-1) (0.65g, 0.99mmol), add 100mL of dichloromethane and 10mL of methanol, stir, and slowly add sodium borohydride (0.50g, 13.2mmol), after the addition, the reaction was stirred at room temperature, the reaction was monitored by TLC, after the reaction was completed, the reaction was quenched with 10% dilute acetic acid aqueous solution, followed by the addition of 150mL of methylene chloride and 100mL of water, the organic phase was separated, anhydrous sodium sulfate Dry, filter, and concentrate the filtrate under reduced pressure to obtain a black solid, and proceed directly to the next reaction;
将上述所得的黑色体转入一个250mL单口圆底烧瓶中,加入150mL二氯甲烷,缓慢搅拌,通入溴化氢气体至饱和,继续搅拌,TLC监控反应,反应结束后,减压浓缩至干,直接进行下一步反应;Transfer the black body obtained above into a 250mL single-necked round-bottom flask, add 150mL of dichloromethane, slowly stir, pass hydrogen bromide gas to saturation, continue stirring, TLC monitor the reaction, after the reaction is completed, concentrate to dryness under reduced pressure , Proceed directly to the next reaction;
将上述所得的浓缩物转入一个500mL三口圆底烧瓶中,称入化合物(IV-6)(0.40g,2.17mmol),加入50mL二氯甲烷,搅拌,冰浴冷却下,滴加30mL三乙胺的二氯甲烷溶液(含三乙胺2mL,14.34mmol),加完后,常温反应,TLC监控反应,反应结束后,减压浓缩至干,得到包含化合物(IIIA-1)的浓缩物,直接用于下步反应。Transfer the concentrate obtained above into a 500mL three-necked round bottom flask, weigh in compound (IV-6) (0.40g, 2.17mmol), add 50mL of dichloromethane, stir, and ice-cooled, add 30mL of triethylbenzene dropwise A dichloromethane solution of amine (containing 2mL of triethylamine, 14.34mmol). After the addition, the reaction was performed at room temperature. The reaction was monitored by TLC. After the reaction was completed, it was concentrated to dryness under reduced pressure to obtain a concentrate containing compound (IIIA-1). Used directly in the next reaction.
第三步:化合物(I-6)的合成Step 3: Synthesis of compound (I-6)
将上步反应所得的浓缩物转入一个500mL三口圆底烧瓶中,加入150mL二氯甲烷,搅拌,接着加入正丁胺(3mL,30.35mmol),继续常温搅拌,TLC监控反应,反应结束后,减压浓缩至干,用硅胶柱层析法纯化,洗脱剂及体积比为:二氯甲烷:丙酮=100:4,得到0.3g化合物(I-6),总收率为48.3%。Transfer the concentrate from the previous reaction to a 500mL three-necked round bottom flask, add 150mL of dichloromethane, stir, then add n-butylamine (3mL, 30.35mmol), continue stirring at room temperature, TLC monitor the reaction, after the reaction, It was concentrated to dryness under reduced pressure and purified by silica gel column chromatography. The eluent and volume ratio were: dichloromethane: acetone = 100: 4, 0.3 g of compound (I-6) was obtained, and the total yield was 48.3%.
化合物(I-6)的表征数据如下:The characterization data of compound (I-6) are as follows:
MS(ESI):M/Z[M+H
+]:778.46
MS (ESI): M / Z [M + H + ]: 778.46
1H NMR(400MHz,CDCl
3)δ-0.37(s,1H),-0.03(s,1H),1.08-1.13(m,5H),1.26-1.29(m,1H),1.58-1.67(m,6H),1.71-1.73(m,3H),1.78-1.82(m,1H),1.92-2.06(m,6H),2.20(s,1H),2.28-2.39(m,3H),2.63-2.67(m,1H),3.24(s,2H),3.58(s,3H),3.66(s,3H),4.04-4.091(m,1H),4.14-4.23(m,2H),4.43-4.48(m,2H),4.51-4.63(m,1H),5.15-5.22(m,1H),5.29-5.32(m,1H),5.71-5.78(m,1H),6.94-7.09(m,2H),8.38(s,1H),8.65(s,1H),8.77(s,1H),8.84(s,1H);
1 H NMR (400MHz, CDCl 3 ) δ-0.37 (s, 1H),-0.03 (s, 1H), 1.08-1.13 (m, 5H), 1.26-1.29 (m, 1H), 1.58-1.67 (m, 6H), 1.71-1.73 (m, 3H), 1.78-1.82 (m, 1H), 1.92-2.06 (m, 6H), 2.20 (s, 1H), 2.28-2.39 (m, 3H), 2.63-2.67 ( m, 1H), 3.24 (s, 2H), 3.58 (s, 3H), 3.66 (s, 3H), 4.04-4.091 (m, 1H), 4.14-4.23 (m, 2H), 4.43-4.48 (m, 2H), 4.51-4.63 (m, 1H), 5.15-5.22 (m, 1H), 5.29-5.32 (m, 1H), 5.71-5.78 (m, 1H), 6.94-7.09 (m, 2H), 8.38 ( s, 1H), 8.65 (s, 1H), 8.77 (s, 1H), 8.84 (s, 1H);
13C NMR(100MHz,CDCl
3)δ10.70,10.91,11.94,14.06,20.75,22.74,23.59,24.18,30.22,31.10,32.19,40.06,48.16,49.29,51.50,53.81,55.37,70.18,70.95,71.51,94.93,99.77,101.45,114.12,115.43,117.15,124.63,129.8,131.06,132.78,133.40,137.01,137.30,140.79,143.29,149.83,152.20,161.18,163.93,167.73,170.25,170.34,172.46,173.59,174.04.
13 C NMR (100 MHz, CDCl 3 ) δ 10.70, 10.91, 11.94, 14.06, 20.75, 22.74, 23.59, 24.18, 30.22, 31.10, 32.19, 40.06, 48.16, 49.29, 51.50, 53.81, 55.37, 70.18, 70.95, 71.51 , 94.93,99.77,101.45,114.12,115.43,117.15,124.63,129.8,131.06,132.78,133.40,137.01,137.30,140.79,143.29,149.83,152.20,161.18,163.93,167.73,170.25,170.34,172.46,173.59,174. .
生物学评价Biological evaluation
实验动物及饲养说明Experimental animals and feeding instructions
品系:SPF级BALB/c裸鼠Strain: SPF grade BALB / c nude mouse
性别:雄性Gender: male
周龄:28-41天Weeks of age: 28-41 days
来源:北京维通利华实验动物技术有限公司Source: Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.
许可证号:SCXK(京)2012-0001License number: SCXK (Beijing) 2012-0001
饲养条件:动物饲养于实验动物房IVC系统中,5只/笼,光照周期10h/14,室内温度20-26℃,相对湿度40-70%,所有笼具、垫料及饮水在使用前均需灭菌,饲料为专用灭菌饲料,笼具、饲料及水每周更换1次,所有更换操作均在超净工作台进行。Feeding conditions: Animals are housed in the IVC system of the experimental animal house, 5 animals / cage, light period 10h / 14, indoor temperature 20-26 ℃, relative humidity 40-70%, all cages, bedding and drinking water are required before use Sterilization, the feed is special sterilized feed, cages, feed and water are replaced once a week, all replacement operations are carried out on the ultra-clean workbench.
测试例1不同给药剂量化合物(I-1)对裸鼠NCI-H460(人肺癌)移植瘤的抑瘤率Test Example 1 Tumor inhibition rate of different doses of compound (I-1) on nude mice transplanted with NCI-H460 (human lung cancer)
NCI-H460细胞于含10%胎牛血清、100U/mL青霉素、100μg/mL链霉素的RPMI-1640完全培养液中,在5%CO
2,37℃,饱和湿度恒温培养箱内培养,取 对数生长期细胞种植于裸鼠右前肢腋下,细胞接种数量2×10
6个。取接种于裸鼠腋下处于快速增殖期的NCI-H460瘤块(3-10代),除去肿瘤体中心坏死组织,将瘤块切成1mm×1mm×1mm的小瘤块,无菌条件下用套管针接种于裸鼠右前肢皮下,待瘤体长至60-120mm
3后结合体重开始分组,共5组,每组8只进行实验。
NCI-H460 cells were cultured in RPMI-1640 complete culture medium containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin in 5% CO 2 , 37 ° C, saturated humidity constant temperature incubator. Cells in the logarithmic growth phase were planted in the axilla of the right forelimb of nude mice, and the number of cells inoculated was 2 × 10 6 cells. Take the NCI-H460 tumor mass (generations 3-10) inoculated in the axillary of nude mice at a rapid proliferation stage, remove the necrotic tissue in the center of the tumor body, and cut the tumor mass into small tumor masses of 1mm × 1mm × 1mm under sterile conditions Use a trocar to inoculate subcutaneously in the right forelimb of nude mice. When the tumor grows to 60-120 mm 3 and then start grouping according to body weight, a total of 5 groups, 8 animals in each group, are tested.
动物分组后,于第二天上午重新称重,拍照,测量瘤体积后,用胰岛素注射针按20mL/kg经尾静脉注射依次上述不同剂量的药物,除HPPH外,其余各组均使用半导体780nm PDT(光动力治疗仪),且于给药0.5h后进行照射治疗,HPPH给药后24h进行照射,使用665nm PDT。动物于照射前5min左右开始用3.5%水合氯醛腹腔注射麻醉(0.1mL/10g),麻醉后用透明胶布固定于泡沫板上,用锡纸包裹裸鼠,只暴露肿瘤部位,以保证在光照过程中光斑不会偏离肿瘤。光纤探头用固定架固定,光斑面积一个平方厘米,光照强度均设为150mw/cm
2,能量密度150J/cm
2,光照时长20min;780nm PDT,探头距瘤体表面约7.0cm,光斑面积一个平方厘米,光照强度均设为100mw/cm
2,光照时长15min。动物光照结束后,继续饲养于IVC系统中,于光照射后的第11天开始,每隔4天测量移植瘤体积,直至第31天实验结束。
After grouping the animals, re-weigh the next morning, take a picture, and after measuring the tumor volume, use the insulin injection needle at 20mL / kg to inject the drugs of different doses in sequence through the tail vein. Except for HPPH, all other groups use semiconductor 780nm PDT (Photodynamic Therapeutic Apparatus), and irradiation treatment was performed 0.5h after administration, and irradiation was performed 24h after HPPH administration, using 665nm PDT. Animals were anesthetized with a 3.5% chloral hydrate intraperitoneal injection (0.1mL / 10g) about 5 minutes before irradiation. After anesthesia, they were fixed on a foam board with transparent tape and wrapped in tin foil to expose only the tumor site to ensure the light process The medium spot will not deviate from the tumor. The optical fiber probe is fixed with a fixed frame, the spot area is one square centimeter, the light intensity is set to 150mw / cm 2 , the energy density is 150J / cm 2 , the illumination time is 20min; 780nm PDT, the probe is about 7.0cm away from the tumor surface, the spot area is one square In centimeters, the light intensity is set to 100mw / cm 2 and the light duration is 15min. After the animals were irradiated, they were kept in the IVC system, and the volume of the transplanted tumor was measured every 4 days from the 11th day after the light irradiation until the end of the experiment on the 31st day.
不同给药剂量化合物(I-1)对NCI-H460移植瘤体积(TV)的影响分别如图1、和表1所示,给药剂量为0.67mg/kg、1.0mg/kg或1.5mg/kg时,化合物(I-1)都能对NCI-H460移植瘤产生良好的抑制作用。The effects of different doses of compound (I-1) on the NCI-H460 transplanted tumor volume (TV) are shown in Figure 1 and Table 1, respectively. The doses are 0.67mg / kg, 1.0mg / kg or 1.5mg / At kg, compound (I-1) can produce a good inhibitory effect on NCI-H460 transplanted tumors.
化合物(I-1)不同给药剂量对NCI-H460移植瘤的抑瘤率如表2所示,其中,抑瘤率(%)计算公式:The inhibition rate of compound (I-1) at different doses on NCI-H460 transplanted tumors is shown in Table 2, where the calculation formula of inhibition rate (%):
抑瘤率(%)=[(模型对照的平均瘤重-实验组的平均瘤重)/实验组的平均瘤重]×100%Tumor inhibition rate (%) = [(average tumor weight of model control-average tumor weight of experimental group) / average tumor weight of experimental group] × 100%
表1不同给药剂量化合物(I-1)对NCI-H460移植瘤体积的影响(单位:mm
3)
Table 1 The effect of different doses of compound (I-1) on the volume of NCI-H460 transplanted tumor (unit: mm 3 )
表2不同给药剂量化合物(I-1)对NCI-H460移植瘤的抑瘤率(%)Table 2 Tumor inhibition rate (%) of different doses of compound (I-1) on NCI-H460 transplanted tumor
组别Group | 平均瘤重/gAverage tumor weight / g | 抑瘤率(%)Tumor inhibition rate (%) |
模型组Model group | 1.5901±0.91721.5901 ± 0.9172 | N/AN / A |
HPPH(1.5mg/kg)HPPH (1.5mg / kg) | 1.2366±0.79391.2366 ± 0.7939 | 22.2322.23 |
化合物(I-1)(0.67mg/kg)Compound (I-1) (0.67mg / kg) | 0.2679±0.21460.2679 ± 0.2146 | 83.1583.15 |
化合物(I-1)(1.0mg/kg)Compound (I-1) (1.0mg / kg) | 0.1422±0.11590.1422 ± 0.1159 | 91.0691.06 |
化合物(I-1)(1.5mg/kg)Compound (I-1) (1.5mg / kg) | 0.0954±0.09580.0954 ± 0.0958 | 94.0194.01 |
由表1中的数据可以看出,在光动力治疗后,化合物(I-1)各治疗组肿瘤体积均逐渐减小,11-15天左右达到最小值,HPPH组同样体现了抑制裸鼠移植瘤的作用,但仅仅使肿瘤生长变得缓慢。同时,化合物(I-1)给药剂量在0.67mg/kg-1.5mg/kg时,体现良好的剂量效应。由表2中的数据可以看出,化合物(I-1)给药剂量为0.67mg/kg-1.5mg/kg时,对比1.5mg/kg剂量的HPPH具有更好的抑制NCI-H460移植瘤的肿瘤生长作用,且抑制率能达到80%以上,化合物(I-1)在抑制NCI-H460移植瘤的肿瘤生长方面具有很大的优势。As can be seen from the data in Table 1, after photodynamic therapy, the tumor volume of each treatment group of compound (I-1) gradually decreased, reaching a minimum value in about 11-15 days, and the HPPH group also reflected the inhibition of nude mouse transplantation. The role of the tumor, but only makes the tumor grow slower. At the same time, when the dose of compound (I-1) is 0.67mg / kg-1.5mg / kg, it shows a good dose effect. It can be seen from the data in Table 2 that when compound (I-1) is administered at a dose of 0.67 mg / kg-1.5 mg / kg, it has better inhibition of NCI-H460 transplanted tumor than HPPH at a dose of 1.5 mg / kg. The tumor growth effect, and the inhibition rate can reach more than 80%, compound (I-1) has a great advantage in inhibiting the tumor growth of NCI-H460 transplanted tumors.
其中HPPH的结构式如下所示:The structural formula of HPPH is as follows:
测试例2本申请化合物(I-1)的药代动力学实验Test Example 2 Pharmacokinetic experiment of compound (I-1) of the present application
NCI-H460细胞于含10%胎牛血清、100U/mL青霉素、100μg/mL链霉素的RPMI-1640完全培养液中,在5%CO
2,37℃,饱和湿度恒温培养箱内培养,取对数生长期细胞种植于裸鼠右前肢腋下,细胞接种数量2×10
6个。取接种于裸鼠腋下处于快速增殖期的NCI-H460瘤块(3-10代),除去肿瘤体中心坏死组织,将瘤块切成1mm×1mm×1mm的小瘤块,无菌条件下用套管针接种于裸鼠右前肢皮下,待瘤体长至80-150mm
3后结合体重开始分组进行实验,使每组小鼠体重肿瘤体积平均值趋于一致。经荷瘤裸鼠,肿瘤体积为80-150mm
3,雄性,体重21-25g。
NCI-H460 cells were cultured in RPMI-1640 complete culture medium containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin in 5% CO 2 , 37 ° C, saturated humidity constant temperature incubator. Cells in the logarithmic growth phase were planted in the axilla of the right forelimb of nude mice, and the number of cells inoculated was 2 × 10 6 cells. Take the NCI-H460 tumor mass (generations 3-10) inoculated in the axillary of nude mice at a rapid proliferation stage, remove the necrotic tissue in the center of the tumor body, and cut the tumor mass into small tumor masses of 1mm × 1mm × 1mm under sterile conditions Use a trocar to inoculate subcutaneously in the right forelimb of nude mice. After the tumor grows to 80-150 mm 3 , combine the body weight to start grouping experiments, so that the average tumor volume of each group of mice tends to be consistent. The tumor-bearing nude mice have a tumor volume of 80-150 mm 3 , a male body weight of 21-25 g.
裸鼠接种荷瘤后,分成3只一组,共10组。常规尾静脉注射给药,心脏穿刺采血,乙二胺四乙酸二钾(EDTA-K2)作为抗凝剂抗凝。分别于化合物(I-1)(3.0mg/kg)给药后第1、2、4、8、12小时和HPPH(0.6mg/kg)给药后5、24、48、96h各时间点杀死裸鼠取得抗凝血浆;进行肿瘤、皮肤和肌肉组织匀浆和甲醇沉淀萃取后,进样高效液相系统进行含量测定,化合物(I-1)在不同组织的药物浓度-时间曲线如图2所示,其相对应时间点的药物浓度见表3;HPPH在不同组织的药物浓度-时间曲线如图3所示,HPPH各组织药物浓度见表4。After inoculated with tumors, nude mice were divided into 3 groups of 10 groups. Conventional tail vein injection, cardiac puncture and blood collection, dipotassium ethylenediaminetetraacetate (EDTA-K2) as anticoagulant. Killed at 1, 24, 48, and 96 hours after administration of compound (I-1) (3.0 mg / kg) at 1, 2, 4, 8, 12 hours, and after HPPH (0.6 mg / kg) administration, respectively Dead nude mice obtained anticoagulated plasma; after tumor, skin and muscle tissue homogenate and methanol precipitation extraction, the sample was injected into a high-performance liquid system for content determination. The drug concentration-time curve of compound (I-1) in different tissues is shown in the figure As shown in 2, the drug concentration at the corresponding time point is shown in Table 3; the drug concentration-time curve of HPPH in different tissues is shown in Figure 3, and the drug concentration of each HPPH tissue is shown in Table 4.
表3化合物(I-1)药代动力学参数的时间-浓度数据Table 3 Time-concentration data of compound (I-1) pharmacokinetic parameters
BLOQ:表示药物含量极低,低于高效液相检测限。BLOQ: It means that the drug content is extremely low, which is lower than the detection limit of high performance liquid phase.
表4 HPPH药代动力学参数的时间-浓度数据Table 4 Time-concentration data of HPPH pharmacokinetic parameters
BLOQ:表示药物含量极低,低于高效液相检测限。BLOQ: It means that the drug content is extremely low, which is lower than the detection limit of high performance liquid phase.
表5 3.0mg/kg剂量的化合物(I-1)在血浆中的药代动力学参数Table 5 Pharmacokinetic parameters of 3.0 mg / kg dose of compound (I-1) in plasma
K el K el | t 1/2 t 1/2 | AUC 0-T AUC 0-T | AUV 0-inf AUV 0-inf | AUMC 0-T AUMC 0-T | AUMC 0-inf AUMC 0-inf | MRT IV MRT IV | ClCl | VD ss VD ss |
0.4530.453 | 1.51.5 | 2435824358 | 2442424424 | 3012030120 | 3105531055 | 1.31.3 | 2.052.05 | 0.1560.156 |
从表3的数据中可以看出,化合物(I-1)在给药3.0mg/kg的剂量浓度下,在给药1小时左右有比较好的组织分布选择性,可减少副作用,减弱了对周围组织的损伤;其在皮肤中检出很少,药物代谢较快,预示着临床治疗后避光周期较短。As can be seen from the data in Table 3, at a dose concentration of 3.0 mg / kg, compound (I-1) has a better tissue distribution selectivity after about 1 hour of administration, which can reduce side effects and weaken the Damage to surrounding tissues; it is rarely detected in the skin and the drug is metabolized faster, which indicates a shorter period of avoiding light after clinical treatment.
从表4的数据中可以看出在注射后24h的时间点,HPPH在肿瘤与皮肤中的含量相近,选择性较差。注射后48小时,仍可在皮肤组织中检出HPPH,意味着HPPH在体内代谢很慢,临床上光动力治疗后需要较长的避光周期。It can be seen from the data in Table 4 that at 24 hours after injection, the content of HPPH in the tumor and skin is similar and the selectivity is poor. 48 hours after the injection, HPPH can still be detected in the skin tissue, which means that HPPH is metabolized slowly in the body, and a longer period of light avoidance is needed after photodynamic therapy in clinic.
对比表3、4中实验数据,化合物(I-1)血浆药物浓度从第1小时的7217ng/ml到第2小时的2600ng/kg,药物浓度降低了约2/3;HPPH的药物浓度从第5小时的1873ng/ml下降到24h的633ng/ml,经过19h药物浓度同样降低了约2/3。按照相同比例换算,HPPH的半衰期(t
1/2)约为化合物(I-1)的19倍。可见,与HPPH相比,化合物(I-1)大幅度缩短了药物半衰期,在临床上可以大幅度减少光动力治疗后的避光时间。
Comparing the experimental data in Tables 3 and 4, the plasma drug concentration of compound (I-1) decreases from 7217 ng / ml at the first hour to 2600 ng / kg at the second hour, and the drug concentration decreases by about 2/3; The 1873ng / ml at 5 hours dropped to 633ng / ml at 24h, and the drug concentration also decreased by about 2/3 after 19h. Converted in the same ratio, the half-life (t 1/2 ) of HPPH is about 19 times that of compound (I-1). It can be seen that, compared with HPPH, Compound (I-1) significantly shortens the half-life of the drug, and can clinically significantly reduce the time for avoiding light after photodynamic therapy.
从表5的数据中可以看出,化合物(I-1)在血浆中的半衰期极短,为1.5h,同时,由图2可以看出,给药约8小时后,各组织中药物浓度极低,由此说明化合物(I-1)在体内各组织中代谢很快。It can be seen from the data in Table 5 that the half-life of compound (I-1) in plasma is extremely short, 1.5 hours. At the same time, as can be seen from FIG. 2, after about 8 hours of administration, the drug concentration in each tissue Low, which indicates that compound (I-1) is rapidly metabolized in various tissues in the body.
根据文献(Molecular Pharmaceutics,2011,8(4):1186-1197)报道,HPPH在小鼠身上尾静脉注射后,需要24小时后进行光动力治疗,治疗后仍需要48h,皮肤荧光量才减至微弱(荧光量与HPPH的含量成正比)。由此可判断,化合物(I-1)在半衰期较短的优势下,其避光周期(避免皮肤中的药物在日光照射下产生皮肤光毒性)也比较短,在光动力治疗中具有很大的优势。According to the literature (Molecular Pharmaceutics, 2011, 8 (4): 1186-1197), after injection of HPPH into the tail vein of mice, photodynamic therapy needs to be performed 24 hours later, and it still takes 48h after treatment to reduce the amount of skin fluorescence to Weak (fluorescence is proportional to the HPPH content). From this, it can be judged that the compound (I-1) has a shorter half-life, and its light avoidance period (to avoid skin phototoxicity of drugs in the skin under sunlight) is relatively short, and it has a great effect in photodynamic therapy. The advantages.
结论:in conclusion:
测试例1中,化合物(I-1)在给药剂量0.67mg/kg-1.5mg/kg之间体现了对裸鼠移植瘤肿瘤抑制效应的量效关系,证明了其有效性;且其在较低剂量时在本测试例中的瘤株上抑瘤率显著高于HPPH组,具有优势。In Test Example 1, the compound (I-1) exemplified the dose-effect relationship of the tumor suppressive effect on nude mice xenografts between 0.67 mg / kg and 1.5 mg / kg, and proved its effectiveness; At lower doses, the tumor inhibition rate of the tumor strains in this test case was significantly higher than that of the HPPH group, which had an advantage.
测试例2中,化合物(I-1)在尾静脉注射3.0mg/kg剂量给药后,其在皮肤、肌肉组织的分布量极少,具有较好选择性。从表3和表4的数据中可以计算得出,HPPH的半衰期(t
1/2)约为化合物(I-1)的19倍,化合物(I-1)相对于HPPH 来说,代谢更快,在临床上可以大幅度减少病人治疗后的避光时间和住院护理时间。
In Test Example 2, after the compound (I-1) was administered at a dose of 3.0 mg / kg in the tail vein, its distribution in the skin and muscle tissue was extremely small, and it had good selectivity. It can be calculated from the data in Tables 3 and 4 that the half-life (t 1/2 ) of HPPH is about 19 times that of compound (I-1), and compound (I-1) is metabolized faster than HPPH In the clinic, it can greatly reduce the time for patients to avoid light and stay in hospital after treatment.
综上所述,本发明化合物与HPPH相比具有优势:本发明化合物具有更好的抑瘤效果和较好的肿瘤选择性;本发明化合物半衰期较短,有益于在临床应用中缩短治疗周期。In summary, the compound of the present invention has advantages over HPPH: the compound of the present invention has better tumor suppressing effect and better tumor selectivity; the compound of the present invention has a shorter half-life, which is beneficial to shorten the treatment cycle in clinical application.
Claims (16)
- 一种式(I)化合物或其立体异构体、或其可药用的盐:A compound of formula (I) or its stereoisomer, or a pharmaceutically acceptable salt thereof:其中,among them,R 1为氢或(CH 2) mCOOH; R 1 is hydrogen or (CH 2 ) m COOH;R 2选自氢、羟基、卤素、C 1-C 6烷基或C 1-C 6烷氧基; R 2 is selected from hydrogen, hydroxyl, halogen, C 1 -C 6 alkyl or C 1 -C 6 alkoxy;R 3,R 4各自独立地选自C 1-C 6烷基; R 3 and R 4 are each independently selected from C 1 -C 6 alkyl;m为1、2、3或4;且m is 1, 2, 3 or 4; andn为1、2、3或4。n is 1, 2, 3 or 4.
- 根据权利要求1所述的化合物或其立体异构体、或其可药用的盐,其为式(II)所示的化合物或其立体异构体、或其可药用的盐:The compound according to claim 1 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which is a compound represented by formula (II) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof:其中,R 2,R 3,R 4,m和n的定义如权利要求1中所述。 Wherein R 2 , R 3 , R 4 , m and n are as defined in claim 1.
- 根据权利要求1所述的化合物或其立体异构体、或其可药用的盐,其为式(III)所示的化合物或其立体异构体、或其可药用的盐:The compound according to claim 1 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which is a compound represented by formula (III) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof:其中,R 2,R 3,R 4和n的定义如权利要求1中所述。 Wherein, R 2 , R 3 , R 4 and n are as defined in claim 1.
- 根据权利要求1-3任一项所述的化合物或其立体异构体、或其可药用的盐,其特征在于,R 3为甲基、乙基、丙基、丁基或己基。 The compound according to any one of claims 1 to 3, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein R 3 is methyl, ethyl, propyl, butyl, or hexyl.
- 根据权利要求1-3任一项所述的化合物或其立体异构体、或其可药用的盐,其特征在于,R 4为甲基、乙基、丙基、丁基或己基。 The compound according to any one of claims 1 to 3, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein R 4 is methyl, ethyl, propyl, butyl, or hexyl.
- 根据权利要求1-3任一项所述的化合物或其立体异构体、或其可药用的盐,其特征在于,所述R 2选自氢、卤素、C 1-C 6烷基或C 1-C 6烷氧基。 The compound according to any one of claims 1-3, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein R 2 is selected from hydrogen, halogen, C 1 -C 6 alkyl or C 1 -C 6 alkoxy.
- 根据权利要求6所述的化合物或者其立体异构体、或其可药用的盐,其特征在于,所述R 2选自氢、氟、甲基或甲氧基。 The compound according to claim 6, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein R 2 is selected from hydrogen, fluorine, methyl or methoxy.
- 一种根据权利要求2所述的式(II)化合物或其立体异构体、或其可药用的盐的制备方法,所述方法包括:A method for preparing a compound of formula (II) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to claim 2, the method comprising:式(IIA)化合物在碱性条件下进行水解反应,然后加酸进行酸化得到式(II)化合物;The compound of formula (IIA) is hydrolyzed under basic conditions, and then acidified by adding acid to obtain the compound of formula (II);其中:R 2,R 3,R 4,m和n的定义如权利要求1中所述。 Wherein: R 2 , R 3 , R 4 , m and n are as defined in claim 1.
- 根据权利要求9所述的制备方法,其特征在于,所述碱性条件所用的碱选自碱金属碳酸盐或碳酸铵,优选碱金属碳酸盐,更优选碳酸钠、碳酸钾或碳酸锂;所述的酸选自无机酸或有机酸,其中,无机酸选自质量分数为1-30%的盐酸或质量分数为1-50%的硫酸,优选质量分数为1-10%的盐酸或质量分数为1-10%的硫酸,有机酸选自C 1-C 4的有机酸,优选甲酸、乙酸或草酸。 The preparation method according to claim 9, wherein the base used in the alkaline condition is selected from alkali metal carbonate or ammonium carbonate, preferably alkali metal carbonate, more preferably sodium carbonate, potassium carbonate or lithium carbonate The acid is selected from inorganic acids or organic acids, wherein the inorganic acid is selected from hydrochloric acid with a mass fraction of 1-30% or sulfuric acid with a mass fraction of 1-50%, preferably hydrochloric acid with a mass fraction of 1-10% or content of 1-10% sulfuric acid, organic acids selected from C 1 -C 4, preferably formic acid, acetic acid or oxalic acid.
- 一种根据权利要求3所述的式(III)化合物或其立体异构体、或其可药用的盐的制备方法,所述方法包括:A method for preparing a compound of formula (III) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to claim 3, the method comprising:式(IIIA)化合物在碱性条件下水解得到式(III)化合物,The compound of formula (IIIA) is hydrolyzed under basic conditions to obtain the compound of formula (III),其中:R 2,R 3,R 4和n的定义如权利要求1中所述。 Wherein: R 2 , R 3 , R 4 and n are as defined in claim 1.
- 根据权利要求11所述的制备方法,其特征在于,所述碱性条件所用的碱选自碱金属碳酸盐、碳酸铵或有机胺,优选碱金属碳酸盐或C 1-C 6的伯胺,更优选碳酸钠、碳酸钾、碳酸锂、正丙胺或丁胺。 The preparation method according to claim 11, characterized in that the base used in the alkaline conditions is selected from alkali metal carbonates, ammonium carbonates or organic amines, preferably alkali metal carbonates or C 1 -C 6 primary The amine is more preferably sodium carbonate, potassium carbonate, lithium carbonate, n-propylamine or butylamine.
- 一种药物组合物,所述的药物组合物含有有效剂量的根据权利要求1-8中任一项所述的化合物或其立体异构体、或其可药用的盐,及其可药用的载体、赋形剂或它们的组合。A pharmaceutical composition containing an effective dose of a compound according to any one of claims 1-8 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable Carrier, excipient or combination thereof.
- 根据权利要求1-8中任一项所述的化合物或其立体异构体、或其可药用的盐,或根据权利要求15所述的药物组合物在制备用于光动力治疗癌症的药物中的应用,其中所述癌症优选为肺癌、前列腺癌、食管癌、黑色素瘤、胆管癌、头颈癌、膀胱癌、胃癌、鼻咽癌、结肠癌中的一种或多种。The compound according to any one of claims 1-8 or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 15 in the preparation of a drug for photodynamic therapy of cancer In the application, wherein the cancer is preferably one or more of lung cancer, prostate cancer, esophageal cancer, melanoma, cholangiocarcinoma, head and neck cancer, bladder cancer, gastric cancer, nasopharyngeal cancer, colon cancer.
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CN111171030A (en) | 2020-05-19 |
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