A kind of N-aromatic alkylamino fern amide compound, preparation method and application thereof

Abstract

The present invention relates to the technical field of derivative synthesis, and specifically to an N-arylalkylamino fernamide compound, a preparation method and an application thereof. The N-arylalkylamino fernamide compound is prepared by reacting fern acid as a substrate with various primary amines containing an aromatic alkyl group under the action of a condensing agent. The anti-tumor activity screening results show that some compounds have good proliferation inhibition activity on human breast cancer cells (MCF-7), human non-small cell lung cancer cells (A549), and human liver cancer cells (HepG2), and the half inhibitory concentration (IC50) of some compounds is less than 10%. 50 ) is less than 10μM, and the anti-tumor activity is better than the positive control drug cisplatin. Among them, N-o-bromophenylethylamino fern amide has the highest inhibitory activity against human liver cancer cells HepG2, IC 50 The value was 1.92±0.22μM, which is expected to be further developed into a new anti-tumor drug candidate.

Description

Technical Field

[0001] The invention relates to the technical field of derivative synthesis, and in particular to an N-aromatic alkylamino fern amide compound, a preparation method and application thereof. Background Art

[0002] In recent years, cancer has become one of the most serious diseases that threaten human health and cause death. Due to the complex characteristics of cancer cells, especially for patients in the middle and late stages of cancer, there is currently no good cure, and the situation of tumor prevention and treatment is becoming increasingly severe.

[0003] Plant secondary metabolites (natural products) are an important source of modern anticancer drug discovery because they are mostly biologically active, low in toxicity and effective, and have good development prospects. A large number of biologically active diterpenoids have been isolated from natural products (Demetzos C, Dimas K S. Labdane-type diterpenes: Chemistry and biological activity [J]. Studies in Natural Products Chemistry, 2001, 25 (01): 235-292.), which have been used to treat a variety of human diseases such as cancer. Medicinal ferns, as a natural product, are often used as medicine in their entirety or in their rhizomes, and most of them contain a variety of active chemical substances such as flavonoids, alkaloids, phenolic acids, and terpenes. Previous studies have shown that many ferns have pharmacological activities, especially their antioxidant, anti-tumor, anti-inflammatory, antibacterial and other aspects, and can be used to treat various human diseases in alternative medicine (Wang Ling, He Zhaorong. Research Progress on Chemical Components of Medicinal Ferns [J]. Chinese Wild Plant Resources, 2006, 025(003): 1-4), which also shows that medicinal ferns have good development prospects in natural product research. Aleuritopteris argentea (SGGmél.) Fée is a medicinal fern, and its main metabolite is a semi-helian diterpenoid compound, namely alepterolic acid. Hiroyuki Ueda et al. (Ageta H, Awata K, Otake Y. Structure of the diterpenoid alepterolic acid from Aleuritopteris argentea, Proceedings of Sympoium on Natural Organic Compounds. 1962, 6: 136-141) isolated diterpenoid alepterolic acid from Aleuritopteris argentea for the first time and identified its structure. Previous studies have shown that diterpenoid alepterolic acid has a variety of biological activities, such as antibacterial, antiparasitic, and antiviral (Trindade RD, Silva J D, Setzer W N. Copaifera of the neotropics: A review of the phytochemistry and pharmacology [J]. International Journal of Molecular Sciences, 2018, 19 (5): 108-1511.).

[0004] The natural product aleptinic acid itself has an unsatisfactory anticancer effect, but by introducing an amide group into aleptinic acid, the obtained compound has better anticancer activity (Zhang S, Feng N, Huang J, et al. Incorporation of amino moiety to aleptinic acid improve activity against cancer cell lines: Synthesis and biological evaluation [J]. Bioorganic Chemistry, 2020, 98: 103756.). Zhang Lei et al. (Zhang Lei, Yu Junjie, Cao Jianguo, et al. Synthesis of a novel aleptinic acid amide derivative and its application in anti-tumor drugs [P]. CN111393318A, 2020.) invented a patent for the synthesis of a novel aleptinic acid amide derivative and its application in anti-tumor drugs, and described in detail the method of aleptinic acid forming amide derivatives with various amine compounds. In the anticancer activity screening, most of the compounds showed significant anti-tumor activity against human prostate cancer cells PC-3, human breast cancer cells MCF-7, human non-small cell lung cancer cells A549, and human cervical cancer cells Hela. However, there are many kinds of amine compounds, and it is particularly important to find more effective anti-tumor drugs. However, in this invention application, there is no attempt to synthesize and evaluate the activity of N-aromatic alkylamino fern amide. There is no study on the inhibitory activity of fern amide derivatives on liver cancer cells.

[0005] In view of the above-mentioned deficiencies, the inventors of the present invention designed and synthesized a series of novel fern amide compounds containing N-aromatic alkylamino groups after a long period of research and practice, and tested their inhibitory activity against several cancer cells. Summary of the invention

[0006] The purpose of the present invention is to solve the problem of how to prepare a fern amide derivative with better anti-tumor activity, and provides an N-aromatic alkylamino fern amide compound, a preparation method and an application thereof.

[0007] In order to achieve the above object, the present invention discloses an N-aromatic alkylamino fern amide compound, the structural formula of which is shown below:

[0008] A primary amine wherein R is an aromatic alkyl group or a pharmaceutically acceptable salt, stereoisomer, solvate or hydrate thereof.

[0009] The pharmaceutically acceptable salts include acetate, benzoic acid, hydrochloride, maleate, fumaric acid, sulfate, benzenesulfonate, succinate and salicylate.

[0010] The R is independently selected from:

[0011]

[0012] The present invention also discloses a method for preparing the above-mentioned N-aromatic alkylamino fern amide compound, and the reaction formula of the preparation method is:

[0013]

[0014] The method comprises the following steps: accurately weighing pteridophytic acid and dissolving it in dichloromethane, then sequentially adding O-(benzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N,N-diisopropylethylamine and amine compounds with different substitutions, stirring the reaction at room temperature, tracking the reaction by thin layer chromatography during the stirring process, adding water to terminate the reaction after the reaction is complete, distilling off the solvent under reduced pressure, separating and purifying the obtained crude product by silica gel column chromatography, and obtaining the target compound.

[0015] The invention also discloses the application of the N-aromatic alkylamino fern amide compound in preparing medicine for treating and / or preventing tumors.

[0016] Compared with the prior art, the beneficial effect of the present invention lies in that: the present invention introduces amine groups of different sizes into fern acid, and systematically tests the anti-proliferation activity of different modifications on cancer cells. The heavy metal complex cisplatin is used as a positive control drug, and three cancer cells, human breast cancer cells (MCF-7), human non-small cell lung cancer cells (A549), and human liver cancer cells (HepG2) are used as experimental subjects. The results show that all compounds have good inhibitory effects on the three cancer cells of MCF-7, A549 and HepG2. The inhibitory activity of most derivatives on MCF-7 is generally greater than their inhibitory activity on A549 and HepG2. Among them, compounds 1o-1s have the best proliferation inhibitory activity on the three cancer cells, among which compound 1r has the best anti-tumor activity, and its IC 20 for HepG2 is 1. 50 =1.92±0.22μM, which is much higher than the positive control drugs cisplatin and fern acid. Therefore, this type of derivative has a good development prospect and is expected to be further developed and applied in the field of anti-tumor drugs. DETAILED DESCRIPTION

[0017] The term "pharmaceutically acceptable salt" refers to salts that are suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reaction, etc., and are commensurate with a reasonable benefit / risk ratio, within the scope of sound medical judgment. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66: 1-19, including acetate, benzoic acid, hydrochloride, maleate, fumaric acid, sulfate, benzenesulfonate, succinate, salicylate, etc. Pharmaceutically acceptable salts of the compounds of the present invention include salts derived from suitable inorganic and organic acids and bases.

[0018] "Disease," "disorder," and "condition" are used interchangeably in this application.

[0019] As used herein, and unless otherwise indicated, the terms "treatment," "treating," "treating," "treating," and "treatment" include actions that occur while a subject has a particular disease, disorder, or condition, that reduce the severity of, or delay or slow the development of, the disease, disorder, or condition ("therapeutic treatment"), and also include actions that occur before a subject develops a particular disease, disorder, or condition ("prophylactic treatment").

[0020] Generally, an "effective amount" of a compound refers to an amount sufficient to cause a target biological response. As will be appreciated by those of ordinary skill in the art, the effective amount of the compounds of the invention may vary depending on factors such as the biological target, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the age, health, and symptoms of the subject. An effective amount includes both therapeutic and prophylactic treatment effective amounts.

[0021] Unless otherwise specified, a "therapeutically effective amount" of a compound as used herein is an amount sufficient to provide a therapeutic benefit in the treatment of a disease, disorder, or condition, or to delay or minimize one or more symptoms associated with the disease, disorder, or condition. A therapeutically effective amount of a compound refers to an amount of a therapeutic agent, used alone or in combination with other therapies, that provides a therapeutic benefit in the treatment of a disease, disorder, or condition. The term "therapeutically effective amount" may include an amount that improves overall treatment, reduces or avoids symptoms or causes of a disease or condition, or enhances the therapeutic efficacy of other therapeutic agents.

[0022] Unless otherwise specified, a "prophylactically effective amount" of a compound as used herein is an amount sufficient to prevent a disease, disorder, or condition, or an amount sufficient to prevent one or more symptoms associated with a disease, disorder, or condition, or an amount to prevent the recurrence of a disease, disorder, or condition. A prophylactically effective amount of a compound refers to an amount of a therapeutic agent used alone or in combination with other agents that provides a prophylactic benefit in the process of preventing a disease, disorder, or condition. The term "prophylactically effective amount" may include an amount that improves overall prevention, or an amount that enhances the prophylactic efficacy of other prophylactic agents.

[0023] The compounds of the present invention may include one or more asymmetric centers and may therefore exist in a variety of stereoisomeric forms, for example, enantiomers and / or diastereoisomeric forms. For example, the compounds of the present invention may be individual enantiomers, diastereomers or geometric isomers (e.g., cis and trans isomers), or may be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. Isomers may be separated from the mixture by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers may be prepared by asymmetric synthesis.

[0024] Those skilled in the art will appreciate that many organic compounds can form complexes with solvents in which they react or from which they precipitate or crystallize. These complexes are referred to as "solvates". When the solvent is water, the complex is referred to as a "hydrate". The present invention encompasses all solvates of the compounds of the present invention.

[0025] On the other hand, the present invention provides a pharmaceutical composition comprising a compound of the present invention (also referred to as an "active ingredient") and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition comprises an effective amount of a compound of the present invention. In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of a compound of the present invention. In some embodiments, the pharmaceutical composition comprises a preventive effective amount of a compound of the present invention.

[0026] The pharmaceutically acceptable excipient used in the present invention refers to a non-toxic carrier, adjuvant or vehicle that does not destroy the pharmacological activity of the compound formulated together. Pharmaceutically acceptable carriers, adjuvants or vehicles that can be used in the composition of the present invention include (but are not limited to) ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffer substances (such as phosphates), glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, silica gel, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and lanolin.

[0027] The present invention also includes a kit (e.g., a pharmaceutical package). The kit provided may include a compound of the invention, other therapeutic agents, and a first and second container (e.g., a vial, an ampoule, a bottle, a syringe, and / or a dispersible package or other suitable container) containing the compound of the invention and other therapeutic agents. In some embodiments, the kit provided may also optionally include a third container containing a pharmaceutical excipient for diluting or suspending the compound of the invention and / or other therapeutic agents. In some embodiments, the compound of the invention and other therapeutic agents provided in the first container and the second container are combined to form a unit dosage form.

[0028] Synthesis method

[0029] Accurately weigh fernic acid (32.0 mg, 0.1 mmol) and dissolve it in 5 mL of dichloromethane. Then add HBTU (46.7 mg, 0.123 mmol), DIPEA (19.4 mg, 0.15 mmol) and amine compounds with different substitutions (0.15 mmol) in sequence, and stir to react at room temperature. During the stirring process, the reaction was tracked by TLC. After the reaction was complete, water was added to terminate the reaction, and the solvent was evaporated under reduced pressure. The crude product was separated and purified by silica gel column chromatography to obtain the target compounds 1a-1x.

[0030] Reagents and reaction conditions: N,N-diisopropylethylamine (DIPEA), dichloromethane (CH 2 Cl 2 ), O-(Benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), amine compounds, room temperature.

[0031] Example 1

[0032] (E)-N-(Furan-2-ylmethyl)-5-((1R,4aS,6R,8aS)-6-hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methylpent-2-enamide (1a)

[0033] White powder, yield 86%; mp = 149.7 ~ 151.1 ℃.

[0034] 1 H NMR (400 MHz, CDCl 3 )δ7.35 (dd, J=1.9, 0.9Hz, 1H, eH), 6.32 (dd, J=3.2, 1.9Hz, 1H, cH), 6.23 (dd, J=3.2, 0.8Hz, 1H, dH), 5.66 (t, J=6.0Hz, 1H , NH), 5.50 (q, J=1.3Hz, 1H, 14-H), 4.85 (d, J=1.5Hz, 1H, 17-H), 4.50 (d, J=1.5Hz, 1H, 17-H), 4.46 (d, J=5.5Hz, 2H, NH-CH 2 ), 3.25 (dd, J=11.7, 4.3Hz, 1H, 3-H), 2.39 (ddd, J=12.7, 4.3, 2.4Hz, 1H, 7-H), 2.23 (ddd, J=14.6, 10.2, 4.1Hz, 1H, 12-H), 2.16 (d, J=1.3Hz, 3H, 16-CH 3 ), 2.00–1.84(m, 2H, 7-H, 12-H), 1.80–1.32(m, 7H, 1-H, 6-CH 2 ,9-H,11-CH 2 , 2-CH 2 ), 1.15 (td, J=13.1, 3.8Hz, 1H, 1-H), 1.07 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.99 (s, 3H, 18-CH 3 ), 0.76(s, 3H, 19-CH 3 ), 0.68 (d, J=0.7Hz, 3H, 20-CH 3 ).

[0035] HRMS(ESI)calcd for C 25 H 37 NO 3 [M+H] + :400.2846, found 400.2839.

[0036] Example 2

[0037] 2(E)-5-((1R,4aS,6R,8aS)-6-Hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methyl-N-(thiophen-2-ylmethyl)pent-2-enamide (1b)

[0038] White powder, yield 83%, mp=162.0~162.6℃.

[0039] 1 H NMR (400MHz, CDCl3) δ7.21 (dd, J=5.0, 1.3Hz, 1H, eH), 6.97 (dd, J=3.5, 1.3Hz, 1H, cH), 6.94 (dd, J=5.1, 3.4Hz, 1H, dH), 5.74 (t, J=5.7Hz, 1H, NH), 5.50 (q, J=1.3Hz, 1H, 14-H), 4.84 (q, J=1.5Hz, 1H, 17-H), 4.63 (d, J=5.6Hz, 2H, a-CH 2 ), 4.49 (d, J=1.8Hz, 1H, 17-H), 3.24 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.39 (ddd, J=12.8, 4. 3, 2.4Hz, 1H, 7-H), 2.23 (ddd, J=13.8, 9.8, 3.9Hz, 1H, 12-H), 2.16 (d, J=1.3Hz, 3H, 16-CH 3 ), 2.00–1.84(m, 2H, 7-H, 12-H), 1.80–1.44(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.36 (td, J=12.9, 4.2Hz, 1H, 6-H), 1.14 (td, J=13.1, 3.8Hz, 1H 1-H), 1.07 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.98 (s, 3H, 18-CH 3 ), 0.76(s, 3H, 19-CH 3 ), 0.67(s, 3H, 20-CH 3 ).

[0040] HRMS(ESI)calcd for C 25 H 37 NO 2 S[M+H] + :416.2618, found 416.2610.

[0041] Example 3

[0042] (E)-N-Benzyl-5-((1R,4aS,6R,8aS)-6-hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methylpent-2-enamide (1c)

[0043] White powder, yield 94%, mp=158.4~160.2℃.

[0044] 1 H NMR (400MHz, CDCl3) δ7.37–7.26 (m, 5H, Ph×5), 5.69 (t, J=5.8Hz, 1H, NH), 5.52 (d, J=1.3Hz, 1H, 1 4-H), 4.84 (q, J=1.6Hz, 1H, 17-H), 4.50 (d, J=1.7Hz, 1H, 17-H), 4.46 (dd, J=5.8, 2.1Hz, 2H, NH-CH 2 -Ph), 3.24 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.39 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.24 (ddd, J=14.0, 9.8, 3.8Hz, 1H, 12-H), 2.17 (d, J=1.3Hz, 3H, 16-CH 3 ), 2.00–1.84(m, 2H, 7&12-H), 1.81–1.45(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.38 (qd, J=12.8, 4.2Hz, 1H, 6-H), 1.15 (td, J=13.2, 3.8Hz, 1H, 1-H), 1.07 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.98 (s, 3H, 18-CH3), 0.76 (s, 3H, 19-CH 3 ), 0.67(s, 3H, 20-CH 3 ).

[0045] HRMS(ESI)calcd for C 27 H 39 NO 2 [M+H] + :410.3054, found 410.3049.

[0046] Example 4

[0047] (E)-5-((1R,4aS,6R,8aS)-6-Hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methyl-N-(1-phenylethyl)pent-2-enamide (1d)

[0048] White powder, yield 89%, mp = 150.9 ~ 152.8 ° C;

[0049] 1 H NMR (400MHz, CDCl3) δ7.36-7.31(m, 4H, Ph*4-H), 7.28-7.25(m, 1H, Ph-H), 5.59(d , J=8.0Hz, 1H, NH), 5.50 (s, 1H, 14-H), 5.16 (p, J=7.1Hz, 1H, aH), 4.85 (s, 1H, 17-H) , 4.50 (s, 1H, 17-H), 3.24 (ddd, J=11.7, 4.5, 1.6Hz, 1H, 3-H), 2.39 (dq, J=10.1, 3.9 Hz, 1H, 7-H), 2.23 (ddd, J=14.1, 10.0, 3.8Hz, 1H, 12-H), 2.16–2.12 (brs, 3H, 16-CH 3 ), 2.00–1.82(m, 2H, 7&12-H), 1.80–1.35(m, 10H, 1-H, 6-H, 9-H, f-CH 3 , 11-CH 2 , 2-CH 2 ), 1.35–1.21 (m, 1H, 6-H), 1.15 (t, J=12.8Hz, 1H, 1-H), 1.07 (dt, J=12.5, 2.5Hz, 1H, 5-H), 0.99 (d, J=1.7Hz, 3H, 18-CH 3 ), 0.76 (d, J=1.0Hz, 3H, 19-CH 3 ), 0.68(s, 3H, 20-CH 3 ).

[0050] HRMS(ESI)calcd for C 28 H 41 NO 2 [M+H] + :424.3210, found 424.3204.

[0051] Example 5

[0052] (E)-5-((1R,4aS,6R,8aS)-6-Hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methyl-N-(4-methylbenzyl)pent-2-enamide (1e)

[0053] White powder, yield 92%, mp = 136.4 ~ 137.9 ℃;

[0054] 1 H NMR (400 MHz, CDCl 3 )δ7.33–7.18(m, 5H, 5×Ph-H), 5.63(d, J=8.2Hz, 1H, NH), 5.51(q, J=1.3Hz, 1H, 14-H), 4.90 (q, J=7.6Hz, 1H, aH), 4.84 (q, J=1.5Hz, 1H, 17-H), 4.50 (d, J=1.7H z, 1H, 17-H), 3.23 (dd, J=11.6, 4.4Hz, 1H, 3-H), 2.38 (ddd, J=12.8, 4.3, 2.4Hz , 1H, 7-H), 2.22 (ddd, J=13.9, 10.4, 3.8Hz, 12-H), 2.12 (d, J=1.3Hz, 3H, 16-CH 3 ), 1.98–1.43(m, 11H, 7&12-H, 1-H, 6-H, 9-H, b-CH 2 , 11-CH 2 , 2-CH 2 ), 1.38 (qd, J=12.8, 4.2Hz, 1H, 6-H), 1.14 (td, J=13.2, 3.8Hz, 1H, 1-H), 1.06 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.98 (s, 3H18-CH 3 ), 0.89 (t, J=7.4Hz, 3H, c-CH 3 ), 0.76(s, 3H, 19-CH 3 ), 0.67(s, 3H, 20-CH 3 ).

[0055] HRMS(ESI)calcd for C 29 H 43 NO 2 [M+H] + :438.3367, found 438.3362.

[0056] Example 6

[0057] (E)-5-((1R,4aS,6R,8aS)-6-Hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methyl-N-(2-methylbenzyl)pent-2-enamide (1f)

[0058] White powder, yield 91%, mp = 158.4 ~ 158.9 ℃;

[0059] 1 H NMR (400 MHz, CDCl 3 )δ7.25–7.1(m, 1H, Ph-H), 7.21–7.14(m, 3H, Ph-H*3), 5.51(s, 1H, 14-H), 5.59(t, J=5.8Hz, 1H, NH), 4.84 (s, 1H, 17-H), 4.50 (s, 1H, 17-H), 4.46 (d, J=5.4Hz, 2H, a-CH 2 ), 3.28–3.19 (m, 1H, 3-H), 2.39 (ddd, J=12.8, 4.3, 2.5Hz, 1H, 7-H), 2.33 (s, 3H, Ph-CH 3 ), 2.23 (ddd, J=14.0, 9.9, 3.7Hz, 1H, 12-H), 2.17 (d, J=1.3Hz, 3H, 16-H), 1.99–1.85 (m, 2H, 7&12-H), 1.80–1.43 (m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.37 (td, J=12.9, 4.3Hz, 1H, 6-H), 1.15 (td, J=13.1, 3.9Hz, 1H, 1-H), 1.07 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.98 (s, 3H 18-CH 3 ), 0.76(s, 3H 19-CH 3 ), 0.67(s, 3H, 20-CH 3 ).

[0060] HRMS(ESI)calcd for C 28 H 41 NO 2 [M+H] + :424.3210, found 424.3205.

[0061] Example 7

[0062] (E)-5-((1R,4aS,6R,8aS)-6-Hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methyl-N-(3-methylbenzyl)pent-2-enamide (1 g)

[0063] White powder, yield 90%, mp=149.0~150.5℃

[0064] 1 H NMR (400MHz, CDCl3) δ7.22 (t, J=7.5Hz, 1H, Ph-H), 7.13–7.08 (m, 2H, Ph-H), 7.07 (d, J=2.0Hz, 1H, Ph-H), 5.65 (s, 1H, NH) , 5.52 (q, J=1.3Hz, 1H, 14-H), 4.85 (q, J=1.6Hz, 1H, 17-H), 4.50 (q, J=1.5Hz, 1H, 17-H), 4.43 (dd, J=5.6, 1.8Hz, 2H, NH-CH 2 -Ph), 3.24 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.39 (ddd, J=12.7, 4.2, 2.4Hz, 1H, 7-H), 2.3 4(s, 3H, Ph-Me), 2.24 (ddd, J=13.8, 9.7, 3.6Hz, 1H, 12-H), 2.17 (d, J=1.2Hz, 3H, 16-CH 3 ), 2.00–1.84(m, 2H, 7-H, 12-H), 1.80–1.44(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.38 (qd, J=12.9, 4.2Hz, 1H, 6-H), 1.15 (td, J=13.1, 3.8Hz, 1H, 1-H), 1.07 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.98 (s, 3H, 18-CH 3 ), 0.76(s, 3H, 19-CH 3 ), 0.67(s, 3H, 20-CH 3 ).

[0065] HRMS(ESI)calcd for C 28 H 41 NO 2 [M+H] + :424.3210, found 424.3202.

[0066] Example 8

[0067] (E)-5-((1R,4aS,6R,8aS)-6-Hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methyl-N-(4-methylbenzyl)pent-2-enamide (1h)

[0068] White powder, yield 85%, mp = 162.4 ~ 164.1 ℃;

[0069] 1 H NMR (400 MHz, CDCl 3 )δ7.19 (d, J=8.0Hz, 2H, Ph-H), 7.14 (d, J=7.9Hz, 2H, Ph-H), 5.62 (s, 1H, NH), 5.5 1 (s, 1H, 14-H), 4.85 (s, 1H, 17-H), 4.50 (s, 1H, 17-H), 4.42 (d, J=4.9Hz, 2H, NH-CH 2 -Ph), 3.25 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.39 (ddd, J=12.7, 4.3, 2.4Hz, 1H, 7-H), 2.3 3(s, 3H, Ph-Me), 2.24 (td, J=10.6, 9.9, 5.1Hz, 1H, 12-H), 2.17 (d, J=1.2Hz, 3H, 16-CH 3 ), 2.00–1.84(m, 2H, 7-H, 12-H), 1.81–1.46(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.38 (qd, J=13.0, 4.1Hz, 1H, 6-H), 1.15 (td, J=13.0, 3.7Hz, 1H, 1-H), 1.07 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.99 (s, 3H, 18-CH 3 ), 0.77(s, 3H, 19-CH 3 ), 0.68(s, 3H, 20-CH 3 ).

[0070] HRMS(ESI)calcd for C 28 H 41 NO 2 [M+H] + :424.3210, found 424.3202.

[0071] Example 9

[0072] (E)-N-(3-Fluorobenzyl)-5-((1R,4aS,6R,8aS)-6-hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methylpent-2-enamide (1i)

[0073] White powder, yield 79%, mp = 148.8 ~ 148.8 ° C;

[0074] 1 H NMR (400MHz, CDCl3) δ7.28 (td, J=7.9, 5.9Hz, 1H, cH), 7.07 (ddq, J=7.6, 1.6, 0.8Hz, 1H, eH), 7.00 (dt, J=9.6, 2.0Hz, 1H, dH), 6.95 (tdd, J=8.4, 2.6, 1.0Hz, 1H, fH), 5.70 (t, J=5.9Hz, 1H, NH), 5.54 (q, J=1.3Hz, 1H, 14-H), 4.85 (q, J=1. 5Hz, 1H, 17-H), 4.50 (d, J=1.6Hz, 1H, 17-H), 4.47 (dd, J=5.9, 2.6Hz, 2H, NH-CH 2 -Ph), 3.25 (dd, J=11.8, 4.3Hz, 1H, 3-H), 2.40 (ddd, J=12.8, 4.3, 2.5Hz, 1H, 7-H), 2.25 (ddd, J=14.0, 10.1, 3.9Hz, 1H, 12-H), 2.18 (d, J=1.2Hz, 3H, 16-CH 3 ), 2.01–1.85(m, 2H, 7-H, 12-H), 1.80–1.329(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.39–1.30 (m, 1H, 6-H), 1.16 (td, J=13.1, 3.8Hz, 1H, 1-H), 1.07 (dd, J=12.5, 2.7Hz, 1H, 5-H), 0.99 (s, 3H, 18-CH 3 ), 0.76(s, 3H, 19-CH 3 ), 0.68 (d, J=0.7Hz, 3H, 20-CH 3 ).

[0075] HRMS(ESI)calcd for C 27 H 38 FNO 2 [M+H] + :428.2959, found 428.2952.

[0076] Example 10

[0077] (E)-N-(2-Chlorobenzyl)-5-((1R,4aS,6R,8aS)-6-hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methylpent-2-enamide (1j)

[0078] White powder, yield 88%, mp = 177.6 ~ 179.6 ℃;

[0079] 1 H NMR (400 MHz, CDCl 3 )δ7.44–7.39 (m, 1H, Ph-H), 7.38–7.33 (m, 1H, Ph-H), 7.25–7.18 (m, 2H, Ph-H), 5.81 (t, J=5.9Hz , 1H, NH), 5.53 (q, J=1.3Hz, 1H, 14-H), 4.85 (q, J=1.5Hz, 1H, 17-H), 4.56 (d, J=6.1Hz, 2H, NH-CH 2 -Ph), 4.50 (q, J=1.5Hz, 1H, 17-H), 3.24 (dd, J=11.7, 4.3Hz, 1H, 3-H), 2.39 (ddd, J=12.7, 4 .2, 2.4Hz, 1H, 7-H), 2.24 (ddd, J=13.8, 10.1, 3.9Hz, 1H, 12-H), 2.15 (d, J=1.2Hz, 3H, 16-CH 3 ), 2.00–1.84(m, 2H, 7-H, 12-H), 1.82–1.38(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.35 (dd, J=11.9, 3.2Hz, 1H, 6-H), 1.20–1.11 (m, 1H, 1-H), 1.07 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.99 (s, 3H, 18-CH 3 ), 0.76(s, 3H, 19-CH 3 ), 0.68(s, 3H, 20-CH 3 ).

[0080] HRMS(ESI)calcd for C 27 H 38 ClNO 2 [M+H] + :444.2664, found 444.2654.

[0081] Embodiment 11

[0082] (E)-N-(3-Fluorobenzyl)-5-((1R,4aS,6R,8aS)-6-hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methylpent-2-enamide (1k)

[0083] White powder, yield 92%, mp = 147.2 ~ 149.5 ℃;

[0084] 1 H NMR (400MHz, CDCl3) δ7.28 (td, J=7.9, 5.9Hz, 1H, Ph-H), 7.06 (dt, J=7.8, 1.2Hz, 1H, Ph-H), 6.99 (dt, J=9.7, 2.1Hz, 1H, Ph-H), 6.95 (td, J=8.5, 2.4Hz, 1 H, Ph-H), 5.74 (t, J=6.0Hz, 1H, NH), 5.54 (q, J=1.3Hz, 1H, 14-H), 4.85 (d, J= 1.5Hz, 1H, 17-H), 4.53–4.48 (m, 1H, 17-H), 4.46 (dd, J=5.9, 2.5Hz, 2H, NH-CH 2 -Ph), 3.24 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.39 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.31–2.19 (m, 1H, 12-H), 2.17 (d, J=1.3Hz, 3H, 16-CH 3 ), 2.00–1.85(m, 2H, 7&12-H), 1.75–1.53(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.38 (qd, J=13.0, 4.1Hz, 1H, 6-H), 1.16 (td, J=13.2, 3.8Hz, 1H, 1-H), 1.07 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.99 (s, 3H, 18-CH 3 ), 0.76(s, 3H, 19-CH 3 ), 0.68(s, 3H, 20-CH 3 ).

[0085] HRMS(ESI)calcd for C 27 H 38 FNO 2 [M+H] + :428.2959, found 428.2953.

[0086] Example 12

[0087] (E)-N-(3-bromobenzyl)-5-((1R,4aS,6R,8aS)-6-hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methylpent-2-enamide (1l)

[0088] White powder, yield 85%, mp = 145.7 ~ 147.2 ° C;

[0089] 1 H NMR (400MHz, CDCl3) δ7.44–7.39 (m, 1H, Ph-H), 7.37 (dt, J=7.4, 1.8Hz, 1H, Ph-H), 7.21 (dt, J=7.7, 1.6Hz, 1H, Ph-H), 7.19–7.15 (m, 1H, Ph-H ), 5.87 (t, J = 6.0Hz, 1H, NH), 5.54 (q, J = 1.3Hz, 1H, 14-H), 4.84 (q, J = 1.5Hz, 1H, 17-H), 4.49 (q, J = 1.4Hz, 1H, 17-H), 4.49–4.34 (m, 2H, NH-CH 2 -Ph), 3.23 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.38 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.24 (m, 1H, 12-H), 2.16 (d, J=1.2Hz, 3H, 16-CH 3 ), 2.00–1.83(m, 2H, 7-H, 12-H), 1.80–1.43(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.37 (qd, J=13.0, 4.4Hz, 1H, 6-H), 1.14 (td, J=13.1, 3.8Hz, 1H, 1-H), 1.06 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.97 (s, 3H, 18-CH 3 ), 0.75(s, 3H, 19-CH 3 ), 0.67(s, 3H, 20-CH 3 ).

[0090] (ESI)calcd for C 27 H 38 BrNO 2 [M+H] + :488.2159, found 488.2149.

[0091] Example 13

[0092] (E)-N-(4-Chlorobenzyl)-5-((1R,4aS,6R,8aS)-6-hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methylpent-2-enamide (1m)

[0093] White powder, yield 87%, mp = 191.4 ~ 193.2 ° C;

[0094] 1 H NMR (400 MHz, CDCl 3 )δ7.33–7.27(m, 2H, Ph-H), 7.25–7.19(m, 2H, Ph-H), 5.67(t, J=5.7Hz, 1H, NH), 5.52(q, J=1.3Hz, 1H , 14-H), 4.85 (q, J=1.5Hz, 1H, 17-H), 4.50 (d, J=1.7Hz, 1H, 17-H), 4.43 (dd, J=5.9, 3.3Hz, 2H, NH-CH 2 -Ph), 3.24 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.39 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.30–2.19 (m, 1H, 12-H), 2.17 (d, J=1.2Hz, 3H, 16-CH 3 ), 2.00–1.84(m, 2H, 7-H, 12-H), 1.81–1.46(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.37 (td, J=12.9, 5.2Hz, 1H, 6-H), 1.15 (td, J=13.1, 3.8Hz, 1H, 1-H), 1.07 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.99 (s, 3H, 18-CH 3 ), 0.76(s, 3H, 19-CH 3 ), 0.68(s, 3H, 20-CH 3 ).

[0095] HRMS(ESI)calcd for C 27 H 38 ClNO 2 [M+H] + :444.2664, found 444.2658.

[0096] Embodiment 14

[0097] (E)-5-((1R,4aS,6R,8aS)-6-Hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-N-(4-methoxybenzyl)-3-methylpent-2-enamide (1n)

[0098] White powder, yield 80%, mp = 165.9-167.7;

[0099] 1 H NMR (400 MHz, CDCl 3 )δ7.24–7.18(m, 2H, 2×Ph-H), 6.89–6.81(m, 2H, 2×Ph-H), 5.65(t, J=5.6Hz, 1H, 14-H), 5.50(q, J= 1.3Hz, 1H, NH), 4.84 (q, J=1.6Hz, 1H, 17-H), 4.49 (d, J=1.7Hz, 1H, 17-H), 4.46–4.33 (m, 2H, NH-CH 2 -Ph), 3.78(s, 3H, OMe-CH 3 ), 3.23 (dd, J = 11.7, 4.4Hz, 1H, 3-H), 2.38 (ddd, J = 12.7, 4.3, 2.5Hz, 1H, 7-H), 2.22 (ddd, J = 13.9, 9.9, 3.9Hz, 1H, 12-H), 2.16 (d, J = 1.3Hz, 3H, 16-CH 3 ), 1.99–1.82(m, 2H, 7&12-H), 1.81–1.30(m, 7H, 1-H, 6-H, 9-H, 12-CH 2 , 2-CH 2 ), 1.28-1.21(m, 1H, 6-H), 1.20–1.02(m, 2H, 1&5-H), 0.98(s, 3H, 18-CH 3 ), 0.76(s, 3H, 19-CH 3 ), 0.67(s, 3H,, 20-CH 3 ).

[0100] HRMS(ESI)calcd for C 28 H 41 NO 3 [M+H] + :440.3159, found 440.3151.

[0101] Embodiment 15

[0102] (E)-5-((1R,4aS,6R,8aS)-6-Hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methyl-N-phenylpent-2-enamide (1o)

[0103] White powder, yield 93%, mp = 138.5 ~ 140.7 ° C;

[0104] 1 H NMR (400 MHz, CDCl 3 )δ7.36–7.27(m, 2H, Ph-H), 7.28–7.17(m, 3H, Ph-H), 5.43(q, J=1.3Hz, 1H, 14-H), 5.38(s, 1 H, NH), 4.84 (q, J=1.6Hz, 1H, 17-H), 4.49 (q, J=1.5Hz, 1H, 17-H), 3.55 (q, J=6.6Hz, 2H, a-CH 2 ), 3.24 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.84 (t, J=7.0Hz, 2H, b-CH 2 ), 2.39 (ddd, J=12.8, 4.4, 2.4Hz, 1H, 7-H), 2.20 (td, J=9.7, 4.9Hz, 1H, 12-H), 2.13 (d, J=1.2Hz, 3H, 16-CH 3 ), 2.01–1.85(m, 2H, 7-H, 12-H), 1.81–1.40(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.44–1.31 (m, 1H, 6-H), 1.14 (td, J=13.1, 3.7Hz, 1H, 1-H), 1.06 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.99 (s, 3H, 18-CH 3 ), 0.76(s, 3H, 19-CH 3 ), 0.67(s, 3H, 20-CH 3 ).

[0105] HRMS(ESI)calcd for C 28 H 41 NO 2 [M+H] + :424.3210, found 424.3203.

[0106] Example 16

[0107] (E)-5-((1R,4aS,6R,8aS)-6-Hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methyl-N-(4-methylphenethyl)pent-2-enamide (1p)

[0108] White powder, yield 91%, mp = 147.3 ~ 147.9 ℃;

[0109] 1 H NMR (400 MHz, CDCl 3 )δ7.16–7.06 (m, 4H, 4×Ph-H), 5.45–5.39 (q, J=1.3Hz, 1H, 14-H), 5.39–5.34 (s, 1H, NH), 4.87–4 .82(q, J=1.6Hz, 1H, 17-H), 4.52–4.47(d, J=1.7Hz, 1H, 17-H), 3.58–3.46 (q, J=6.7Hz, 2H, a-CH 2 ), 3.29–3.20 (dd, J=4.2, 11.9Hz, 1H, 3-H), 2.83–2.75 (t, J=6.9Hz, 2H, b-CH 2 ), 2.44–2.35 (ddd, J=2.5, 4.3, 12.8Hz, 1H, 7-H), 2.35–2.31 (s, 3H, 16-CH 3 ), 2.27–2.15 (ddd, J=3.8, 9.7, 13.8Hz, 1H, 12-H), 2.20–2.10 (d, J=1.3Hz, 3H, Ph-Me-CH 3 ), 2.01–1.82(m, 2H, 7&12-H), 1.81–1.44(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.40–1.32 (m, 1H, 6-H), 1.21–1.09 (td, J=3.8, 13.2Hz, 1H, 1-H), 1.11–1.03 (dd, J=2.8, 12.5Hz, 1H, 5-H), 1.01–0.97 (s, 3H, 18-CH 3 ), 0.79–0.74(s, 3H, 19-CH 3 ), 0.70–0.66(s, 3H, 20-CH 3 ).

[0110] HRMS(ESI)calcd for C 29 H 43 NO 2 [M+H] +:438.3367, found 438.3362.

[0111] Embodiment 17

[0112] (E)-N-(2-Fluorophenethyl)-5-((1R,4aS,6R,8aS)-6-hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methylpent-2-enamide (1q)

[0113] White powder, yield 87%, mp = 143.8 ~ 145.2 ° C;

[0114] 1 H NMR (400 MHz, CDCl 3 )δ7.21 (tdt, J=7.4, 3.5, 1.5Hz, 2H, Ph-H), 7.08 (td, J=7.4, 1.2Hz, 1H, Ph-H), 7.03 (ddd, J=9.9, 8.6, 1.3Hz, 1H, Ph -H), 5.47–5.43 (m, 2H, 14-H, NH), 4.85 (q, J=1.5Hz, 1H, 17-H), 4.50 (d, J=1.7Hz, 1H, 17-H), 3.59–3.46 (m, 2H, a-CH 2 ), 3.25 (dd, J=11.7, 4.3Hz, 1H, 3-H), 2.88 (t, J=7.0Hz, 2H, b-CH 2 ), 2.39 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.21 (ddd, J=14.0, 9.6, 4.1Hz, 1H, 12-H), 2.12 (d, J=1.3Hz, 3H, 16-CH 3 ), 2.04–1.83(m, 2H, 7&12-H), 1.81–1.43(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.37 (qd, J=12.9, 4.2Hz, 1H, 6-H), 1.15 (td, J=13.1, 3.8Hz, 1H, 1-H), 1.07 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.99 (s, 3H, 18-CH 3 ), 0.77(s, 3H, 19-CH 3 ), 0.68(s, 3H, 20-CH 3 ).

[0115] HRMS(ESI)calcd for C 28 H 40FNO 2 [M+H] + :442.3116, found 442.3108.

[0116] Embodiment 18

[0117] (E)-N-(2-Bromophenethyl)-5-((1R,4aS,6R,8aS)-6-hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methylpent-2-enamide (1r)

[0118] White powder, yield 82%, mp = 139.6 ~ 139.8 ℃;

[0119] 1 H NMR (400 MHz, CDCl 3 )δ7.53 (d, J=7.9Hz, 1H, Ph-H), 7.25–7.21 (m, 2H, Ph-H), 7.08 (dt, J=8.9, 4.5Hz, 1H, Ph-H), 5.57 (s, 1H, NH), 5. 45 (q, J=1.3Hz, 1H, 14-H), 4.84 (t, J=1.5Hz, 1H, 17-H), 4.49 (d, J=1.7Hz, 1H, 17-H), 3.54 (q, J=6.8Hz, 2H, a-CH 2 ), 3.23 (dd, J=11.8, 4.3Hz, 1H, 3-H), 2.97 (t, J=7.0Hz, 2H, (b-CH 2 ), 2.38 (ddd, J = 12.7, 4.3, 2.4Hz, 1H, 7-H), 2.21 (ddd, J = 13.8, 9.7, 3.8Hz, 1H, 12-H), 2.13 (d, J = 1.4Hz, 3H, 16-CH 3 ), 1.97–1.83(m, 2H, 7&12-H), 1.78–1.43(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.37 (qd, J=12.9, 4.2Hz, 1H, 6-H), 1.13 (td, J=13.2, 3.8Hz, 1H, 1-H), 1.06 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.98 (s, 3H, 18-CH 3 ), 0.75(s, 3H, 19-CH 3 ), 0.67(s, 3H, 20-CH 3 ).

[0120] HRMS(ESI)calcd for C 28 H 40 BrNO 2 [M+H] + :502.2315, found 502.2307.

[0121] Embodiment 19

[0122] (E)-5-((1R,4aS,6R,8aS)-6-Hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-N-(3-methoxyphenethyl)-3-methylpent-2-enamide (1s)

[0123] White powder, yield 79%, mp = 153.8 ~ 155.5 ° C;

[0124] 1 H NMR (400 MHz, CDCl 3 )δ7.22(t, J=7.8Hz, 1H, Ph-H), 6.81–6.72(m, 3H, 3×Ph-H), 5.63 (d, J=8.2Hz, 1H, NH), 5.30 (s, 1H, 14-H) 4.84 (q, J=1.6Hz, 1H, 17-H), 4.49 (d, J=1.8Hz, 1H, 17-H), 3.79 (s, 3H, MeO-CH 3 ), 3.61–3.48 (m, 2H, a-CH 2 ), 3.24 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.81 (t, J=6.9Hz, 2H, b-CH 2 ), 2.38 (ddd, J=12.8, 4.2, 2.4Hz, 1H, 7-H), 2.26–2.19 (m, 1H, 12-H), 2.12 (d, J=1.3Hz, 3H, 16-CH 3 ), 2.01–1.83(m, 2H, 7&12-H), 1.81-1.52(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.52–1.49(s, 1H, 6-H), 1.40–1.30(m, 2H, 1&5-H), 0.98(s, 3H, 18-CH 3 ), 0.76(s, 3H, 19-CH 3 ), 0.67(s, 3H, 20-CH 3 ).

[0125] HRMS(ESI)calcd for C29 H 43 NO 3 [M+H] + :454.3316, found 454.3312.

[0126] Embodiment 20

[0127] (E)-5-((1R,4aS,6R,8aS)-6-Hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methyl-N-(4-phenylbutyl)pent-2-enamide (1t)

[0128] 1 H NMR (400 MHz, CDCl 3 )δ7.32–7.23 (m, 2H, Ph*2-H), 7.22–7.13 (m, 3H, Ph*3-H), 5.46 (q, J=1.3Hz, 1H, 14-H), 5.33 (t, J =5.8Hz, 1H, NH), 4.88–4.83 (m, 1H, 17-H), 4.50 (d, J = 1.8Hz, 1H, 17-H), 3.31 (d, J = 7.2Hz, 2H, a-CH 2 ), 3.24 (dd, J=11.9, 4.6Hz, 1H, 3-H), 2.63 (t, J=7.5Hz, 2H, d-CH 2 ), 2.40 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.22 (ddd, J=14.0, 10.1, 3.9Hz, 1H, 12-H), 2.13 (d, J=1.2Hz, 3H, 16-CH 3 ), 2.06-1.83(m, 2H, 7&12-H), 1.82-1.44(m, 11H, 1-H, 6-H, 9-H, 2-CH 2 , 11-CH 2 , c-CH 2 , b-CH 2 ), 1.38 (qd, J=12.8, 4.3Hz, 1H, 6-H), 1.15 (td, J=13.2, 3.9Hz, 1H, 1-H), 1.07 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.99 (s, 3H, 18-CH 3 ), 0.77(s, 3H, 19-CH 3 ), 0.68(s, 3H, 20-CH 3 ).

[0129] HRMS(ESI)calcd for C 30 H45 NO 2 [M+H] + :452.3523, found 452.3517.

[0130] Embodiment 21

[0131] (E)-5-((1R,4aS,6R,8aS)-6-Hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methyl-N-(naphthalen-1-ylmethyl)pent-2-enamide (1u)

[0132] White powder, yield 94%, mp>250℃;

[0133] 1 H NMR (400 MHz, CDCl 3 )δ8.04(dq, J=8.6, 1.0Hz, 1H Ph-H), 7.88 (dd, J=7.7, 1.8Hz, 1H, Ph-H), 7.82 (dt, J=7.6, 1.2Hz, 1H, Ph-H), 7.60–7.49 (m, 2H, Ph-H×2), 7.47– 7.40 (m, 2H, Ph-H×2), 5.60 (t, J=5.5Hz, 1H, NH), 5.48 (q, J=1.3Hz, 1H, 14-H), 4.92 (dd, J=5.6, 1.1Hz, 2H, NH-CH 2 -Ph), 4.82 (q, J=1.5Hz, 1H, 17-H), 4.47 (d, J=1.6Hz, 1H, 17-H), 3.22 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.36 (ddd, J=12.8, 4.4, 2.5Hz, 1H, 7-H), 2.23 (td, J=10.6, 9.9, 5.1Hz, 1H, 12-H), 2.19 (d, J=1.2Hz, 3H, 16-CH 3 ), 1.98–1.81(m, 2H, 7&12-H), 1.79–1.42(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.36 (qd, J=13.0, 4.2Hz, 1H, 6-H), 1.13 (td, J=13.2, 3.8Hz, 1H, 1-H), 1.05 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.99 (s, 3H, 18-CH 3 ), 0.75(s, 3H, 19-CH 3 ), 0.68 (m, 3H, 20-CH 3 ).

[0134] HRMS(ESI)calcd for C 31 H 41 NO 2 [M+H] + :460.3210, found 460.3205.

[0135] Embodiment 22

[0136] (E)-5-((1R,4aS,6R,8aS))-6-Hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methyl-N-(naphthalen-2-ylmethyl)pent-2-enamide (1v)

[0137] White powder, yield 92%, mp = 146.2 ~ 148.0 ℃;

[0138] 1 H NMR (400 MHz, CDCl 3 )δ7.85–7.77 (m, 3H, Ph-H×3), 7.73 (d, J=1.7Hz, 1H, Ph-H), 7.52–7.42 (m, 2H, Ph-H×2), 7.42 (dd, J=8.5, 1.8H z, 1H, Ph-H), 5.72 (s, 1H, NH), 5.55 (q, J=1.3Hz, 1H, 14-H), 4.88–4.82 (m, 1H, 17-H), 4.67–4.59 (m, 2H, NH-CH 2 -Ph), 4.51 (s, 1H, 17-H), 3.24 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.39 (ddd, J=12. 8, 4.3, 2.5Hz, 1H, 7-H), 2.31–2.21 (m, 1H, 12-H), 2.19 (d, J=1.3Hz, 3H, 16-CH 3 ), 2.01–1.86(m, 2H, 7&12-H), 1.86–1.28(m, 7H, 1-H, 6-H, 9-H, 11-CH 2 , 2-CH 2 ), 1.39 (qd, J=12.8, 4.2Hz, 1H, 6-H), 1.15 (td, J=13.2, 3.8Hz, 1H, 1-H), 1.07 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.99 (s, 3H, 18-CH 3 ), 0.76(s, 3H, 18-CH 3 ), 0.68(s, 3H, 18-CH 3 ).

[0139] HRMS(ESI)calcd for C 31 H 41 NO 2 [M+H] + :460.3210, found 460.3204.

[0140] Embodiment 23

[0141] (E)-N-(adamantan-1-yl)-5-((1R,4aS,6R,8aS)-6-hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methylpent-2-enamide (1w)

[0142] White powder, yield 89%, mp = 203.2 ~ 205.2 ° C;

[0143] 1 H NMR (400 MHz, CDCl 3 )δ5.42 (d, J=1.9Hz, 1H, 14-H), 5.08 (s, 1H, NH), 4.84 (d, J=1.9Hz, 1H, 17-H), 4.50 (d, J=1.8Hz, 1H, 17-H), 3.24 (dd, J=11.7, 4 .4Hz, 1H, 3-H), 2.39 (ddd, J=12.8, 4.3, 2.4Hz, 1H, 7-H), 2.20 (ddt, J=14.0, 9.7, 5.1Hz, 1H, 12-H), 2.10 (d, J=1.2Hz, 3H, 16-CH 3 ), 2.08–2.04(m, 3H), 2.08–2.04(d, J=3.2Hz, 3H, cH*3), 2.03–1.99(d, J=2.9Hz, 6H, b-CH 2 *3), 1.94 (td, J=12.9, 5.0Hz, 1H), 1.89–1.69 (m, 4H), 1.65–1.43 (m, 5H), 1.37 (qd, J=12.8, 4.3Hz, 1 H, 6-H), 1.15 (t, J=12.8Hz, 1H, 1-H), 1.07 (dt, J=12.5, 2.5Hz, 1H, 5-H), 0.99 (d, J=1.7Hz, 3H, 18-CH 3 ), 0.76 (d, J=1.0Hz, 3H, 19-CH 3 ), 0.68(s, 3H, 20-CH 3 ).

[0144] HRMS(ESI)calcd for C30 H 47 NO 2 [M+H] + :454.3680, found 454.3672.

[0145] Embodiment 24

[0146] (E)-N-(3,5-dimethyladamantan-1-yl)-5-((1R,4aS,6R,8aS)-6-hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)-3-methylpent-2-enamide (1x)

[0147] White powder, yield 83%, mp = 129.5 ~ 131.2 ° C;

[0148] 1 H NMR (400 MHz, CDCl 3 )δ5.41 (dd, J=2.6, 1.1Hz, 1H, 14-H), 5.08 (s, 1H, NH), 4.85 (q, J=1.6Hz, 1H, 17-H), 4.50 (d, J=1.7Hz, 1H, 17-H), 3 .25 (dd, J=11.7, 4.4Hz, 1H, 3-H), 2.39 (ddd, J=12.7, 4.2, 2.4Hz, 1H, 7-H), 2.20 (ddd, J=14.0, 9.5, 4.0Hz, 1H, 12- H), 2.14 (q, J=3.1Hz, 1H), 2.11 (d, J=1.3Hz, 3H, 16-H), 1.95 (td, J=13.0, 5.1Hz, 1H), 1.86 (dd, J=12.6, 2.8Hz, 3H ), 1.80–1.69 (m, 3H), 1.64–1.25 (m, 13H), 1.22–1.11 (m, 3H), 1.08 (dd, J=12.5, 2.8Hz, 1H, 5-H), 0.99 (s, 3H, 18-CH 3 ), 0.84(s, 6H, 2×h-CH 3 ), 0.77(s, 3H, 19-CH 3 ), 0.68(s, 3H, 20-CH 3 ).

[0149] HRMS(ESI)calcd for C 32 H 51 NO 2 [M+H] + :482.3993, found 482.3990.

[0150] The compounds of the present invention may include one or more asymmetric centers and may therefore exist in a variety of stereoisomeric forms, for example, enantiomers and / or diastereoisomeric forms. For example, the compounds of the present invention may be individual enantiomers, diastereomers or geometric isomers (e.g., cis and trans isomers), or may be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. Isomers may be separated from the mixture by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers may be prepared by asymmetric synthesis.

[0151] Those skilled in the art will appreciate that many organic compounds can form complexes with solvents in which they react or from which they precipitate or crystallize. These complexes are referred to as "solvates". When the solvent is water, the complex is referred to as a "hydrate". The present invention encompasses all solvates of the compounds of the present invention.

[0152] On the other hand, the present invention provides a pharmaceutical composition comprising a compound of the present invention (also referred to as an "active ingredient") and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition comprises an effective amount of a compound of the present invention. In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of a compound of the present invention. In some embodiments, the pharmaceutical composition comprises a preventive effective amount of a compound of the present invention.

[0153] The pharmaceutically acceptable excipient used in the present invention refers to a non-toxic carrier, adjuvant or vehicle that does not destroy the pharmacological activity of the compound formulated together. Pharmaceutically acceptable carriers, adjuvants or vehicles that can be used in the composition of the present invention include (but are not limited to) ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffer substances (such as phosphates), glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, silica gel, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and lanolin.

[0154] The present invention also includes a kit (e.g., a pharmaceutical package). The kit provided may include a compound of the invention, other therapeutic agents, and a first and second container (e.g., a vial, an ampoule, a bottle, a syringe, and / or a dispersible package or other suitable container) containing the compound of the invention and other therapeutic agents. In some embodiments, the kit provided may also optionally include a third container containing a pharmaceutical excipient for diluting or suspending the compound of the invention and / or other therapeutic agents. In some embodiments, the compound of the invention and other therapeutic agents provided in the first container and the second container are combined to form a unit dosage form.

[0155] The following formulation examples illustrate representative pharmaceutical compositions that can be prepared according to the present invention. However, the present invention is not limited to the following pharmaceutical compositions.

[0156] Exemplary Formulation - Tablets: The compound of the invention in dry powder form can be mixed with a dry gel binder in a weight ratio of about 1:2. A small amount of magnesium stearate is added as a lubricant. The mixture is formed into 0.1-40 mg tablets in a tablet press.

[0157] Exemplary Formulation - Capsules: A compound of the invention in dry powder form can be mixed with a starch diluent in a weight ratio of about 1: 1. The mixture is filled into capsules (each capsule contains 100 mg of active compound).

[0158] Exemplary formulation-liquid: The compounds of the present invention (125 mg) can be mixed with sucrose (1.75 g) and xanthan gum (4 mg), and the resulting mixture can be blended, sieved, and then mixed with an aqueous solution of pre-prepared microcrystalline cellulose and sodium carboxymethyl cellulose (11:89, 50 mg). Sodium benzoate (10 mg), flavoring agents and coloring agents are diluted with water and added under stirring. Then, sufficient water can be added to obtain a total volume of 5 mL.

[0159] Exemplary Formulations - Injectable: The compounds of the invention may be dissolved or suspended in a buffered sterile saline injectable aqueous medium to a concentration of about 10 mg / mL.

[0160] Drug administration

[0161] Pharmaceutical composition provided by the invention can be administered by many ways, including but not limited to: oral administration, parenteral administration, inhalation administration, topical administration, rectal administration, nasal administration, oral administration, vaginal administration, administration by implant or other modes of administration. For example, parenteral administration used herein includes subcutaneous administration, intradermal administration, intravenous administration, intramuscular administration, intraarticular administration, intraarterial administration, intrasynovial cavity administration, intrasternal administration, intrathecal administration, intralesional administration and intracranial injection or infusion technology.

[0162] Generally, an effective amount of the compounds provided herein is administered. The amount of the compound actually administered can be determined by a physician according to the relevant circumstances, including the condition being treated, the route of administration selected, the compound actually administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, and the like.

[0163] When used to prevent the conditions described herein, the compounds provided herein are administered to a subject at risk of developing the condition, typically based on the advice of a physician and under the supervision of a physician, at dosage levels as described above. Subjects at risk of developing a specific condition typically include those with a family history of the condition, or those identified by genetic testing or screening as being particularly susceptible to developing the condition.

[0164] The pharmaceutical compositions provided by the present invention can also be administered for a long time ("chronic administration"). Long-term administration refers to administration of a compound or its pharmaceutical composition over a long period of time, for example, 3 months, 6 months, 1 year, 2 years, 3 years, 5 years, etc., or administration can be continued indefinitely, for example, for the rest of the subject's life. In some embodiments, long-term administration is intended to provide a constant level of the compound in the blood over a long period of time, for example, within a therapeutic window.

[0165] Various methods of administration can be used to further deliver the pharmaceutical composition of the present invention. For example, in some embodiments, the pharmaceutical composition can be administered by push injection, for example, in order to raise the concentration of the compound in the blood to an effective level. The push dose depends on the target systemic level of the active component through the body, for example, the intramuscular or subcutaneous push dose slowly releases the active component, and the push injection (for example, by IV intravenous drip) delivered directly to the vein can be delivered more quickly, so that the concentration of the active component in the blood is quickly increased to an effective level. In other embodiments, the pharmaceutical composition can be given in a continuous infusion form, for example, by IV intravenous drip, so as to provide a steady-state concentration of the active component in the subject's body. In addition, in other embodiments, the pharmaceutical composition of the push dose can be first given, and then continuous infusion.

[0166] Oral compositions can be in the form of bulk liquid solutions or suspensions or bulk powders. However, more generally, in order to facilitate accurate dosing, the composition is provided in unit dosage form. The term "unit dosage form" refers to a physical discrete unit suitable as a unit dose for human patients and other mammals, each unit containing a predetermined amount of active substances suitable for producing the desired therapeutic effect and a suitable pharmaceutical excipient. Typical unit dosage forms include pre-filled, pre-measured ampoules or syringes of liquid compositions, or pills, tablets, capsules, etc. in the case of solid compositions. In such compositions, the compound is usually a relatively small component (about 0.1 to about 50% by weight, or preferably about 1 to about 40% by weight), and the remainder is various carriers or excipients and processing aids useful for forming the desired administration form.

[0167] For oral dosage, a typical regimen is one to five oral doses per day, especially two to four oral doses, typically three oral doses. Using these dosing modes, each dose provides about 0.01 to about 100 mg / kg of the compound of the invention.

[0168] In order to provide blood levels similar to those using an injectable dose, or lower blood levels than using an injectable dose, a transdermal dose is generally selected in an amount of about 0.01 to about 20% by weight, preferably about 0.1 to about 20% by weight, preferably about 0.1 to about 10% by weight, and more preferably about 0.5 to about 15% by weight.

[0169] From about 1 to about 120 hours, especially 24 to 96 hours, the injected dosage level is in the range of about 0.1 mg / kg / hour to at least 10 mg / kg / hour. In order to obtain sufficient steady state levels, a preload bolus of about 0.1 mg / kg to about 10 mg / kg or more may also be given. For a 40 to 80 kg human patient, the maximum total dose may not exceed about 2 g / day.

[0170] Liquid forms suitable for oral administration may include a suitable aqueous or non-aqueous carrier and buffers, suspending and dispersing agents, colorants, flavoring agents, etc. Solid forms may include, for example, any of the following components, or compounds of a similar nature: binders, such as microcrystalline cellulose, tragacanth, or gelatin; excipients, such as starch or lactose; disintegrants, such as alginic acid, Primogel, or corn starch; lubricants, such as magnesium stearate; glidants, such as colloidal silicon dioxide; sweeteners, such as sucrose or saccharin; or flavoring agents, such as peppermint, methyl salicylate, or orange flavoring.

[0171] Injectable compositions are typically based on injectable sterile saline or phosphate buffered saline, or other injectable excipients known in the art. As previously mentioned, in such compositions, the active compound is typically a minor component, often about 0.05 to 10% by weight, and the remainder is an injectable excipient, etc.

[0172] Typically, transdermal compositions are formulated as topical ointments or creams containing active ingredients. When formulated as an ointment, the active ingredient is typically combined with a paraffin or water-miscible ointment base. Alternatively, the active ingredient can be formulated as an ointment together with, for example, an oil-in-water cream base. Such transdermal formulations are well known in the art and typically include other components for enhancing the stable skin penetration of the active ingredient or formulation. All such known transdermal formulations and components are included within the scope provided by the invention.

[0173] The compounds of the invention may also be administered by transdermal means. Thus, transdermal administration may be achieved using patches of the reservoir or porous membrane type, or a variety of solid matrices.

[0174] The above components of compositions for oral administration, injection or topical administration are merely representative.

[0175] The compounds of the invention may also be administered in sustained release form or from a sustained release delivery system.

[0176] The present invention also relates to pharmaceutically acceptable formulations of the compounds of the present invention. In one embodiment, the formulation comprises water. In another embodiment, the formulation comprises a cyclodextrin derivative.

[0177] Cytotoxicity test of the compounds of the present invention

[0178] Media and reagents

[0179] MCF-7, A549, and HepG2 cells were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA). MCF-7, A549, and HepG2 cells were cultured in DMEM high-glucose medium from HyClone GE Healthcare Life Sciences. The medium was supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 1% antibiotics (penicillin and streptomycin). All cells were cultured at 37°C in a 5% CO 2 cultured in an incubator.

[0180] Cytotoxicity assay

[0181] According to the counting results, cells in the logarithmic growth phase were inoculated in a 96-well plate at 3000-5000 cells per well (5000-10000 cells for HepG2), with 100 μL of single cell suspension per well. A blank group and a control group were set up, and the edge wells were filled with sterile PBS. After 24 hours of cell culture, the culture medium was discarded, and 100 μL of compounds of different concentrations were added to each well of the 96-well plate, with 3 replicates for each concentration. After 72 hours of drug treatment, 10 μL of 5 mg / mL MTT solution was added to each well and incubated in CO 2 After incubation at 37°C for 2 h in an incubator, the medium was aspirated. Then 150 μL of DMSO was added to each well to dissolve the formazan and the mixture was shaken evenly. The absorbance was measured at a wavelength of 490 nm using an ELISA reader. The half-maximal inhibitory concentration (IC) of the compound was calculated using Graphpad Prism 8 software. 50 The calculation formula of compound inhibition rate is: cell inhibition rate (%) = (OD control - OD drug addition) / (OD control - OD blank) × 100%.

[0182] The experimental results are shown in Table 1. By introducing amine groups of different sizes into fern acid, the anti-proliferative activity of different modifications on cancer cells was systematically tested. The heavy metal complex cisplatin was used as a positive control drug, and three cancer cells, human breast cancer cells (MCF-7), human non-small cell lung cancer cells (A549), and human liver cancer cells (HepG2) were used as experimental subjects. The results (Table 1) showed that all compounds had good inhibitory effects on the three cancer cells MCF-7, A549 and HepG2. The inhibitory activity of most derivatives on MCF-7 is generally greater than their inhibitory activity on A549 and HepG2. Among them, compounds 1o-1s have the best proliferation inhibitory activity on the three cancer cells, and compound 1r has the best anti-tumor activity, and its IC against HepG2 is 50 =1.92±0.22μM, which is much higher than the positive control drugs cisplatin and fern acid. The above results show that this type of derivative has a good development prospect and is expected to be further developed and applied in the field of anti-tumor drugs.

[0183] Table 1 Cytotoxicity data (IC 50 , μM)

[0184]

[0185]

[0186] a Human non-small cell lung cancer cells A549; b Human breast cancer cell line MCF-7; c Human hepatoma cell line HepG2; IC50 (μM): half inhibitory concentration of cells; d Positive control drug; e Precipitate was formed during the MTT assay.

[0187] The above description is only a preferred embodiment of the present invention, which is only illustrative and not restrictive of the present invention. Those skilled in the art understand that many changes, modifications, and even equivalences may be made to the present invention within the spirit and scope defined by the claims of the present invention, but all of them will fall within the scope of protection of the present invention.

Claims

1. Use of an N-aromatic alkylamino fern amide compound in the preparation of a drug for inhibiting the proliferation of human liver cancer cells HepG2, characterized in that: The structural formula of the N-aromatic alkylamino fern amide compound is The R is:

2. The use of an N-aromatic alkylamino fern amide compound as claimed in claim 1 in the preparation of a drug for inhibiting the proliferation of human liver cancer cells HepG2, characterized in that: The pharmaceutically acceptable salts are acetate, benzoic acid, hydrochloride, maleate, fumaric acid, sulfate, benzenesulfonate, succinate and salicylate.

3. Use of an N-aromatic alkylamino fern amide compound as claimed in claim 1 in the preparation of a drug for inhibiting the proliferation of human liver cancer cells HepG2, characterized in that: The reaction formula of the preparation method of the N-aromatic alkylamino fern amide compound is:

4. Use of an N-aromatic alkylamino fern amide compound as claimed in claim 1 in the preparation of a drug for inhibiting the proliferation of human liver cancer cells HepG2, characterized in that: The preparation method of the N-aromatic alkylamino fern amide compound comprises the following steps: accurately weighing fern acid and dissolving it in dichloromethane, then sequentially adding O-(benzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N,N-diisopropylethylamine and amine compounds with different substitutions, stirring the reaction at room temperature, tracking the reaction by thin layer chromatography during the stirring process, adding water to terminate the reaction after the reaction is complete, evaporating the solvent under reduced pressure, and separating and purifying the obtained crude product by silica gel column chromatography to obtain the target compound.

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