Nitric oxide donor type carbazole derivatives and their use in the treatment of neoplastic diseases

Abstract

The application belongs to the field of natural medicine and medicinal chemistry, and discloses a class of nitric oxide donor type pyrano-carbazole alkaloid derivatives, a preparation method of the nitric oxide donor type pyrano-carbazole alkaloid derivatives, and a pharmaceutical composition and application of the nitric oxide donor type pyrano-carbazole alkaloid derivatives. Specifically, the application discloses carbazole alkaloids as shown in general formula I and II. The alkaloids are prepared by an artificial synthesis method, contain a pharmaceutical composition, and are used in preparation of medicines for treating tumor diseases.

Description

Technical Field

[0001] This invention relates to the fields of natural medicines and medicinal chemistry, specifically to a class of nitric oxide donor-type pyranocarbazole alkaloid derivatives. The invention also discloses methods for preparing such derivatives, pharmaceutical compositions thereof, and their uses in antitumor applications. Background Technology

[0002] Carbazole is a tricyclic compound consisting of two six-membered benzene rings fused to either side of a five-membered nitrogen-containing ring. It is abundant in the leaves, fruits, roots, and bark of plants in the Rutaceae family. The carbazole ring has been widely used as an effective framework for drug discovery, and its derivatives can trigger various cancer cell death pathways. For example, Ellipticine, Celiptium, and Alectinib are carbazole-based anticancer drugs approved for chemotherapy, demonstrating the potential of carbazole derivatives as candidate anticancer drugs. Previous research by the inventors found that introducing a nitrogen-containing fragment at the 3-position of pyranocarbazole can enhance the antitumor activity of pyranocarbazole alkaloids, such as compound 3, which exhibits IC50 activity against various tumor cells. 50 The values ​​ranged from 1.16 to 4.96 μM (Ke Liu, Yingda Zang, Bioorg. Med. Chem. Lett., 2021, 33: 127739), exhibiting good antitumor cytotoxic activity.

[0003]

[0004] Nitric oxide has a dual role in antitumor activity, primarily depending on the concentration, timing, and site of action of its formation. Sustained low concentrations of nitric oxide can promote tumor cell growth, while high concentrations can exert antitumor activity through cytotoxicity. Furazolidone, as a nitric oxide donor, can continuously release high concentrations of nitric oxide in vivo (Kerwin JF, Heller M., Med. Res. Rev., 1994, 14(1):23.). Nitric oxide donor-drug conjugates often exhibit synergistic effects with the parent drug, thus attracting considerable attention in new drug research.

[0005] To obtain compounds with superior antitumor activity compared to compound 3, we conducted structural modification studies on pyranocarbazole alkaloids. This invention discloses a class of furazolidone nitric oxide donor-type pyranocarbazole alkaloid derivatives with medicinal value and their pharmaceutically acceptable salts; no such compounds have been reported to date. Summary of the Invention

[0006] The technical problem solved by this invention is to provide a class of compounds of general formula I and general formula II, and their uses. Pharmacological experiments have shown that the compounds of this invention have significant apoptotic effects on various tumor cells. Therefore, general formulas I and II of this invention can be used for the prevention and / or treatment of tumor diseases.

[0007] To solve the technical problem of this invention, the present invention provides the following technical solution:

[0008] The first aspect of the present invention provides a compound of general formula I or a pharmaceutically acceptable salt thereof:

[0009]

[0010] R1 and R2 are independent of each other and can be selected from H, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, CH2CH2CH2CH3, CH2CH(CH3)2, CH(CH3)CH2CH3, t-Bu, OCH3, OCH2CH3, OCH2CH2CH3, CF3, and OCF3, respectively.

[0011] R3 is selected from H, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, CH2CH2CH2CH3, CH2CH(CH3)2, CH(CH3)CH2CH3;

[0012] X is selected from substituted amino groups, wherein the substituents can be selected from straight-chain or branched C-chain groups. 1-6 Alkyl, C 3-6 Cycloalkyl groups or substituents together with nitrogen atoms form a five-, six-, or seven-membered saturated heterocycle containing 1-3 heteroatoms. Substituents may be present on the heterocycle, and these substituents can be selected from -OH, -F, -Cl, Br, -COOH, straight-chain or branched C atoms. 1-6 Alkyl, C 3-6 Cycloalkyl, straight-chain or branched C 1-6 Alkoxy, substituted or unsubstituted phenyl groups, wherein the substituents on the phenyl group may be selected from -OH, -F, -Cl, Br, -COOH, straight-chain or branched C. 1-6 Alkyl groups, -CHF2, -CF3, -CN, -NO2, -OCF3, -ONO2, -NH2, or straight-chain or branched C atoms 1-6 Alkyl group.

[0013] The first aspect of the present invention also provides a compound having the structure of Formula II or a pharmaceutically acceptable salt thereof.

[0014]

[0015] R1 and R2 are independent of each other and can be selected from H, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, CH2CH2CH2CH3, CH2CH(CH3)2, CH(CH3)CH2CH3, t-Bu, OCH3, OCH2CH3, OCH2CH2CH3, CF3, and OCF3, respectively.

[0016] R3 is selected from H, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, CH2CH2CH2CH3, CH2CH(CH3)2, CH(CH3)CH2CH3;

[0017] X is selected from substituted amino groups, wherein the substituents can be selected from straight-chain or branched C-chain groups. 1-6 Alkyl, C 3-6 Cycloalkyl groups or substituents together with nitrogen atoms form a five-, six-, or seven-membered saturated heterocycle containing 1-3 heteroatoms. Substituents may be present on the heterocycle, and these substituents can be selected from -OH, -F, -Cl, Br, -COOH, straight-chain or branched C atoms. 1-6 Alkyl, C 3-6 Cycloalkyl, straight-chain or branched C 1-6 Alkoxy, substituted or unsubstituted phenyl groups, wherein the substituents on the phenyl group may be selected from -OH, -F, -Cl, Br, -COOH, straight-chain or branched C. 1-6 Alkyl groups, -CHF2, -CF3, -CN, -NO2, -OCF3, -ONO2, -NH2, or straight-chain or branched C atoms 1-6 Alkyl group.

[0018] The compounds mentioned are selected from:

[0019]

[0020] The second aspect of this technical solution is to provide a method for preparing the compound described in the first aspect.

[0021] The compounds represented by general formula I of this invention can be prepared by the following methods:

[0022]

[0023] Condition a: reactants HXOH (X as in claim 1), 30% NaOH, solvent tetrahydrofuran, reaction at room temperature; Condition b: catalysts EDCI and HOBt, solvent dichloromethane. The preparation methods for intermediates 1 and 2 are described in J. Med. Chem. 2011, 54, 3251–3259 and Molecules 2021, 26, 1303.

[0024] The compounds represented by general formula II of this invention can be prepared by the following methods:

[0025]

[0026] Among them, condition a is the same as above; condition c: reactant XH (X is selected from substituted amino groups, the same as above), reducing agent is sodium borohydride acetate, catalyst is acetic acid, solvent is anhydrous tetrahydrofuran, room temperature.

[0027] A third aspect of the present invention is to provide a pharmaceutical composition, characterized in that the pharmaceutical composition comprises the compounds described in FN-1 to 7, CY-1 to 3, CY-5, and CY-7 as active ingredients, or pharmaceutically acceptable salts thereof, and a pharmacodynamically acceptable carrier or excipient.

[0028] Use of the compounds described in FN-1~7, CY-1~3, CY-5, CY-7 or their pharmaceutically acceptable salts in the preparation of medicaments for the prevention and / or treatment of tumor diseases.

[0029] Another aspect of the present invention relates to pharmaceutical compositions in which the compounds of the present invention are used as active ingredients. These pharmaceutical compositions are prepared according to methods known in the art. They can be formulated into any dosage form suitable for human or animal use by combining the compounds of the present invention with one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants. The content of the compounds of the present invention in their pharmaceutical compositions is typically 0.1-95% by weight.

[0030] The compounds of this invention or pharmaceutical compositions containing them can be administered in unit dose form via enteral or non-enteric routes, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum, etc.

[0031] Dosage forms can be liquid, solid, or semi-solid. Liquid dosage forms can include solutions (including true solutions and colloidal solutions), emulsions (including o / w, w / o, and double emulsions), suspensions, injections (including aqueous injections, powder injections, and infusions), eye drops, nasal drops, lotions, and liniments, etc.; solid dosage forms can include tablets (including regular tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, and orally disintegrating tablets), capsules (including hard capsules, soft capsules, and enteric-coated capsules), granules, powders, microcapsules, pellets, suppositories, films, patches, aerosols, and sprays, etc.; semi-solid dosage forms can include ointments, gels, and pastes, etc.

[0032] The compounds of this invention can be formulated into conventional formulations, sustained-release formulations, controlled-release formulations, targeted formulations, and various microparticle delivery systems.

[0033] To formulate the compounds of the present invention into tablets, a wide variety of excipients known in the art can be used, including diluents, binders, wetting agents, disintegrants, lubricants, and flow aids. Diluents can be starch, dextrin, sucrose, glucose, lactose, mannose, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, dicalcium phosphate, calcium carbonate, etc.; wetting agents can be water, ethanol, isopropanol, etc.; binders can be starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic paste, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfonate, etc.; lubricants and flow aids can be talc, silica, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0034] Tablets can also be further processed into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer and multilayer tablets.

[0035] To formulate the drug delivery unit into capsules, the active ingredient, the compound of the present invention, can be mixed with a diluent and a disintegrant, and the mixture can be placed directly into hard or soft capsules. Alternatively, the active ingredient, the compound of the present invention, can be first formed into granules or microspheres with a diluent, binder, and disintegrant, and then placed into hard or soft capsules. Various diluents, binders, wetting agents, disintegrants, and disintegrants used to prepare tablets of the compound of the present invention can also be used to prepare capsules of the compound of the present invention.

[0036] To prepare the compounds of this invention into injectable formulations, water, ethanol, isopropanol, propylene glycol, or mixtures thereof can be used as solvents, and appropriate amounts of commonly used solubilizers, co-solvents, pH adjusters, and osmotic pressure adjusters can be added. Solubilizers or co-solvents can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc. pH adjusters can be phosphates, acetates, hydrochloric acid, sodium hydroxide, etc.; osmotic pressure adjusters can be sodium chloride, mannitol, glucose, phosphates, acetates, etc. If preparing lyophilized powder injections, mannitol, glucose, etc., can also be added as supporting agents.

[0037] In addition, colorants, preservatives, flavorings, tasters or other additives may be added to pharmaceutical preparations if necessary.

[0038] To achieve the purpose of medication and enhance the therapeutic effect, the drug or drug composition of the present invention can be administered using any known method of administration.

[0039] The dosage of the pharmaceutical compositions of the present invention can vary widely depending on the nature and severity of the disease to be prevented or treated, the individual condition of the patient or animal, the route of administration, and the dosage form. Generally, the suitable daily dose range of the compounds of the present invention is 0.001-150 mg / kg body weight, preferably 0.1-100 mg / kg body weight, more preferably 1-60 mg / kg body weight, and most preferably 2-30 mg / kg body weight. The above doses can be administered as a single dose unit or divided into several dose units, depending on the physician's clinical experience and the administration regimen, including the use of other treatment methods.

[0040] The compounds or compositions of the present invention can be taken alone or in combination with other therapeutic or symptomatic drugs. When the compounds of the present invention have a synergistic effect with other therapeutic drugs, their dosage should be adjusted according to the actual situation.

[0041] The fourth aspect of the present invention is to provide the use of the compound described in the first aspect or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention and / or treatment of tumor diseases.

[0042] The compounds of this invention, or their pharmaceutically acceptable salts, can be used in the pharmaceutical field. For example, in vivo pharmacodynamic experiments for the preparation of anti-breast cancer drugs show that compound FN-1 of this invention, at an oral dose of 150 mg / kg, exhibits significant anti-breast cancer activity, with a tumor inhibition rate of approximately 50.91%. Preliminary oral acute toxicity experiments indicate that the median lethal dose (LD50) of compound FN-1 of this invention is... 50 The concentration was greater than 2 g / kg. These results indicate that FN-1 is a compound with high safety and anti-breast cancer activity. Attached Figure Description

[0043] Figure 1 These are tumor tissue images from an in vivo activity assay of compound FN-1 against mouse EMT6 breast cancer cells. Detailed Implementation

[0044] Chemical Experiment

[0045] Examples 1-7 can be synthesized according to the following route:

[0046]

[0047] Preparation of (FN-1) in Example 1

[0048] J-1 (25 mg, 1.5 eq) was dissolved in 2 mL of tetrahydrofuran, and CZA-1 (96 mg, 1 eq), EDCI (98 mg, 2 eq), and HOBt (69 mg, 2 eq) were added sequentially. The mixture was stirred overnight, and the reaction was quenched dropwise with water. The mixture was extracted with ethyl acetate-water, and the ethyl acetate layer was developed using a scraper (petroleum ether:acetone = 2:1). 87 mg of a yellow powder was obtained, with a yield of 53.0%. 1 H NMR (500MHz, Acetone-d6) δ8.25(s,1H),8.09(dt,J=26.1,11.7Hz,5H),7.82(t,J=7.7Hz,1H),7.73(t,J=7.8Hz,3H),7.43(d,J=8.5Hz,1H) ,7.36(d,J=8.5Hz,1H),6.92(dd,J=16.9,12.7Hz,2H),5.85(d,J=9.8Hz,1H),3.85(t,J=5.5Hz,2H),1.53(s,6H),1.42(s,9H),1.29(s,1H). 13 C NMR (101MHz, Acetone) δ167.5,160.2,151.1,143.3,139.5,139.1,139.0,137.2,136.6,130.7,130.4,129.6,12 4.1,123.8,120.2,119.7,118.9,118.2,116.9,116.6,111.6,111.2,105.7,77.5,71.2,38.7,35.2,32.3,27.9.

[0049] HRESIMS m / z = 643.22 [M+H] +

[0050] Example 2 Preparation of (FN-2)

[0051] J-1 (100 mg, 1.5 eq) was dissolved in 3 mL of tetrahydrofuran. CZA-1 (63 mg, 1 eq), EDCI (64 mg, 2 eq), and HOBt (45 mg, 2 eq) were added sequentially. The mixture was stirred overnight, and the reaction was quenched dropwise with water. Extraction was performed with ethyl acetate-water, and the ethyl acetate layer was developed using a scraper (petroleum ether:acetone = 2:1). 69 mg of a yellow powder was obtained, with a yield of 62.7%. 1H NMR (500MHz, Acetone-d6) δ8.22(s,1H),8.12(t,J=7.7Hz,2H),8.07(t,J=2.2Hz,1H),8.04(dd,J=15.7,5 .1Hz,1H),7.83(dt,J=15.3,8.2Hz,1H),7.73(q,J=7.6Hz,2H),7.42(dd,J=8.6,2.0Hz,1H),7.35(d,J=8. 5Hz,1H),6.92(d,J=9.8Hz,1H),6.84(dd,J=15.7,4.0Hz,1H),5.84(d,J=9.8Hz,1H),4.61–4.46(m,2H),2 .81(s,1H),1.95(s,1H),1.53–1.46(m,7H),1.41(s,9H),1.39(d,J=6.5Hz,2H),1.28(s,1H),1.19(s,1H). 13 C NMR (101MHz, Acetone) δ167.6,160.3,151.1,143.3,139.1,137.0,136.6,136.6,130.7,130.7,130.4,129. 6,129.5,123.8,119.8,118.2,116.6,116.6,111.2,111.2,105.7,77.5,74.3,35.2,32.3,27.9,27.8,17.5.

[0052] HRESIMS m / z = 657.23 [M+H] +

[0053] Preparation of (FN-3) in Example 3

[0054] J-3 (140 mg, 2 eq) was dissolved in 3 mL of tetrahydrofuran, and CZA-1 (77 mg, 1 eq), EDCI (79 mg, 2 eq), and HOBt (56 mg, 2 eq) were added sequentially. The mixture was stirred overnight, and the reaction was quenched dropwise with water. The mixture was extracted with ethyl acetate-water, and the ethyl acetate layer was developed using a scraper (petroleum ether:acetone = 2:1). 93 mg of a yellow powder was obtained, with a yield of 64.6%. 1H NMR (500MHz, Acetone-d6) δ8.13–8.05(m,4H),7.89(t,J=7.5Hz,1H),7.77(t,J=7.8Hz,2H),7.46–7.37(m,2H),7.36(d,J=8.2H z,1H),6.94(d,J=9.8Hz,1H),5.85(d,J=9.8Hz,1H),4.39(d,J=6.1Hz,2H),2.78(s,2H),2.09(s,2H),1.52(s,6H),1.41(s,9H). 13 C NMR (101MHz, Acetone) δ166.3,160.2,151.0,143.3,139.6,139.5,139.2,139.1,138.9,136.7,130.8,130.2,129.4,124.3,12 3.8,120.3,119.0,118.9,118.3,117.4,116.7,116.1,111.4,111.2,111.1,105.7,105.6,77.4,75.8,36.7,35.2,32.3,27.9.

[0055] HRESIMS m / z = 697.27 [M+H] +

[0056] Preparation of (FN-4) in Example 4

[0057] J-4 (140 mg, 2 eq) was dissolved in 3 mL of tetrahydrofuran. CZA-1 (81 mg, 1 eq), EDCI (83 mg, 2 eq), and HOBt (58 mg, 2 eq) were added sequentially. The mixture was stirred overnight, and the reaction was quenched dropwise with water. Extraction was performed with ethyl acetate-water, and the ethyl acetate layer was developed using a scraper (petroleum ether:acetone = 2:1). 91 mg of a yellow powder was obtained, with a yield of 61.9%. 1H NMR(500MHz, Acetone-d6)δ8.40(s,1H),8.16–8.03(m,5H),7.71(t,J=7.8Hz,2H),7 .43(d,J=8.7Hz,1H),7.36(d,J=8.5Hz,1H),7.22(d,J=15.5Hz,1H),6.94(d,J=9.8Hz ,1H),5.86(d,J=9.8Hz,1H),4.60(dt,J=10.0,5.3Hz,2H),3.96(s,1H),3.84(q,J=8 .2Hz,1H),2.80(d,J=16.8Hz,4H),2.09(s,2H),1.54(d,J=6.6Hz,7H),1.41(s,11H). 13 C NMR (101MHz, Acetone) δ166.5,160.4,151.3,143.4,138.8,136.6,130.7,130.2,129.4,123.8,121.0,118.3,117 .8,117.1,116.8,111.2,111.2,105.7,77.5,72.0,56.3,48.1,35.2,32.3,28.0,25.0.HRESIMSm / z=683.25[M+H] +

[0058] Preparation of (FN-5) in Example 5

[0059] J-5 (260 mg, 2 eq) was dissolved in 5 mL of tetrahydrofuran, and CZA-1 (150 mg, 1 eq), EDCI (154 mg, 2 eq), and HOBt (108 mg, 2 eq) were added sequentially. The mixture was stirred overnight, and the reaction was quenched dropwise with water. The mixture was extracted with ethyl acetate-water, and the ethyl acetate layer was developed using a scraper (petroleum ether:acetone = 2:1). 183 mg of a yellow powder was obtained, with a yield of 67.0%. 1 H NMR (500MHz, Acetone-d6) δ8.40(s,1H),8.15–8.04(m,4H),7.90(t,J=7.6Hz,1H),7.77(t,J=7.8Hz,2H),7.43(dd,J=8.5,1.9Hz,1H),7.36(dd,J =12.0,3.5Hz,2H),6.94(d,J=9.7Hz,1H),5.86(d,J=9.8Hz,1H),5.28(dt ,J=7.0,3.5Hz,1H),2.79(s,2H),2.09(s,2H),1.54(s,6H),1.42(s,9H). 13C NMR (101MHz, Acetone) δ166.5,159.2,151.0,143.3,139.7,139.5,139.2,139.1,139.0,136.7,130.8,130.3,129.5,12 4.3,123.8,120.3,119.0,118.9,118.3,117.4,116.7,115.7,111.6,105.7,105.6,78.5,77.5,44.2,35.2,32.3,27.9.

[0060] HRESIMS m / z = 683.25 [M+H] +

[0061] Preparation of (FN-6) in Example 6

[0062] J-6 (110 mg, 2 eq) was dissolved in 3 mL of tetrahydrofuran, and CZA-1 (69 mg, 1 eq), EDCI (71 mg, 2 eq), and HOBt (50 mg, 2 eq) were added sequentially. The mixture was stirred overnight, and the reaction was quenched dropwise with water. The mixture was extracted with ethyl acetate-water, and the ethyl acetate layer was developed using a scraper (petroleum ether:acetone = 2:1). 83 mg of a yellow powder was obtained, with a yield of 68.6%. 1 H NMR (500MHz, Acetone-d6) δ8.25(s,1H),8.18–8.12(m,2H),8.09(dt,J=6.8,3.7Hz,2H),7.88–7.80(m,1H),7.76(q,J=7.7Hz,2H),7.45(dd,J=8. 5,2.0Hz,1H),7.38(d,J=8.6Hz,1H),6.95(d,J=9.8Hz,1H),6.87(dd,J=1 5.7, 4.0Hz, 1H), 5.87 (d, J = 9.8Hz, 1H), 1.59–1.48 (m, 8H), 1.44 (s, 12H). 13 CNMR(101MHz,Acetone)δ166.8,160.3,151.1,143.3,139.6,139.5,139.1,139.1,136.6,130.7,130.7, 130.7,130.7,130.4,130.4,129.6,129.6,129.5,129.5,123.8,123.8,119.8,118.2,32.3,27.9,27.8.

[0063] HRESIMS m / z = 657.24 [M+H] +

[0064] Preparation of (FN-7) in Example 7

[0065] J-7 (133 mg, 2 eq) was dissolved in 3 mL of tetrahydrofuran, and CZA-1 (71 mg, 1 eq), EDCI (72 mg, 2 eq), and HOBt (51 mg, 2 eq) were added sequentially. The mixture was stirred overnight, and the reaction was quenched dropwise with water. The mixture was extracted with ethyl acetate-water, and the ethyl acetate layer was developed using a scraper (petroleum ether:acetone = 2:1). 85 mg of a yellow powder was obtained, with a yield of 63.0%. 1 H NMR(500MHz, Acetone-d6)δ8.40(d,J=3.8Hz,1H),8.14–8.10(m,2H),8.09(dd,J=14.9,4.4Hz,2H),8.00–7.92( m,1H),7.90(s,1H),7.78(dt,J=12.0,5.7Hz,2H),7.72(dd,J=8.5,4.1Hz,1H),7.55(dt,J=11.9,5.8Hz,1H),7.4 4(s,2H),7.39–7.28(m,2H),6.97–6.91(m,1H),5.89–5.82(m,1H),4.65(q,J=4.6Hz,2H),3.74(s,5H),2.96(q, J=4.9Hz,2H),1.57–1.47(m,6H),1.46(s,1H),1.42(t,J=5.5Hz,10H),1.32–1.27(m,1H),1.20(d,J=3.7Hz,1H). 13 C NMR (101MHz, Acetone) δ166.4,160.3,151.0,143.3,139.5,139.3,139.0,139.0,136.7,130.8,130.3,129.4,128.0,125.2 ,124.2,123.8,120.2,120.1,118.9,118.3,117.3,116.7,115.7,111.2,110.3,105.6,77.4,70.2,56.8,35.2,32.3,27.9.

[0066] HRESIMS m / z = 712.28 [M+H] +

[0067] Examples 8-12 can be synthesized according to the following route:

[0068]

[0069] Example 8 Preparation (CY-1)

[0070] J-1 (25 mg, 2 eq) was dissolved in 2 mL of tetrahydrofuran, and CZB (15 mg, 1 eq) and AcOH (5 μL, 2 eq) were added sequentially. The mixture was stirred at room temperature for 1 h, and then NaBH(OAc)3 (19 mg, 2 eq) was added. The mixture was stirred overnight, and the reaction was quenched dropwise with water. The mixture was extracted with ethyl acetate-water, and the ethyl acetate layer was developed using a scraping agent (acetone:petroleum ether = 2:1). 19 mg of a yellow powder was obtained, with a yield of 71.9%. 1 H NMR (700MHz, Acetone-d6) δ8.10–8.01(m,5H),7.79–7.71(m,3H),7.64–7.61(m,2H),7.40(dd,J=8.4,1 .9Hz,1H),7.36(d,J=8.6Hz,1H),6.95(d,J=9.8Hz,1H),5.80(d,J=9.8Hz,1H),4.68(t,J=5.3Hz,2H),4 .16(s,2H),3.68(dt,J=16.2,5.6Hz,2H),3.47(t,J=5.5Hz,1H),3.31–3.25(m,2H),1.50(s,5H),1.47( d,J=7.6Hz,2H),1.40(d,J=1.7Hz,2·H),1.39(s,2H),1.38(s,6H),1.38(s,2H),1.37(d,J=2.1Hz,1H). 13 C NMR (176MHz, Acetone) δ206.4,206.3,160.2,150.3,142.8,139.2,136.9,136.0,134.0,130.8,130.6,130. 5,130.1,129.5,124.2,123.2,122.1,118.5,116.4,111.0,105.6,71.5,60.8,60.5,48.9,46.9,35.2,32.3.

[0071] HRESIMS m / z = 603.23 [M+H] +

[0072] Example 9 Preparation of (CY-2)

[0073] J-2 (100 mg, 2 eq) was dissolved in 3 mL of tetrahydrofuran, and CZB (56 mg, 1 eq) and AcOH (19 μL, 2 eq) were added sequentially. The mixture was stirred at room temperature for 1 h, and then NaBH(OAc)3 (70 mg, 2 eq) was added. The mixture was stirred overnight, and the reaction was quenched dropwise with water. The mixture was extracted with ethyl acetate-water, and the ethyl acetate layer was developed using a scraper (acetone:petroleum ether = 2:1). 75 mg of a yellow powder was obtained, with a yield of 72.8%. 1HNMR(500MHz, Acetone-d6)δ8.06(d,J=7.7Hz,4H),8.03(d,J=3.6Hz,3H),7.94(s,2H),7.64–7.5 6(m,3H),7.35(d,J=6.5Hz,2H),6.94(dd,J=9.8,2.5Hz,2H),5.80(dt,J=9.9,3.3Hz,2H),4.09(s ,1H),4.03(s,1H),1.57–1.45(m,14H),1.43(d,J=24.9Hz,9H),1.39(d,J=6.6Hz,11H),1.36(dd, J=6.6,3.1Hz,13H),1.20(s,8H),1.16(t,J=6.7Hz,3H),0.96(d,J=6.7Hz,1H),0.91–0.81(m,5H). 13 C NMR(151MHz,Acetone)δ138.7,136.0,134.1,130.4,129.8,129.7,129.5,128.6,127 .0,124.5,116.5,112.1,111.1,35.7,32.4,32.0,28.0,27.8,25.7,24.3,23.4,14.4.

[0074] HRESIMS m / z = 617.24 [M+H] +

[0075] Example 10 Preparation (CY-3)

[0076] J-3 (140 mg, 2 eq) was dissolved in 3 mL of tetrahydrofuran, and CZB (69 mg, 1 eq) and AcOH (24 μL, 2 eq) were added sequentially. The mixture was stirred at room temperature for 1 h, and then NaBH(OAc)3 (87 mg, 2 eq) was added. The mixture was stirred overnight, and the reaction was quenched dropwise with water. The mixture was extracted with ethyl acetate-water, and the ethyl acetate layer was developed using a scraper (acetone:petroleum ether = 2:1). 82 mg of a yellow powder was obtained, with a yield of 60.3%. 1 HNMR (500MHz, Acetone-d6) δ8.12–8.05(m,4H),7.84(t,J=7.5Hz,1H),7.74(t,J=7.7Hz,2H),7.44–7.33(m,2H),6.95(d,J =9.8Hz,1H),5.82(d,J=9.8Hz,1H),4.36(d,J=6.7Hz,2H),3.45(d,J=6.3Hz,1H),1.98(s,3H),1.51(s,6H),1.40(s,11H). 13C NMR (151MHz, Acetone) δ150.6,143.2,137.3,135.9,135.8,130.9,130.8,130.4,130.2,129.5,128.4,127.0,126.6,125.2,125.2,124 .1,117.8,116.5,112.4,110.9,107.8,78.4,77.7,66.4,60.9,60.1,56.1,55.6,55.3,37.7,36.4,35.3,32.7,32.0,28.5,23.4,14.4.

[0077] HRESIMS m / z = 657.27 [M+H] +

[0078] Example 11 Preparation (CY-5)

[0079] J-5 (260 mg, 2 eq) was dissolved in 5 mL of tetrahydrofuran, and CZB (133 mg, 1 eq) and AcOH (46 μL, 2 eq) were added sequentially. The mixture was stirred at room temperature for 1 h, and then NaBH(OAc)3 (169 mg, 2 eq) was added. The mixture was stirred overnight, and the reaction was quenched dropwise with water. The mixture was extracted with ethyl acetate-water, and the ethyl acetate layer was developed using a scraper (acetone:petroleum ether = 2:1). 152 mg of a yellow powder was obtained, with a yield of 59.1%. 1 HNMR (500MHz, Acetone-d6) δ8.06(d,J=7.0Hz,3H),8.01(s,1H),7.85(t,J=7.6Hz,1H),7.71(t,J=7.8Hz,2H),7.43–7.32(m,2H),6.9 4(d,J=9.8Hz,1H),5.80(d,J=9.8Hz,1H),5.02(s,1H),3.80(s,1H),2.14(s,2H),1.96(s,2H),1.50(s,6H),1.42(s,9H),1.20(s,1H). 13 C NMR(101MHz,Acetone)δ159.3,150.6,142.8,139.3,139.3,139.2,136.6,130.7,130.0,129.4,124.3,124.3 ,123.2,118.7,118.2,118.1,116.4,111.5,111.0,110.9,105.5,105.5,76.9,56.4,50.2,35.2,32.4,27.9.

[0080] HRESIMS m / z = 643.26 [M+H]+

[0081] Example 12 Preparation (CY-7)

[0082] J-7 (110 mg, 2 eq) was dissolved in 3 mL of tetrahydrofuran, and CZB (61 mg, 1 eq) and AcOH (21 μL, 2 eq) were added sequentially. The mixture was stirred at room temperature for 1 h, and then NaBH(OAc)3 (78 mg, 2 eq) was added. The mixture was stirred overnight, and the reaction was quenched dropwise with water. The mixture was extracted with ethyl acetate-water, and the ethyl acetate layer was developed using a scraper (acetone:petroleum ether = 2:1). 68 mg of a yellow powder was obtained, with a yield of 60.2%. 1 HNMR (500MHz, Acetone-d6) δ8.08(s,3H),8.03(d,J=7.5Hz,2H),7.71(t,J=7.5Hz,1H),7.64(t,J=7.7Hz,2H),7.41(d,J=10.0Hz,1H) ,7.36(d,J=8.6Hz,1H),6.94(d,J=9.8Hz,1H),5.81(d,J=9.8Hz,1H),4.57(t,J=5.2Hz,2H),1.97(s,3H),1.50(s,7H),1.41(s,11H). 13 CNMR(101MHz,Acetone)δ160.2,150.8,143.0,139.3,139.2,136.6,130.7,130.1,129.3, 124.2,123.4,118.4,116.6,111.0,105.5,77.3,70.2,56.6,56.1,53.0,35.2,32.4,27.9.

[0083] HRESIMS m / z = 672.29 [M+H] +

[0084] Pharmacological experiments

[0085] Example 1: Cytotoxic Activity Test

[0086] 1. Experimental materials: Tetramethylazazole salt (MTT, produced by Serva, prepared on-site with fresh 1640 culture medium), H460 (human lung cancer cells), HeLa (cervical cancer cells), MDA-MB-231 (human breast cancer cells), BIORAD 550 microplate reader.

[0087] 2. Experimental Methods

[0088] (1) Cell culture: RRMI1640 medium containing 10% bovine fetal serum, 100 U / mL penicillin and 100 mg / L was used. Cells were passaged in a 37℃, 5% CO2 saturated humidity incubator. Logarithmic growth cycle cells were selected for the experiment.

[0089] (2) MTT assay: Logarithmic growth phase cells were taken, digested, and thoroughly pipetted into a single-cell suspension. After counting, the suspension was diluted to 1×10⁻⁶ cells. 4 Cells / mL were seeded into 96-well plates at 100 μL / well. Each sample had 4-5 concentration levels. Then, 100 μL of culture medium for each concentration level was added to each well, with three replicates per concentration level. An equal volume of solvent was added to the control group. The 96-well plates were incubated at 37°C with 5% CO2 for 96 hours. The culture medium was discarded, and freshly prepared serum-free medium containing 0.20 mg / mL MTT was added to each well. Incubation continued at 37°C for 4 hours, followed by centrifugation to remove the supernatant. 150 μL of DMSO was added to each well to dissolve the formazan precipitate, and the mixture was gently shaken for 5 minutes to ensure complete dissolution. The optical density at 570 nm was measured using a BIORAD 550 microplate reader. The tumor cell growth inhibition rate was calculated using the following formula. A metric curve was obtained by plotting the drug concentration against the tumor cell growth inhibition rate. The half-maximal inhibitory concentration (IC50) of the drug was then read from the curve. 50 )value.

[0090] Tumor cell growth inhibition rate (%) = (1 - value measured in experimental wells / value measured in control wells) × 100%

[0091] 3. Experimental results: as shown in Table 1.

[0092] Table 1 Tumor cytotoxic activity

[0093]

[0094] Example 2: In vivo antitumor efficacy of FN-1 against mouse breast cancer EMT6

[0095] 1. Experimental materials: test compound FN-1; paclitaxel (positive control); BALB / c mice, weighing 18-20g, SPF grade, female, provided by the Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences.

[0096] 2. Experimental methods:

[0097] (1) Procedure: After homogenizing the passaged EMT6 tumor cells with a homogenizer, wash them twice with sterile physiological saline, count the cells, and adjust the cell concentration to 1.26 × 10⁻⁶ cells with physiological saline. 70.2 ml of cell suspension was diluted and injected into the right axilla of mice. The day after injection, animals were randomly divided into groups of 6. After weighing, the test compound was administered once daily for 14 days. Animals were then sacrificed, and tumor tissue was harvested and weighed. The tumor inhibition rate was calculated to evaluate the antitumor activity.

[0098] (2) Grouping: blank control group, positive drug control group (Taxol, administered once every three days) 24mg / kg (intraperitoneal); FN-1 100mg / kg (gavage), FN-1 150mg / kg (gavage).

[0099] (3) Calculation method:

[0100] Relative tumor proliferation rate T / C (%): T / C% = T / C × 100%. (T: tumor weight in the treatment group; C: tumor weight in the negative control group).

[0101] Tumor proliferation inhibition rate (TGI) (%): TGI = (1 - T / C) × 100. (T: tumor weight in the treatment group; C: tumor weight in the negative control group).

[0102] 3. Experimental Results:

[0103] Fourteen days after administration, the tumors of the treated animals were weighed. The effect of the compound on mouse EMT6 is described in [reference needed]. Figure 1 And Table 2.

[0104] Table 2. Growth inhibitory effect of compound FN-1 on EMT6 subcutaneous allogeneic xenograft tumors

[0105]

[0106] 1. TTEST, * P<0.05, ** P<0.01

[0107] 2. NA: Not applicable

[0108] Example 3: Preliminary oral acute toxicity test of FN-1

[0109] 1. Experimental materials: test compound FN-1; mouse weighing scale; gavage injection (size 12); half male and half female ICR mice, weighing 18-20g, purchased from Beijing Huafukang Biotechnology Co., Ltd.

[0110] 2. Experimental Methods: Compound FN-1 was dissolved in CMC-Na to prepare suspensions at doses of 1 g / kg and 2 g / kg. After administration by gavage once to each group, the weight changes and survival of mice were observed over 14 days.

[0111] 3. Experimental Results: During the observation period, the mice's body weight did not change significantly, and no significant toxic reactions were observed. The survival rate reached 100%. This indicates that the compound has low toxicity, with a low LD50. 50 >2g / kg.

Claims

1. A compound having the structure of Formula I or a pharmaceutically acceptable salt thereof, R1 and R2 are independent of each other and are selected from H, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, CH2CH2CH2CH3, CH2CH(CH3)2, CH(CH3)CH2CH3, t-Bu, OCH3, OCH2CH3, OCH2CH2CH3, CF3, and OCF3, respectively. R3 is selected from H, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, CH2CH2CH2CH3, CH2CH(CH3)2, CH(CH3)CH2CH3; X is selected from substituted amino groups, where... Substituents are selected from straight-chain or branched C-chains. 1-6 Alkyl, C 3-6 Cycloalkyl groups or substituents together with nitrogen atoms form a five-, six-, or seven-membered saturated heterocycle containing 1-3 heteroatoms. The heterocycle has substituents selected from -OH, -F, -Cl, Br, -COOH, straight-chain or branched C atoms. 1-6 Alkyl, C 3-6 Cycloalkyl, straight-chain or branched C 1-6 Alkoxy, substituted or unsubstituted phenyl groups, wherein the substituents on the phenyl group are selected from -OH, -F, -Cl, Br, -COOH, straight-chain or branched C. 1-6 Alkyl groups, -CHF2, -CF3, -CN, -NO2, -OCF3, -ONO2, -NH2, or straight-chain or branched C atoms 1-6 Alkyl group.

2. Compounds having the structure of Formula II or pharmaceutically acceptable salts thereof, R1 and R2 are independent of each other and are selected from H, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, CH2CH2CH2CH3, CH2CH(CH3)2, CH(CH3)CH2CH3, t-Bu, OCH3, OCH2CH3, OCH2CH2CH3, CF3, and OCF3, respectively. R3 is selected from H, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, CH2CH2CH2CH3, CH2CH(CH3)2, CH(CH3)CH2CH3; X is selected from substituted amino groups, where... Substituents are selected from straight-chain or branched C-chains. 1-6 Alkyl, C 3-6 Cycloalkyl groups or substituents together with nitrogen atoms form a five-, six-, or seven-membered saturated heterocycle containing 1-3 heteroatoms. The heterocycle has substituents selected from -OH, -F, -Cl, Br, -COOH, straight-chain or branched C atoms. 1-6 Alkyl, C 3-6 Cycloalkyl, straight-chain or branched C 1-6 Alkoxy, substituted or unsubstituted phenyl groups, wherein the substituents on the phenyl group are selected from -OH, -F, -Cl, Br, -COOH, straight-chain or branched C. 1-6 Alkyl groups, -CHF2, -CF3, -CN, -NO2, -OCF3, -ONO2, -NH2, or straight-chain or branched C atoms 1-6 Alkyl group.

3. The pyranocarbazole alkaloid compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 or 2, wherein the compound is selected from the following:

4. A pharmaceutical composition comprising, as an active ingredient, a compound of any one of claims 1-3 or a pharmaceutically acceptable salt thereof, and a pharmacodynamically acceptable carrier or excipient.

5. The use of the compound according to any one of claims 1-3 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of tumor diseases.

Images