A dipeptide compound with anticancer activity and preparation method thereof

Abstract

The present invention discloses a dipeptide compound with anticancer activity and a preparation method thereof. The dipeptide compound is a compound shown in formula I. The compound has anticancer activity, and in particular shows obvious anticancer activity against lung cancer, colon cancer, cervical cancer and / or liver cancer.

Description

Technical Field

[0001] The invention belongs to the field of pharmaceutical chemistry, and specifically relates to a dipeptide compound with anticancer activity and a preparation method thereof. Background Art

[0002] Secondary metabolites produced by microorganisms are an important source for new drug discovery. In 2015, Mark Bronstrup, Dominic Hoepfner, and others reported the isolation and identification of nannocystin-like natural products from the fermentation broth of two different strains of a rare myxobacterium, Nannocyctis sp., ST201196 and MB1016 (angew.chem.int.ed.2015,54,10145-10148;

[0003] angew.chem.int.ed.2015,54,10149-10154), the main representative compound is nannocystin A, and the absolute stereo configuration of the molecule was determined by nuclear magnetic resonance (NMR) technology, molecular dynamics calculation, chemical degradation and X-ray crystallography. Nannocystin natural products have a 21-membered macrocyclic structure, and its skeleton is composed of two fragments of polyketide and tripeptide connected end to end. It is a hybrid compound catalyzed by polymer ketase-nonribosomal peptide synthetase (PKS-NRPS). Nannocystin natural products show extensive and excellent cancer cell proliferation inhibition activity (its IC50 value ranges from 0.5μm to 5nm), among which nannocystin A has good anticancer activity against MDA-MB231, MDA-A1, PBL, HCT116, P30 and HL60 cells, and the IC50 values ​​are all at a low nanomolar concentration level, especially for drug-resistant breast cancer cells mda-a1. The IC50 of this molecule for ordinary breast cancer cells mda-mb231 and drug-resistant breast cancer cells mda-a1ic50 is 12nm, while docetaxel has a poor inhibitory effect on mda-a1 (ic50=570nm). After conducting biological research on nannocystin A, Dr. Dominic Hoepfner and others believed that eukaryotic translation elongation factor Ια (eEFΙα) is its target. The eEFΙα target is different from the targets of other common anticancer drugs paclitaxel and cisplatin. However, due to the limited source of nannocystin natural products, it is difficult to obtain sufficient amounts of compounds for subsequent pharmacological and toxicological research by separation and extraction. In addition, the synthesis is difficult, the production cost is high, and it is not easy to obtain. Therefore, it is necessary to simplify the molecule and explore its further drugability. Summary of the invention

[0004] The purpose of the present invention is to provide a dipeptide compound with anti-cancer activity, which is a compound of formula I.

[0005] A compound represented by formula I or a pharmaceutically acceptable salt thereof of the present invention,

[0006]

[0007] In the formula,

[0008] R1 is H or NHMe or NH2 optionally substituted by C1-C4 alkyl,

[0009] R2 is H, C1-C6 straight or branched alkyl, or aryl,

[0010] Alternatively, R1 and R2 may form a carbocyclic group containing 5-6 ring members, of which at most two may be optionally replaced by heteroatoms independently selected from oxygen and nitrogen, preferably one may be optionally replaced by a nitrogen heteroatom. The carbocyclic group is preferably an aromatic ring group, a heteroaryl group, a cycloalkyl group, a heterocycloalkyl group, an arylcycloalkyl group or an arylheterocycloalkyl group, preferably, the aryl group is a phenyl group, the heteroaryl group is a pyrazole group or a pyridine group, the heterocycloalkyl group is a piperidinyl group or a pyrrolyl group, and the arylheterocycloalkyl group is a benzopiperidine group.

[0011] Preferably, in the compound of formula I of the present invention, the aromatic ring formed by R1 and R2 is phenyl or phenyl substituted by C1-C4 alkyl, and the heterocyclic ring surrounded by R1 and R2 is pyridine, piperidine, benzopiperidine or

[0012] Preferably, in the compound of formula I of the present invention, the C1-C4 alkyl group is methyl, ethyl, methylethyl, methylpropyl, isopropyl or isobutyl.

[0013] More preferably, the compound of formula I of the present invention is selected from the following compounds or pharmaceutically acceptable salts thereof:

[0014]

[0015]

[0016] The present invention also provides a method for preparing the compound represented by formula I, comprising the following steps:

[0017] 1) reacting a compound of formula II (i.e., compound 2.1 in the example) with a compound of formula III or its acyl chloride to obtain a compound of formula IV;

[0018] 2) preparing a compound of formula VI by reacting compound IV with a compound of formula V;

[0019] 3) reacting the compound of formula V with silane to obtain a compound of formula I;

[0020]

[0021] In the formula, R1 and R2 have the same definitions as in claim 1.

[0022] In the above reaction formula, the compound of formula III can specifically be the compound shown in 2-2, 3-1, 4-1, 5-1, 6-1, 7-1, 8-1, 9-1, 10-1 in Example 2-10 or its acyl chloride.

[0023] The acyl chloride is:

[0024]

[0025] The above method of the present invention, when R1 is NH2 or a substituted amino group, further comprises the steps of amino protection and deprotection, wherein the amino protection group is tert-butyloxycarbonyl (BOC) or benzyloxycarbonyl (CBz).

[0026] On the other hand, the present invention provides a pharmaceutical composition, comprising a compound represented by Formula I or a pharmaceutically acceptable salt thereof or a compound represented by Formula 2-9 or a pharmaceutically acceptable salt thereof and a pharmaceutical excipient. The composition is in the form of an oral solid preparation, such as a tablet, a capsule, a soft capsule, etc., or an injection, such as an injection solution, a powder injection, etc. The excipient is a commonly used pharmaceutical excipient in the art, usually a filler, a disintegrant, a binder, a lubricant, etc.

[0027] The compounds of the present invention can be formulated into a variety of oral and parenteral dosage forms for administration, including transdermal and rectal administration. It will be recognized by those skilled in the art that the following dosage forms may contain a compound of formula I or a pharmaceutically acceptable salt or solvate of the corresponding compound of formula I.

[0028] The present invention also includes pharmaceutical preparations comprising a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, diluent or excipient. The pharmaceutically acceptable carrier may be a solid or a liquid. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories and dispersible granules. A solid carrier may be one or more substances which may also act as a diluent, flavoring agent, binder, preservative, tablet disintegrant or encapsulating material.

[0029] In powders, the carrier is a finely divided solid such as talc or starch, which is mixed with the finely divided active ingredient. In tablets, the active ingredient is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.

[0030] The formulation of the present invention preferably contains from about 5% to about 80% or more of the active compound. Suitable carriers include magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low melting wax, cocoa butter, and the like. A preferred oral dosage form is a capsule, which includes a formulation of the active compound with an encapsulating material as a carrier providing a capsule, wherein the active component, with or without other carriers, is surrounded by a carrier, thereby being associated with it. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

[0031] Liquid form preparations include solutions, suspensions and emulsions, such as water or water / propylene glycol solutions. For parenteral injection, liquid preparations can be formulated as solutions in aqueous polyethylene glycol solution, isotonic saline, 5% aqueous dextrose, and the like. Aqueous solutions suitable for oral use can be prepared by dissolving the active ingredient in water and adding suitable colorants, flavoring agents, stabilizers, and thickeners as required. Aqueous suspensions suitable for oral use can be prepared by dispersing the finely divided active ingredient in water and mixing with a viscous material, such as a natural or synthetic gum, resin, methylcellulose, sodium carboxymethylcellulose, or other well-known suspending agents.

[0032] The pharmaceutical preparations of the present invention are preferably in unit dosage form. In such dosage forms, the preparation is subdivided into unit doses containing appropriate quantities of the active ingredient. The unit dosage form can be a packaged preparation containing discrete quantities of the preparation, such as small packages of tablets, capsules, and powders in vials or ampoules. Furthermore, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.

[0033] The therapeutically effective dose of the compound of formula I will vary from about 0.01 mg / kg to about 100 mg / kg body weight per day. A typical adult dose will be about 0.1 mg to about 1000 mg per day. Depending on the specific application and the effectiveness of the active ingredient, the amount of the active ingredient in the unit dose preparation may vary from about 1 mg to about 500 mg. If necessary, the composition may also contain other compatible therapeutic agents. A dose of about 1 to about 500 mg per day is given to a subject who needs to be treated with the compound of formula I, in a single or multiple dose within 24 hours. This type of treatment can be repeated at consecutive intervals for the necessary length of time.

[0034] In another aspect, the present invention also provides use of the compound represented by Formula I or a pharmaceutically acceptable salt thereof or the compound represented by Formula 2-9 or a pharmaceutically acceptable salt thereof in the manufacture of a drug for treating tumors.

[0035] Preferably, in the above-mentioned use of the present invention, the tumor is selected from one or more of lung cancer, colon cancer, cervical cancer and liver cancer.

[0036] The pharmaceutically acceptable salts of the compound represented by formula I of the present invention include hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, toluenesulfonate, citrate, maleate and fumarate.

[0037] The compounds of formula I of the present invention, specifically including the compounds of formula 2-9, have inhibitory activity against A549, HCT116, Hela and Sk-Hep-1 cells, and their IC 50 The value is no more than 10nm. It shows strong anticancer activity. DETAILED DESCRIPTION

[0038] The following examples are representative and are used to further understand the essence of the present invention, but are not intended to limit the scope of the present invention in any way.

[0039] Example 1: Synthesis of Compound 1

[0040]

[0041] Step 1: Synthesis of compound 1-2

[0042] At 0°C, add Boc2O (20.1 mL, 88 mmol) and Et3N (20.2 mL, 146 mmol) to a solution of D-serine methyl ester hydrochloride (11.34 g, 73 mmol) in THF (150 mL). Warm to room temperature and stir for 18 h. After concentration, add H2O (150 mL) and EtOAc (150 mL). The aqueous phase was extracted with EtOAc (100 mL × 4), and the organic phases were combined and washed with 1N HCl (100 mL), saturated NaHCO3 solution (100 mL), and saturated NaCl solution (100 mL), respectively. Add anhydrous Na2SO4 to dry, filter, and concentrate to obtain a colorless liquid 1-2 (15.03 g, 94%), R f = 0.8 (EtOAc; PMA). This compound was used directly in the second step.

[0043] Step 2: Synthesis of Compound 1-3

[0044] At -15°C, MeMgBr (3M in THF, 87mL, 260mmol) was slowly added dropwise to a solution of compound 1-2 (9.49g, 43.3mmol) in THF (150mL). After stirring for 12h under this condition, the reaction was quenched with saturated NH4Cl solution (80mL) and H2O (80mL). The aqueous phase was extracted with EtOAc (200mL×3), and the organic phases were combined, dried over anhydrous Na2SO4, filtered, concentrated, and column chromatographed (DCM / MeOH=20 / 1) to obtain a light yellow oily liquid 1-3 (6.26g, 66%). This compound was used directly in the third step.

[0045] Step 3: Synthesis of Compounds 1-4

[0046] At 0°C, DMP (18.18 g, 43 mmol) was added to a solution of compound 1-3 (6.26 g, 28.6 mmol) in DCM (150 mL). After warming to room temperature and stirring for 1 h, the reaction was quenched with saturated Na2SO3 solution (100 mL) and saturated NaHCO3 (100 mL) solution in an ice-water bath, and then warmed to room temperature and stirred until the solution was clear. The aqueous phase was extracted with EtOAc (100 mL×4), dried with anhydrous Na2SO4, filtered, concentrated, and column chromatographed (petroleum ether / EtOAc=2 / 1) to obtain a colorless liquid 1-4 (4.67 g, 75%). This compound was used directly in the fourth step.

[0047] Step 4: Synthesis of Compound 1-5

[0048] PhI(OAc)2(10.39g, 32.3mmol) and NaBr(4.42g, 43.0mmol) were added to a solution of compound 1-4(4.67g, 21.5mmol) in MeOH(150mL) at 0℃. After warming to room temperature and stirring for 12h, the reaction was quenched with H2O(100mL). The aqueous phase was extracted with DCM(150mL×3), and the organic phases were combined, dried over anhydrous Na2SO4, filtered, concentrated, and column chromatographed (petroleum ether / EtOAc=2 / 1) to obtain brown-yellow oily liquid 1-5(3.66g, 69%). This compound was used directly in the fifth step.

[0049] Step 5: Synthesis of Compound 1

[0050] At 0°C, TFA (1.0 mL, 11.0 mmol) was slowly added dropwise to a DCM (50 mL) solution of compound 1-5 (1.20 g, 5.5 mmol). After stirring for 1 h under this condition, DCM (50 mL) was added after concentration, and the pH value was adjusted to about 9 with dilute ammonia water. The aqueous phase was extracted with DCM (200 mL × 3), and the organic phases were combined, dried with anhydrous Na2SO4, filtered, and concentrated to obtain a light yellow oily liquid as compound 1 (722.7 mg, 90%). f =0.2 (EtOAc; ninhydrin); 1 H NMR (400MHz, CDCl3): δ3.75(s,3H),3.33(s,1H),1.30(s,3H),1.11(s,3H).

[0051] Example 2: Synthesis of Compound 2

[0052]

[0053] Step 1: Synthesis of compound 2-3

[0054] Weigh HBTU (6.68 g, 20.0 mmol) and DMAP (1221.1 mg, 10.0 mmol) and dissolve them in DCM (70 mL), add DIPEA (3.60 mL, 20.0 mmol), add compound 2-1 (2.2 g, 10.0 mmol) and 2-2 (5.00 g, 20.0 mmol) in DCM (30 mL) to the above solution at 0°C, warm to room temperature and stir for 10 h, filter with diatomaceous earth, concentrate, and column chromatography (petroleum ether / EtOAc=5 / 1) to obtain colorless oily liquid 2-3 (3.45 g, 85%). This compound is used directly in the second step.

[0055] Step 2: Synthesis of Compound 2-4

[0056] At 0°C, a solution of LiOH.H2O (428.1 mg, 10.2 mmol) in H2O (50 mL) was added dropwise to a solution of compound 2-3 (1.38 g, 3.4 mmol) in DME (50 mL), and then the mixture was heated to room temperature and stirred for 10 h. The pH value was adjusted to about 2 with 1N HCl solution, and then extracted with EtOAc (100 mL × 3), dried with anhydrous Na2SO4, filtered, concentrated, and column chromatographed (petroleumether / EtOAc = 1 / 1) to obtain a white solid 2-4 (1.11 g, 83%). This compound was used directly in the third step.

[0057] Step 3: Synthesis of Compound 2-5

[0058] EDCI (615.5 mg, 3.2 mmol) and HOAt (437.0 mg, 3.2 mmol) were weighed and dissolved in DCM (15 mL), and DIPEA (0.7 mL, 4.3 mmol) was added. Compound 1 (187.5 mg, 1.28 mmol) and compound 2-4 (419.0 mg, 1.07 mmol) in DCM (15 mL) were added to the above solution at 0°C, stirred in an ice-water bath for 1 h, warmed to room temperature and reacted for 10 h, concentrated, and column chromatographed (petroleum ether / EtOAc=1.5 / 1) to obtain a light yellow oily liquid 2-5 (390.7 mg, 70%). This compound was used directly in the fourth step.

[0059] Step 4: Synthesis of Compound 2

[0060] At 0°C, TESOTf (0.27 mL, 1.2 mmol) and 2,6-lutidine (0.28 mL, 2.4 mmol) were added to a DCM (5 mL) solution of compound 2-5 (104.5 mg, 0.2 mmol). After stirring for 4 h under this condition, the reaction was quenched with a saturated NaHCO3 solution (15 mL), and the aqueous phase was extracted with DCM (20 mL×4), dried over anhydrous Na2SO4, filtered, concentrated, and column chromatographed (petroleum ether / EtOAc=1 / 1) to obtain a colorless liquid compound 2 (75.8 mg, 77%). 1 H NMR (400MHz, CDCl3) δ7.54(d,J=7.8Hz,1H),7.26–7.17(m,5H),6.85(d,J=8.7Hz,1H),4.71(dd ,J=15.0,8.1Hz,1H),3.66(s,3H),3.22(dd,J=14.1,6.8Hz,1H),3.01(dd,J=14.1,8.3Hz,1H),2 .75(d,J=4.6Hz,1H),2.07(s,3H),1.73(ddt,J=9.2,6.9,4.5Hz,1H),1.56(s,2H),1.47–1.37( m,2H),1.30(s,3H),1.08(s,3H),0.94–0.88(m,12H),0.85(t,J=7.4Hz,3H),0.59–0.52(m,6H).

[0061] Example 3: Synthesis of Compound 3

[0062]

[0063] In this example, except that 3-1 was used to replace 2-2 in the first step of Example 2 in the first step, the remaining steps and methods were referred to Example 2, and compound 3 (19.0 mg, total yield of four steps 31%) was obtained through four steps of transformation. 1 H NMR (400MHz, CDCl3) δ7.55(d,J=7.7Hz,1H),7.26(s,5H),6.89(d,J=8.8Hz,1H),4.72(dd,J=15.0 ,8.1Hz,1H),4.38(d,J=8.8Hz,1H),3.66(s,3H),3.23(dd,J=14.1,6.7Hz,1H),3.01(dd,J=14.2, 8.5Hz,1H),2.69(d,J=4.5Hz,1H),2.06(s,3H),2.05–2.00(m,1H),1.31(s,3H),1.26(s,2H),1.1 0(s,3H),0.96–0.93(m,5H),0.91(d,J=7.9Hz,7H),0.85(d,J=6.9Hz,3H),0.55(q,J=7.9Hz,6H).

[0064] Example 4 Synthesis of Compound 4

[0065]

[0066] In this example, except that 4-1 was used in the first step to replace 2-2 in the first step of Example 2, the remaining steps and methods were the same as those of Example 2, and compound 4 (18.1 mg, total yield of four steps: 27%) was obtained through four steps of transformation. 1H NMR(400MHz,Chloroform-d)δ7.82(d,J=8.2Hz,1H),7.29–7.21(m,5H),6.69(d ,J=8.8Hz,1H),4.70(td,J=8.1,6.4Hz,1H),4.40(d,J=8.8Hz,1H),3.68(s,3H), 3.21(d,J=3.9Hz,1H),3.16–3.01(m,2H),2.23(pd,J=6.9,3.8Hz,1H),1.32(s, 3H), 1.21 (s, 3H), 0.94–0.89 (m, 12H), 0.60 (d, J = 6.9Hz, 3H), 0.58–0.52 (m, 6H).

[0067] Example 5 Synthesis of Compound 5

[0068]

[0069] In this example, except that 5-1 was used to replace 2-2 in the first step of Example 2 in the first step, the remaining steps and methods were referred to Example 2, and compound 5 (29.0 mg, total yield of four steps 14%) was obtained through four steps of transformation. 1 H NMR (400MHz, CDCl3) δ7.59(d,J=7.7Hz,1H),7.25(s,5H),6.88(d,J=8.8Hz,1H),4.77(d d,J=15.0,8.1Hz,1H),4.41(d,J=8.8Hz,1H),3.69(s,3H),3.33(dd,J=14.1,6.7Hz,1H), 3.11(dd,J=14.2,8.5Hz,1H),2.74(d,J=4.5Hz,1H),2.13(s,3H),2.09–2.12(m,1H),1. 36(s,3H),1.16(s,3H),0.99–0.95(m,6H),0.91(t,J=7.9Hz,9H),0.85(d,J=6.9Hz,3H).

[0070] Example 6 Synthesis of Compound 6

[0071]

[0072] In this example, except that 6-1 was used to replace 2-2 in the first step of Example 2 in the first step, the remaining steps and methods were referred to Example 2, and compound 6 (19.7 mg, total yield of four steps 13%) was obtained through four steps of transformation. 1 H NMR(400MHz,Chloroform-d)δ7.62(d,J=8.1Hz,1H),7.18(dd,J=12.8,1.9Hz,5H),6.5 6(d,J=8.8Hz,1H),4.61(dd,J=8.0,6.8Hz,1H),4.32(d,J=8.9Hz,1H),3.60(s,3H),3.1 4(d,J=20.3Hz,1H),3.08(t,J=7.1Hz,1H),2.99(dd,J=14.0,7.6Hz,1H),2.23(s,3H), 1.24(d,J=2.3Hz,3H),1.14(s,3H),0.85(t,J=8.0Hz,9H),0.47(dd,J=7.9,2.6Hz,6H).

[0073] Embodiment 7 Synthesis of Compound 7

[0074]

[0075] In this example, except that 7-1 was used to replace 2-2 in the first step of Example 2 in the first step, the remaining steps and methods were referred to Example 2, and compound 7 (31.9 mg, total yield of four steps 27%) was obtained through four steps of transformation. 1 H NMR(400MHz,Chloroform-d)δ7.32(d,J=8.0Hz,1H),7.27–7.19(m,5H),6.64(d,J=8.9Hz,1H),4. 67(qd,J=7.7,6.1Hz,1H),4.35(dd,J=8.9,4.3Hz,1H),3.66(d,J=4.8Hz,3H),3.22–3.10(m,2H),3 .06–2.95(m,1H),2.66–2.53(m,1H),1.92–1.77(m,2H),1.77–1.62(m,1H),1.38(ddt,J=10.1,6. 4,3.2Hz,2H),1.30(s,3H),1.19(s,3H),0.90(td,J=7.9,2.5Hz,9H),0.53(qd,J=7.9,1.7Hz,6H).

[0076] Example 8 Synthesis of Compound 8

[0077]

[0078] In this example, except that 8-1 was used to replace 2-2 in the first step of Example 2 in the first step, the remaining steps and methods were referred to Example 2, and compound 8 (18.9 mg, total yield of four steps 11%) was obtained through four steps of transformation. 1 H NMR(400MHz,Chloroform-d)δ7.82(d,J=8.0Hz,1H),7.28–7.11(m,9H),6.57(d,J=8. 8Hz,1H),4.67(q,J=7.3Hz,1H),4.36(dd,J=8.9,6.0Hz,1H),3.68(d,J=10.5Hz,3H),3 .53(ddd,J=10.6,5.4,1.7Hz,1H),3.29–2.93(m,4H),2.80(dd,J=16.4,10.5Hz,1H),2 .02(s,2H),1.27(s,3H),1.15(s,3H),0.89(dd,J=7.9,4.4Hz,9H),0.56–0.50(m,6H).

[0079] Example 9 Synthesis of Compound 9

[0080]

[0081] In this example, the specific operation of the first step is as follows: weigh compound 2-1 (3.38 g, 16.5 mmol) and DMAP (185.0 mg, 1.5 mmol) and add them to a reaction bottle, add DCM (100 mL), add NEt3 (6.3 mL, 45.0 mmol) after cooling in an ice-water bath, add 9-1 (2.74 g, 15.0 mmol) to the above solution at 0°C, warm to room temperature and stir for 10 hours, concentrate, column chromatography (petroleum ether / EtOAc=5 / 1) to obtain a light yellow oily liquid 9-2 (3.32 g, 67%). Except that in the first step, 9-1 is used to replace 2-2 in the first step of Example 2, the remaining steps and methods refer to Example 2, and compound 9 (8.3 mg, total yield of four steps 8%) is obtained through four steps of transformation. 1 H NMR(400MHz,Chloroform-d)δ7.67(d,J=8.0Hz,1H),7.28–7.11(m,7H),6.61(d,J=8.8Hz,1H),4.52(d,J=7.3Hz,1H),4.31(dd,J=8.9, 6.0Hz,1H),3.73(s,3H),2.35(s,9H),2.32(d,J=8.9Hz,2H),1.25(s,3H),1.17(s,3H),0.89(dd,J=7.9,4.4Hz,9H),0.56–0.50(m,6H).

[0082] Example 10 Synthesis of Compound 10

[0083]

[0084] In this example, the specific operation of the first step is as follows: weigh compound 2-1 (1.1 g, 5.0 mmol) and DMAP (49.0 mg, 0.4 mmol) and add them to a reaction bottle, add DCM (40 mL), add NEt3 (1.7 mL, 12.0 mmol) after cooling in an ice-water bath, add 10-1 (0.81 g, 4.0 mmol) to the above solution at 0°C, warm to room temperature and stir for 10 hours, concentrate, column chromatography (petroleum ether / EtOAc=5 / 1) to obtain a light yellow oily liquid 9-2 (682.9 mg, 50%). Except that 10-1 is used to replace 2-2 in the first step of Example 2 in the first step in Example 2, the remaining steps and methods refer to Example 2, and compound 10 (9.4 mg, total yield of four steps 9%) is obtained through four steps of transformation. 1H NMR(400MHz,Chloroform-d)δ7.59(d,J=8.0Hz,1H),7.27–7.10(m,5H),6.63(d,J=8.8Hz,1H),4.49(d,J=7.3Hz,1H),4.33(dd,J=8.9 ,6.0Hz,1H),3.71(s,3H),1.31(m,3H),1.27(s,3H),1.16(s,3H),1.02-0.97(m,10H),0.89(dd,J=7.9,4.4Hz,9H),0.56–0.50(m,6H).

[0085] Example 11 Activity test data:

[0086] A549, HCT116, Hela and Sk-Hep-1 cells were purchased from Wuhan Sepno Life Science Co., Ltd. The cells were cultured in an incubator for 24 hours according to the ATCC standard method, and then the above compounds (DMSO solution) were added, and then continued to be incubated in the incubator for 24 hours for detection using the CCK8 method. 1-Methoxy-5-methylphenolazine methyl sulfate (1-Methoxy PMS) was purchased from Beijing Bailingwei Technology Co., Ltd. The results of the biological activity test are shown in Tables 1-4 below:

[0087] Table 1 Experimental study on the inhibition of A549 cell proliferation by compounds

[0088] Compound <![CDATA[IC 50 / nM]]> 1 23 2 10.0 3 4.3 4 7.9 5 1.1 6 4.3 7 5.6 8 7.1 9 6.9 10 2.2

[0089] Table 2 Experimental study on the inhibition of HCT-116 cell proliferation by compounds

[0090] Compound <![CDATA[IC 50 / nM]]> 1 33.9 2 2.7 3 3.1 4 5.0 5 5.6 6 5.9 7 5.5 8 5.8 9 4.2 10 3.2

[0091] Table 3 Inhibition of Hela cell proliferation by compounds

[0092] Compound <![CDATA[IC 50 / nM]]> 1 21.9 2 2.7 3 3.0 4 5.1 5 2.5 6 6.1 7 5.5 8 6.9 9 6.7 10 5.5

[0093] Table 4 Experiment on the inhibition of Sk-Hep-1 cell proliferation by compounds

[0094] Compound <![CDATA[IC 50 / nM]]> 1 44.9 2 3.9 3 2.6 4 6.6 5 3.2 6 4.8 7 6.9 8 7.2 9 5.7 10 6.2

[0095] The results in Tables 1-4 above show that the compounds of Formula 2-10 of the present invention inhibit the activity of A549, HCT116, Hela and Sk-Hep-1 cancer cells IC 50 Below 10 nM, it exhibited strong anticancer activity.

Claims

1. A compound of formula I, In the formula, R1 is -NHMe or NH2, R2 is H, methyl, ethyl, methylpropyl or isopropyl.

2. The compound of formula I according to claim 1, selected from the following compounds:

3. A compound selected from the following compounds:

4. A pharmaceutical composition comprising any one of the compounds of claims 1 to 3 or a pharmaceutically acceptable salt thereof and a pharmaceutical excipient.

5. Use of the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof in the manufacture of a drug for treating tumors, wherein the tumor is selected from one or more of lung cancer, colon cancer, cervical cancer and liver cancer.

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