CN114507256A

CN114507256A - Chiral isomer of Rudexiluwei process, preparation method and application thereof

Info

Publication number: CN114507256A
Application number: CN202011278875.XA
Authority: CN
Inventors: 方治坤; 朱阳; 周伟; 姚长亮; 翟雄; 张�荣; 李玉成; 邓宇; 毕光庆; 夏广兵; 江云兵; 夏广新; 柯樱
Original assignee: Shanghai Pharmaceuticals Holding Co Ltd
Current assignee: Shanghai Pharmaceuticals Holding Co Ltd
Priority date: 2020-11-16
Filing date: 2020-11-16
Publication date: 2022-05-17

Abstract

The invention discloses a chiral isomer of a Rudexiwei process, a preparation method and application thereof. The invention provides a compound shown as a formula I and a compound shown as a formula II. In addition, the preparation method of the compound shown as the formula I and the compound shown as the formula II has the advantages of simple reaction, mild condition, high yield, no high-low temperature extreme reaction condition, no use of high-risk virulent substances, simple operation, high chiral purity, no special requirement, stable process, suitability for laboratory synthesis and industrial production, and wide industrial application prospect.

Description

Rudexiwei process chiral isomer, preparation method and application thereof

Technical Field

The invention relates to the field of drug synthesis, and in particular relates to a chiral isomer of a Rudexiluwei process, a preparation method and application thereof.

Background

Reidesciclovir was developed by Reinecke, USA, for the treatment of Ebola virus, and the latter novel coronavirus outbreak was used as a clinical drug for new coronavirus, which had been approved by FDA in USA for marketing in 2020 for the treatment of new coronavirus. Nature,2016, Doi:10.1038/Nature17180(Nature,2016,531(7594):381-385.) provides a process for the preparation of Reidcisvir, a second generation synthetic method of Gilidard Remdesivir. The synthetic route is as follows:

preparation methods and crystal form patents of Reidevir are reported in Jilidd Chinese patents CN110636884A and CN 103052631A. In order to study the preparation process of the Reidesciclovir, the chiral impurities generated by the process need to be studied.

Journal of Medicinal Chemistry 2017,60,5,1648-1661 and US20160122374 report a synthesis method of a chiral isomer RDV-ISO-2 of Reidesciclovir, in which an intermediate C-3' debenzylate is reacted with phosphoryl chloride F to obtain a product G (containing R, S configuration, S type is Reidesciclovir, R type is RDV-ISO-2), and the product S type is Reidesciclovir and R type is RDV-ISO-2 by chiral preparative HPLC, the route is as follows:

the preparation method of the chiral isomer RDV-ISO-2 of the Radexilawir adopts the expensive chiral HPLC separation, and the final yield of the reaction is only about 20 percent and still needs to be further improved. No synthesis of the remaining important chiral isomers has been reported in the literature, nor the use of chiral isomers of ridciclovir.

Disclosure of Invention

The invention provides a chiral isomer of a Reidesvir process, a preparation method and application thereof, in particular a compound shown as a formula I and a compound shown as a formula II, which are different from the prior art, and unexpectedly discovers that the compound shown as the formula I and the compound shown as the formula II have the effects of inhibiting Hela cells, COLO 205 cells and U-87MG cells, and the invention also discovers that the compound shown as the formula III also has the effects of inhibiting Hela cells, COLO 205 cells and U-87MG cells for the first time. In addition, the preparation method of the compound shown as the formula I and the compound shown as the formula II has the advantages of simple reaction, mild condition, high yield, no high-low temperature extreme reaction condition, no use of high-risk virulent substances, simple operation, high chiral purity, no special requirement, stable process, suitability for laboratory synthesis and industrial production, and wide industrial application prospect.

The invention provides a compound shown as a formula I or a formula II shown as follows, or a pharmaceutically acceptable salt thereof:

the invention also provides a preparation method of the compound shown in the formula I, which comprises the following steps: in a solvent, in the presence of acid, carrying out deprotection reaction shown as the following on a compound E to obtain a compound shown as a formula I;

the conditions and procedures for the deprotection reaction may be those conventional in the art, and the following conditions are particularly preferred in the present invention:

in the deprotection reaction, the solvent can be a solvent which is conventional in the art, and preferably, the solvent is one or more of an ether solvent, a nitrile solvent and a ketone solvent; more preferably, the solvent is one or more of tetrahydrofuran, acetonitrile, 1, 4-dioxane and acetone.

In the deprotection reaction, the acid for the deprotection reaction can be an acid conventionally used in the deprotection reactions of this type in the art, and preferably, the acid is one or more of hydrochloric acid, sulfuric acid and phosphoric acid.

In the deprotection reaction, the reaction temperature can be the temperature conventional in the deprotection reactions in the field, and preferably, the reaction temperature is 0-30 ℃; more preferably, the reaction temperature is 0-5 ℃ or 20-25 ℃.

In the deprotection reaction, the compound E disappears as a reaction end point, preferably, the reaction time is 2 to 5 hours, and more preferably, the reaction time is 3 hours.

The preparation method of the compound shown in the formula I can further comprise the following steps: in a solvent, in the presence of a catalyst and alkali, carrying out substitution reaction on a compound C and a compound D as shown in the specification to obtain a compound E;

the conditions and procedures of the substitution reaction may be those conventional in the art, and the following conditions are particularly preferred in the present invention:

in the substitution reaction, the solvent can be a solvent conventional in the substitution reaction in the field, preferably an anhydrous solvent, preferably an ether solvent and/or a nitrile solvent, and more preferably, the solvent is one or more of acetonitrile, 1, 4-dioxane and tetrahydrofuran.

In the substitution reaction, the catalyst can be a catalyst conventional in the substitution reaction of the type in the field, preferably, the catalyst is magnesium chloride, and more preferably, the catalyst is anhydrous magnesium chloride.

In the substitution reaction, the base can be a base conventionally used in the substitution reaction of this type in the art, and preferably, the base is N, N-diisopropylethylamine.

In the substitution reaction, the molar ratio of the compound C to the compound D may be a conventional ratio of the substitution reaction in the field, and is preferably 1: 1-1: 1.5, more preferably 1:1.1 or 1: 1.2.

In the substitution reaction, the reaction temperature may be a reaction temperature conventional in the substitution reaction of this kind in the art, preferably, the reaction temperature is 10 ℃ to 50 ℃, more preferably, the reaction temperature is 35 ℃ to 45 ℃.

In the substitution reaction, the compound C disappears as a reaction end point, preferably, the reaction time is 1 hour to 5 hours, and more preferably, the reaction time is 3 hours.

The preparation method of the compound shown in the formula I can further comprise the following steps: in a solvent, in the presence of acid, carrying out condensation reaction on a compound C-3 and 2, 2-dimethoxypropane as shown in the specification to obtain a compound C;

the conditions and steps of the condensation reaction may be those conventional in the art, and the following conditions are particularly preferred in the present invention:

in the condensation reaction, the solvent can be a solvent conventional in the condensation reaction of the type in the field, preferably, the solvent is a ketone solvent, and more preferably, the solvent is acetone.

In the condensation reaction, the acid may be an acid conventionally used in such condensation reactions in the art, and preferably, the acid is sulfuric acid.

In the condensation reaction, the reaction temperature may be a temperature that is conventional in the field of such condensation reactions, preferably, the reaction temperature is 35 to 60 ℃, more preferably, the reaction temperature is 40 to 50 ℃, and most preferably, the reaction temperature is 45 ℃.

In the condensation reaction, the condensation reaction takes disappearance of the compound C-3 as a reaction end point, and preferably, the reaction time is 1 to 2 hours.

The preparation method of the compound shown in the formula I can further comprise the following steps: in a solvent, in the presence of a debenzylation reagent, performing debenzylation reaction on the compound C-2 as shown in the specification to obtain a compound C-3;

the conditions and steps of the debenzylation reaction may be those conventional in the art, and the following conditions are particularly preferred in the present invention:

in the debenzylation, the solvent may be a solvent conventionally used in such debenzylation in the art, preferably, the solvent is one or more of a haloalkane-type solvent, a nitrile-type solvent and an ether-type solvent, more preferably, the solvent is one or more of dichloromethane, acetonitrile, 1, 4-dioxane and tetrahydrofuran, and most preferably, the solvent is dichloromethane.

In the debenzylation, the debenzylation reagent can be a reagent conventionally used in the debenzylation in the field, and preferably, the debenzylation reagent is BCl₃And/or BBr₃More preferably, the debenzylation reagent is BCl₃。

In the debenzylation reaction, the molar ratio of the compound C-2 to the debenzylation reagent can be a conventional ratio of the debenzylation reaction in the field, and is preferably 1: 4-1: 5.

In the debenzylation reaction, the reaction temperature can be the reaction temperature which is conventional in the debenzylation reaction of the type in the field, preferably, the reaction temperature is-70 ℃ to-40 ℃, more preferably, the reaction temperature is-65 ℃,60 ℃ or-50 ℃.

In the debenzylation, the debenzylation takes the compound C-2 hours as a reaction end point, and preferably, the reaction time is 2 to 5 hours.

After the debenzylation reaction is finished, the method preferably comprises post-treatment, and the post-treatment preferably comprises the following steps: adding methanol and NaHCO₃And water, adjusting the pH value to 8, and separating by preparative HPLC to obtain the compound C-3.

The preparation method of the compound shown in the formula I can further comprise the following steps: adding TMSOTf and TMSCN into a solvent at 0 ℃, and carrying out cyanation reaction on the compound C-1 at 15-20 ℃ to obtain a compound C-2;

in the cyanation reaction, the solvent may be a solvent conventional in the cyanation reaction in the art, preferably, the solvent is one or more of a haloalkane solvent, a nitrile solvent and an ether solvent, more preferably, the solvent is one or more of dichloromethane, acetonitrile, 1, 4-dioxane and tetrahydrofuran, and most preferably, the solvent is dichloromethane.

In the cyanation reaction, the cyanation reaction may not contain trifluoromethanesulfonic acid.

In the cyanation reaction, the cyanation reaction may comprise only the solvent, the TMSOTf, the TMSCN, and the compound C-1.

In the cyanation reaction, the molar ratio of the compound C-1 to the TMSCN can be a conventional ratio in the cyanation reaction in the field, and is preferably 1: 4-1: 5.

In the cyanation reaction, the molar ratio of the compound C-1 to the TMSOTf can be a conventional ratio of the cyanation reaction in the field, and is preferably 1: 4-1: 4.2.

In one embodiment, the cyanation reaction preferably comprises the steps of: adding TMSCN in the presence of a compound C-1 and a solvent, and then adding TMSOTf to carry out the cyanation reaction to obtain the compound C-2.

In one embodiment, the cyanation reaction preferably comprises the steps of: and in the presence of a compound C-1 and a solvent, adding TMSCN with the amount of 4 equivalents relative to the compound C-1, then adding TMSOTf with the amount of 2-2.1 equivalents relative to the compound C-1, and then supplementing TMSOTf with the amount of 2-2.1 equivalents relative to the compound C-1 to carry out the cyanation reaction to obtain the compound C-2.

In the cyanation reaction, the cyanation reaction is performed with compound C-1 hour as a reaction end point, preferably, the reaction time is 2 hours to 5 hours, and more preferably, the reaction time is 2.5 hours or 4.5 hours.

The preparation method of the compound shown in the formula I can further comprise the following steps: in a solvent, adding a compound A in the presence of B, TMSCl and PhMgCl to carry out a format reaction as described below to obtain a compound C-1;

the conditions and procedures of the format reaction described may be those conventional in the art, and the following conditions are particularly preferred in the present invention:

in the grignard reaction, the solvent may be a solvent conventionally used in grignard reactions in the art, preferably, the solvent is an ether solvent, and more preferably, the solvent is tetrahydrofuran.

In the Grignard reaction, the reaction temperature may be a temperature conventional in the Grignard reaction in the art, and preferably, the reaction temperature is-40 ℃ to 0 ℃, preferably-5 ℃,5 ℃ or-20 ℃.

In the grignard reaction, the raw material of the grignard reaction also preferably comprises iPrMgCl; the molar ratio of the iPrMgCl to the compound B is preferably 1:1.

In the Grignard reaction, the Grignard reaction takes disappearance of the compound A as a reaction end point, preferably, the reaction time is 0.5 to 2 hours, and more preferably, the reaction time is 1 hour.

The invention also provides a preparation method of the compound shown as the formula II, which comprises the following steps: in a solvent, in the presence of an acid, the compound E' is subjected to deprotection reaction as described below, and then the compound is shown as a formula II;

the conditions and procedures for the deprotection reaction may be as described above.

The preparation method of the compound shown in the formula II can further comprise the following steps: in a solvent, in the presence of a catalyst and alkali, carrying out substitution reaction on a compound C and a compound D 'as shown in the specification to obtain a compound E';

the conditions and procedures for the substitution reaction may be as described above;

the preparation method of the compound C can be as described above.

The invention also provides a preparation method of the compound C-2, which comprises the following steps: in a solvent, in the presence of TMSOTf and TMSCN, carrying out a cyanation reaction on the compound C-1 as shown in the specification to obtain a compound C-2;

the cyanation reaction conditions and operation may be as described above.

The present invention also provides a compound C:

the present invention also provides a compound E:

the invention also provides a compound E':

the invention also provides a pharmaceutical composition, which comprises one or more of the substance X, the substance Y and the substance Z, and pharmaceutic adjuvants; the substance X is the compound shown in the formula I or the pharmaceutically acceptable salt thereof, the substance Y is the compound shown in the formula II or the pharmaceutically acceptable salt thereof, and the substance Z is the compound shown in the formula III or the pharmaceutically acceptable salt thereof. The amount of substance X, substance Y or substance Z may be a therapeutically effective amount;

the invention also provides an application of the substance X or the pharmaceutical composition in preparing a cell proliferation inhibitor; the cell is one or more of Hela cell, COLO 205 cell and U-87MG cell.

The invention also provides an application of the substance X or the pharmaceutical composition in preparing a medicament for treating and/or preventing cancers, wherein the cancers are one or more of cervical cancer, colon cancer and glioma.

The invention also provides an application of the substance Y or the pharmaceutical composition in preparing a cell proliferation inhibitor; the cell is one or more of Hela cell, COLO 205 cell and U-87MG cell.

The invention also provides an application of the substance Y or the pharmaceutical composition in preparing a medicament for treating and/or preventing cancers, wherein the cancers are one or more of cervical cancer, colon cancer and glioma.

The invention also provides an application of the substance Z or the pharmaceutical composition in preparing a cell proliferation inhibitor; the cell is one or more of Hela cell, COLO 205 cell and U-87MG cell.

The invention also provides an application of the substance Z or the pharmaceutical composition in preparing a medicament for treating and/or preventing cancers, wherein the cancers are one or more of cervical cancer, colon cancer and glioma.

In the above-mentioned applications, the cell proliferation inhibitor can be used in a mammalian organism; also useful in vitro, primarily for experimental purposes, for example: the kit can be used as a standard sample or a control sample for comparison, or can be prepared into a kit according to the conventional method in the field, so as to provide rapid detection for the cell inhibition effect. Unless otherwise indicated, the following terms appearing in the present invention have the following meanings:

the medicinal auxiliary materials can be auxiliary materials widely adopted in the field of medicine production. The excipients are used primarily to provide a safe, stable and functional pharmaceutical composition and may also provide methods for dissolving the active ingredient at a desired rate or for promoting the effective absorption of the active ingredient after administration of the composition by a subject. The pharmaceutical excipients may be inert fillers or provide a function such as stabilizing the overall pH of the composition or preventing degradation of the active ingredients of the composition. The pharmaceutical excipients may include one or more of the following excipients: buffers, chelating agents, preservatives, co-solvents, stabilizers, excipients and surfactant colorants, flavors and sweeteners.

The term "pharmaceutically acceptable salt" refers to salts prepared from the compounds of the present invention with relatively nontoxic, pharmaceutically acceptable acids or bases. When compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral forms of such compounds with a sufficient amount of a pharmaceutically acceptable base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to: lithium salt, sodium salt, potassium salt, calcium salt, aluminum salt, magnesium salt, zinc salt, bismuth salt, ammonium salt, and diethanolamine salt. When compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of a pharmaceutically acceptable acid in neat solution or in a suitable inert solvent. The pharmaceutically acceptable acids include inorganic acids including, but not limited to: hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, phosphoric acid, phosphorous acid, sulfuric acid, and the like. The pharmaceutically acceptable acids include organic acids including, but not limited to: acetic acid, propionic acid, oxalic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, salicylic acid, tartaric acid, methanesulfonic acid, isonicotinic acid, acid citric acid, oleic acid, tannic acid, pantothenic acid, hydrogen tartrate, ascorbic acid, gentisic acid, fumaric acid, gluconic acid, saccharic acid, formic acid, ethanesulfonic acid, pamoic acid (i.e. 4, 4' -methylene-bis (3-hydroxy-2-naphthoic acid)), amino acids (e.g. glutamic acid, arginine), and the like. When the compounds of the present invention contain relatively acidic and relatively basic functional groups, they may be converted to base addition salts or acid addition salts. See in particular Berge et al, "Pharmaceutical Salts", Journal of Pharmaceutical Science 66:1-19(1977), or, Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P.Heinrich Stahl and Camile G.Wermuth, ed., Wiley-VCH, 2002).

The term "treatment" or its equivalent when used with reference to, for example, cancer, refers to a procedure or process for reducing or eliminating the number of cancer cells in a patient or alleviating the symptoms of cancer. "treating" cancer or another proliferative disorder does not necessarily mean that the cancer cells or other disorder will actually be eliminated, that the number of cells or disorders will actually be reduced or that the symptoms of the cancer or other disorder will actually be alleviated. Generally, methods for treating cancer are performed even with a low likelihood of success, but are still considered to induce an overall beneficial course of action, given the patient's medical history and estimated survival expectations.

The term "prevention" refers to a reduced risk of acquiring or developing a disease or disorder.

The above preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention without departing from the common general knowledge in the art.

The reagents and starting materials used in the present invention are commercially available.

The positive progress effects of the invention are as follows: the compound shown in the formula I, the compound shown in the formula II and the compound shown in the formula III can be used as a standard substance or a reference substance for controlling the quality of a Reidesciclovir medicament, and have the effects of inhibiting Hela cells, COLO 205 cells and U-87MG cells. In addition, the preparation method disclosed by the invention is simple in reaction, mild in condition, high in yield, free of high-temperature and low-temperature extreme reaction conditions, free of high-risk and highly toxic substances, simple to operate, high in chiral purity, free of special requirements, stable in process, suitable for laboratory synthesis and industrial production, and wide in industrial application prospect.

Detailed Description

The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The present invention will be described in further detail with reference to examples. These specific examples are only illustrative of the embodiments chosen for the purpose of this application and are intended to be included within the scope of the present application in combination with the prior art improvements.

The implementation conditions used in the examples can be further adjusted according to specific requirements, and the implementation conditions not indicated are generally the conditions in routine experiments. The chemical reagents used in the examples are all commercially available chemical reagents.

EXAMPLE 1 preparation of a hydroxy compound of formula C-1

To a 5L reaction flask was added compound B iodide (50g,192.3mmol,1.0eq), tetrahydrofuran (1.25L) was added and stirred to dissolve, argon was replaced three times, TMSCl (48.8mL,384.6mmol,2.0eq) was added dropwise, stirring was carried out at room temperature for 40 minutes, cooling was carried out to 0 ℃, PhMgCl (192.5mL,384.6mmol,2.0eq, 2M in THF) was added dropwise, and stirring was continued for 1 hour after the addition was completed. Slowly adding iPrMgCl & LiCl (200mL,192.3mmol, 1eq) dropwise under the condition of controlling the internal temperature to be 0-5 ℃, stirring for about 30 minutes after the addition is finished, cooling to-20 ℃, adding a compound A (88g,211.53mmol,1.1eq, in 250mL of THF) dropwise, continuing stirring for reaction for 1 hour, sampling TLC (detection by TLC (a developing agent is isopropyl acetate), controlling the temperature to be about 0 ℃ after the reaction is finished, slowly adding methanol (200mL), quenching the reaction by acetic acid (200mL) and water (200mL), concentrating under reduced pressure, adding ethyl acetate (2L) and 1N HCl (2L), and respectively using saturated NaHCO to obtain an organic phase₃(1 L.times.2) and saturated brine (1 L.times.2) were washed twice, dried over anhydrous sodium sulfate, suction-filtered, and the filtrate was concentrated under reduced pressure to give a pale oily liquid, which was passed through a silica gel column to give C-1 hydroxy compound (50.6g, yield: 47.7%) as a white solid.

Compound C-1:

¹H NMR(400MHz,DMSO-d₆)δ8.049(brs,1H),7.985(s,1H),7.362-7.226(m,11H),7.157-7.006(m,3H),7.157-7.126(m,2H),6.941(d,J＝4.8Hz,1H),5.385(d,J＝5.6Hz,1H),5.033(d,J＝5.2Hz,1H),4.595-4.554(m,2H),4.495-4.447(m,4H),4.0142-4.001(m,1H),3.927(dd,J＝5.9,4.4Hz,1H),3.688(dd,J＝10.1,3.4Hz,1H),3.472(dd,J＝10.0,6.4Hz,1H)；¹³C-NMR(100MHz,DMSO-d₆)δ187.893,155.786,148.900,138.605,138.105,128.570,128.194,128.172,128.100,128.090,127.776,127.706,127.668,127.553,127.492,127.424,127.397,127.339,127.222,127.147,127.043,126.963,118.499,117.425,102.162,81.870,80.790,72.421,72.243,71.643,71.340,69.341；MS(ESI):m/z 553.1[M+H]⁺.

EXAMPLE 2 preparation of Compound C-2 nitrile

Adding a compound C-1 hydroxyl substance (40g,0.0724mmol,1.0eq) into a 5L reaction bottle, adding dichloromethane (1.2L), stirring for dissolving, replacing argon for three times, cooling to 0 ℃, adding TMSCN (28.74g,0.2897mol,4.0eq), stirring for 30 minutes, dropwise adding TMSOTf (33.8g,0.1521mol,2.1eq), reacting at 0 ℃ for 2 hours, supplementing TMSOTf (33.8g,0.1521mol,2.1eq) for 1 hour, heating to 15 ℃ for reacting for 2 hours, cooling to 0 ℃, adding 2.4L for diluting, pouring the reaction solution into 3.6L saturated NaHCO₃The reaction was quenched with solution and the aqueous phase was extracted once more with 2L of dichloromethane. The organic phases were combined, washed once with saturated brine, concentrated to give a crude product, and crystallized from isopropyl alcohol and n-heptane to give C-2 nitrile as a white solid (35.1g, yield: 86.3%).

Compound C-2:

¹H NMR(400MHz,DMSO-d₆)δ7.931-7.881(m,3H),7.367-7.252(m,15H),6.947(d,J＝4.5Hz,1H),6.874(d,J＝4.5Hz,1H),6.758(dd,J＝4.5,5.9Hz,1H)，6.660(dd,J＝4.5,5.8Hz,1H),4.921-4.710(m,2H),4.646-4.503(m,4H),4.472-4.386(m,1H),4.140-4.028(m,1H),3.808-3.580(m,2H)；¹³C-NMR(100MHz,DMSO-d6):δ155.530,155.453,148.029,147.832,138.167,138.079,137.938,137.689,137.364,137.311,128.250,128.159,128.127,128.095,127.911,127.843,127.732,127.703,127.504,127.390,126.963,122.632,121.959,116.758,116.720,115.608,110.480,100.780,81.884,80.408,80.369,79.236,78.288,77.611,76.940,76.260,73.108,72.324,72.293,72.265,71.891,71.448,68.779,68.695；MS(ESI):m/z 562.1[M+H]⁺.

comparative example 2 preparation of Compound C-2 nitrile

Adding a compound C-1 hydroxyl compound (1.1g,2mmol,1.0eq) into a 500mL reaction bottle, adding dichloromethane (50mL), stirring for dissolving, replacing argon for three times, cooling to 0 ℃, adding TMSCN (0.794g,8mmol,4.0eq), stirring for 30 minutes, slowly adding TMSOTf (2g,9mmol,4.5eq) dropwise, reacting at 30 ℃ for 1 hour, TLC (petroleum ether: ethyl acetate ═ 1:2) shows that the reaction is finished but some impurities are generated, cooling to 0 ℃, adding 100mL dichloromethane for dilution, pouring the reaction solution into 100mL saturated NaHCO₃The reaction was quenched with solution and the aqueous phase was extracted once more with 100mL of dichloromethane. The combined organic phases were washed once with saturated brine and concentrated to give the crude product, which was chromatographed on silica gel with a gradient elution of 0-75% ethyl acetate and n-heptane to give C-2 nitrile as a white solid (0.62g, yield: 55%).

EXAMPLE 3 preparation of Compound C-3

Adding C-2(10g,0.0178mol,1.0eq) dissolved in dichloromethane (320mL) in a 1L reaction bottle, cooling to-65 ℃ under the protection of argon, dropwise adding 1M boron trichloride hexane solution (89mL,0.089mol,5.0eq), controlling the internal temperature at about-50 ℃, completing the addition within about 25 minutes and reacting at about-65 ℃ for about 2 hours, finishing the reaction, adding methanol (100mL), adding NaHCO₃(12g) And water (20mL), stirring and naturally raising the temperature to 0 ℃, measuring the pH value at 8, stirring for 20 minutes, filtering, concentrating the filtrate to obtain a crude product, performing preparative HPLC separation on the crude product by using C18, and separating the crude product by using H₂O (0.1% TFA), gradient elution with 10-60% MeOH afforded compound C-3' (debenzylate, Redcixivir intermediate) as a white solid (2.3g, yield: 44.4%), compound C-3 as a white solid (2.4g, yield: 46.3%).

Compound C-3':

¹H NMR(400MHz,water-d2):δ8.13(s,1H),7.42(d,J＝4.9Hz,1H),7.16(d,J＝4.9Hz,1H),4.94(d,J＝5.4Hz,1H),4.43(td,J＝4.8,3.2Hz,1H),4.38-4.31(m,1H),3.88(dd,J＝12.8,3.2Hz,1H),3.80(dd,J＝12.8,4.8Hz,1H)；¹³C-NMR(100MHz,DMSO-d₆):δ155.634,147.872,123.887,117.344,116.540,110.788,100.816,85.444,78.574,74.245,70.087,60.962；MS(ESI):m/z 292.1[M+H]⁺.

compound C-3:

¹H NMR(400MHz,water-d2):δ8.11(s,1H),7.43(d,J＝4.8Hz,1H),7.10(d,J＝4.8Hz,1H),5.14(d,J＝4.4Hz,1H),4.75(dd,J＝9.0,4.5Hz,1H),4.33-4.26(m,1H),4.07(dd,J＝12.9,2.1Hz,1H),3.88(dd,J＝12.9,4.5Hz,1H)；¹³C-NMR(100MHz,water-d2):δ148.833,136.031,128.111,116.941,113.587,112.978,109.950,83.305,77.589,76.376,70.608,60.261；MS(ESI):m/z 292.1[M+H]⁺.

the structural data of the compound C-3' Rudexiluwei intermediate obtained from CN103052631A in the preparation method are consistent with the data reported in Nature,2016, Doi:10.1038/Nature17180(Nature,2016,531(7594): 381-385).

EXAMPLE 4 preparation of Compound C

A1000 mL reaction vessel was charged with Compound C-3(6.8g,0.02336mmol,1.0eq) and acetone (200mL), and 2, 2-dimethoxypropane (12.16g,0.1168mol,5.0eq) and concentrated sulfuric acid (2mL) were added, and after stirring at room temperature for 0.5 hour, the temperature was raised to 45 ℃. The reaction was allowed to cool to room temperature for 1 hour, followed by the addition of solid sodium bicarbonate (7g) and water (7 mL). The mixture was stirred for 15 minutes and concentrated under reduced pressure. The crude product obtained by concentration was added with water (50mL) and ethyl acetate (150 mL). After stirring and standing, the lower aqueous phase was separated and extracted with ethyl acetate (2X 50 mL). The organic phases were combined, dried over sodium sulfate, concentrated under reduced pressure, and dried in vacuo to give Compound C (7.52g, yield: 97.2%).

Compound C:

¹H NMR(400MHz,DMSO-d₆)δ7.92(s,1H),7.81(s,2H),6.89(d,J＝4.5Hz,1H),6.68(d,J＝4.5Hz,1H),5.50(d,J＝5.8Hz,1H),5.21(t,J＝5.0Hz,1H),5.04(dd,J＝5.8,0.7Hz,1H),4.43(t,J＝6.1Hz,1H),3.71(t,J＝5.3Hz,2H),1.17(s,3H),0.89(s,3H)；¹³C-NMR(100MHz,DMSO-d₆):δ155.364,147.687,122.727,118.573,115.442,112.515,108.992,100.726,86.651,85.772,82.688,79.520,60.455,25.464,24.973；MS(ESI):m/z 332.2[M+H]⁺.

EXAMPLE 5 preparation of Compound E (esterified product)

A100 mL reaction flask was charged with Compound C (1.1g,0.00332mol,1.0eq) and Compound D (1.8g,0.004mol,1.2eq), anhydrous acetonitrile (16mL) was added, the mixture was dissolved, anhydrous magnesium chloride was added at room temperature, the mixture was stirred and warmed to 50 ℃ for 20 minutes, DIPEA (1.074g,0.0093mol,2.5eq) was added dropwise, the mixture was allowed to react for 1 hour, cooled to room temperature, diluted with ethyl acetate (50mL), washed with 5% citric acid (20 mL. times.2), a saturated ammonium chloride solution (20mL), a 5% aqueous potassium carbonate solution (20 mL. times.2), a saturated sodium chloride (20mL) was washed once, dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure to a white solid, and then passed through a column to obtain Compound E (1.6g, yield: 75.1%) as an oil.

Compound E:

¹H NMR(400MHz,methanol-d₄)δ7.83(s,1H),7.38(t,J＝7.8Hz,2H),7.30(dd,J＝8.8,1.1Hz,2H),7.21(t,J＝7.3Hz,1H),6.88(d,J＝4.5Hz,1H),6.77(d,J＝4.5Hz,1H),5.59(d,J＝5.8Hz,1H),5.08(dd,J＝5.8,1.2Hz,1H),4.70(t,J＝5.2Hz,1H),4.40(ddd,J＝19.5,11.1,5.7Hz,2H),4.26–4.03(m,3H),1.43–1.24(m,11H),1.22(s,3H),0.96(s,3H),0.90(t,J＝7.5Hz,6H)；¹³C-NMR(100MHz,methanol-d₄):δ175.079,175.027,156.998,152.219,152.150,147.879,130.831,126.235,124.900,121.534,121.485,119.359,116.868,115.019,110.914,102.399,87.862,86.294,84.041,81.730,68.286,67.141,67.086,51.642,41.777,33.034,30.142,26.037,25.339,24.342,24.306,23.730,20.697,20.629,14.411,11.368；³¹P-NMR(162MHz,methanol-d4):δ3.480(s)；MS(ESI):m/z 642.9[M+H]⁺.

EXAMPLE 6 preparation of RDV-ISO-1 (i.e., the Compound of formula I)

The esterified compound E (1.8g,0.0028mol) and tetrahydrofuran (20mL) are put into a 250mL reaction bottle, stirred and dissolved, cooled to 0-5 ℃, dropwise added with concentrated hydrochloric acid (4mL), and heated to 20-25 ℃ for 5 hours of heat preservation reaction. 20mL of water was added and the pH was adjusted to 7-8 with saturated sodium bicarbonate solution. Ethyl acetate (50mL) was added, followed by stirring to separate the organic phase, which was washed with an aqueous solution of sodium chloride (20 mL. times.2), dried over anhydrous sodium sulfate, filtered, concentrated to dryness under reduced pressure, and the crude product was subjected to column chromatography to give RDV-ISO-1(1.26g, yield: 75%).

Compound RDV-ISO-1:

¹H NMR(400MHz,methanol-d₄)δ7.84(s,1H),7.42–7.32(m,2H),7.32–7.27(m,2H),7.18(td,J＝7.3,0.8Hz,1H),6.90(d,J＝4.5Hz,1H),6.80(d,J＝4.5Hz,1H),5.00(d,J＝4.3Hz,1H),4.60(dd,J＝8.6,4.3Hz,2H),4.48(dd,J＝9.2,6.0Hz,1H),4.38–4.28(m,2H),4.06(tdd,J＝16.5,10.3,5.9Hz,3H),1.54–1.43(m,1H),1.43–1.26(m,8H),0.86(td,J＝7.5,2.4Hz,6H).¹³C-NMR(100MHz,methanol-d₄):δ175.13,175.06,156.95,152.23,152.16,148.07,130.73,126.08,124.62,121.54,121.49,118.77,116.87,111.91,102.65,82.56,82.48,79.70,77.21,72.66,68.25,67.41,67.35,51.56,41.64,24.23,24.18,20.78,20.72,11.35,11.29；³¹P-NMR(162MHz,methanol-d4):δ3.641(s)；MS(ESI):m/z 602.9[M+H]⁺.

example 7 preparation Synthesis of E-ISO-2 (esterified R70%, S30%)

A100 mL reaction flask was charged with C '(1 g,0.003mol,1.0eq) and D' (1.62g,0.0036mol,1.2eq, R70%, S30%), anhydrous acetonitrile (16mL) was added, the mixture was dissolved, anhydrous magnesium chloride was added at room temperature, the mixture was stirred and warmed to 50 ℃ for 20 minutes, DIPEA (0.969g,0.0075mol,2.5eq) was added dropwise, the reaction was allowed to keep warm for 1 hour, the mixture was cooled to room temperature, ethyl acetate (50mL) was added for dilution, and the mixture was washed with 5% citric acid (20 mL. times.2), saturated ammonium chloride solution (20mL), 5% aqueous potassium carbonate (20 mL. times.2), saturated sodium chloride (20mL) was washed once, anhydrous magnesium sulfate was dried, filtered, the organic phase was concentrated under reduced pressure to an oil, and E-ISO-2(1.5g, yield: 77.3%) was obtained as a white solid.

Compound E-ISO-2:

¹H NMR(400MHz,methanol-d₄)δ7.86(s,1H),7.26(dd,J＝15.6,7.8Hz,2H),7.18–7.02(m,3H),6.89(dt,J＝4.6,3.9Hz,2H),5.26(dd,J＝63.7,6.6Hz,1H),4.97(ddd,J＝15.6,6.6,3.3Hz,1H),4.67–4.50(m,1H),4.40–4.25(m,2H),4.19–3.78(m,3H),1.70(d,J＝2.2Hz,3H),1.54–1.20(m,12H),0.87(t,J＝7.5Hz,6H)；¹³C-NMR(100MHz,methanol-d₄):δ175.14,175.09,174.96,174.90,157.26,152.15,152.08,148.52,148.49,130.68,126.20,126.10,124.72,124.61,121.40,121.37,121.36,121.32,118.34,118.31,117.80,117.62,117.06,117.02,112.35,112.28,102.46,85.70,84.97,84.94,84.89,84.85,83.09,83.03,82.73,82.60,68.12,68.07,67.03,66.97,66.95,66.90,51.61,51.48,41.75,41.73,26.52,26.43,25.54,25.45,24.24,24.22,20.38,20.31,11.33,11.30；³¹P-NMR(162MHz,methanol-d4):δ3.533(s),3.219(s)；MS(ESI):m/z 665.0[M+Na]⁺.

EXAMPLE 8 preparation Synthesis of RDV-ISO-2

Putting the esterified substance E-ISO-2(1g,0.00156mol) and tetrahydrofuran (16mL) into a 100mL reaction bottle, stirring to dissolve, cooling to 0-5 ℃, dropwise adding concentrated hydrochloric acid (4mL), finishing dripping, heating to 20-25 ℃, and keeping the temperature for reaction for 5 hours. 20mL of water was added and the pH was adjusted to 7-8 with saturated sodium bicarbonate solution. Ethyl acetate (50mL) was added and the mixture was stirred, the organic phase was separated, washed with aqueous sodium chloride (20 mL. times.2), dried over anhydrous sodium sulfate, filtered, concentrated to dryness under reduced pressure, and the crude product was isolated by preparative HPLC to give RDV-ISO-2(492mg, yield: 52.6%).

Compound RDV-ISO-2:

¹H NMR(400MHz,methanol-d₄)δ7.85(s,1H),7.28(dd,J＝10.0,5.7Hz,2H),7.21–7.08(m,3H),6.91(dd,J＝10.7,4.6Hz,2H),4.79(d,J＝5.5Hz,1H),4.53–4.36(m,2H),4.36–4.26(m,1H),4.21(t,J＝5.4Hz,1H),4.00(qd,J＝10.9,5.7Hz,2H),3.86(dq,J＝14.3,7.1Hz,1H),1.46(dq,J＝12.1,6.0Hz,1H),1.39–1.27(m,4H),1.25(dd,J＝7.1,1.0Hz,3H),0.86(t,J＝7.5Hz,6H)；¹³C-NMR(100MHz,methanol-d₄):δ175.15,175.11,157.26,152.17,152.10,148.34,130.68,126.11,125.54,121.43,121.39,118.00,117.64,112.37,102.63,84.61,84.53,81.09,75.68,71.55,68.09,66.83,66.77,51.61,41.73,24.23,24.21,20.45,20.38,11.33,11.30；³¹P-NMR(162MHz,methanol-d₄):δ3.545(s)；MS(ESI):m/z602.9[M+H]⁺.

EXAMPLE 9 preparation of Compound E' (esterified product)

A100 mL reaction flask was charged with Compound C (1.1g,0.00332mol,1.0eq) and Compound D '(R, about 70%; S, about 30%) (1.8g,0.004mol,1.2eq), anhydrous acetonitrile (16mL) was added, the mixture was dissolved, anhydrous magnesium chloride was added at room temperature, the mixture was stirred and warmed to 50 ℃ for 20 minutes, DIPEA (1.074g,0.0093mol,2.5eq) was added dropwise, the mixture was allowed to keep warm for 1 hour, cooled to room temperature, diluted with ethyl acetate (50mL), washed with 5% citric acid (20 mL. times.2), saturated ammonium chloride solution (20mL), 5% aqueous potassium carbonate (20 mL. times.2), saturated sodium chloride (20mL) was washed once, anhydrous magnesium sulfate was dried, filtered, and concentrated under reduced pressure to an oil to obtain a white solid Compound E' (1.5g, yield: 70%) on the column.

A compound E':

¹H NMR(400MHz,methanol-d₄)δ7.85(s,1H),7.45–7.35(m,2H),7.35–7.27(m,2H),7.23(t,J＝7.8Hz,1H),6.88(t,J＝4.4Hz,1H),6.76(t,J＝4.8Hz,1H),5.28(d,J＝5.7Hz,1H),4.96(dd,J＝5.7,1.0Hz,1H),4.78–4.66(m,1H),4.42(t,J＝5.1Hz,2H),4.17–4.00(m,3H),1.52(dd,J＝12.3,6.2Hz,1H),1.45–1.15(m,11H),0.96(s,1H),0.90(td,J＝7.2,2.5Hz,7H)；¹³C-NMR(100MHz,methanol-d₄):δ175.33,175.28,156.99,152.31,152.24,147.87,147.82,130.90,130.83,126.37,126.24,125.19,124.91,121.64,121.59,121.53,121.49,119.44,119.36,116.86,115.02,114.84,110.92,110.82,102.35,87.87,86.38,86.29,86.19,86.09,84.07,81.96,81.73,68.29,68.18,67.38,67.32,67.14,61.53,51.69,51.65,41.82,41.78,26.04,25.88,25.34,25.18,24.32,20.63,20.57,14.46,11.36；³¹P-NMR(162MHz,methanol-d4):δ3.705(s),3.482(s)；MS(ESI):m/z665.0[M+Na]⁺.

EXAMPLE 10 preparation of RDV-ISO-3 (i.e., the compound of formula II)

The esterified compound E' (1g,0.00156mol) and tetrahydrofuran (16mL) are put into a 100mL reaction flask, stirred, dissolved and cleared, cooled to 0-5 ℃, dropwise added with concentrated hydrochloric acid (4mL), and heated to 20-25 ℃ for 5 hours of heat preservation reaction. 20mL of water was added and the pH was adjusted to 7-8 with saturated sodium bicarbonate solution. Ethyl acetate (50mL) was added, stirring was carried out, the organic phase was separated, washed with aqueous sodium chloride solution (20 mL. times.2), dried over anhydrous sodium sulfate, filtered, concentrated to dryness under reduced pressure, and the crude product was isolated by preparative HPLC to give RDV-ISO-3(528mg, yield: 56.3%).

Compound RDV-ISO-3:

¹H NMR(400MHz,methanol-d₄)δ7.86(s,1H),7.40–7.33(m,2H),7.32–7.25(m,2H),7.20(td,J＝7.5,0.9Hz,1H),6.90(d,J＝4.5Hz,1H),6.80(d,J＝4.5Hz,1H),4.97(d,J＝4.2Hz,1H),4.60(dd,J＝8.7,4.3Hz,1H),4.57–4.50(m,1H),4.35–4.24(m,2H),4.11(tt,J＝14.2,7.1Hz,1H),4.01(d,J＝5.7Hz,2H),1.46(dq,J＝12.1,6.0Hz,1H),1.42–1.25(m,8H),0.85(td,J＝7.4,1.8Hz,6H)；

¹³C-NMR(100MHz,methanol-d₄):δ175.34,175.29,157.01,152.23,152.16,148.06,130.72,126.16,124.76,121.72,121.67,118.75,116.92,111.85,102.67,82.50,82.41,79.81,77.32,72.15,68.07,66.13,66.08,51.57,41.73,24.22,24.21,20.71,20.64,11.32,11.30；³¹P-NMR(162MHz,methanol-d4):δ3.654(s)；MS(ESI):m/z 603.3[M+H]⁺.

EXAMPLE 11 Effect test

The three process impurities RDV-ISO-1 (formula I), RDV-ISO-2 (formula III) and RDV-ISO-3 (formula II) of the Rudeseivir prepared by the invention have obvious inhibitory activity on Hela (human cervical carcinoma cells), COLO 205 (human colon carcinoma cells), U-87MG (human brain astrocytoma cells) and the like. The results are as follows:

the cytotoxicity detection method comprises the following steps:

concentration of drug treatment

The compound is prepared into 10 by DMSO^-2And (4) subpackaging a small amount of the stock solution of M and storing at-80 ℃ for later use. In the experiment, the initial concentration of the compound to be detected is set as 100uM, 10 gradient dilution concentrations are selected, the gradient dilution multiple is 3 times, and the compound is detected in 2 multiple wells per concentration. The Control group without drug was a corresponding medium solution without drug containing 0.2% DMSO.

CTG detection method

Hela (human cervical carcinoma cells), COLO 205 (human colon cancer cells) and U-87MG (human brain astrocytoma cells, namely glioma cells) in logarithmic growth phase are respectively taken and planted in 384-well plates according to certain cell density. The cell culture plate was placed in 5% CO at 37 deg.C₂Culturing for 24h under the condition of (1). The compounds diluted in gradient were added separately to the wells at 37 ℃ with 5% CO₂For 72 h. The cell culture plate was removed from the incubator and allowed to equilibrate at room temperature for 30 minutes. Add 50. mu.l of the mixture to each well after equilibration at room temperature

Shaking the Luminecent Cell vitality Assay reaction solution at 500rpm for 2 minutes at room temperature, and centrifuging the reaction solution at 1000rpm for 1 minute; after equilibration at room temperature for 10 minutes, the fluorescence signal was measured. The in vitro inhibitory activity of the compounds was calculated according to the following formula:

inhibition (%) - (signal value control-signal value administration)/signal value control × 100%. And the 50% inhibitory concentration (IC 50) was calculated by the LOGIT method based on the inhibition ratio of each concentration.

Note: rf IC50 and Abs IC50 represent relative and absolute half-maximal inhibition, respectively.

From the data in the above table, the IC50 value of RDV-ISO-2 on Hela (human cervical cancer cells), COLO 205 (human colon cancer cells) and U-87MG (human brain astrocytoma cells) is significantly lower than that of RDV-ISO-1 and RDV-ISO-3, and has better inhibitory activity. All three compounds have potential anticancer activity and have further research value.

Claims

1. A compound of formula I or formula II, or a pharmaceutically acceptable salt thereof:

2. a preparation method of a compound shown in a formula I or a compound shown in a formula II is characterized in that,

the preparation method of the compound shown as the formula I comprises the following steps: in a solvent, in the presence of acid, carrying out deprotection reaction on the compound E as shown in the specification to obtain a compound as shown in a formula I;

the preparation method of the compound shown as the formula II comprises the following steps: in a solvent, in the presence of an acid, the compound E' is subjected to deprotection reaction as described below, and then the compound is shown as a formula II;

3. the method of claim 2, wherein the method of preparing the compound of formula I further comprises the steps of: in a solvent, in the presence of a catalyst and alkali, carrying out substitution reaction on a compound C and a compound D as shown in the specification to obtain a compound E;

the catalyst is preferably a magnesium chloride (mgci),

the alkali is preferably N, N-diisopropylethylamine;

the preparation method of the compound shown in the formula I can further comprise the following steps:

in a solvent, in the presence of acid, carrying out condensation reaction on a compound C-3 and 2, 2-dimethoxypropane as shown in the specification to obtain a compound C;

in a solvent, in the presence of a debenzylation reagent, performing debenzylation reaction on the compound C-2 as shown in the specification to obtain a compound C-3;

the debenzylation reagent is preferably BCl₃And/or BBr₃；

adding TMSOTf and TMSCN into a solvent at 0 ℃, carrying out cyanation reaction on the compound C-1 at 15-20 ℃ to obtain a compound C-2,

in a solvent, adding a compound A in the presence of B, TMSCl and PhMgCl to carry out a format reaction as described below to obtain a compound C-1;

4. the method according to claim 3, wherein in the cyanation reaction, the solvent is one or more of a haloalkane-type solvent, a nitrile-type solvent and an ether-type solvent, such as one or more of dichloromethane, acetonitrile, 1, 4-dioxane and tetrahydrofuran;

and/or in the cyanation reaction, the molar ratio of the compound C-1 to the TMSCN is 1: 4-1: 5;

and/or, the cyanation reaction does not comprise trifluoromethanesulfonic acid;

and/or, said cyanation reaction comprises only said solvent, said TMSOTf, said TMSCN, and said Compound C-1;

and/or, in the cyanation reaction, the molar ratio of the compound C-1 to the TMSOTf is 1: 4-1: 4.2;

and/or, in the cyanation reaction, the following steps are included: adding TMSCN in the presence of a compound C-1 and a solvent, and then adding TMSOTf to carry out the cyanation reaction to obtain a compound C-2; preferably, the cyanation reaction comprises the following steps: adding TMSCN with the amount of 4-5 equivalents relative to the amount of a compound C-1 in the presence of the compound C-1 and a solvent, adding TMSOTf with the amount of 2-2.1 equivalents relative to the amount of the compound C-1, and supplementing TMSOTf with the amount of 2-2.1 equivalents relative to the amount of the compound C-1 to perform the cyanation reaction to obtain a compound C-2;

and/or, in the cyanation reaction, the reaction time is 2 to 5 hours.

5. The method of claim 2, wherein the method of preparing the compound of formula II further comprises the steps of: in a solvent, in the presence of a catalyst and alkali, carrying out substitution reaction on a compound C and a compound D 'as shown in the specification to obtain a compound E';

the catalyst is preferably a magnesium chloride (MgCl) catalyst,

the alkali is preferably N, N-diisopropylethylamine;

the preparation method of the compound shown in the formula II can further comprise the following steps:

said debenzylationThe base reagent is preferably BCl₃And/or BBr₃；

in a solvent, in the presence of a compound B, TMSCl and PhMgCl, carrying out a format reaction on a compound A as described below to obtain a compound C-1;

6. a compound C, compound E or compound E':

7. a pharmaceutical composition comprising one or more of substance X, substance Y and substance Z, together with a pharmaceutically acceptable adjuvant; the substance X is a compound shown as a formula I or a pharmaceutically acceptable salt thereof, the substance Y is a compound shown as a formula II or a pharmaceutically acceptable salt thereof, and the substance Z is a compound shown as a formula III or a pharmaceutically acceptable salt thereof; the amount of substance X, substance Y or substance Z may be a therapeutically effective amount;

8. use of substance X, substance Y, substance Z or a pharmaceutical composition according to claim 7 for the preparation of an inhibitor of cell proliferation; the substance X, substance Y and substance Z are defined as in claim 7, and the cells are one or more of Hela cells, COLO 205 cells and U-87MG cells.

9. Use of substance X, substance Y, substance Z or a pharmaceutical composition according to claim 7 for the manufacture of a medicament for the treatment and/or prophylaxis of cancer, said cancer being one or more of cervical cancer, colon cancer and glioma, said substance X, substance Y and substance Z being as defined in claim 7.

10. A method for preparing a compound C-2, which is characterized by comprising the following steps: adding TMSOTf and TMSCN into a solvent at 0 ℃, and carrying out cyanation reaction on the compound C-1 at 15-20 ℃ to obtain a compound C-2;

the cyanation reaction conditions and steps can be as described in claim 4.