Nitric oxide generator and preparation method and application thereof

Abstract

The invention discloses a nitric oxide generator as well as a preparation method and application thereof, an (iso) quinoline structure, monoalkyl aniline and a nitric oxide donor are connected to an aza-pyrrolidine parent structure, and the I-type near-infrared nitric oxide generator is constructed. The material can be used for near-infrared two-window fluorescence imaging guided tumor photo-thermal therapy, gas therapy and I-type photodynamic synergistic therapy, and the inhibition effect on tumor growth is achieved. After the nitric oxide generator releases nitric oxide, obviously enhanced near-infrared two-window fluorescence signals, photoacoustic signals and photothermal signals are shown, and growth of tumor cells can be effectively inhibited.

Description

Technical Field

[0001] The invention relates to the fields of biochemical materials and organic optoelectronic information materials, and in particular to a nitric oxide generator and a preparation method and application thereof. Background Art

[0002] Light-regulated tumor treatment mainly includes photothermal therapy, chemotherapy, photodynamic therapy, etc. In recent years, many light-regulated near-infrared agents have been reported, and have shown good application prospects in the field of tumor treatment. At present, there are many studies on photodynamic therapy that rely on oxygen to produce singlet oxygen. However, the abnormal metabolism of tumor cells leads to hypoxia in the tumor site, which seriously weakens the effect of photodynamic therapy. In addition, the abnormal metabolism of tumor cells causes blood vessels in the tumor site to rupture and block, which is not conducive to the uniform distribution of agents in the tumor site, further reducing the photodynamic therapy effect of the tumor. Thermal therapy can effectively promote blood circulation and promote the distribution of agents in the tumor site, thereby effectively improving the treatment of tumors.

[0003] In type I photodynamic therapy, the production of ·OH does not rely on oxygen and has a good therapeutic effect on hypoxic tumors. However, due to the need for energy level matching, photosensitizers used in type I photodynamic therapy for hypoxic tumors are usually excited by visible light. Compared with short wavelengths, long wavelengths have deeper penetration depths and cause less damage to the body. Therefore, it is very necessary to design and synthesize easily available nitric oxide generators for near-infrared two-window fluorescence imaging-guided tumor photothermal therapy, gas therapy, and type I photodynamic therapy.

[0004] In order to solve the problems in the prior art, the present invention connects an (iso)quinoline structure, a monoalkylaniline, and a nitric oxide donor to an azapyrrolidine parent structure to construct a nitric oxide generator. The release of nitric oxide (for tumor gas therapy) is utilized to activate its near-infrared two-window fluorescence imaging performance, thereby guiding tumor photothermal therapy and type I photodynamic synergistic therapy, thereby achieving an inhibitory effect on tumor growth.

[0005] The present invention provides a nitric oxide generator that can be excited by near-infrared light and release NO to achieve photothermal therapy, gas therapy, and type I photodynamic tumor therapy under the guidance of near-infrared two-window fluorescence imaging. After the nitric oxide generator releases nitric oxide, it shows significantly enhanced near-infrared two-region fluorescence signals, photoacoustic signals, and photothermal signals, and can effectively inhibit the growth of tumor cells. Summary of the invention

[0006] In view of this, the present invention provides a nitric oxide generator and a preparation method and application thereof.

[0007] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0008] A nitric oxide generator, wherein the nitric oxide generator has any one of the structures of Formulas I to III:

[0009]

[0010] Wherein, Z1 and Z2 are either C or N, and Z1 and Z2 are different;

[0011] Z3 and Z4 are either C or N, and Z3 and Z4 are different.

[0012] A method for preparing a nitric oxide generator comprises the following steps:

[0013] S1, compound 1' and compound 2' react in a mixed solution of water and alcohol of sodium hydroxide to obtain compound 3;

[0014] S2, compound 3' reacts with nitromethane to obtain compound 4';

[0015] S3, compound 4' reacts in a mixed solution of alcohol and ammonium acetate to obtain compound 5';

[0016] S4, compound 5' undergoes coordination reaction with boron trifluoride ether to obtain compound 6';

[0017] S5, compound 6' reacts with sodium nitrite to obtain the structure shown in formula I;

[0018] Among them, compound 1' is Compound 2' is Compound 3' is Compound 4' is Compound 5' is Compound 6' is

[0019] Preferably, in step S1, the mixed solution of sodium hydroxide in water and alcohol is H2O:ROH=1:10, and equimolar amounts of compound 1' and compound 2' undergo addition elimination reaction at room temperature for 3-24 hours to obtain compound 3'.

[0020] Preferably, in step S2, the reaction is carried out in methanol or ethanol, ROH weakly alkaline diethylamine solution, and the reaction temperature is 65-85°C; in step S3, the reaction temperature is 85-100°C; in step S4, the reaction is carried out under weak alkaline conditions; in step S5, the reaction is carried out under acidic conditions.

[0021] Preferably, the method further comprises the following steps:

[0022] SA, compound 1' and compound 2-1' react in a mixed solution of water and alcohol of sodium hydroxide to obtain compound 3-1';

[0023] SB, compound 3-1' and nitromethane react to obtain compound 4-1';

[0024] SC and compound 4-1' react in a mixed solution of alcohol and ammonium acetate to obtain compound 5-1';

[0025] SD, compound 5-1' and boron trifluoride ether undergo coordination reaction to obtain compound 6-1';

[0026] SE, compound 6-1' and sodium nitrite react to obtain the structure shown in formula II;

[0027] Among them, compound 1' is Compound 2-1' is Compound 3-1' is Compound 4-1' is Compound 5-1' is Compound 6-1' is

[0028] A nitric oxide generator and a method for preparing the nitric oxide generator are used in preparing drugs for treating tumors.

[0029] Preferably, the nitric oxide generator releases nitric oxide through near-infrared light stimulation to achieve gas therapy for tumors.

[0030] Preferably, the nitric oxide generator generates reactive oxygen species (including type I reactive oxygen species) through near-infrared light stimulation for type I photodynamic therapy of tumors.

[0031] Preferably, the phototherapy agent produced by releasing nitric oxide exhibits near-infrared second-window fluorescence emission and significantly enhanced photoacoustic signals and photothermal conversion performance, and can be used for near-infrared second-window fluorescence imaging-guided photothermal therapy of hypoxic tumors.

[0032] Preferably, the application is:

[0033] The phototherapy agent produced by releasing nitric oxide exhibits significantly enhanced near-infrared second-window fluorescence emission, photoacoustic signal and photothermal conversion performance, and can be used for near-infrared second-window fluorescence imaging-guided photothermal therapy of hypoxic tumors.

[0034] Preferably, the application method is:

[0035] The nitric oxide generator is used for photothermal therapy and type I photodynamic therapy of hypoxic tumors. Under near-infrared light excitation, the nitric oxide generator generates heat and OH, which are used for photothermal therapy and type I photodynamic therapy of tumors guided by near-infrared two-window fluorescence imaging.

[0036] Compared with the prior art, the present invention has achieved the following technical effects:

[0037] The nitric oxide generator of the present invention is a near-infrared light-excitable (iso)quinoline structure combined with monoalkylaniline and nitric oxide donor connected to an azapyrrolidine skeleton; under the irradiation of near-infrared light, the nitric oxide generator can produce toxic substances such as reactive oxygen through a photoinduced electron transfer mechanism to kill tumor cells, including type I photodynamic therapy for tumors; under the irradiation of a long-wavelength light source, the phototherapy agent exhibits significantly enhanced photothermal effects and photothermal, photoacoustic, and fluorescent signals; the increase in temperature can also promote the circulation of blood in the tumor site, further promote the uniform distribution of the nitric oxide generator in the tumor site, and improve the effects of photodynamic and photothermal therapy of tumors. This has important guiding significance for the design and synthesis of efficient nitric oxide generators. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 is the mass spectrum of A in Example 2 of the present invention;

[0039] Figure 2 is the absorption spectrum of A in Example 2 of the present invention;

[0040] Figure 3 is the fluorescence spectrum of A in Example 2 of the present invention;

[0041] Figure 4 This is a proof that after the compound A in Example 1 of the present invention releases nitric oxide, a near-infrared fluorophore with obvious red-shifted absorption is generated;

[0042] Figure 5 This is a proof that compound A in Example 1 of the present invention generates a fluorophore with near-infrared second window emission after releasing nitric oxide;

[0043] Figure 6 This is a proof that A in Example 1 of the present invention releases active oxygen under near-infrared light;

[0044] Figure 7 This is a proof that A in Example 1 of the present invention releases NO by using a commercial NO probe (Griess Reagent) under near-infrared light;

[0045] Figure 8 It is proved in Example 1 of the present invention that compound A has type I active oxygen (·OH) generation; DETAILED DESCRIPTION

[0046] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0047] Embodiment 1:

[0048] The present invention discloses a nitric oxide generator, which has any one of the structures of formulas I to III:

[0049]

[0050] Wherein, Z1 and Z2 are either C or N, and Z1 and Z2 are different;

[0051] Z3 and Z4 are either C or N, and Z3 and Z4 are different.

[0052] The present invention also discloses a method for preparing a nitric oxide generator, comprising the following steps:

[0053] Equimolar amounts of S1, compound 1' and compound 2' are added and eliminated in a mixed solution of sodium hydroxide in water and alcohol with H2O:ROH=1:10 at room temperature for 3-24 hours to obtain compound 3'.

[0054] S2, compound 3' and nitromethane are reacted in methanol or ethanol, ROH weakly alkaline diethylamine solution, the reaction temperature is 65-85 ° C to obtain compound 4';

[0055] S3, compound 4' is reacted in a mixed solution of alcohol and ammonium acetate at 85-100°C to obtain compound 5';

[0056] S4, compound 5' undergoes coordination reaction with boron trifluoride ether under weak base conditions to obtain compound 6';

[0057] S5, compound 6' and sodium nitrite react under acidic conditions to obtain the structure shown in formula I;

[0058] Among them, compound 1' is Compound 2' is Compound 3' is Compound 4' is Compound 5' is Compound 6' is

[0059] The following steps are also included:

[0060] SA, compound 1' and compound 2-1' react in a mixed solution of water and alcohol of sodium hydroxide to obtain compound 3-1';

[0061] SB, compound 3-1' and nitromethane react to obtain compound 4-1';

[0062] SC and compound 4-1' react in a mixed solution of alcohol and ammonium acetate to obtain compound 5-1';

[0063] SD, compound 5-1' and boron trifluoride ether undergo coordination reaction to obtain compound 6-1';

[0064] SE, compound 6-1' and sodium nitrite react to obtain the structure shown in formula II;

[0065] Among them, compound 1' is Compound 2-1' is Compound 3-1' is Compound 4-1' is Compound 5-1' is Compound 6-1' is

[0066] The nitric oxide generator releases nitric oxide through near-infrared light stimulation to achieve gas therapy for tumors.

[0067] The nitric oxide generator produces reactive oxygen species (including type I reactive oxygen species) through near-infrared light stimulation for type I photodynamic therapy of tumors.

[0068] The phototherapy agent produced by releasing nitric oxide exhibits significantly enhanced near-infrared second-window fluorescence emission, photoacoustic signal and photothermal conversion performance, and can be used for near-infrared second-window fluorescence imaging-guided photothermal therapy of hypoxic tumors.

[0069] The application method is:

[0070] Under near-infrared light excitation conditions, the nitric oxide generator produces heat, nitric oxide and ·OH, and simultaneously produces near-infrared two-window fluorescence signals and significantly enhanced photoacoustic signals and photothermal properties, which can be used for near-infrared two-window fluorescence imaging-guided tumor photothermal therapy, type I photodynamic therapy and gas therapy.

[0071] Embodiment 2:

[0072] The chemical structure and synthesis route of nitric oxide generator I are as follows:

[0073]

[0074] in,

[0075] Synthesis method of compound 3:

[0076] To a mixture of 3.82 g of p-diethylaminoacetophenone (about 20 mmol), 3.14 g (about 20 mmol) of 4-quinolinecarboxaldehyde and 30 mL of ethanol solution, 10 mL of sodium hydride solution (containing 2.00 g of sodium hydroxide) was slowly added. After reacting at room temperature for 12 h, 5.17 g of compound 3 (yield of about 78.5%) was obtained by column chromatography.

[0077] Synthesis method of compound 4:

[0078] A mixture of 3.31 g of compound 3 (about 10 mmol), 15 mL of nitromethane, 8 mL of diethylamine and 20 mL of ethanol was reacted under reflux at 75° C. overnight, and 2.74 g of compound 4 (yield of about 88%) was obtained by column chromatography.

[0079] Synthesis method of compound 5:

[0080] A mixture of compound 4, 50 g of ammonium acetate solid and 20 mL of ethanol was heated to 90° C. and refluxed for 24 h. The reaction solution was filtered under reduced pressure and washed with cold ethanol to obtain compound 5 (yield of about 20.1%), which was directly used in the next step reaction.

[0081] Synthesis method of compound 6:

[0082] In a mixture of 0.139 g (0.2 mmol) of compound 5, 15 mL of dry dichloromethane, and 2 mL of diisopropylethylamine, 3 mL of boron trifluoride etherate was slowly added and reacted at room temperature for 4 hours. After the reaction was completed, the excess boron trifluoride etherate was quenched with ethanol, the low boiling point solvent was removed under low pressure, and the solid was filtered under reduced pressure and washed with cold ethanol. The target compound A was finally obtained by separation on a silica gel plate with a yield of about 13.1%.

[0083] Experiments have shown that under the irradiation of longer wavelength near-infrared light, the target material can not only produce NO for gas therapy of tumors, but also show obvious photothermal conversion ability and photoacoustic, photothermal, and fluorescence signals, and can dilate blood vessels to improve the uniform distribution of materials in the tumor site. It can be used for tumor photothermal therapy and gas therapy under the guidance of near-infrared two-window multi-mode imaging (photothermal imaging, fluorescence imaging).

[0084] The above description is only a preferred embodiment of the present invention and does not limit the technical scope of the present invention. Therefore, any slight modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention are still within the scope of the technical solution of the present invention.

Claims

1. A nitric oxide generator, characterized in that: The nitric oxide generator has any one of the structures of Formulas I to III: Wherein, Z1 and Z2 are either C or N, and Z1 and Z2 are different; Z3 and Z4 are either C or N, and Z3 and Z4 are different.

2. A method for preparing a nitric oxide generator, characterized in that: The following steps are involved: S1, compound 1' and compound 2' react in a mixed solution of water and alcohol of sodium hydroxide to obtain compound 3; S2, compound 3' reacts with nitromethane to obtain compound 4'; S3, compound 4' reacts in a mixed solution of alcohol and ammonium acetate to obtain compound 5'; S4, compound 5' undergoes coordination reaction with boron trifluoride ether to obtain compound 6'; S5, compound 6' reacts with sodium nitrite to obtain the structure shown in formula I; Among them, compound 1' is Compound 2' is Compound 3' is Compound 4' is Compound 5' is Compound 6' is 3. The method for preparing a nitric oxide generator according to claim 2, characterized in that: In step S1, the mixed solution of water and alcohol of sodium hydroxide is H2O:ROH=1:10, and equimolar amounts of compound 1' and compound 2' undergo addition elimination reaction at room temperature for 3-24 hours to obtain compound 3'.

4. The method for preparing a nitric oxide generator according to claim 2, characterized in that: In the step S2, the reaction is carried out in a weakly alkaline diethylamine solution of methanol or ethanol and ROH at a reaction temperature of 65-85°C; in the step S3, the reaction temperature is 85-100°C; in the step S4, the reaction is carried out under weakly alkaline conditions; in the step S5, the reaction is carried out under acidic conditions.

5. The method for preparing a nitric oxide generator according to claim 2, characterized in that: The following steps are also included: SA, compound 1' and compound 2-1' react in a mixed solution of water and alcohol of sodium hydroxide to obtain compound 3-1'; SB, compound 3-1' and nitromethane react to obtain compound 4-1'; SC and compound 4-1' react in a mixed solution of alcohol and ammonium acetate to obtain compound 5-1'; SD, compound 5-1' and boron trifluoride ether undergo coordination reaction to obtain compound 6-1'; SE, compound 6-1' and sodium nitrite react to obtain the structure shown in formula II; Among them, compound 1' is Compound 2-1' is Compound 3-1' is Compound 4-1' is Compound 5-1' is Compound 6-1' is 6. Use of the nitric oxide generator according to claim 1 and the method for preparing a nitric oxide generator according to claims 2-5 in preparing drugs for treating tumors.

7. The use according to claim 6, characterized in that: The nitric oxide generator releases nitric oxide through near-infrared light stimulation to achieve gas therapy for tumors.

8. The use according to claim 6, characterized in that: The nitric oxide generator generates reactive oxygen through near-infrared light stimulation and is used for type I photodynamic therapy of tumors.

9. The use according to claim 6, characterized in that: The phototherapy agent produced by releasing nitric oxide exhibits significantly enhanced near-infrared second-window fluorescence emission, photoacoustic signal and photothermal conversion performance, and can be used for near-infrared second-window fluorescence imaging-guided photothermal therapy of hypoxic tumors.

10. The use according to claim 8, characterized in that: The application is: The nitric oxide generator is used for photothermal therapy of hypoxic tumors and type I photodynamic therapy. Under near-infrared light excitation conditions, the nitric oxide generator produces nitric oxide, heat, and ·OH, which are used for tumor photothermal therapy, gas therapy, and type I photodynamic therapy guided by near-infrared two-window fluorescence imaging.

Images