Sildenafil hapten, sildenafil artificial antigen and preparation method of sildenafil artificial antigen

Abstract

The invention discloses a sildenafil hapten, a sildenafil artificial antigen and a preparation method of the sildenafil artificial antigen and is intended to provide a sildenafil hapten, a sildenafilartificial antigen and a preparation method of the sildenafil artificial antigen. Connection arm structures and active groups for coupling macromolecules are introduced in the hapten at the premise ofretaining the basic structure of sildenafil, and therefore, coupling of sildenafil with macromolecules is benefited, molecular structure of sildenafil itself and the basic structure of sildenafil having low molecular weight are fully exposed after coupling, and recognition of animal bodies due to macromolecular covering of sildenafil is avoided. The hapten and a protein are coupled to prepare theartificial antigen through an active esterification process, the coupling ratio may reach 10.7, the artificial antigen is suitable for immunoassay of sildenafil and has the molecular structural formula that is shown in the description.

Description

technical field

[0001] The invention discloses a hapten and an artificial antigen, and in particular relates to a sildenafil hapten and an artificial antigen. The invention also discloses a preparation method of the sildenafil hapten and the artificial antigen. Background technique

[0002] Sildenafil (Sildenafil), that is, Sildenafil Citrate Tablets, also known as Viagra, commonly known as "Viagra", molecular formula: C22H30N6O4S, CAS registration number is 139755-83-2, can effectively improve the hardness of penile erection, used for Treats erectile dysfunction (ED, also known as impotence) in men. The drug has been on the market for more than 20 years and has made revolutionary progress in the treatment of ED. Oral drugs represented by it are recommended as the first-line treatment of ED by the medical guidelines of many countries and regions such as the United States, Europe, China, and Japan.

[0003] The most common side effects of sildenafil are: headache, flushing,...

Examples

preparation example Construction

[0038] In the embodiment provided by the present invention, the preparation method of the hapten shown in formula I comprises steps:

[0039] After mixing 1g of sildenafil with 5-13g of cyclic anhydride, dissolve it in 30g of 1,4-dioxane, and then add 5 drops of triethylamine dropwise, so that the pH of the reaction solution is 7.5, 40- 50 ℃ airtight stirring reaction 10-12h. After the reaction finished, the reaction solution was poured into water, extracted 3 times with 100g ethyl acetate, the combined extracts were washed with anhydrous Na 2 SO 4 After drying and concentrating, a solid is obtained, and the solid product obtained by recrystallization from 75% ethanol is the hapten of sildenafil.

[0040] In the embodiment provided by the present invention, the preparation method of sildenafil artificial antigen as shown in formula II comprises steps:

[0041] 1) The hapten shown in formula I, N-hydroxysuccinimide (NHS), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), ...

Embodiment 1

[0049] The preparation of embodiment 1 sildenafil hapten (n=1)

[0050] After mixing 1g of sildenafil and 10g of succinic anhydride, dissolve it in 30g of 1,4-dioxane, then add 5 drops of triethylamine drop by drop, so that the pH of the reaction solution is 7.5, and keep stirring at 40°C Reaction 12h. After the reaction finished, the reaction solution was poured into water, extracted 3 times with 100g ethyl acetate, the combined extracts were washed with anhydrous Na 2 SO 4 After drying and concentrating, a solid is obtained, and the solid product obtained by recrystallization from 75% ethanol is a sildenafil derivative (hapten).

[0051] The structure of the resulting product is characterized by: 1 H NMR (400MHz, DMSO) δ8.33(s, J=7.9Hz, 1H), 8.01(d, J=7.6Hz, 1H), 7.27(d, J=7.6Hz, 1H), 4.08(m, J =3.8Hz, 2H), 4.00(s, J=4.4Hz, 4H), 3.06(t, J=4.8Hz, 4H), 2.63(t, J=4.7Hz, 2H), 2.49(t, J=3.4 Hz, 2H), 2.40(t, J=4.2Hz, 2H), 2.35(t, J=4.8Hz, 4H), 2.18(s, J=4.6Hz, 3H), 1.72(m, J=...

Embodiment 2

[0052] The preparation of embodiment 2 sildenafil hapten (n=2)

[0053] After mixing 1g of sildenafil and 5g of glutaric anhydride, dissolve it in 30g of 1,4-dioxane, then add 3 drops of triethylamine drop by drop, so that the pH of the reaction solution is 7.5, and seal it at 50°C The reaction was stirred for 10 h. After the reaction was over, the reaction solution was poured into water, extracted 3 times with 100 g of ethyl acetate, and the combined extracts were washed with anhydrous Na 2 SO 4 After drying and concentrating, a solid is obtained, and the solid product obtained by recrystallization from 75% ethanol is a sildenafil derivative (hapten).

[0054] The structure of the resulting product is characterized by: 1 H NMR (400MHz, DMSO) δ8.30(s, J=7.8Hz, 1H), 8.10(d, J=8.6Hz, 1H), 7.10(d, J=7.2Hz, 1H), 4.10(m, J =3.8Hz, 2H), 3.98(s, J=4.0Hz, 4H), 3.12(t, J=5.6Hz, 4H), 2.48(t, J=3.3Hz, 2H), 2.39(t, J=3.7 Hz, 2H), 2.30(t, J=4.8Hz, 4H), 2.26(m, J=3.4Hz, 2H), 2.15(s, J=4...

technical field

[0001] The invention discloses a hapten and an artificial antigen, and in particular relates to a sildenafil hapten and an artificial antigen. The invention also discloses a preparation method of the sildenafil hapten and the artificial antigen. Background technique

[0002] Sildenafil (Sildenafil), that is, Sildenafil Citrate Tablets, also known as Viagra, commonly known as "Viagra", molecular formula: C22H30N6O4S, CAS registration number is 139755-83-2, can effectively improve the hardness of penile erection, used for Treats erectile dysfunction (ED, also known as impotence) in men. The drug has been on the market for more than 20 years and has made revolutionary progress in the treatment of ED. Oral drugs represented by it are recommended as the first-line treatment of ED by the medical guidelines of many countries and regions such as the United States, Europe, China, and Japan.

[0003] The most common side effects of sildenafil are: headache, flushing,...

Examples

preparation example Construction

[0038] In the embodiment provided by the present invention, the preparation method of the hapten shown in formula I comprises steps:

[0039] After mixing 1g of sildenafil with 5-13g of cyclic anhydride, dissolve it in 30g of 1,4-dioxane, and then add 5 drops of triethylamine dropwise, so that the pH of the reaction solution is 7.5, 40- 50 ℃ airtight stirring reaction 10-12h. After the reaction finished, the reaction solution was poured into water, extracted 3 times with 100g ethyl acetate, the combined extracts were washed with anhydrous Na 2 SO 4 After drying and concentrating, a solid is obtained, and the solid product obtained by recrystallization from 75% ethanol is the hapten of sildenafil.

[0040] In the embodiment provided by the present invention, the preparation method of sildenafil artificial antigen as shown in formula II comprises steps:

[0041] 1) The hapten shown in formula I, N-hydroxysuccinimide (NHS), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), ...

Embodiment 1

[0049] The preparation of embodiment 1 sildenafil hapten (n=1)

[0050] After mixing 1g of sildenafil and 10g of succinic anhydride, dissolve it in 30g of 1,4-dioxane, then add 5 drops of triethylamine drop by drop, so that the pH of the reaction solution is 7.5, and keep stirring at 40°C Reaction 12h. After the reaction finished, the reaction solution was poured into water, extracted 3 times with 100g ethyl acetate, the combined extracts were washed with anhydrous Na 2 SO 4 After drying and concentrating, a solid is obtained, and the solid product obtained by recrystallization from 75% ethanol is a sildenafil derivative (hapten).

[0051] The structure of the resulting product is characterized by: 1 H NMR (400MHz, DMSO) δ8.33(s, J=7.9Hz, 1H), 8.01(d, J=7.6Hz, 1H), 7.27(d, J=7.6Hz, 1H), 4.08(m, J =3.8Hz, 2H), 4.00(s, J=4.4Hz, 4H), 3.06(t, J=4.8Hz, 4H), 2.63(t, J=4.7Hz, 2H), 2.49(t, J=3.4 Hz, 2H), 2.40(t, J=4.2Hz, 2H), 2.35(t, J=4.8Hz, 4H), 2.18(s, J=4.6Hz, 3H), 1.72(m, J=...

Embodiment 2

[0052] The preparation of embodiment 2 sildenafil hapten (n=2)

[0053] After mixing 1g of sildenafil and 5g of glutaric anhydride, dissolve it in 30g of 1,4-dioxane, then add 3 drops of triethylamine drop by drop, so that the pH of the reaction solution is 7.5, and seal it at 50°C The reaction was stirred for 10 h. After the reaction was over, the reaction solution was poured into water, extracted 3 times with 100 g of ethyl acetate, and the combined extracts were washed with anhydrous Na 2 SO 4 After drying and concentrating, a solid is obtained, and the solid product obtained by recrystallization from 75% ethanol is a sildenafil derivative (hapten).

[0054] The structure of the resulting product is characterized by: 1 H NMR (400MHz, DMSO) δ8.30(s, J=7.8Hz, 1H), 8.10(d, J=8.6Hz, 1H), 7.10(d, J=7.2Hz, 1H), 4.10(m, J =3.8Hz, 2H), 3.98(s, J=4.0Hz, 4H), 3.12(t, J=5.6Hz, 4H), 2.48(t, J=3.3Hz, 2H), 2.39(t, J=3.7 Hz, 2H), 2.30(t, J=4.8Hz, 4H), 2.26(m, J=3.4Hz, 2H), 2.15(s, J=4...

Images