CN111606917A

CN111606917A - Abietane compound with C-ring-fused lactone ring novel skeleton and preparation method and application thereof

Info

Publication number: CN111606917A
Application number: CN202010490069.2A
Authority: CN
Inventors: 周长新; 曾林伟; 侯阿芳; 甘礼社; 高玉丽
Original assignee: Hangzhou Jisong Pharmaceutical Technology Co ltd
Current assignee: Hangzhou Hecheng Pharmaceutical Technology Co.,Ltd.
Priority date: 2020-06-02
Filing date: 2020-06-02
Publication date: 2020-09-01
Anticipated expiration: 2040-06-02
Also published as: CN111606917B

Abstract

The invention discloses a abietane compound with a novel C-ring-fused lactone ring framework, which is prepared by taking abietane diterpene quinone compounds as raw materials and condensing with acid anhydride under the action of a metal catalyst. The non-planarity of the constructed novel abietane compounds is effectively improved, the novel abietane compounds have novelty in structure, and the corresponding biological activity is remarkably improved, so that the compounds are prompted to have greater druggability.

Description

Abietane compound with C-ring-fused lactone ring novel skeleton and preparation method and application thereof

Technical Field

The invention relates to the field of medicinal chemistry, in particular to a abietane compound with a novel skeleton of a C-ring-fused lactone ring, a preparation method and application thereof.

Background

Abietane diterpenoid compounds the first abietane diterpenoid compound abietic acid was first discovered by Ellingson from Colophonium (Colophony) at the beginning of the 19 th century (Ellingson, E.O. Abietic acid and solids of materials. journal of the American Chemical society, 1914,36, 325. 335.), and it was shown to have a variety of pharmacological effects against bacteria, gastric acid secretion, etc. (San Feiliano, A. Gordalliza, M.; Salinero, M.A. et al. Abietic acids: sources, biological activities, and botanical, plant a medical.1993, 59,485-90.). And then has attracted the attention of people. In nature, abietane diterpene mainly exists in plants of Labiatae, Pinaceae, Compositae, Euphorbiaceae, Leguminosae, Celastraceae, etc., especially in Labiatae, and its basic skeleton is hydrogenated phenanthrene structure. In the labiatae family, the plants are most widely distributed in the sage and rabdosia.

Later, scholars at home and abroad successively discover that the similar compounds also have the effects of resisting tumors, inflammation, viruses and the like. Among the compounds, researches are more intensive on diterpenoid compounds in salvia plants, the main structure type of the diterpenoid compounds is abietane diterpenoid quinine compounds, and many of the diterpenoid quinine compounds have more remarkable physiological activities. Pharmacological experiments prove that the compounds have a plurality of effects such as anti-tumor (Zhang, Y.; Jiang, P.X.; Ye, M.et al. Tanshinones: sources, pharmacological activities-peptides. International Journal of Molecular sciences.2012,13, 13621.), anti-inflammation, crown expansion, free radical elimination, platelet aggregation and the like (Jiang, Z.Q.; Gao, W.; Huang, L.Q. Tanshinones, clinical pharmacological compositions in Salvimamitorrhiza. Fronti. in pharmacological chemistry.2019, 10, 1-14.). In order to better exert the drug effect and improve the drug potency, a plurality of scholars at home and abroad carry out structural modification, and some compounds are widely applied clinically through simple structural modification. Such as tanshinone IIA sodium sulfonate, is a product of tanshinone IIA introduced with sulfonic group in D ring, and is clinically used for preventing and treating coronary heart disease and angina pectoris.

At present, the structural modification methods of the compounds are summarized into two types: i.e., modifications to the A and D rings. This is because, in the case where the ring a is alicyclic, the benzylic position is more active, and there are many reaction sites, and various types of substituents can be introduced at different positions; and the furan ring of the D ring, hydrogen is more active site, for example, the tanshinone IIA sodium sulfonate is prepared, and the water solubility is obviously improved. However, at present, the two structural modification effects are not ideal, the activity improvement of the compounds is limited, and researches show that the drug effect of tanshinone IIA sodium sulfonate is even lower than that of tanshinone IIA, and presumably because the sodium sulfonate is an ionic compound, the compound is difficult to pass through a biomembrane structural system based on lipid and protein components (tanshinone IIA and tanshinone IIA sodium sulfonate in vivo pharmacokinetics and pharmacodynamics such as shiyuan, li xiying, royal tianyi, and the like, and Chinese journal of pharmacy 2009,7(3), 143-153.). The tanshinone IIA sodium sulfonate has the greatest advantage of being a water-soluble compound, can be injected for administration, and is a drug approved in the 80 th century in China to come into the market. However, in modern times, this strategy to increase water solubility at the expense of drug efficacy has not been commonly used. Furthermore, these structural modification methods are also limited to two conditions: (1) the A ring must be alicyclic; (2) the presence of a D-furan ring is essential.

Through research, the applicant finds that the antibacterial and antitumor activity of the compound can be remarkably improved by carrying out structural modification on a C-ring o-quinone fragment (Zeng, L.W.; Zhou, C.X.; et al. design, synthesis, and antibacterial and antitumor activities of new tanshinone IIA esters. Natural product research,2016,30, 2662-shaped 2668. Hou Aromata, Zanling, Ganmeisha et al. design synthesis of danshenxintong C-cyclic phenol ester derivatives and research on the antitumor activity. modern applied pharmacology in China, 2016,33, 1396-shaped 1402.), so that structural modification of the C-ring has important scientific significance for improving the pharmaceutical activity of the compound, but the C-ring of the compound is just difficult to modify due to the blunt framework. Through long-term intensive research, the applicant successfully develops a modification method for synthesizing a novel C-ring framework by taking abietane-type diterpenoid quinone as a precursor, obtains a novel structure of a C-ring-fused lactone ring, and realizes the construction of a novel framework with greater potency of the compound.

Disclosure of Invention

Aiming at the defects in the prior art, the invention aims to provide the abietane compound with the novel skeleton of the C-ring-fused lactone ring, effectively improves the non-planarity of the compound, has novelty in structure, and obviously improves the corresponding biological activity, thereby prompting that the compound has greater potential for drug development; secondly, the preparation method of the abietane compound which takes abietane diterpenoid quinone as a precursor to synthesize a new C-ring framework is provided and applied to clinical application of relevant medicine.

In order to achieve the first object, the invention provides the following technical scheme:

the abietane compound with C-ring lactone ring skeleton includes A, B, C tricyclic skeleton structure and has the following structural formulas:

the compound of the structural type shown in the formula I, wherein n is 0-2.

In the compound with the structure shown in the formulas I, II and III, the ring A can be alicyclic or aromatic ring, and the ring B and the ring C are aromatic rings.

When in use, the compound has the structure shown as the following formula I, II and III or pharmaceutically acceptable salts thereof. The medicinal salt of the compound with the structure shown in the formula I, II and III refers to a medicinal salt generated by the reaction of side chain carboxyl of lactone ring and alkali.

The pharmaceutically acceptable salt in the "pharmaceutically acceptable salt of the compound having a structure represented by formula I, II, III" is not particularly limited, and is in the form of a salt which does not cause a pharmacological side effect and is conventionally used, and includes salts formed by reacting the compound having a structure represented by formula I, II, III with any inorganic or organic base, such as sodium salts, potassium salts, magnesium salts, ammonium salts, and the like. For example, it may be salified with an inorganic base such as sodium bicarbonate, sodium carbonate, sodium hydroxide, potassium carbonate, potassium hydroxide, magnesium hydroxide, preferably sodium bicarbonate, sodium carbonate and potassium carbonate; salts with organic bases such as methylamine, ethylamine, ethanolamine, ethylenediamine, dimethylamine, trimethylamine, diethylamine, triethylamine, propylamine, butylamine are also possible, dimethylamine and diethylamine being preferred.

In the compound with the structure shown in the formulas I, II and III, the substituent on the 4-position of the ring A is alkyl or hydrogen;

when the ring A is an alicyclic ring, an alicyclic ring having a gem-dimethyl substitution at the 4-position is preferred; when ring A is an aromatic ring, the aromatic ring substituted with a methyl group at the 4-position is preferred.

R₁、R₂Is alkyl or hydrogen, the substituent outside the C ring is alkyl, and a furan ring or dihydrofuran ring combined structure is also formed; when the exocyclic substituent is alkyl, preferably R1 is isopropyl; when the C ring is exocyclic, a fused structure of furan ring or dihydrofuran ring is preferable. I.e. when the substituent R is at the outer 13-position of the C ring₁R in the 14-position in the case of isopropyl₂Is hydrogen; when the isopropyl end at position 13 is oxidized, it may be condensed with position 14 to form a furan ring or a dihydrofuran ring structure.

The compound with the structures of the formulas I, II and III has a basic structure in the structure of the type I, and the structures of the types II and III are obtained by converting the structure of the type I. I.e. when n ═ 0, the class I structure is converted to a class II structure by further reaction; when n ═ 1, the class I structure is converted to a class III structure by further reaction.

The invention also provides a preparation method of the abietane compound with the novel skeleton of the C-ring-fused lactone ring, which comprises the following steps in sequence:

(1) synthesis of compound with structure in class I

Putting raw materials of abietane diterpene quinone compounds and acid anhydride into a reaction bottle according to a molar ratio of 1: 1-1: 1.5, adding a solvent, DMAP and simple substance reducing metal, heating under the protection of nitrogen, monitoring by TLC to react completely, filtering, evaporating filtrate to dryness, and then recrystallizing or carrying out column chromatography to obtain the target product with the I-type structure.

(2) Synthesis of compounds with II and III structures

Dissolving the compound with the I-type structure, adding the compound into a reaction bottle, adding DCC, DMAP and organic base, heating under the protection of nitrogen, monitoring by TLC to complete the reaction, filtering, evaporating the filtrate to dryness, and then performing recrystallization or column chromatography to obtain target products with II-type and III-type structures respectively according to different n values.

The four compounds prepared by the reaction are racemes, namely the benzyl carbon of the C ring is a chiral center and comprises two configurations of R and S, and a single optically active body can be obtained by chiral resolution.

The raw materials and reagents in the steps (1) and (2), wherein the abietane-type diterpene quinone is selected from compounds with an o-quinone structure type, preferably o-quinone compounds in salvia plants, more preferably tanshinone I, tanshinone IIA, cryptotanshinone and tanshinone in salvia miltiorrhiza in salvia plants; the acid anhydride is selected from cyclic dianhydride, preferably succinic anhydride, glutaric anhydride, adipic anhydride; the simple reducing metal is selected from magnesium powder, iron powder and zinc powder; the organic base is selected from triethylamine and N, N-Diisopropylethylamine (DIPEA).

The reaction mechanism of the abietane derivative for constructing the novel skeleton of the C-ring fused lactone ring is that the reaction of tanshinone IIA and succinic anhydride under the action of DMAP and zinc powder is shown as follows:

the reaction of tanshinone IIA and succinic anhydride under the action of DMAP and zinc powder is shown as the following '9' supposing that in the whole reaction system, the reaction system is slightly alkaline, and a five-membered lactone ring is relatively long in side chain, so that the lactone ring is relatively easy to open, and the carboxylic acid at the other end can form ester with phenolic hydroxyl to form a relatively stable heptatomic lactone ring, so that a product is obtained.

In the reaction, firstly, the 10-bit carbonyl group of tanshinone IIA is reduced by zinc powder, and is subjected to acylation reaction with succinic anhydride activated by DMAP, and after DMAP leaves, zinc is removed in the form of zinc hydroxide under the action of a small amount of water. The isomerized ester carbonyl is then condensed with the carbonyl at the 11 position, and aromatized to form the product after removal of 1 molecule of water.

The reaction mechanism of converting the I structure into the II and III structures through further reaction is that tanshinone IIA and glutaric anhydride react in the presence of an alkaline catalyst and a condensing agent as follows:

in the whole reaction system, the reaction system is slightly alkaline, the five-membered lactone ring is relatively long in side chain, the lactone ring is relatively easy to open, and the carboxylic acid at the other end can form ester with the phenolic hydroxyl group to form a relatively stable seven-membered lactone ring, so that the product is obtained.

The invention also provides a medicine for treating tumors and cardiovascular and cerebrovascular diseases, which comprises the abietane compound with the new skeleton of the C-ring-fused lactone ring.

In one embodiment of the invention, the abietane derivative with the new skeleton of C-ring-fused lactone ring can be added into the medicine separately; the new skeleton derivative and other known active components of antitumor and cardio-cerebral vascular treating medicine may be also added together.

In another embodiment of the invention, in the anti-tumor and cardiovascular and cerebrovascular disease treatment drug, the content of the abietane diterpene derivatives as active ingredients is in the range of 0.1-99.9%. Preferably 50% to 99.9%.

The content of the derivative in the medicament can be selected by those skilled in the art according to the dosage, dosage form and administration mode of the medicament.

The preparation of the anti-tumor and cardiovascular and cerebrovascular disease treatment drug can be tablets, capsules, orally disintegrating tablets, dripping pills, sprays, aerosols, injections and the like.

The invention has the beneficial effects that firstly, the invention provides a novel abietane compound with a novel skeleton of C-ring-fused lactone ring, a derivative thereof, pharmaceutically acceptable salts thereof and a preparation method thereof; secondly, the compounds have better anti-tumor and cardiovascular and cerebrovascular activities. In vitro experiments show that the compounds have good inhibitory activity on human lung adenocarcinoma cells A549; has obvious protective effect on myocardial cells H9C 2. In vivo animal experiments show that the compounds have good tumor inhibition effect on lung cancer tumor-bearing mice after intraperitoneal injection administration; the compound can obviously reduce the cerebral infarction range of a middle cerebral artery occlusion Model (MCAO) of a rat, has good protection effect on focal cerebral ischemic injury, and shows that the compound has important application value in clinic.

Drawings

FIG. 1 is an X single crystal diffractogram of Compound 1.

Fig. 2 is a hydrogen spectrum of compound 2.

Fig. 3 is a carbon spectrum of compound 2.

Fig. 4 is a hydrogen spectrum of compound 3.

Fig. 5 is a carbon spectrum of compound 3.

FIG. 6 is a hydrogen spectrum of Compound 4.

FIG. 7 is a carbon spectrum of Compound 4.

Fig. 8 is a hydrogen spectrum of compound 5.

Fig. 9 is a carbon spectrum of compound 5.

FIG. 10 is a mass spectrum of Compound 6.

FIG. 11 is a carbon spectrum of Compound 6.

Fig. 12 is a hydrogen spectrum of compound 7.

Fig. 13 is a high resolution mass spectrum of compound 7.

Figure 14 is a high resolution mass spectrum of compound 8.

Fig. 15 is a high resolution mass spectrum of compound 9.

Detailed Description

Example 1

Weighing 10g (34mmol) of tanshinone IIA raw material into a reaction bottle, adding 100ml tetrahydrofuran, adding 3.4g (34mmol) of succinic anhydride, 9.5g of DMAP and 6.6g of zinc powder while stirring, heating and refluxing under the protection of nitrogen, stirring for 3 hours, and monitoring the reaction completion by TLC. The zinc powder is removed by filtration, the filtrate is evaporated to dryness and then recrystallized by methanol, and the product (compound 1) is obtained by drying, wherein the yield is 75 percent. The chemical structure is confirmed by spectrum analysis and X-single crystal diffraction as follows:

spectral data for compound 1 are as follows:

ESI–MS:m/z＝379.6[M+H]⁺。

¹H NMR(600MHz,DMSO-d₆)12.43(s,1H),8.03(d,J＝8.8Hz,1H),7.91(d, J＝1.2Hz,1H),7.69(d,J＝8.8Hz,1H),4.54(t,J＝4.6Hz,1H),3.38–3.31(m,2H), 3.25(dd,J＝17.2,5.6Hz,1H),3.14(dd,J＝17.2,5.6Hz,1H),2.35(d,J＝1.2Hz,3H), 1.92–1.79(m,2H),1.77–1.64(m,2H),1.35(s,3H),1.34(s,3H)；¹³C NMR(150 MHz,DMSO-d₆)176.61,170.37,147.32,145.55,142.61,142.48,130.18,126.09, 118.64,118.29,117.14,116.21,114.67,114.50,38.50,37.32,35.35,33.69,30.89, 30.82,28.87,18.52,9.06.

example 2

Weighing 10g (34mmol) of tanshinone IIA raw material in a reaction bottle, adding 100ml tetrahydrofuran,

glutaric anhydride 3.88g (34mmol), DMAP 7.8g and iron powder 5.9g were added with stirring, heated under reflux under nitrogen, stirred for 6 hours and then monitored by TLC for completion of the reaction. Iron powder is removed by filtration, the filtrate is evaporated to dryness and then recrystallized by acetone, and the product (compound 1) is obtained by drying, wherein the yield is 81 percent. The chemical structure was confirmed by spectroscopic analysis as follows:

spectral data for compound 2 are as follows:

ESI-MS:m/z＝391.30[M-H]^-.

¹H NMR(500MHz,Acetone-d₆)8.09(d,J＝8.8Hz,1H),7.80(d,J＝1.2Hz, 1H),7.72(d,J＝8.8Hz,1H),4.39(dd,J＝8.5,3.5Hz,1H),3.43(t,J＝6.4Hz,2H), 2.47(d,J＝1.0Hz,3H),2.44–2.31(m,3H),1.94–1.87(m,2H),1.75(dd,J＝12.1, 6.2Hz,2H),1.38(d,J＝3.7Hz,6H).¹³C NMR(125MHz,Acetone-d₆)177.93, 173.64,149.72,147.32,144.27,143.80,132.32,127.67,121.07,120.24,118.85, 118.47,116.73,116.69,42.35,39.25,35.28,32.06,32.02,30.64,30.30,28.44, 20.31,9.92.

example 3

Weighing 1.0g (2.55mmol) of compound 2 raw material in a reaction bottle, adding 30ml dichloromethane, adding 1.2g DMAP, 0.6g DCC and 0.2ml triethylamine with stirring, heating and refluxing under the protection of nitrogen, stirring for 5 hours, and monitoring the reaction completion by TLC. The reaction was quenched with ice water, extracted with dichloromethane (20 ml. times.3), combined, concentrated in the organic phase, and isolated on silica gel column to give 0.59 g of the product (compound 3) in 59% yield. The chemical structure was confirmed by spectroscopic analysis as follows:

spectral data for compound 3 are as follows:

ESI-MS:m/z＝391.28]M-H]^-.

¹H NMR(500MHz,Acetone-d₆)10.66(1H,br.s),8.11(d,J＝8.7Hz,1H),7.79 (s,1H),7.63(d,J＝8.7Hz,1H),4.55(dd,J＝9.7,2.9Hz,1H),3.42–3.39(m,1H),3.26 (d,J＝16.3Hz,1H),2.62–2.47(m,2H),2.42–2.36(m,4H),2.02–1.95(m,2H),1.39(s, 3H),1.36(s,3H)；¹³C NMR(125MHz,Acetone-d₆)178.19,173.58,153.91,147.34, 145.34,142.88,132.30,128.25,125.80,119.55,118.83,115.83,115.11,112.19, 45.62,39.50,35.49,32.54,31.87,30.67,30.09,29.61,20.56,9.17。

example 4

500mg of danshenxintong was weighed into 100ml of a three-necked flask containing 20ml of dioxane, and 370mg of succinic anhydride, 450mg of DMAP and 300mg of magnesium powder were added under stirring and refluxed under the protection of nitrogen. After stirring for 4 hours, the reaction was monitored by TLC for completion. Filtering to remove magnesium powder, evaporating the filtrate to dryness, and adding petroleum ether: ethyl acetate 1:1 was recrystallized and dried to give 568mg of the product (Compound 4) in 88% yield. The chemical structure was confirmed by spectroscopic analysis as follows:

spectral data for compound 4 are as follows:

ESI-MS:m/z＝367.51[M+H]⁺.

¹H NMR(500MHz,DMSO-d₆)12.46(s,1H),7.66(d,J＝8.8Hz,1H), 7.53(s,1H),7.50(d,J＝8.8Hz,1H),4.37(t,J＝4.5Hz,1H),3.28(dd,J＝9.5, 6.2Hz,2H),3.15(dd,J＝4.4,2.2Hz,2H),3.06(hep,J＝6.8Hz,1H),1.89–1. 77(m,2H),1.66(dd,J＝7.5,4.1Hz,2H),1.30(s,6H),1.29(d,J＝6.8Hz,3 H),1.26(d,J＝6.8Hz,3H)；¹³C NMR(125MHz,DMSO-d₆)177.27,171.13, 149.70,142.54,141.81,133.13,129.21,125.85,125.76,121.23,119.74,117.6 7,39.11,38.03,34.95,34.07,31.27,31.23,29.30,29.19,23.77,23.39,19.14。

example 5

500mg of Salvianolic acid was weighed into a 100ml three-necked flask containing 30ml of tetrahydrofuran, 422mg of glutaric anhydride, 530mg of DMAP and 400mg of zinc powder were added with stirring, and heated under reflux under nitrogen. After stirring for 8 hours, TLC monitored the reaction for completion. The zinc powder was removed by filtration, and the filtrate was evaporated to dryness and passed through a silica gel column to isolate 475mg of the product (compound 4) with a yield of 71%. The chemical structure was confirmed by spectroscopic analysis as follows:

spectral data for compound 5 are as follows:

ESI-MS:m/z＝379.27[M-H]^-.

¹H NMR(500MHz,Acetone-d₆)7.67(d,J＝8.8Hz,1H),7.59(s,1H), 7.52(d,J＝8.8Hz,1H),4.18(dd,J＝8.2,3.3Hz,1H),3.35(t,J＝6.4Hz,2H), 3.26(dt,J＝13.7,6.8Hz,1H),2.56–2.47(m,2H),2.37–2.27(m,2H),1.91–1. 86(m,2H),1.71(dd,J＝12.8,7.0Hz,2H),1.41(d,J＝6.8Hz,3H),1.34(s,6 H),1.30(d,J＝6.8Hz,3H)；¹³C NMR(125MHz,DMSO-d₆)177.93,173.71, 150.74,143.79,143.26,134.92,130.80,126.91,126.88,122.63,121.27,119.29, 42.29,39.27,35.03,31.84,31.74,30.42,30.39,29.99,27.24,24.53,23.80,20. 28。

example 6

Weighing 5g (18mmol) of tanshinone I raw material into a reaction bottle, adding 100ml of tetrahydrofuran, adding 2.7g (27mmol) of succinic anhydride, 5.8g of DMAP and 6.7g of zinc powder while stirring, heating and refluxing under the protection of nitrogen, stirring for 6 hours, and monitoring the reaction completion by TLC. The zinc powder is removed by filtration, the filtrate is evaporated to dryness and then recrystallized by acetone, and the product (compound 6) is obtained by drying, 5.9g, and the yield is 91%. The chemical structure was confirmed by spectroscopic analysis as follows:

spectral data for compound 6 are as follows:

ESI–MS:m/z＝361.23[M+H]⁺。

¹H NMR(500MHz,DMSO-d₆)12.49(s,1H),9.16(d,J＝8.5Hz,1H),8.30(d, J＝9.2Hz,1H),8.21(d,J＝9.2Hz,1H),8.08(s,1H),7.70(t,J＝8.5Hz,1H),7.58(d, J＝8.5Hz,1H),4.80(t,J＝4.6Hz,1H),3.35(dd,J＝17.1,4.6Hz,1H),3.18(dd,J＝ 17.1,4.6Hz,1H),2.77(s,3H),2.43(s,3H)；¹³C NMR(150MHz,DMSO-d₆)177.03,171.00,148.02,146.44,144.27,134.73,130.22,128.06,127.88,127.03, 124.73,124.35,120.81,118.32,118.09,116.65,115.10,112.69,39.78,36.01,19.73, 9.62。

example 7

Weighing 1.0g (2.65mmol) of the compound 1 raw material in a reaction bottle, adding 50ml of dichloromethane, adding 1.3g of DMAP, 0.8g of DCC and 0.3ml of triethylamine while stirring, heating and refluxing under the protection of nitrogen, stirring for 5 hours, and monitoring the reaction completion by TLC. The reaction was quenched with ice water, extracted with dichloromethane (20 ml. times.3), combined and the organic phases concentrated and isolated on silica gel column to give 0.66 g of the product (compound 7) in 66% yield. The chemical structure was confirmed by spectroscopic analysis as follows:

spectral data for compound 7 are as follows:

HRESI-MS:m/z＝379.1551[M+H]⁺；779.2909[2M+Na]⁺。

¹H NMR(500MHz,DMSO-d₆)12.49(br.s,1H),8.03(d,J＝8.8Hz,1H),7.93 (d,J＝1.3Hz,1H),7.70(d,J＝8.8Hz,1H),4.59(dd,J＝5.4,3.8Hz,1H),3.25(dd, J＝17.4,3.8Hz,1H),3.14(dd,J＝17.4,3.8Hz,1H),2.35(d,J＝1.2Hz,3H),1.86–1.84 (m,2H),1.70–1.68(m,2H),1.34(s,3H),1.33(s,3H)；¹³C NMR(125MHz, DMSO-d₆)177.32,171.06,147.93,146.18,143.28,143.12,130.80,126.77,119.24, 118.94,117.79,116.83,115.34,115.15,39.78,37.92,35.95,34.33,31.52,31.45, 29.52,19.14,9.72.

example 8

Weighing 10g (34mmol) of tanshinone IIA raw material in a reaction bottle, adding 100ml tetrahydrofuran, adding 4.36g (34mmol) of adipic anhydride, 8.8g of DMAP and 6.9g of iron powder while stirring, heating and refluxing under the protection of nitrogen, stirring for 6 hours, and monitoring the reaction completion by TLC. Iron powder is removed by filtration, the filtrate is evaporated to dryness and then recrystallized by acetone, and the product (compound 1) is obtained by drying, wherein the yield is 56 percent. The chemical structure was confirmed by spectroscopic analysis as follows:

spectral data for compound 8 are as follows:

HRESI-MS:m/z＝429.0694[M+Na]⁺

¹H NMR(500MHz,Acetone-d₆)12.45(br.s,1H),8.07(d,J＝8.8Hz,1H), 7.82(d,J＝1.2Hz,1H),7.75(d,J＝8.8Hz,1H),4.46(dd,J＝6.8,4.4Hz,1H),3.25 (t,J＝6.7Hz,2H),2.44(d,J＝1.0Hz,3H),2.41–2.29(m,4H),1.92–1.88(m,2H), 1.77(m,2H),1.36(s,3H),1.34(s,3H).¹³C NMR(125MHz,Acetone-d₆) 177.91,173.62,149.70,147.29,144.26,143.77,132.30,127.63,121.03,120.21, 118.82,118.44,116.71,116.66,42.33,39.23,35.26,32.03,32.01,30.63,30.48, 30.29,28.41,20.28,9.88.

example 9

Weighing 10g (34mmol) of cryptotanshinone raw material into a reaction bottle, adding 100ml of tetrahydrofuran, adding 3.4g (34mmol) of succinic anhydride, 9.5g of DMAP and 6.6g of zinc powder while stirring, heating and refluxing under the protection of nitrogen, stirring for 6 hours, and monitoring the reaction completion by TLC. The zinc powder is removed by filtration, the filtrate is dried by distillation and recrystallized by methanol, and the product (compound 1) is obtained by drying, with the yield of 68 percent. The chemical structure was confirmed by spectroscopic analysis as follows:

spectral data for compound 9 are as follows:

HRESI-MS:m/z＝381.1706[M+H]⁺,403.1527[M+Na]⁺,783.3165[2M+Na]⁺

¹H NMR(600MHz,DMSO-d₆)12.41(s,1H),7.72(d,J＝8.4Hz,1H),7.41(d, J＝8.4Hz,1H),4.82(dd,J＝8.8,6.8Hz,1H),4.39(t,J＝3.8Hz,1H),4.28(t,J＝8.8,6.2 Hz,1H),3.57(1H,m),3.32-3.28(m,2H),1.92–1.79(m,2H),1.77–1.64(m,2H), 1.32(d,J＝6.8Hz,3H),1.29(s,3H)；¹³C NMR(150MHz,DMSO-d₆)175.69, 171.36,152.32,144.67,135.98,128.76,126.24,122.57,118.34,117.81,116.33, 114.50,71.61,37.50,34.42,34.29,32.93,32.88,30.88,29.69,28.36,19.62,18.73.

example 10: anti-tumor effect

1 in vitro anti-tumor activity experiment on human lung adenocarcinoma cell A549

1.1 Experimental methods

Digesting the monolayer cultured cells to prepare a single cell suspension, counting the cells to 3 × 10 per well⁴Cells were seeded in well-prepared 96-well culture plates at 200uL per well volume. The plates were placed at 37 ℃ in 5% CO₂Culturing in an incubator for 24 h. After changing, different concentrations of test compound 4 (compounds dissolved in DMSO and diluted in PBS, test compound concentrations of 0.5. mu.M, 1. mu.M, 2. mu.M, 4. mu.M, 8. mu.M, 16. mu.M, 32. mu.M and 64. mu.M, respectively) were added at 37 ℃ with 5% CO₂And culturing in an incubator for 24 hours. MTT was added to 96-well plates at 10uL per well and the reaction was carried out in an incubator for 4 h. The supernatant was aspirated, 150uL of DMSO was added to each well, and the plate was placed on a shaker at room temperature and shaken for 10min to allow complete dissolution. The wavelength of 492nm was selected, the light absorption value (OD value) of each well was measured using a microplate reader, and the IC was calculated₅₀The value is obtained.

1.2 results of the experiment

The results show that the antitumor activity of the compound 4 in the table 1 on A549 is obviously improved compared with that of the parent compound danshenxintong, and the IC of the compound is₅₀5.5. mu.M.

Table 1 IC of compound 4 to a549₅₀(μM)

2 in vivo experiment on mice bearing Lung cancer

2.1 preparation of mouse Lewis Lung cancer model

The mouse Lewis lung cancer cell is cultured in RPMI-1640 complete culture solution (containing 10% fetal calf serum, 100u/ml penicillin and 100u/ml streptomycin) by conventional method, digested and passaged by 0.25% trypsin solution, and cultured at 37 deg.C saturation humidity and 5% CO₂Collecting tumor cells in logarithmic growth phase, centrifuging for 3 min at 1500 rpm, washing cell precipitate with serum-free and antibiotic-free medium, counting trypan blue cells, and regulating cell concentration to 1 × 10 with serum-free and antibiotic-free medium⁷/ml。

Selecting 40 healthy C57BL/6J mice, inoculating the tumor cells into subcutaneous tissues of hind limbs on the left and right sides of the miceEach mouse was inoculated with 100ul of 1 × 10⁶And (3) preparing tumor cells for later use after tumor formation (after 7 days, the inoculated part can touch tumor tissues, and tumor masses with the diameter of 1.5-2 cm can be seen after 3 weeks).

2.2 Effect on mouse Lewis Lung cancer

The mice were randomly divided into 5 groups of 8 mice each: the negative control group is injected with 0.05ml/10g of normal saline every day in the abdominal cavity; low, Medium, and high dose groups of Compound 1 (1-L, 1-M, 1-H): performing intraperitoneal injection at 2.5mg/kg, 5mg/kg and 10mg/kg every day; positive control drug 0.05ml/10g of 0.5mg/ml liquid medicine was administered intraperitoneally every other day along with platinum group (Cisplatin), mice were sacrificed on day 15 after inoculation, tumor masses were weighed, and percent tumor inhibition was calculated.

2.3. Results of the experiment

The results are shown in Table 2. As can be seen from the experimental results, Lewis lung carcinoma began to develop in each group of animals one week after inoculation and then entered the rapid growth phase. At the end of the experiment, the Lewis lung cancer volume of each experimental group animal is reduced to different degrees. The compound 1 has obvious inhibition effect on the tumor weight of Lewis lung cancer. Wherein the tumor weight and the tumor weight index of the mice in the high-dose group are obviously lower than those of the control group (P < 0.01).

TABLE 2 Effect of Compound 1 on mouse Lewis Lung carcinoma (II)

n＝8)

Note: comparison with negative control group: p <0.05, P <0.01

Example 11: protective effect on H9C2 cardiac muscle cells

The

compounds

2 and 3 are prepared into three concentrations of 10 μ M, 5 μ M and 2.5 μ M, the parent compound tanshinone IIA is prepared into 5 μ M, H9C2 cells are prepared into 1 × 10⁴After 24 hours for each cell/ml plate, serum and high sugar type medium were removed and replaced with sugar-free serum containing different drug concentrations. After 6 hours of hypoxia induction and 12 hours of reoxygenation, supernatant is sucked for measuring LDH activity, and MTT is used for incubation for 4 hours for measuring the cell survival rate. All dataStatistical analysis was performed with t-text.

The results show that compounds 2 and 3 can increase the survival rate of H9C2 cells and reduce the activity of cellular LDH within the set concentration, have a relatively obvious protective effect on myocardial cells, and are superior to the parent compound tanshinone IIA (Table 3) at the same concentration.

TABLE 3 protective Effect of

Compounds

2 and 3 on H9C2 cells

Note: comparison with negative control group: p <0.05, P <0.01

Example 12: protective effect on Middle Cerebral Artery Occlusion (MCAO) injury of rat

1. Experimental methods and grouping: an intravascular thrombus line blocking method is adopted to prepare a rat Middle Cerebral Artery Occlusion (MCAO) model, tail vein administration is carried out, and the cerebral infarction range is measured. 60 healthy male Sprague Dawley rats weighing 230-280 g were randomly divided into 7 groups, namely a sham operation group, a positive control group (nimodipine, 2mg/kg), a model group, a compound 7 low dose group (7-L, 5mg/kg), a medium dose group (7-M, 10mg/kg), a high dose group (7-H, 20mg/kg) and a parent compound tanshinone I group (20 mg/kg). 1 day before operation, the rat is injected into abdominal cavity; the next day before 30 minutes, the medicine is taken again; the third dose was administered 4 hours after the operation.

2. And (3) measuring the cerebral infarction range:

after 24 hours of operation, the animal is cut off the head and the brain is taken out, the olfactory bulb, the cerebellum and the lower brainstem are removed, and then the coronaries are cut into 4 pieces in total of 5 pieces. 5 pieces of brain tissue are placed in 0.25% TTC solution, incubated for 30min at 37 deg.C in dark, and stained to obtain red normal tissue and white infarcted part. Calculating the infarct area by using a morphological analysis system, and comparing. The results are shown in Table 4.

TABLE 4 Effect of Compound 6 on the cerebral infarct size in rats with focal cerebral ischemia

Group of	n (number)	Infarct size (%)
			Model set	10	39.16±2.74
Positive control group (nimodipine)	10	20.47±5.91**
			6-H	10	22.36±6.81**
6-M	10	25.13±5.86*
			6-L	10	29.64±11.59
Parent compound tanshinone I	10	31.29±8.94

Note: comparison with model groups: p <0.05, P < 0.01.

The results show that the middle and high dose groups of the compound 6 can obviously reduce the cerebral infarction range, have good protective effect on the mouse focal cerebral ischemic injury, and have almost no effect on the parent compound tanshinone I.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention.

Claims

1. The abietane compound with C-ring lactone ring skeleton includes A, B, C tricyclic skeleton structure and has the following structural formulas:

wherein n is 0 to 2.

2. The abietane compound having a novel skeleton containing a C-ring-fused lactone ring as claimed in claim 1, wherein the a ring is an alicyclic ring or an aromatic ring, and both the B ring and the C ring are aromatic rings; the substituent on the 4-position of the A ring is alkyl or hydrogen;

preferably, when the A ring is alicyclic, the substituent at the 4-position is gem-dimethyl; when the A ring is aromatic, the substituent at the 4-position is methyl.

3. The compound of claim 2, wherein R is a abietane compound having a novel skeleton containing a C-ring-fused lactone ring₁、R₂Is alkyl or hydrogen, the substituent outside the C ring is alkyl, and a furan ring or dihydrofuran ring combined structure is also formed;

preferably, the substituent R is at the 13-position outside the C ring₁R in the 14-position in the case of isopropyl₂Is hydrogen; when the 13-isopropyl group is oxidized, it may be condensed with the 14-positionFuran ring or dihydrofuran ring structures.

4. The abietane compound with a novel C-ring-fused lactone ring skeleton as claimed in claim 1, wherein the structure of group I is a basic structure, and the structures of group II and III are both obtained by converting the structure of group I, i.e. when n is 0, the structure of group I is converted into the structure of group II by further reaction; when n ═ 1, the class I structure is converted to a class III structure by further reaction.

5. The abietane compound with a novel C-ring-fused lactone ring skeleton as claimed in claim 4, which is prepared from abietane-type diterpene quinone compounds and acid anhydride, and has the following reaction equation:

6. the process for preparing the abietane compound having a novel skeleton containing a C-ring-fused lactone ring as claimed in any one of claims 1 to 5, which comprises the following steps:

(1) putting raw materials of abietane diterpene quinone compounds and anhydride into a reaction bottle according to a molar ratio of 1: 1-1: 1.5, adding a solvent, DMAP and simple substance reducing metal, heating under the protection of nitrogen, monitoring by TLC (thin layer chromatography) for complete reaction, filtering, evaporating filtrate to dryness, and then recrystallizing or carrying out column chromatography to obtain a target product with a class I structure;

(2) dissolving the compound with the I-type structure, adding the compound into a reaction bottle, adding DCC, DMAP and organic base, heating under the protection of nitrogen, monitoring by TLC to complete the reaction, filtering, evaporating the filtrate to dryness, and then performing recrystallization or column chromatography to obtain target products with II-type and III-type structures respectively according to different n values.

7. The method for preparing the abietane-based compound having a novel C-ring-fused lactone ring skeleton according to claim 6, wherein the abietane-based diterpene quinone is selected from compounds having an o-quinone structure type, preferably, the abietane-based diterpene quinone is an o-quinone compound in a Salvia species, more preferably, the abietane-based diterpene quinone is tanshinone I, tanshinone IIA, cryptotanshinone, tanshinone;

the acid anhydride is selected from cyclic dianhydride, and preferably, the acid anhydride is succinic anhydride, glutaric anhydride or adipic anhydride.

8. The process according to claim 6, wherein the simple reducing metal is selected from the group consisting of magnesium powder, iron powder and zinc powder;

the organic base is selected from triethylamine and N, N-diisopropylethylamine.

9. The use of the abietane compounds with a novel C-ring-fused lactone ring skeleton as claimed in any one of claims 1 to 5, wherein the abietane compounds, their derivatives and their pharmaceutically acceptable salts are used for preparing anti-tumor and cardiovascular and cerebrovascular therapeutic drugs.

10. A pharmaceutical composition comprising the abietane compound having a novel skeleton with a C-ring-fused lactone ring, its derivative, and its pharmaceutically acceptable salt of any one of claims 1 to 5, and a pharmaceutically acceptable excipient, wherein the pharmaceutical composition is in the form of tablet, capsule, orally disintegrating tablet, dripping pill, spray, aerosol, or injection.