CN111763236B

CN111763236B - A kind of Xenopus venom carbamate derivative and its application

Info

Publication number: CN111763236B
Application number: CN202010585479.5A
Authority: CN
Inventors: 高波; 罗川; 缪震元; 吴岳林; 庄春林
Original assignee: Anhui China Resources Jinchan Pharmaceutical Co Ltd
Current assignee: Anhui China Resources Jinchan Pharmaceutical Co Ltd
Priority date: 2020-06-24
Filing date: 2020-06-24
Publication date: 2021-10-22
Anticipated expiration: 2040-06-24
Also published as: CN111763236A

Abstract

The invention discloses a arenobufagin carbamate derivative or a medicinal salt thereof, and the structure of the arenobufagin carbamate derivative or the medicinal salt is shown as a general formula (1):

Description

Arenobufagin carbamate derivatives and application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a arenobufagin carbamate derivative and application thereof in medicines.

Background

The Arenobufagin is a toad venom lactone compound extracted from toad venom, and has the following chemical structure:

research shows that the arenobufagin has stronger anti-tumor activity, has the effects of inhibiting adhesion, migration and invasion of liver cancer HepG2 cells and angiogenesis (Chinese pharmacological report, 2011, 27; 19-23), and can also obviously inhibit apoptosis of the tumor cells induced by the apoptosis pathway of liver cancer SMMC-7721 mitochondria (Chinese traditional medicine emergency, 2013, 22: 1845 and 1883). Therefore, the synthesis of the arenobufagin derivative, the study of the structure-activity relationship of the arenobufagin derivative and the discovery of the arenobufagin derivative with high efficiency and low toxicity have important significance.

At present, the research on the structural modification of the bufogenin is relatively few, a plurality of patents report the structural modification of another natural product bufalin with antitumor activity extracted from toads, and WO2011085641A1 discloses a class of bufalin derivatives and the application thereof in treating cancers. Patent application with publication number CN102532235A reports a class of bufogenin derivatives, preparation methods thereof, and uses of compositions containing the derivatives. Patent applications with publication numbers of CN102656179A, CN103980337A and CN110483608A and patent application with publication number of CN103980338A disclose bufalin derivatives, pharmaceutical compositions and uses thereof, wherein 3-hydroxy is esterified to obtain a series of derivatives.

Although the arenobufagin has excellent antitumor activity, the toxicity is relatively high, so structural modification is necessary on the basis of the toxicity, the activity of the compound is enhanced, the toxicity is reduced, and a new antitumor drug is discovered from the structural modification.

Disclosure of Invention

The invention aims to provide a arenobufagin carbamate derivative.

The second purpose of the invention is to provide the application of the arenobufagin carbamate derivatives in preparing antitumor drugs.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

the first aspect of the invention provides a arenobufagin carbamate derivative or a medicinal salt thereof, the structure of which is shown as a general formula (1):

in the formula 1, the reaction mixture is,

R₁、R₂and C or heteroatom O form a ring of C3-C8;

or, R₁、R₂Each independently selected from C1-C20 alkyl;

or, R₁Is hydrogen, R₂Selected from the group consisting of cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclopentane in which at least one hydrogen of the alkyl group is replaced by an amino group, and a group up to the alkyl groupA cyclohexane group in which at least one hydrogen is substituted by an amino group, a cycloheptane group in which at least one hydrogen of the alkyl group is substituted by an amino group, a cyclooctane group in which at least one hydrogen of the alkyl group is substituted by an amino group, a cyclohexyl group in which at least one hydrogen of the alkyl group is substituted by an amino group, a cycloheptyl group in which the cyclohexyl group is substituted by a cyclohexyl group, a cycloheptyl group in which the cycloheptyl group is substituted by an amino group, a cycloheptyl group in which the cycloheptyl group is substituted by a cyclohexyl group, a cycloheptyl group in which at least one hydrogen of the alkyl group is substituted by an amino group, a cycloheptyl group is substituted by an amino group, a cyclohexyl group,

In the above-mentioned manner, the first and second substrates are,

R₃is methyl, ethyl, n-propyl or n-butyl,

R₄is methyl, ethyl, n-propyl or n-butyl,

n is 1, 2, 3, 4.

More preferably, in said formula 1, R₁、R₂The ring which forms C3-C8 with C or heteroatom O is one of the following structures:

or, R₁、R₂Each independently selected from methyl, ethyl, n-propyl;

or, R₁Is hydrogen, R₂Selected from the group consisting of cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclohexane, and mixtures thereof,

One kind of (1).

Most preferably, the structure of the arenobufagin carbamate derivative or the pharmaceutically acceptable salt thereof is one of the following structures:

in the definitions of formula 1 given above, the terms used in the collection are generally defined as follows:

the term C3-C8 ring such as pyrrolidinyl, azacycloalkyl, 4-amino azacycloalkyl, and the like.

The term alkyl refers to a straight or branched chain saturated aliphatic hydrocarbon group containing 1 to 20 carbon atoms, for example: methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, and the like.

The compounds of the present invention can be prepared in the form of pharmaceutically acceptable salts according to conventional methods.

The medicinal salt is organic acid or inorganic acid salt.

The inorganic acid refers to hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid, nitric acid and the like.

The organic acid is acetic acid, malic acid, maleic acid, citric acid, fumaric acid, tartaric acid, succinic acid, lactic acid, p-toluenesulfonic acid, salicylic acid or oxalic acid.

The second aspect of the invention provides an application of the arenobufagin carbamate derivative or the pharmaceutical salt thereof in preparing an anti-tumor drug.

The compound prepared by the invention has anti-tumor activity and can be used for treating tumors, such as cancers of esophagus, stomach, intestine, rectum, oral cavity, pharynx, larynx, lung, colon, breast, uterus, endometrium, ovary, prostate, testis, bladder, kidney, liver, pancreas, bone, connective tissue, skin, eye, brain, central nervous system and the like, thyroid cancer, leukemia, Hodgkin's disease, lymphoma, myeloma and the like.

The compound prepared by the invention can be used for preparing anti-tumor drugs, and especially has an inhibiting effect on human lung cancer A549, human colon cancer cell HCT116, human breast cancer MDA-MB-231 and human liver cancer cell Bel7404, so that the compound can be used as drugs for resisting colon cancer, breast cancer, lung cancer and liver cancer.

In a third aspect, the invention provides a pharmaceutical composition comprising the said arenobufagin carbamate derivative or its pharmaceutically acceptable salt or pharmaceutically acceptable carrier. The pharmaceutical composition may be in solid form or in liquid form.

The fourth aspect of the invention provides an application of the pharmaceutical composition in preparing an anti-tumor medicament.

Due to the adoption of the technical scheme, the invention has the following advantages and beneficial effects:

the arenobufagin carbamate derivative has the activity of inhibiting human lung adenocarcinoma A549, human colon cancer cell HCT116, human breast cancer MDA-MB-231 and human liver cancer cell Bel7404, and can be used for preparing anti-tumor medicaments. The invention provides a lead compound for developing new anti-tumor drugs and opens up a new way for further research and development of new anti-tumor drugs.

Detailed Description

In order to more clearly illustrate the invention, the invention is further described below in connection with preferred embodiments. It is to be understood by persons skilled in the art that the following detailed description is illustrative and not restrictive, and is not to be taken as limiting the scope of the invention.

Example 1

Preparation of Compound 1a

Reacting (3S,5R,8R,9S,10S,11S,13R,14S,17R) -11, 14-dihydro-10, 13-dimethyl-12-oxo-17- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H-cyclopentyl [ a]Phenanthren-3-yl (4-nitrophenyl) carbonate (0.70g, 1.60mmol), prepared according to the patent application published under CN110483608A, was dissolved in 5mL of dichloromethane, 21mg of triethylamine and 18mg of piperidine were added and reacted at room temperature overnight. After the reaction was completed, the reaction mixture was washed with water, a saturated sodium carbonate solution and a saturated sodium chloride solution, respectively, and dried over anhydrous sodium sulfate, the solvent was evaporated, and column chromatography purification was performed to obtain 20mg of a white solid 1a in a yield of 54%.¹H NMR(501MHz,Chloroform-d)δ：7.74(dd,J＝9.9,2.5Hz,1H),7.41(s,1H),6.31(d,J＝9.7Hz,1H),5.01(s,1H),4.35(dd,J＝10.9,3.5Hz,1H),4.13(t,J＝8.4Hz,1H),3.85(d,J＝3.6Hz,1H),3.44(s,4H),2.47(d,J＝13.8Hz,1H),2.15–1.93(m,3H),1.78(q,J＝8.3,4.9Hz,5H),1.69(s,2H),1.55(s,6H).1.43–1.24(m,7H),1.21(s,3H),0.94(s,3H)。

Example 2

Preparation of Compound 1b

Compound 1b was obtained as a white solid in 55.6% yield according to the procedure in example 1, substituting diethylamine for piperidine.¹H NMR(501MHz,Chloroform-d)δ：7.73(dd,J＝9.6,2.5Hz,1H),7.41(s,1H),6.31(d,J＝9.8Hz,1H),5.02(s,1H),4.35(dd,J＝11.1,3.6Hz,1H),4.13(dd,J＝9.5,7.1Hz,1H),3.85(d,J＝3.6Hz,1H),3.30(s,4H),2.49(d,J＝14.1Hz,1H),2.16–1.91(m,4H),1.88–1.72(m,5H),1.42–1.25(m,8H),1.21(s,3H),1.16(t,J＝7.2Hz,6H),0.94(s,3H).

Example 3

Preparation of Compound 1c

Compound 1c was obtained as a white solid in 34.6% yield according to the procedure in example 1, substituting cyclopentylamine for piperidine.¹H NMR(501MHz,Chloroform-d)δ：7.74(dd,J＝9.8,2.6Hz,1H),7.41(d,J＝2.6Hz,1H),6.31(d,J＝9.7Hz,1H),4.98(s,1H),4.63(d,J＝7.4Hz,1H),4.34(dd,J＝11.1,3.6Hz,1H),4.13(dd,J＝9.6,7.0Hz,1H),3.98(s,1H),3.84(d,J＝3.6Hz,1H),2.43(d,J＝14.0Hz,1H),2.16–1.91(m,4H),1.91–1.73(m,7H),1.72–1.66(m,4H),1.60–1.25(m,10H),1.20(s,3H),0.94(s,3H).

Example 4

Preparation of Compound 1d

Following the procedure of example 1, pyrrolidine was used in place of piperidine to give compound as a white solidProduct 1d, yield 60.7%.¹H NMR(501MHz,Chloroform-d)δ：7.74(d,J＝9.7Hz,1H),7.41(s,1H),6.31(t,J＝7.7Hz,1H),5.01(s,1H),4.43–4.25(m,1H),4.13(d,J＝9.6Hz,1H),3.85(s,1H),3.39(s,4H),2.46(d,J＝14.1Hz,1H),2.07(s,2H),1.96–1.73(m,11H),1.39–1.25(m,8H),1.20(s,3H),0.94(s,3H).

Example 5

Preparation of Compound 1e

According to the procedure of example 1, cyclohexylamine was used instead of piperidine to give compound 1e as a white solid in a yield of 64.7%.¹H NMR(300MHz,Chloroform-d)δ：7.73(dd,J＝9.7,2.6Hz,1H),7.45–7.37(m,1H),6.37–6.23(m,1H),4.98(s,1H),4.54(s,1H),4.34(d,J＝11.0Hz,1H),4.20–4.07(m,1H),3.83(s,1H),3.48(s,1H),2.41(t,J＝13.7Hz,1H),2.17–1.89(m,4H),1.88–1.49(m,14H),1.47–1.25(m,9H),1.20(s,3H),0.94(s,3H).

Example 6

Preparation of Compound 1f

According to the procedure of example 1, morpholine was used instead of piperidine to give compound 1f as a white solid in 49% yield.¹H NMR(501MHz,DMSO-d6)δ：7.83(dd,J＝9.8,2.6Hz,1H),7.60(dd,J＝2.7,1.2Hz,1H),6.34(dd,J＝9.7,1.1Hz,1H),5.00(s,1H),4.86(s,1H),4.57(d,J＝4.8Hz,1H),4.29(dd,J＝11.1,4.8Hz,1H),3.98(dd,J＝9.6,6.9Hz,1H),3.55(t,J＝4.9Hz,4H),3.35(s,4H),2.45(s,1H),2.00(t,J＝11.7Hz,1H),1.86(p,J＝14.1Hz,4H),1.79–1.56(m,5H),1.43(d,J＝14.5Hz,1H),1.33–1.15(m,5H),1.10(s,3H),0.81(s,3H).

Example 7

Preparation of Compound 1g

According to the procedure of example 1, trans-1, 4-cyclohexanediamine was used in place of piperidine to give 1g of a white solid compound in a yield of 46.7%.¹H NMR(501MHz,DMSO-d6)δ：7.82(dd,J＝9.7,2.5Hz,1H),7.59(d,J＝2.5Hz,1H),6.86(d,J＝7.9Hz,1H),6.34(d,J＝9.8Hz,1H),4.99(s,1H),4.78(s,1H),4.55(d,J＝4.7Hz,2H),4.28(dd,J＝10.8,5.0Hz,1H),3.98(t,J＝8.3Hz,1H),3.16(s,2H),2.42(d,J＝12.9Hz,1H),1.99(t,J＝11.6Hz,2H),1.93–1.52(m,13H),1.41(d,J＝13.5Hz,1H),1.25(td,J＝22.3,11.1Hz,9H),1.09(s,3H),0.81(s,3H).

Example 8

Preparation of Compound 1h

The procedure of example 1 was followed, using 4-aminopiperidine instead of piperidine, to give compound 1h as a white solid in 61.4% yield.¹H NMR(501MHz,DMSO-d6)δ：7.83(dd,J＝9.8,2.6Hz,1H),7.59(d,J＝2.5Hz,1H),6.34(d,J＝9.7Hz,1H),5.01(s,1H),4.83(d,J＝4.4Hz,1H),4.57(d,J＝4.8Hz,1H),4.29(dd,J＝10.8,3.6Hz,1H),3.97(dt,J＝13.4,6.6Hz,3H),3.35(s,4H),3.05(d,J＝11.5Hz,1H),2.86(s,2H),2.00(t,J＝12.1Hz,1H),1.93–1.79(m,5H),1.79–1.56(m,5H),1.43(d,J＝14.6Hz,1H),1.25(ddt,J＝35.1,23.6,13.0Hz,7H),1.11(s,3H),0.81(s,3H).

Example 9

Preparation of Compound 1i

According to the method of example 1, L-alanine ethyl ester hydrochloride instead of piperidine, obtained white solid compound 1i, yield 48.2%.¹H NMR(501MHz,Chloroform-d)δ：7.75(dd,J＝9.8,2.6Hz,1H),7.41(d,J＝2.6Hz,1H),6.29(d,J＝9.7Hz,1H),5.33–5.21(m,1H),4.98(s,1H),4.33(dd,J＝11.1,3.7Hz,2H),4.21(q,J＝7.1Hz,2H),4.12(dt,J＝13.0,5.0Hz,1H),3.84(d,J＝3.6Hz,1H),2.42(d,J＝14.1Hz,1H),2.28(s,1H),2.13–1.90(m,4H),1.82–1.70(m,6H),1.51(d,J＝11.7Hz,1H),1.46–1.24(m,10H),1.19(s,3H),0.93(s,3H).

Example 10

Preparation of Compound 1j

The procedure of example 1 was followed, substituting 3-hydroxyazetidine hydrochloride for piperidine, to give compound 1j as a white solid in 45.3% yield.¹H NMR(501MHz,Chloroform-d)δ：7.74(dd,J＝9.8,2.6Hz,1H),7.46–7.38(m,1H),6.31(dd,J＝9.7,1.1Hz,1H),4.96(t,J＝2.8Hz,1H),4.66(d,J＝5.9Hz,1H),4.34(dd,J＝11.2,3.6Hz,1H),4.30–4.20(m,2H),4.13(dd,J＝9.6,6.9Hz,1H),3.94–3.82(m,3H),2.49–2.38(m,2H),2.07(qd,J＝12.3,10.7,3.9Hz,2H),1.65(s,1H),1.94(d,J＝15.8Hz,2H),1.88–1.69(m,6H),1.51(d,J＝12.9Hz,1H),1.43–1.26(m,5H),1.20(s,3H),0.94(s,3H).

Example 11

Preparation of Compound 1k

According to the procedure of example 1, N- (2-aminoethyl) morpholine was used in place of piperidine to give compound 1k as a white solid in 34.9% yield.¹H NMR(501MHz,Chloroform-d)δ：7.73(dd,J＝9.8,2.6Hz,1H),7.41(d,J＝2.6Hz,1H),6.31(d,J＝9.7Hz,1H),5.04(d,J＝53.3Hz,2H),4.35(dd,J＝11.1,3.6Hz,1H),4.13(dd,J＝9.7,6.9Hz,1H),3.85(d,J＝3.6Hz,1H),3.74(t,J＝4.7Hz,4H),3.37–3.22(m,2H),2.50(dd,J＝12.4,6.2Hz,6H),2.15–1.91(m,4H),1.89–1.73(m,6H),1.47–1.24(m,8H),1.21(s,3H),0.94(s,3H).

Example 12

Preparation of Compound 1l

According to the method of example 1, N-aminopropylmorpholine was used in place of piperidine to give 1l of compound as a white solid in 48.9% yield.¹H NMR(501MHz,Chloroform-d)δ：7.75(dd,J＝9.8,2.6Hz,1H),7.41(d,J＝2.6Hz,1H),6.30(d,J＝9.7Hz,1H),5.77(t,J＝5.5Hz,1H),4.96(s,1H),4.34(d,J＝11.1Hz,1H),4.12(dd,J＝9.6,7.1Hz,1H),3.75(t,J＝4.7Hz,4H),3.27(hept,J＝7.0Hz,2H),2.61–2.37(m,8H),2.14–2.01(m,3H),1.95(td,J＝14.2,11.8,7.4Hz,1H),1.75(ddd,J＝25.8,12.2,6.0Hz,9H),1.51(d,J＝12.9Hz,1H),1.44–1.24(m,5H),1.19(s,3H),0.93(s,3H).

Example 13

Preparation of Compound 1m

According to the procedure of example 1, 1- (2-aminoethyl) pyrrolidine was used in place of piperidine to give compound 1m as a white solid in a yield of 64.7%.¹H NMR(501MHz,DMSO-d6)δ：7.82(dd,J＝9.8,2.6Hz,1H),7.59(d,J＝2.5Hz,1H),7.16(d,J＝5.9Hz,1H),6.38–6.28(m,1H),5.03(s,1H),4.86–4.76(m,1H),4.56(d,J＝4.7Hz,1H),4.28(dd,J＝11.2,4.0Hz,1H),3.97(dd,J＝9.6,7.0Hz,1H),3.25(q,J＝6.4Hz,2H),2.96(d,J＝26.9Hz,6H),2.44(dd,J＝13.6,3.4Hz,1H),2.07–1.94(m,1H),1.92–1.78(m,8H),1.72(dd,J＝19.2,13.2Hz,2H),1.48–1.39(m,1H),1.37–1.15(m,8H),1.09(s,3H),0.81(s,3H).

Example 14

In vitro antitumor Activity test of Compounds prepared in examples 1 to 13 of the present invention

The compound of the invention is subjected to a tumor cell proliferation inhibition test by adopting a conventional MTT method (such as New drug pharmacology research method, 2007: 242-243, compiled by Luqiu army).

The cell strain is selected from A549 (human lung adenocarcinoma cell) HCT116 (human intestinal cancer cell), MDA-MB-231 (human breast cancer cell) and Bel7404 (human hepatoma cell), and is frozen and passaged by a pharmacological laboratory of Shanghai pharmaceutical industry institute. The culture solution is DMEM + 10% FBS + double antibody.

In vitro activity test: the concentration of the added solution in each hole of a 96-hole plate is 4-5 multiplied by 10⁴Cell suspension 100. mu.L/mL, at 37 ℃ in 5% CO₂In the incubator. After 24h, the sample solution was added at 10. mu.L/well in duplicate wells at 37 ℃ with 5% CO₂The reaction is carried out for 72 hours. Adding 20 mu L of 5mg/ml MTT solution into each hole, reacting for 4h, adding a dissolving solution into each hole, placing the holes with the concentration of 100 mu L in an incubator, dissolving, and measuring the OD value of 570nm by using a full-wavelength multifunctional microplate reader. The Inhibition Rate (IR) and half-maximal Inhibition Concentration (IC) were calculated by Excel software₅₀)。

The inhibition rate (IR%) of the drug on cell growth was calculated according to the following formula

IR (%). times (1-mean OD of sample group/mean OD of control group) × 100%

The results are shown in table 1, wherein the samples refer to the arenobufagin carbamate derivatives prepared in the corresponding examples.

In vitro antitumor Activity of the Compounds of Table 1

The experimental results show that the compounds prepared in the embodiments 1 to 13 have good antitumor activity, most of the compounds are higher than the marketed drug doxorubicin and show excellent activity on lung cancer, breast cancer, colon cancer and liver cancer cell lines, so that the compounds and salts thereof can be used for preparing antitumor drugs.

Although the present invention has been described with reference to a preferred embodiment, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.

Claims

1. a class of Xenopusin carbamate derivative or its medicinal salt, it is characterized in that, the structure of described Xenopusin carbamate derivative is a kind of following structure:

2. The carbamate derivative of Xenopusin according to claim 1 or a medicinal salt thereof, wherein the medicinal salt is an organic acid or an inorganic acid salt.

3. Xenopusin carbamate derivative or its medicinal salt according to claim 2, is characterized in that, described inorganic acid refers to hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid or nitric acid.

4. Xenopusin carbamate derivative or its medicinal salt according to claim 2, is characterized in that, described organic acid refers to acetic acid, malic acid, maleic acid, citric acid, fumaric acid , tartaric acid, succinic acid, lactic acid, p-toluenesulfonic acid, salicylic acid or oxalic acid.

5. the application of the Xenopusin carbamate derivative or its medicinal salt according to any one of claims 1 to 4 in the preparation of antitumor drugs, it is characterized in that, described tumor is mammary gland, liver cancer occurred.

6 . A pharmaceutical composition, characterized in that it comprises the xenobufacin carbamate derivative according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. 7 .

7 . The application of the pharmaceutical composition according to claim 6 in the preparation of anti-tumor drugs, wherein the tumor is breast cancer or liver cancer. 8 .