CN107298686B - A kind of psoralen amine derivant and purposes - Google Patents

Abstract

The present invention relates to a kind of psoralen amine derivatives and its application. The derivatives use resorcinol as the starting material, and under the action of sulfuric acid, first condense with ethyl chloroacetoacetate to obtain 4-chloromethyl ‑7‑hydroxycoumarin (intermediate 1); intermediate 1 is then reacted with five different secondary amines to give 4‑substituted aminomethyl‑7‑hydroxycoumarin 2 (intermediate 2a‑2e) ; Next, four different side chains are introduced at the 7-position of intermediate 2 to obtain intermediate 3 (3a-3t); finally, intermediate 3 is ring-closed in potassium hydroxide ethanol solution to form psoralen, The psoralen amine derivative 4 (4a‑4t) was finally obtained. And the obtained 20 psoralen amine derivatives were tested for melanogenesis and tyrosinase activity in mouse B16 cells, the results: compounds 4c-4h, 4j-4p and 4s-4t co- Fifteen psoralen amine derivatives are used in the clinical preparation of medicines for treating vitiligo.

Description

A kind of psoralen amine derivative and its application

technical field

The present invention relates to a kind of psoralen amine derivatives and its application. After the anti-vitiligo activity screening of such compounds, 15 compounds including 4c-4h, 4j-4p and 4s-4t can be used in the clinical preparation of medicine for treating vitiligo .

Background technique

Vitiligo is a common spontaneous or idiopathic depigmentation skin disease, known as one of the three chronic skin diseases in the world, afflicting more than 50 million patients worldwide. The incidence rate varies from 0.1% to 8% in different regions and different races in the world. The prevalence rate of the general population in my country is about 0.56%. About half of the patients develop the disease before the age of 20, and the prevalence rate of males and females is equal. Vitiligo mainly manifests as white spots on the skin and mucous membranes and gray/white hairs. Western medicine believes that vitiligo is caused by the dysfunction and loss of the tyrosinase system in the melanocytes of the skin and hair follicles.

Psoralen is the dried and mature fruit of Psoralea corylifolia L., an annual herbaceous plant of the leguminous family. The main classic drug for the treatment of vitiligo in Uighur medicine. Uyghur medicine believes that vitiligo is mainly caused by abnormal mucous temperament. During treatment, it is advocated to remove abnormal body fluids and correct abnormal temperament, so as to restore the natural force of the body. Psoraleae has the effects of stemming and generating heat, clearing abnormal mucus, purifying blood and detoxifying, increasing pigment, killing and repelling intestinal worms, etc. In addition, psoralen also has various biological activities such as antibacterial, estrogen-like effects, anti-tumor, anti-oxidation, immune regulation and anti-depression.

At present, the medicines and preparations with plant psoralen as the main component are mainly used to treat vitiligo clinically: such as compound psoralen granule, psoralen injection, compound psoralen tincture, Qubai Babusi tablet, compound card Li cumin tincture (Uyghur drug name Vialu), etc. Although Uyghur medicine has a remarkable effect and advantages in treating vitiligo, its material basis research is weak, and its mechanism of action and metabolic process in vivo and in vitro are not clear, which seriously restricts its secondary development.

After years of research, researchers at home and abroad have isolated psoralens (furanocoumarins), flavonoids, terpenes, and a small amount of lipids and volatile oils from Psoralea corylifolia L.. Among them, psoralen compounds (furanocoumarins) are the main chemical components, and the research scope is the most extensive.

Psoralen compounds (Psoralen) are currently the most commonly used photosensitive drugs for clinical treatment of vitiligo, but they must be treated with sunlight or long-wave ultraviolet (UVA) irradiation, so this treatment method is called PUVA (Psoralen+UVA). Commonly used are 8-methoxypsoralen (8-MOP) and 5-methoxypsoralen (5-MOP), and later artificially synthesized trimethylpsoralen (TMP), as shown in the structure shown. In the treatment of vitiligo, PUVA can activate the activity of tyrosinase, catalyze the synthesis of melanin, promote the division and movement of melanocytes, finally increase the synthesis of melanin, and gradually restore the color of leukoplakia.

Structures of three commonly used psoralen compounds

After searching, the relevant documents are:

[1]Kruger C., Schallreuter K.U.A review of the worldwide prevalence of vitiligo in children/adolescents and adults[J].Int.J.Dermatol.,2012,51,1206-1212.

[2] Wang Xiaoyan. Epidemiological survey of vitiligo in six provinces and cities in China [M]. Chinese Journal of Dermatology, 2010, 463-466.

[3] Alikhan A., Felsten L.M., Daly M., Petronic-Rosic V. Vitiligo: A comprehensive overview Part I. Introduction, epidemiology, quality of life, diagnosis, differential diagnosis, associations, histopathology, etiology, and work-up [J ].J.Am.Acad.Dermatol.,2011,65,473-491.

[4] Arcos-Burgos M., Parodi E., Salgar M., Bedoya E., Builes J., Jaramillo D., Ceballos G., Uribe A., Rivera N., Rivera D., Fonseca I., Camargo M. .,PalacioG.Vitiligo:complex segregation and linkage disequilibrium analyzes with respect to microsatellite loci spanning the HLA[J].Hum.Genet.,2002,110,334-342.

[5]Alain Vitiligo as an inflammatory skin disorder: a therapeutic perspective[J].Pigm.Cell Melanoma Res.,2012,25,9-13.

[6] Hirschhorn J.N., Lohmueller K., Byrne E., Hirschhorn K. A comprehensive review of genetic association studies [J]. Genet. Med., 2002, 4, 45-61.

[7] Schallreuter K.U., Krüger C., Rokos H., Hasse S., Zothner C., Panske A. Basic research confirms coexistence of acquired blaschkolinear Vitiligo and acrofacial vitiligo [J]. .

[8] Ezzedine K., Eleftheriadou V., Whitton M., van Geel N. Vitiligo. Lancet [J]. 2015, in press, Doi: 10.1016/S0140-6736(14) 60763-7.

[9] Tursun Uful, Slav Aibai, Refuhati Saimaiti, Julaiti Abduwaiti, Ilkal Baktia. Uyghur medicine maturation agent Observation on curative effect of 166 cases of advanced vitiligo treated with scavenger and scavenger[J]. Pharmacology and Clinic of Traditional Chinese Medicine, 2012, 28, 161-162.

[10] Xia Muxinuer Xiaogaiti, Slav Aibai, Rena Kasim. Current status of drug treatment of vitiligo [J]. Chinese Journal of Ethnic Medicine, 2011, 8, 62-64.

[11] Huang Jian, Zhao Luhua, Zou Qiaogen. Advances in Chemical Constituents and Pharmacology of Psoralen [J]. Progress in Pharmacy, 2000, 24, 212-214.

[12] Yin S., Fan C.Q., Wang Y., Dong L., Yue J.M.Antibacterial prenylflavone derivatives from Psoralea corylifolia and their structure-activity relationship study[J].Bioorg.Med.Chem.,2004,12,4387-4392 .

[13] Zhang C.Z., Wang S.X., Zhang Y., Chen J.P., Liang X.M. In vitroestrogenic activities of Chinese medicinal plants traditionally used for the management of menopausal symptoms[J].J.Ethnopharmacol.,2005,98,295-300.

[14]Bapat K., Chintalwar G.J., Pandey U., Thakur V.S., Sarma H.D., Samuel G., Pillai M.R, Chattopadhyay S., Venkatesh M. Preparation and in vitroevaluation of radio iodinated bakuchiol as an anti tumor agent[J]. Appl. Radiat. Isot., 2005, 62, 389-393.

[15] Guo J.N., Wen X.C., Wu H., Li Q.H., Bi K. Antioxidants from a Chinese medicinal herb-Psoralea corylifolia[J]. Food Chem., 2005, 91, 287-292.

[16] Latha P.G., Evans D.A., Panikkar K.R., Jayavardhanan K.K. Immunomodulatory and antitumour properties of Psoralea corylifolia seeds [J]. Fitoterapia, 2000, 71, 223-231.

[17] Chen Y., Wang H.D., Xia X., Kung H.F., Pan Y., Kong L.D. Behavioral and biochemical studies of total furocoumarins from seeds of Psoralea corylifolia in the chronic mild stress model of depression in mice [J]. Phytomedicine.2007 ,14,523-529.

[18]Yue F.W., Ya N.L., Wen X., Dong M.Y., Yan Z., Xiu M.G., Yan T.X., AiD.Q.A UPLC–MS/MS method for in vivo and in vitro pharmacokinetic studies of psoralenoside, isopsoralenoside, psoralen and isopsoralen from Psoraleacorylifolia extract[J].J.Ethnopharmacol.,2014,151,609-617.

[19]Ruan B., Kong L.Y., Takaya Y., Niwa M.Studies on the chemical constituents of Psoralea corylifolia L[J].J.Asian Nat.Prod.Res.,2007,9,41-44.

[20] Hsu Y.T., Wu C.J., Chen J.M., Yang Y.C., Wang S.Y. The presence of three isoflavonoid compounds in Psoralea corylifolia [J]. J. Chromatogr. Sci., 2001, 39, 441-444.

[21] Amit N.S., Nutan M., Devanand P., Fulzele. Determination of isoflavone content and antioxidant activity in Psoralea corylifolia L. calluscultures [J]. Food Chem., 2010, 118, 128-132.

[22] Cheng Z.W., Cai X.F., Dat N.T., Hong S.S., Han A.R., Seo E.K., Hwang B.Y., Nan J.X., Lee D.H., Lee J.J. Bisbakuchiols A and B, novel dimericmeroterpenoids from Psoralea corylifolia[J]. Tetrahedron Lett. ,2007,48,8861-8864.

[23] Njoo M.D., Westerhof W. Vitiligo. Pathogenesis and treatment [J]. Am. J. Clin. Dermatol., 2001, 2, 167-181.

Based on the comprehensive analysis of relevant patents and documents at home and abroad and the previous research work of our research group, the present invention further transforms and modifies the active compound psoralen in the plant, and converts different secondary substances through amination reaction. Amines were introduced into psoralen molecules to improve its druggability, and the effects of these amine derivatives on melanogenesis and tyrosinase activity in mouse B16 cells were studied. A new anti-vitiligo drug with clear action mechanism.

Contents of the invention

The object of the present invention is to provide a kind of psoralen amine derivatives and its application. The derivatives take resorcinol as the starting material, and under the action of sulfuric acid, first condense with ethyl chloroacetoacetate to obtain 4 -chloromethyl-7-hydroxycoumarin (intermediate 1); then, intermediate 1 is reacted with five different secondary amines to obtain 4-substituted aminomethyl-7-hydroxycoumarin 2 (intermediate 2a-2e); Next, four different side chains are introduced at the 7-position of intermediate 2 to obtain intermediate 3 (3a-3t); finally, intermediate 3 is closed in potassium hydroxide ethanol solution to form a complementary psoralen, finally obtained psoralen amine derivatives 4 (4a-4t). And the obtained 20 psoralen amine derivatives were tested for melanogenesis and tyrosinase activity in mouse B16 cells, the results: compounds 4c-4h, 4j-4p and 4s-4t co- All the 15 psoralen amine derivatives can be used in the clinical preparation of medicines for treating vitiligo.

A kind of psoralen amine derivative according to the present invention, the derivative structure is:

in:

Compound 4a is 3-methyl-5-piperidinylmethyl-7H-fur[3,2-g]benzopyran-7-one;

Compound 4b is 3,4-dimethyl-5-piperidinylmethyl-7H-fur[3,2-g]benzopyran-7-one;

Compound 4c is 4-piperidinylmethyl-6,7,8,9-tetrahydro-2H-benzofur[3,2-g]benzopyran-7-one;

Compound 4d is 3-phenyl-5-piperidinylmethyl-7H-furo[3,2-g]benzopyran-7-one;

Compound 4e is 3-methyl-5-morpholinomethyl-7H-fur[3,2-g]benzopyran-7-one;

Compound 4f is 3,4-dimethyl-5-morpholinomethyl-7H-fur[3,2-g]benzopyran-7-one;

Compound 4g is 4-morpholinomethyl-6,7,8,9-tetrahydro-2H-benzofur[3,2-g]benzopyran-7-one;

Compound 4h is 3-phenyl-5-morpholinomethyl-7H-fur[3,2-g]benzopyran-7-one;

Compound 4i is 3-methyl-5-(4-methylpiperazine)methyl-7H-furo[3,2-g]benzopyran-7-one;

Compound 4j is 3,4-dimethyl-5-(4-methylpiperazine)methyl-7H-fur[3,2-g]benzopyran-7-one;

Compound 4k is 4-(4-methylpiperazine)methyl-6,7,8,9-tetrahydro-2H-benzofur[3,2-g]benzopyran-7-one;

Compound 4l is 3-phenyl-5-(4-methylpiperazine)methyl-7H-fur[3,2-g]benzopyran-7-one;

Compound 4m is 3-methyl-5-diethylaminomethyl-7H-fur[3,2-g]benzopyran-7-one;

Compound 4n is 3,4-dimethyl-5-diethylaminomethyl-7H-fur[3,2-g]benzopyran-7-one;

Compound 4o is 4-diethylaminomethyl-6,7,8,9-tetrahydro-2H-benzofur[3,2-g]benzopyran-7-one;

Compound 4p is 3-phenyl-5-diethylaminomethyl-7H-fur[3,2-g]benzopyran-7-one;

Compound 4q is 3-methyl-5-dipropylaminomethyl-7H-fur[3,2-g]benzopyran-7-one;

Compound 4r is 3,4-dimethyl-5-dipropylaminomethyl-7H-fur[3,2-g]benzopyran-7-one;

Compound 4s is 4-dipropylaminomethyl-6,7,8,9-tetrahydro-2H-benzofur[3,2-g]benzopyran-7-one;

Compound 4t is 3-phenyl-5-dipropylaminomethyl-7H-furo[3,2-g]benzopyran-7-one.

Among the psoralen amine derivatives, 15 compounds including 4c-4h, 4j-4p and 4s-4t are used for clinical preparation of medicine for treating vitiligo.

A kind of psoralen ester derivative of the present invention, its structure is as shown in general formula (I):

A kind of psoralen amine derivatives described in the present invention, the 20 psoralen amine derivatives that obtain have been carried out in the mouse B16 cell melanogenesis and the action test of tyrosinase activity, promote The results of melanin synthesis showed that: compared with the negative control, all 20 derivatives could promote the production of melanin in B16 cells, and the promoting effects ranged from 119.96% to 200.85%; compared with the positive control, compound 4c- 15 compounds including 4h, 4j-4p, and 4s-4t have better effects on melanin production than the positive control, and compounds 4h, 4l and 4p have more than 1.4 times the positive control on melanogenesis;

The results of activating tyrosinase activity showed that compared with the negative control, all compounds except 4b, 4p, 4q and 4r could activate tyrosinase activity, and the activation effect ranged from 114.78% to 178.88%; Compared with the positive control, compounds 4g and 4j-4m have a better activation effect on tyrosinase activity than the positive control, and the activation effect of compound 4j on tyrosinase activity is more than 1.3 times that of the positive control.

In summary, compounds 4c-4h, 4j-4p and 4s-4t, a total of 15 psoralen amine derivatives, can be used in the clinical preparation of drugs for the treatment of vitiligo.

Novel psoralen amine derivatives and purposes of the present invention, wherein the preparation method of psoralen amine derivatives, according to the following steps:

Preparation of Intermediate 1:

a. Under ice bath conditions, dissolve resorcinol in an appropriate amount of dry concentrated sulfuric acid, stir until completely dissolved, then slowly add ethyl 4-chloroacetoacetate dropwise to keep the temperature at 0°C, after the addition is complete , moved to room temperature for reaction, stirred overnight, poured the reaction solution into ice water, stirred thoroughly, and after the solid was fully precipitated, suctioned and dried to obtain 4-chloromethyl-7-hydroxycoumarin (intermediate 1);

Preparation of Intermediate 2:

b. Dissolve piperidine/or morpholine, 4-methylpiperazine/or diethylamine or dipropylamine and the intermediate 1 obtained in step a in dry tetrahydrofuran, heat to reflux for reaction, and after all the raw materials disappear, Concentrate under reduced pressure, and the residue is eluted by gradient column chromatography, the eluent is chloroform:methanol with a volume ratio of 30:1-10:1, and intermediate 2 is obtained in sequence;

Preparation of intermediate 3:

c. Dissolve potassium carbonate and the intermediate 2 obtained in step b in 20 mL of acetone, and add chloroacetone, 3-chloro-2-butanone, bromocyclohexane or α-bromoacetophenone, and reflux the reaction Until all the raw materials disappeared, the reaction solution was filtered, concentrated under reduced pressure, and the residue was eluted by column chromatography gradient, the eluent was chloroform:methanol with a volume ratio of 40:1-15:1, and intermediate 3 was obtained in sequence;

Preparation of compound 4:

Dissolve the intermediate 3 obtained in step c in absolute ethanol, add an ethanol solution with a concentration of 4% potassium hydroxide, and heat to reflux for reaction. After all the raw materials disappear, concentrate under reduced pressure, and wash the residue by gradient column chromatography. The eluent is chloroform:methanol with a volume ratio of 50:1-10:1, and compound 4 is finally obtained.

The chemical reaction formula is:

3a-3d, 4a-4d: R ₁ , R ₂ =(CH ₂ ) ₅ 3a, 4a: R = CH ₃ , R' = H; 3b, 4b: R = CH ₃ , R' = CH ₃ ; 3c, 4c: R, R' = (CH ₂ ) ₅ ; 3d, 4d: R = Ph, R' = H

3e-3h, 4e-4h: R ₁ , R ₂ =(CH ₂ CH ₂ ) ₂ O 3e, 4e: R = CH ₃ , R' = H; 3f, 4f: R = CH ₃ , R' = CH ₃ ; 3g, 4g: R, R'=(CH ₂ ) ₅ ; 3h, 4h: R=Ph, R'=H

3i-3l, 4i-4l: R ₁ , R ₂ =N(CH ₂ CH ₂ ) ₂ NCH ₃ 3i, 4i: R = CH ₃ , R' = H; 3j, 4j: R = CH ₃ , R' = CH ₃ ; 3k, 4k: R, R' = (CH ₂ ) ₅ ; 3l, 4l: R = Ph, R' = H

3m-3p, 4m-4p: R ₁ =R ₂ =CH ₂ CH ₃ 3m, 4m: R = CH ₃ , R' = H; 3n, 4n: R = CH ₃ , R' = CH ₃ ; 3o, 4o : R, R'=(CH ₂ ) ₅ ; 3p, 4p: R=Ph, R'=H

3q-3t, 4q-4t: R ₁ =R ₂ =CH ₂ CH ₂ CH ₃ 3q, 4q: R = CH ₃ , R' = H; 3r, 4r: R = CH ₃ , R' = CH ₃ ; 3s ,4s:R,R'=(CH ₂ ) ₅ ; 3t,4t:R=Ph,R'=H

(i) ethyl chloroacetoacetate, concentrated sulfuric acid, 0°C-room temperature (ii) for 2a: piperidine, THF, reflux; for 2b: morpholine, THF, reflux; for 2c: N-methylpiperazine, THF , reflux; for 2d: diethylamine, tetrahydrofuran, reflux; for 2e: dipropylamine, tetrahydrofuran, reflux; (iii) for 3a, 3e, 3i, 3m, 3q: chloroacetone, potassium carbonate, acetone reflux; for 3b, 3f, 3j, 3n, 3r: 2-chlorobutanone, potassium carbonate, acetone, reflux; for 3c, 3g, 3k, 3o, 3s: bromocyclohexane, potassium carbonate, acetone, reflux; for 3d, 3h, 3l, 3p, 3t: bromoacetophenone, potassium carbonate, acetone, reflux (iv) 4% potassium hydroxide ethanol solution, ethanol, reflux.

Wherein R and R are respectively piperidinyl, morpholinyl, _{4 -} methylpiperazinyl, diethylamino, dipropylamino; R and R' are respectively methyl, hydrogen; methyl, methyl; Cyclohexyl; phenyl, hydrogen.

Description of drawings

Fig. 1 is the graph that psoralen amine derivative 4a-4t of the present invention is to melanin synthesis in B16 cell;

Fig. 2 is a graph showing the effects of psoralen amine derivatives 4a-4t of the present invention on tyrosinase activity in B16 cells.

Detailed ways

The present invention is further described according to the examples, but the present invention is not limited to these examples;

Reagents: All reagents are of commercially available analytical grade;

Embodiment 1 (preparation of intermediate 1-intermediate 3)

Preparation of Intermediate 1:

Under ice-bath conditions, dissolve 1.10g of resorcinol in an appropriate amount of dry concentrated sulfuric acid, stir until completely dissolved, then slowly add 1.62mL of ethyl 4-chloroacetoacetate dropwise to keep the temperature at 0°C, drop After the addition, move to room temperature to react, stir overnight, pour the reaction solution into ice water, stir thoroughly, and after the solid is fully precipitated, filter and dry to obtain 0.188g of intermediate 1, named 4-chloromethyl-7- Hydroxycoumarin;

Preparation of Intermediate 2:

Dissolve 0.50mL of piperidine and 0.21g of intermediate 1 in dry tetrahydrofuran, heat to reflux for reaction, after all the raw materials disappear, concentrate under reduced pressure, and elute the residue by column chromatography gradient, the eluent is volume ratio 25:1 chloroform:methanol, to obtain 0.233g intermediate 2a, 4-piperidylmethyl-7-hydroxycoumarin;

Dissolve 0.44mL of morpholine and 0.21g of intermediate 1 in dry tetrahydrofuran, heat to reflux for reaction, after all the raw materials disappear, concentrate under reduced pressure, and elute the residue by column chromatography gradient, the eluent is volume ratio 10:1 chloroform: methanol, to obtain 0.240g intermediate 2b, 4-morpholinomethyl-7-hydroxycoumarin;

Dissolve 0.55mL of 4-methylpiperazine and 0.21g of intermediate 1 in dry tetrahydrofuran, heat to reflux for reaction, and after all the raw materials disappear, concentrate under reduced pressure, and elute the residue by gradient column chromatography. Agent is chloroform of volume ratio 30:1: Methanol, obtains 0.252g intermediate 2c, 4-(4-methylpiperazine) methyl-7-hydroxycoumarin;

Dissolve 0.51mL of diethylamine and 0.21g of intermediate 1 in dry tetrahydrofuran, heat to reflux for reaction, and after all the raw materials disappear, concentrate under reduced pressure, and elute the residue by gradient column chromatography. The eluent is volume Ratio of 30:1 chloroform: methanol, to obtain 0.211g intermediate 2d, 4-diethylaminomethyl-7-hydroxycoumarin;

Dissolve 0.68mL of di-n-propylamine and 0.21g of intermediate 1 in dry tetrahydrofuran, heat to reflux for reaction, and after all the raw materials disappear, concentrate under reduced pressure, and elute the residue by gradient column chromatography. The eluent is volume Ratio of 30:1 chloroform: methanol, to obtain 0.254g intermediate 2e, 4-dipropylaminomethyl-7-hydroxycoumarin;

Preparation of intermediate 3:

Dissolve 0.69g of potassium carbonate and 0.26g of intermediate 2a in 20mL of acetone, add 0.10mL of chloroacetone, reflux the reaction until all the raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and elute the residue by column chromatography gradient , the eluent is chloroform with a volume ratio of 30:1: methanol, to obtain 0.29g of intermediate 3a, 4-piperidinylmethyl-7-(2-oxopropoxy)coumarin;

Dissolve 0.69g of potassium carbonate and 0.26g of intermediate 2a in 20mL of acetone, add 0.12mL of 3-chloro-2-butanone, reflux until all raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and use Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 30:1, to obtain 0.303g of intermediate 3b, 4-piperidinylmethyl-7-(3-oxobutan-2-yl)oxy base coumarin;

Dissolve 0.69g of potassium carbonate and 0.26g of intermediate 2a in 20mL of acetone, add 0.14mL of 2-bromocyclohexanone, reflux the reaction until all the raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and use the column layer Analysis gradient elution, eluent is chloroform:methanol of volume ratio 20:1, obtains 0.328g intermediate 3c, 4-piperidinylmethyl-7-(2-oxocyclohexyl)oxycoumarin;

Dissolve 0.69g of potassium carbonate and 0.26g of intermediate 2a in 20mL of acetone, add 0.24g of α-bromoacetophenone, reflux the reaction until all the raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and use the column Chromatographic gradient elution, the eluent is chloroform:methanol with a volume ratio of 30:1, to obtain 0.350 g of intermediate 3d, 4-piperidinylmethyl-7-(2-oxophenethyl)oxycoumarin ;

Dissolve 0.69g of potassium carbonate and 0.26g of intermediate 2b in 20mL of acetone, add 10mL of chloroacetone, reflux until all the raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and elute the residue by gradient column chromatography. The eluent is chloroform with a volume ratio of 25:1: methanol, to obtain 0.289g of intermediate 3e, 4-morpholinomethyl-7-(2-oxopropoxy)coumarin;

Dissolve 0.69g of potassium carbonate and 0.26g of intermediate 2b in 20mL of acetone, add 0.12mL of 3-chloro-2-butanone, reflux the reaction until all raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and use Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 20:1, to obtain 0.300 g of intermediate 3f, 4-morpholinomethyl-7-(3-oxobutan-2-yl)oxy base coumarin;

Dissolve 0.69g of potassium carbonate and 0.26g of intermediate 2b in 20mL of acetone, add 0.14mL of 2-bromocyclohexanone, reflux the reaction until all raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and remove the residue by column layer Analysis gradient elution, eluent is the chloroform of volume ratio 25:1: Methanol, obtains 0.325g intermediate 3g, 4-morpholinomethyl-7-(2-oxocyclohexyl)oxycoumarin;

Dissolve 0.69g of potassium carbonate and 0.26g of intermediate 2b in 20mL of acetone, add 0.24g of α-bromoacetophenone, reflux the reaction until all the raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and use the column Chromatographic gradient elution, the eluent is chloroform:methanol with a volume ratio of 20:1, to obtain 0.361g of intermediate 3h, 4-morpholinomethyl-7-(2-oxophenethyl)oxycoumarin ;

Dissolve 0.69g of potassium carbonate and 0.27g of intermediate 2c in 20mL of acetone, add 10mL of chloroacetone, reflux the reaction until all the raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and elute the residue by gradient column chromatography. The eluent was chloroform:methanol with a volume ratio of 20:1, to obtain 0.297g of intermediate 3i, 4-(4-methylpiperazine)-7-(2-oxopropoxy)coumarin;

Dissolve 0.69g of potassium carbonate and 0.27g of intermediate 2c in 20mL of acetone, add 0.12mL of 3-chloro-2-butanone, reflux the reaction until all raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and use Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 20:1, to obtain 0.312g of intermediate 3j, 4-(4-methylpiperazine)-7-(3-oxobutane-2 -yl)oxycoumarin;

Dissolve 0.69g of potassium carbonate and 0.26g of intermediate 2c in 20mL of acetone, add 0.14mL of 2-bromocyclohexanone, reflux the reaction until all raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and use the column layer Analytical gradient elution, the eluent is chloroform: methanol with a volume ratio of 20:1, to obtain 0.355g of intermediate 3k, 4-(4-methylpiperazine)-7-(2-oxocyclohexyl)oxycyclohexyl Soybean;

Dissolve 0.69g of potassium carbonate and 0.26g of intermediate 2c in 20mL of acetone, add 0.24g of α-bromoacetophenone, reflux the reaction until all raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and use the column Chromatographic gradient elution, the eluent is chloroform:methanol with a volume ratio of 25:1, and 0.365 g of intermediate 3l, 4-(4-methylpiperazine)-7-(2-oxophenethyl)oxy base coumarin;

Dissolve 0.69g of potassium carbonate and 0.25g of intermediate 2d in 20mL of acetone, add 0.10mL of chloroacetone, reflux the reaction until all the raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and elute the residue by column chromatography gradient , the eluent is chloroform with a volume ratio of 30:1: methanol, to obtain 0.286g intermediate 3m, 4-diethylaminomethyl-7-(2-oxopropoxy)coumarin;

Dissolve 0.69g of potassium carbonate and 0.25g of intermediate 2d in 20mL of acetone, add 0.12mL of 3-chloro-2-butanone, reflux the reaction until all raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and use Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 20:1, to obtain 0.293g of intermediate 3n, 4-diethylaminomethyl-7-(3-oxobutan-2-yl ) oxycoumarin;

Dissolve 0.69g of potassium carbonate and 0.25g of intermediate 2d in 20mL of acetone, add 0.14mL of 2-bromocyclohexanone, reflux the reaction until all the raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and use the column layer Analytical gradient elution, the eluent is chloroform: methanol with a volume ratio of 30:1, to obtain 0.314g intermediate 3o, 4-diethylaminomethyl-7-(2-oxocyclohexyl)oxycoumarin ;

Dissolve 0.69g of potassium carbonate and 0.25g of intermediate 2d in 20mL of acetone, add 0.24g of α-bromoacetophenone, reflux the reaction until all the raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and use the column Chromatographic gradient elution, the eluent is chloroform:methanol with a volume ratio of 30:1, to obtain 0.333g of intermediate 3p, 4-diethylaminomethyl-7-(2-oxophenethyl)oxyphenamine Soybean;

Dissolve 0.69g of potassium carbonate and 0.28g of intermediate 2e in 20mL of acetone, add 0.10mL of chloroacetone, reflux the reaction until all the raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and elute the residue by column chromatography gradient , the eluent is chloroform with a volume ratio of 15:1: methanol, to obtain 0.296g of intermediate 3q, 4-dipropylaminomethyl-7-(2-oxopropoxy)coumarin;

Dissolve 0.69g of potassium carbonate and 0.28g of intermediate 2e in 20mL of acetone, add 0.12mL of 3-chloro-2-butanone, reflux the reaction until all raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and use Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 25:1, to obtain 0.315g intermediate 3r, 4-dipropylaminomethyl-7-(3-oxobutan-2-yl) Oxycoumarin;

Dissolve 0.69g of potassium carbonate and 0.28g of intermediate 2e in 20mL of acetone, add 0.14mL of 2-bromocyclohexanone, reflux the reaction until all raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and use the column layer Analysis gradient elution, eluent is the chloroform of volume ratio 20:1: Methanol, obtains 0.343g intermediate 3s, 4-dipropylaminomethyl-7-(2-oxocyclohexyl)oxycoumarin;

Dissolve 0.69g of potassium carbonate and 0.28g of intermediate 2e in 20mL of acetone, add 0.24g of α-bromoacetophenone, reflux the reaction until all raw materials disappear, filter the reaction solution, concentrate under reduced pressure, and use the column Chromatographic gradient elution, the eluent is chloroform:methanol with a volume ratio of 25:1, to obtain 0.367g of intermediate 3t, 4-dipropylaminomethyl-7-(2-oxophenethyl)oxycoumarin white.

Example 2

Preparation of compound 4a:

Dissolve 0.315g of intermediate 3a obtained in Example 1 in absolute ethanol, add 7mL of ethanol solution with a concentration of 4% potassium hydroxide, heat to reflux reaction, after all the raw materials disappear, concentrate under reduced pressure, and adopt the residue Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 15:1, and finally 0.267g of compound 4a, 3-methyl-5-piperidinylmethyl-7H-furan[3,2-g] was obtained Benzopyran-7-one;

NMR data of 3-methyl-5-piperidinylmethyl-7H-furo[3,2-g]benzopyran-7-one (compound 4a):

¹ H NMR (400MHz, CDCl ₃ )δ7.98(s,1H),7.46(s,1H),7.42(s,1H),6.54(s,1H),3.69(s,2H),2.59-2, 46(m,4H),2.29(s,3H),1.68-1.58(m,4H),1.54-1.45(s,2H).

¹³ C NMR (101MHz, CDCl ₃ ) δ161.55, 156.69, 152.11, 143.18, 126.44, 115.88, 115.17, 110.18, 99.91, 60.13, 55.17, 26.13, 24.19, 8.01.

Example 3

Preparation of compound 4b:

Dissolve 0.329g of intermediate 3b obtained in Example 1 in dehydrated ethanol, add 7mL of ethanol solution with a concentration of 4% potassium hydroxide, heat to reflux reaction, after all the raw materials disappear, concentrate under reduced pressure, and adopt Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 15:1, and finally 0.280 g of compound 4b, 3,4-dimethyl-5-piperidinylmethyl-7H-furan[3,2 -g] benzopyran-7-one;

NMR data of 3,4-dimethyl-5-piperidinylmethyl-7H-furo[3,2-g]benzopyran-7-one (compound 4b):

¹ H NMR (400MHz, CDCl ₃ )δ7.82(s,1H),7.33(s,1H),6.52(s,1H),3.68(s,2H),2.60-2.46(m,4H),2.40( s,3H),2.19(s,3H),1.70-1.58(m,4H),1.54-1.45(m,2H).

¹³ C NMR (101MHz, CDCl ₃ ) δ161.90, 155.42, 152.76, 151.60, 127.96, 114.80, 113.80, 112.56, 109.81, 99.16, 60.02, 55.19, 26.15, 24.23, 12.10, 8.04.

Example 4

Preparation of compound 4c:

Dissolve 0.355g of intermediate 3c obtained in Example 1 in absolute ethanol, add 7mL of ethanol solution with a concentration of 4% potassium hydroxide, heat to reflux reaction, after all the raw materials disappear, concentrate under reduced pressure, and adopt the residue Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 15:1, and finally 0.305g of compound 4c, 4-piperidinylmethyl-6,7,8,9-tetrahydro-2H-benzo Furan[3,2-g]benzopyran-7-one;

NMR data of 4-piperidinylmethyl-6,7,8,9-tetrahydro-2H-benzofuro[3,2-g]benzopyran-7-one (compound 4c):

¹ H NMR (400MHz, CDCl ₃ )δ7.79(s,1H),7.36(s,1H),6.52(s,1H),3.66(s,2H),2.79-2.71(m,2H),2.69- 2.61(m,2H),2.57-2.46(m,4H),2.00-1.83(m,4H),1.67-1.57(m,4H),1.52-1.43(m,2H).

¹³ C NMR (101MHz, CDCl ₃ ) δ161.63, 156.22, 155.82, 151.29, 126.23, 114.60, 113.39, 112.76, 99.41, 59.69, 54.96, 25.83, 23.93, 23.39, 22.68, 22.432, 20.3

Example 5

Preparation of compound 4d:

Dissolve 0.377g of intermediate 3d obtained in Example 1 in dehydrated ethanol, add 7mL of ethanol solution with a concentration of 4% potassium hydroxide, heat to reflux reaction, after all the raw materials disappear, concentrate under reduced pressure, and adopt the residue Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 15:1, and finally 0.327g of compound 4d, 3-phenyl-5-piperidinylmethyl-7H-furan[3,2-g] was obtained Benzopyran-7-one;

NMR data of 3-phenyl-5-piperidinylmethyl-7H-furo[3,2-g]benzopyran-7-one (compound 4d):

¹ H NMR (400MHz, CDCl ₃ ) δ8.48(s, 1H), 7.84(s, 1H), 7.67(d, J=7.8Hz, 2H), 7.56-7.47(m, 3H), 7.43(t, J＝7.3Hz,1H),6.49(s,1H),3.65(s,2H),2.55-2.45(m,4H),1.68-1.59(m,5H),1.54-1.45(m,3H).

¹³ C NMR (101MHz, CDCl ₃ ) δ161.47, 157.10, 153.14, 152.20, 142.81, 131.33, 129.30, 128.11, 127.60, 123.74, 122.41, 117.06, 115.83, 113.46, 100.987, 24.64.

Example 6

Preparation of compound 4e:

Dissolve 0.317g of intermediate 3e obtained in Example 1 in absolute ethanol, add 7mL of ethanol solution with a concentration of 4% potassium hydroxide, heat to reflux reaction, after all the raw materials disappear, concentrate under reduced pressure, and use the residue Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 50:1, and finally 0.260 g of compound 4e, 3-methyl-5-morpholinomethyl-7H-furan[3,2-g] was obtained Benzopyran-7-one;

NMR data of 3-methyl-5-morpholinomethyl-7H-furo[3,2-g]benzopyran-7-one (compound 4e):

¹ H NMR (400MHz, CDCl ₃ )δ7.92(s,1H),7.47(s,1H),7.42(s,1H),6.54(s,1H),3.76(t,(t,J＝4.3Hz ,4H),3.74(s,2H),2.61(t,J=4.3Hz,4H),2.29(s,3H).

¹³ C NMR (101 MHz, CDCl ₃ ) δ 161.48, 156.71, 152.00, 143.30, 126.49, 115.78, 114.91, 113.09, 99.99, 67.10, 59.68, 54.03, 7.98.

Example 7

Preparation of compound 4f:

Dissolve 0.331g of intermediate 3f obtained in Example 1 in absolute ethanol, add 7mL of ethanol solution with a concentration of 4% potassium hydroxide, heat to reflux reaction, after all the raw materials disappear, concentrate under reduced pressure, and adopt Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 50:1, and finally 0.284g of compound 4f, 3,4-dimethyl-5-morpholinomethyl-7H-furan[3,2 -g] benzopyran-7-one;

NMR data of 3,4-dimethyl-5-morpholinomethyl-7H-furo[3,2-g]benzopyran-7-one (compound 4f):

¹ H NMR (400MHz, CDCl ₃ )δ7.77(s,1H),7.34(s,1H),6.53(s,1H),3.77(t,J＝4.2Hz,4H),3.74(s,2H) ,2.61(t,J=4.2Hz,4H),2.40(s,3H),2.19(s,3H).

¹³ C NMR (101 MHz, CDCl ₃ ) δ161.68, 155.50, 152.94, 151.60, 128.05, 114.52, 113.60, 112.76, 109.75, 99.28, 67.10, 59.64, 54.05, 12.10, 8.04.

Example 8

Preparation of compound 4g:

The 0.357g intermediate 3g obtained in Example 1 was dissolved in absolute ethanol, and 7mL of ethanol solution of 4% potassium hydroxide was added, heated to reflux reaction, after all the raw materials disappeared, concentrated under reduced pressure, and the residue was used Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 50:1, and finally 0.302g of compound 4g, 4-morpholinomethyl-6,7,8,9-tetrahydro-2H-benzo Furan[3,2-g]benzopyran-7-one;

NMR data of 4-morpholinomethyl-6,7,8,9-tetrahydro-2H-benzofuro[3,2-g]benzopyran-7-one (compound 4g):

¹ H NMR (400MHz, CDCl ₃ )δ7.76(s,1H),7.37(s,1H),6.53(s,1H),3.76(t,J=4.2Hz,4H),3.71(s,2H) ,2.78-2.72(m,2H),2.67-2.62(m,2H),2.59(t,J＝4.2Hz,4H),2.00-1.89(m,4H).

¹³ C NMR (101MHz, CDCl ₃ ) δ161.68, 155.50, 152.94, 151.60, 128.05, 114.52, 113.60, 112.76, 109.75, 99.28, 67.10, 59.64, 54.05, 31.07, 29.98, 12.10, 8.

Example 9

Preparation of compound 4h:

Dissolve 0.379g of intermediate 3h obtained in Example 1 in dehydrated ethanol, add 7mL of ethanol solution with a concentration of 4% potassium hydroxide, heat to reflux reaction, after all raw materials disappear, concentrate under reduced pressure, and adopt Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 50:1, and finally 0.323g of compound 4h, 3-phenyl-5-morpholinomethyl-7H-furan[3,2-g] was obtained Benzopyran-7-one;

NMR data of 3-phenyl-5-morpholinomethyl-7H-furo[3,2-g]benzopyran-7-one (compound 4h):

¹ H NMR (400MHz, CDCl ₃ ) δ8.39(s, 1H), 7.85(s, 1H), 7.66(d, J=7.1Hz, 2H), 7.57-7.50(m, 3H), 7.45(t, J＝7.3Hz, 1H), 6.53(s, 1H), 3.76(t, J＝4.0Hz, 4H), 3.71(s, 2H), 2.59(t, J＝4.0Hz, 4H).

¹³ C NMR (101MHz, CDCl ₃ ) δ161.24, 157.18, 152.21, 142.99, 131.22, 129.83, 129.34, 128.23), 127.64, 123.95, 122.42, 116.68, 115.53, 113.51, 100.18.341, 60.67

Example 11

Preparation of compound 4i:

Dissolve 0.330g of intermediate 3i obtained in Example 1 in dehydrated ethanol, add 7mL of ethanol solution with a concentration of 4% potassium hydroxide, heat to reflux reaction, after all the raw materials disappear, concentrate under reduced pressure, and adopt Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 30:1, and finally 0.276g of compound 4i, 3-methyl-5-(4-methylpiperazine)methyl-7H-furan[ 3,2-g]benzopyran-7-one;

NMR data of 3-methyl-5-(4-methylpiperazine)methyl-7H-furo[3,2-g]benzopyran-7-one (compound 4i):

¹ H NMR (400MHz, CDCl ₃ ) δ7.90(s,1H),7.46(s,1H),7.42(s,1H),6.55(s,1H),3.74(s,2H),2.74-2.45( m,8H),2.34(s,3H),2.29(s,3H).

¹³ C NMR (101 MHz, CDCl ₃ ) δ161.56, 156.71, 152.39, 152.11, 143.26, 126.48, 115.83, 114.86, 112.86, 99.99, 59.06, 55.19, 53.34, 45.93, 8.01.

Example 11

Preparation of compound 4j:

Dissolve 0.344g of intermediate 3j obtained in Example 1 in dehydrated ethanol, add 7mL of ethanol solution with a concentration of 4% potassium hydroxide, heat to reflux reaction, after all the raw materials disappear, concentrate under reduced pressure, and adopt Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 30:1, and finally 0.291g of compound 4j, 3,4-dimethyl-5-(4-methylpiperazine)methyl-7H was obtained - Furo[3,2-g]benzopyran-7-one;

NMR data of 3,4-dimethyl-5-(4-methylpiperazine)methyl-7H-furo[3,2-g]benzopyran-7-one (compound 4j):

¹ H NMR (400MHz, CDCl ₃ )δ7.75(s,1H),7.33(s,1H),6.53(s,1H),3.72(s,2H),2.72-2.42(m,8H),2.40( s,3H),2.31(s,3H),2.19(s,3H).

¹³ C NMR (101MHz, CDCl ₃ ) δ161.81, 155.46, 152.85, 152.63, 151.59, 127.99, 114.61, 113.57, 112.47, 109.77, 99.22, 59.09, 55.26, 53.52, 45.06, 12.09, 8.8

Example 12

Preparation of compound 4k:

Dissolve 0.370g of intermediate 3k obtained in Example 1 in dehydrated ethanol, add 7mL of ethanol solution with a concentration of 4% potassium hydroxide, heat to reflux reaction, after all the raw materials disappear, concentrate under reduced pressure, and adopt the residue Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 20:1, and finally 0.320 g of compound 4k, 4-(4-methylpiperazine)methyl-6,7,8,9-tetra Hydrogen-2H-benzofuro[3,2-g]benzopyran-7-one;

NMR of 4-(4-methylpiperazine)methyl-6,7,8,9-tetrahydro-2H-benzofuro[3,2-g]benzopyran-7-one (compound 4k) data:

¹ H NMR (400MHz, CDCl ₃ )δ7.72(s,1H),7.37(s,1H),6.53(s,1H),3.72(s,2H),2.80-2.48(m,12H),2.35( s,3H),2.01-1.83(m,4H).

¹³ C NMR (101MHz, CDCl ₃ ) δ161.81, 156.46, 156.00, 152.59, 151.47, 126.41, 114.62, 113.26, 112.88, 112.50, 99.63, 59.00, 55.17, 53.36, 45.93, 23.487, 2.22

Example 13

Preparation of compound 4l:

Dissolve 0.392g of intermediate 3l obtained in Example 1 in dehydrated ethanol, add 7mL of ethanol solution with a concentration of 4% potassium hydroxide, heat to reflux reaction, after all the raw materials disappear, concentrate under reduced pressure, and use the residue Column chromatography gradient elution, the eluent is chloroform: methanol with a volume ratio of 20:1, and finally 0.351g of compound 4l, 3-phenyl-5-(4-methylpiperazine)methyl-7H-furan[ 3,2-g]benzopyran-7-one;

NMR data of 3-phenyl-5-(4-methylpiperazine)methyl-7H-furo[3,2-g]benzopyran-7-one (compound 4l):

¹ H NMR (400MHz, CDCl ₃ ) δ8.40(s, 1H), 7.87(s, 1H), 7.67(d, J=7.2Hz, 2H), 7.58-7.51(m, 3H), 7.47(t, J＝7.2Hz,1H),6.54(s,1H),3.72(s,2H),2.73-2.44(m,8H),2.36(s,3H).

¹³ C NMR (101MHz, CDCl ₃ ) δ161.34, 158.07, 151.89, 143.20, 131.16, 130.03, 129.42, 128.50, 127.73, 124.21, 122.70, 117.13, 115.43, 114.157, 901.87, 569.5

Example 14

Preparation of compound 4m:

0.303g of intermediate 3m obtained in Example 1 was dissolved in dehydrated ethanol, and 7mL of ethanol solution of 4% potassium hydroxide was added, heated to reflux reaction, after all the raw materials disappeared, concentrated under reduced pressure, and the residue was used Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 10:1, and finally 0.252g of compound 4m, 3-methyl-5-diethylaminomethyl-7H-furo[3,2- g] benzopyran-7-one;

NMR data of 3-methyl-5-diethylaminomethyl-7H-furo[3,2-g]benzopyran-7-one (compound 4m):

¹ H NMR (400MHz, CDCl ₃ )δ7.99(s,1H),7.46(s,1H),7.42(s,1H),6.61(s,1H),3.80(s,2H),2.65(q, J＝7.1Hz, 4H), 2.29(s, 3H), 1.11(t, J＝7.1Hz, 6H).

¹³ C NMR (101 MHz, CDCl ₃ ) δ156.48, 151.86, 142.99, 140.24, 128.84, 126.24, 115.69, 114.83, 110.67, 99.73, 54.76, 47.72, 11.92, 7.82.

Example 15

Preparation of compound 4n:

0.317g of intermediate 3n obtained in Example 1 was dissolved in absolute ethanol, and 7mL of ethanol solution of 4% potassium hydroxide was added, heated to reflux reaction, after all the raw materials disappeared, concentrated under reduced pressure, and the residue was used Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 10:1, and finally 0.270 g of compound 4n, 3,4-dimethyl-5-diethylaminomethyl-7H-furan[3 ,2-g]benzopyran-7-one;

NMR data of 3,4-dimethyl-5-diethylaminomethyl-7H-furo[3,2-g]benzopyran-7-one (compound 4n):

¹ H NMR (400MHz, CDCl ₃ )δ7.81(s,1H),7.32(s,1H),6.59(s,1H),3.77(s,2H),2.62(q,J=7.1Hz,4H) ,2.39(s,3H),2.18(s,3H),1.09(t,J=7.1Hz,6H).

¹³ C NMR (101MHz, CDCl ₃ ) δ162.04, 155.35, 154.74, 152.69, 151.51, 131.03, 128.97, 127.87, 113.61, 112.46, 99.10, 55.03, 47.91, 12.22.12.07, 8.00.

Example 16

Preparation of compound 4o:

Dissolve 0.343g of intermediate 3o obtained in Example 1 in dehydrated ethanol, add 7mL of ethanol solution with a concentration of 4% potassium hydroxide, heat to reflux reaction, after all the raw materials disappear, concentrate under reduced pressure, and adopt Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 10:1, and finally 0.298g of compound 4o, 4-diethylaminomethyl-6,7,8,9-tetrahydro-2H- Benzofuran[3,2-g]benzopyran-7-one;

NMR data of 4-diethylaminomethyl-6,7,8,9-tetrahydro-2H-benzofuro[3,2-g]benzopyran-7-one (compound 4o):

¹ H NMR (400MHz, CDCl ₃ )δ7.79(s,1H),7.35(s,1H),6.59(s,1H),3.76(s,2H),2.78-2.71(m,2H),2.68- 2.56(m,6H),2.02-1.81(m,4H),1.08(t,J=7.1Hz,6H).

¹³ C NMR (101MHz, CDCl ₃ ) δ162.05, 156.29, 155.90, 154.71, 151.40, 126.28, 114.82, 113.28, 112.89, 112.52, 99.52, 54.97, 47.90, 23.56, 22.86, 202.579, 21.2

Example 17

Preparation of compound 4p:

0.365g of intermediate 3p obtained in Example 1 was dissolved in absolute ethanol, and 7mL of ethanol solution of 4% potassium hydroxide was added, heated to reflux reaction, after all the raw materials disappeared, concentrated under reduced pressure, and the residue was used Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 10:1, and finally 0.317g of compound 4p, 3-phenyl-5-diethylaminomethyl-7H-furo[3,2- g] benzopyran-7-one;

NMR data of 3-phenyl-5-diethylaminomethyl-7H-furo[3,2-g]benzopyran-7-one (compound 4p):

¹ H NMR (400MHz, CDCl ₃ ) δ8.42(s, 1H), 7.84(s, 1H), 7.66(d, J=7.4Hz, 2H), 7.55-7.48(m, 3H), 7.42(t, J=7.2Hz, 1H), 6.58(s, 1H), 3.77(s, 2H), 2.61(q, J=7.0Hz, 4H), 1.09(t, J=7.0Hz, 6H).

¹³ C NMR (101MHz, CCDCl ₃ ) δ161.50, 157.12, 152.18, 142.87, 131.30, 129.31, 128.14, 127.67, 123.82, 122.47, 116.67, 115.79, 113.77, 100.22, 55.396, 1.47

Example 18

Preparation of compound 4q:

Dissolve 0.331g of intermediate 3q obtained in Example 1 in dehydrated ethanol, add 7mL of ethanol solution with a concentration of 4% potassium hydroxide, heat to reflux reaction, after all the raw materials disappear, concentrate under reduced pressure, and adopt Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 15:1, and finally 0.277g of compound 4q, 3-methyl-5-dipropylaminomethyl-7H-furo[3,2-g ]benzopyran-7-one;

NMR data of 3-methyl-5-dipropylaminomethyl-7H-furo[3,2-g]benzopyran-7-one (compound 4q):

¹ H NMR (400MHz, CDCl ₃ )δ8.02(s,1H),7.45(s,1H),7.40(s,1H),6.59(s,1H),3.79(s,2H),2.49(t, J＝7.2Hz, 4H), 2.27(s, 3H), 1.53(h, J＝7.3Hz, 4H), 0.89(t, J＝7.3Hz, 7H).

¹³ C NMR (101MHz, CDCl ₃ ) δ161.78, 156.63, 152.03, 143.12, 131.04, 128.97, 126.33, 115.85, 115.26, 113.03, 99.81, 56.77, 56.58, 20.42, 12.04, 7.98.

Example 19

Preparation of compound 4r:

Dissolve 0.345g of intermediate 3r obtained in Example 1 in absolute ethanol, add 7mL of ethanol solution with a concentration of 4% potassium hydroxide, heat to reflux reaction, after all the raw materials disappear, concentrate under reduced pressure, and adopt Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 15:1, and finally 0.302g of compound 4r, 3,4-dimethyl-5-dipropylaminomethyl-7H-furan[3, 2-g] benzopyran-7-one;

NMR data of 3,4-dimethyl-5-dipropylaminomethyl-7H-furo[3,2-g]benzopyran-7-one (compound 4r):

¹ H NMR (400MHz, CDCl ₃ )δ7.86(s,1H),7.33(s,1H),6.57(s,1H),3.77(s,2H),2.48(t,J＝7.3Hz,4H) ,2.40(s,3H),2.18(s,3H),1.52(h,J=7.3Hz,4H),0.89(t,J=7.3Hz,7H).

¹³ C NMR (101MHz, CDCl ₃ ) δ162.03, 155.39, 152.69, 151.53, 131.05, 128.99, 113.95, 112.60, 109.81, 99.10, 56.81, 56.61, 19.31, 12.09, 12.06, 8.04.

Example 20

Preparation of compound 4s:

Dissolve 0.371g of intermediate 3s obtained in Example 1 in dehydrated ethanol, add 7mL of ethanol solution with a concentration of 4% potassium hydroxide, heat to reflux reaction, after all raw materials disappear, concentrate under reduced pressure, and adopt Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 10:1, and finally 0.328g of compound 4s, 4-dipropylaminomethyl-6,7,8,9-tetrahydro-2H-benzene was obtained Furo[3,2-g]benzopyran-7-one;

NMR data of 4-dipropylaminomethyl-6,7,8,9-tetrahydro-2H-benzofuro[3,2-g]benzopyran-7-one (compound 4s):

¹ H NMR (400MHz, CDCl ₃ )δ7.78(s,1H),7.34(s,1H),6.57(s,1H),3.75(s,2H),2.76-2.69(m,2H),2.62- 2.55(m, 2H), 2.47(t, J=7.2Hz, 4H), 2.00-1.80(m, 4H), 1.50(h, J=7.1Hz, 4H), 0.87(t, J=7.2Hz, 7H ).

¹³ C NMR (101MHz, CDCl ₃ ) δ162.20, 156.43, 156.10, 155.01, 151.69, 126.37, 115.08, 113.28, 113.00, 112.33, 100.21, 56.79, 56.54, 23.56, 22.86, 212.459, 2

Example 21

Preparation of compound 4t:

0.393g of intermediate 3t obtained in Example 1 was dissolved in absolute ethanol, and 7mL of ethanol solution of 4% potassium hydroxide was added, heated to reflux reaction, after all the raw materials disappeared, concentrated under reduced pressure, and the residue was used Column chromatography gradient elution, the eluent is chloroform:methanol with a volume ratio of 15:1, and finally 0.342g of compound 4t, 3-phenyl-5-dipropylaminomethyl-7H-furo[3,2-g ]benzopyran-7-one;

NMR data of 3-phenyl-5-dipropylaminomethyl-7H-furo[3,2-g]benzopyran-7-one (compound 4t):

¹ H NMR (400MHz, CDCl ₃ ) δ8.36(s, 1H), 7.83(s, 1H), 7.65(d, J=7.4Hz, 2H), 7.56-7.47(m, 3H), 7.43(t, J＝7.3Hz, 1H), 6.59(s, 1H), 3.77(s, 2H), 2.48(t, J＝7.2Hz, 4H), 1.52(h, J＝7.3Hz, 4H), 0.85(t, J＝7.3Hz,7H).

¹³ C NMR (101MHz, CDCl ₃ ) δ161.61, 156.40, 151.78, 143.07, 130.85, 129.42, 127.95, 127.80, 124.15, 122.67, 116.77, 115.75, 113.78, 99.70, 56.80, 196.57.

Example 22

Determination of the effect of psoralen amine derivatives (4a-4t) as shown in the structural formula of the present invention on melanin synthesis and tyrosinase activity in B16 cells:

Screening models: mouse B16 melanoma cells, human A375 melanoma cells;

Cell source: provided by the Cell Bank of the Chinese Academy of Sciences;

(1) Cytotoxicity test (MTT method):

The melanocytes in the logarithmic growth phase were inoculated in a 96-well plate, the number of cells per well was 5×103/well, and 8 replicate wells were set up for each concentration. After 24 hours (after the cells adhered), different concentrations of After 48 hours, the cells were treated, 10 μL of MTT (concentration: 5 mg/mL) was added to each well, and cultured in a 37°C incubator for 4 hours. After taking out the 96-well plate, 150 μL of DMSO was added to each well and shaken for 10 minutes until the crystals were completely dissolved. Measure the absorbance (A570) at 570nm with a microplate reader, the wells without drug addition are used as the control group, and the cell activity is calculated by 100%, while the experimental data of the drug-adding group is repeated three times at least;

(2) Protein quantification was determined by Bradford method:

Completely dissolve the protein standard (5mg/mL BSA), take 10μL and dilute to 100μL, so that the final concentration is 0.5mg/mL. Which solution should the protein sample be in, and what solution should be used to dilute the standard, but for simplicity, the standard can also be diluted with 0.9% NaCl or PBS; 2, 4, 8, 12, 16, and 20 μL were added to the 96-well plate, and the standard diluent was added to make up all the standards to 20 μL; an appropriate volume of sample was added to the sample well of the 96-well plate, and the standard diluent was added. Make up to 20 μL; add 200 μL Bradford staining solution to each well, gently blow and mix with a sample gun (be careful not to make air bubbles to affect the reading) and place at room temperature for 3-5min; measure A595 with a microplate reader; calculate the concentration in the sample according to the standard curve. protein concentration;

(3) Melanin content is measured by alkali digestion method:

Inoculate the melanocytes in the logarithmic growth phase in a 6cm culture dish at a concentration of 5×105 cells/ml, and add 5mL of cell suspension to each well; 12 hours after inoculation and after the cells are completely adhered to the wall, the drug is used at a concentration of 1, 5, 10, 20, 50, and 100 μg/mL; cells were collected after 72 hours; 200 μL of 1M NaOH solution was added without breaking the cells, and after 2 hours in a water bath at 80 ° C, the absorbance at a wavelength of 470 nm was measured ( A470 value), the unmedicated group is compared with the drug group as the control group;

(4) Tyrosinase activity was measured by L-Dopa staining method:

Melanocytes in the logarithmic growth phase were inoculated in 12-well plates at a concentration of 2×105 cells/mL, and 1 mL of cell suspension was added to each well; 12 hours after inoculation and after the cells were completely adhered to the wall, the drug was used at a concentration of 1, 5, 10, 20, 50 and 100 μg/mL; collect the cells after 48 hours; add 200 μL of Tris-0.1% Triton-100 solution to each EP tube after the cells are processed, and put them in a refrigerator at -20°C for 30min- Lyse for 1 hour; determine the protein content by the Bradford method; adjust the protein content of the sample to a uniform concentration; add 100 μL of 0.1% L-DOPA to each reaction system and put it in an incubator with a temperature of 37 ° C; measure the absorbance at 475 nm after incubation for 2 hours (A475), the group without medication is compared with the medication group as a control group.

Results: Compounds 4c-4h, 4j-4p and 4s-4t, a total of 15 derivatives of psoralen amines, can be used in clinical preparation of drugs for the treatment of vitiligo.

Claims (2)

1. a kind of psoralen amine derivant, it is characterised in that such derivant structure is：

Wherein：

Compound 4b is 3,4- dimethyl -5- piperidine methyl -7H- Fu Nans [3,2-g]Chromene -7- ketone；

Compound 4c is 4- piperidine methyl -6,7,8,9- tetrahydrochysene -2H- Ben Bingfunans [3,2-g]Chromene -7- ketone；

Compound 4f is 3,4- dimethyl -5- morpholine methyl -7H- Fu Nans [3,2-g]Chromene -7- ketone；

Compound 4g is 4- morpholine methyl -6,7,8,9- tetrahydrochysene -2H- Ben Bingfunans [3,2-g]Chromene -7- ketone；

Compound 4i is 3- methyl -5- (4- methyl piperazines) methyl -7H- Fu Nans [3,2-g]Chromene -7- ketone；

Compound 4j is 3,4- dimethyl -5- (4- methyl piperazines) methyl -7H- Fu Nans [3,2-g]Chromene -7- ketone；

Compound 4k is 4- (4- methyl piperazines) methyl -6,7,8,9- tetrahydrochysene -2H- Ben Bingfunans [3,2-g]Chromene -7- Ketone；

Compound 4l is 3- phenyl -5- (4- methyl piperazines) methyl -7H- Fu Nans [3,2-g]Chromene -7- ketone；

Compound 4m is 3- methyl -5- dimethylamino methyl -7H- Fu Nans [3,2-g]Chromene -7- ketone；

Compound 4n is 3,4- dimethyl -5- dimethylamino methyl -7H- Fu Nans [3,2-g]Chromene -7- ketone；

Compound 4o is 4- dimethylamino methyl -6,7,8,9- tetrahydrochysene -2H- Ben Bingfunans [3,2-g]Chromene -7- ketone；

Compound 4p is 3- phenyl -5- dimethylamino methyl -7H- Fu Nans [3,2-g]Chromene -7- ketone；

Compound 4q is 3- methyl -5- dipropyl aminomethyl -7H- furans;3,2-g]Chromene -7- ketone；

Compound 4r is 3,4- dimethyl -5- dipropyl aminomethyl -7H- furans;3,2-g]Chromene -7- ketone；

Compound 4s is 4- dipropyl aminomethyl -6,7,8,9- tetrahydrochysene -2H- benzofurans;3,2-g]Chromene -7- ketone；

Compound 4t is 3- phenyl -5- dipropyl aminomethyl -7H- furans;3,2-g]Chromene -7- ketone.

2. in a kind of psoralen amine derivant as described in claim 1,4c, 4f-4g, 4j-4p and 4s-4t totally 12 changes Close the purposes in drug of the object for clinically preparing treatment leucoderma.