CN106749445B - epirubicin hydrochloride intermediate compound III - Google Patents

Abstract

The invention provides a novel intermediate and a novel route for synthesizing epirubicin hydrochloride by utilizing the intermediate. The method is simple, cheap and efficient, and simultaneously solves the problems that an intermediate generated by the existing epirubicin hydrochloride synthetic route is unstable and easy to decompose when meeting water, and the yield of the whole route is low; meanwhile, a removable selective protection means is adopted, so that the produced impurities are few, the purity is high, and the yield is high.

Description

Epirubicin hydrochloride intermediate compound III

Technical Field

The invention belongs to the field of chemical synthesis, and particularly relates to an epirubicin hydrochloride intermediate compound III.

background

Epirubicin hydrochloride is also called epirubicin hydrochloride, belongs to anthraquinone antibiotics, and has the chemical name: (8S,10S) -10- [ (3 '-amino-2', 3 ', 6' -trideoxy-. alpha. -L-arabinopyranosyl) -O- ] -6,8, 11-trihydroxy-8-hydroxyacetyl-1-methoxy-7, 8,9, 10-tetrahydrotetracene-5, 12-dione hydrochloride having the following structure:

Is an isomer of adriamycin, and has the action mechanism of directly inserting between DNA nuclear alkali pairs, interfering the transcription process and preventing the formation of tumor cell mRNA, thereby inhibiting the synthesis of DNA and RNA. In addition, epirubicin hydrochloride also has an inhibitory effect on topoisomerase II. Is a cell cycle nonspecific drug, and is effective for various transplantation tumors. The clinical curative effect is equal to or slightly higher than that of adriamycin, but the adriamycin has small toxicity to heart, and single medicine has broad-spectrum inhibition effect on various tumors and can be used for breast cancer, malignant lymphoma, soft tissue sarcoma and gastric cancer. It also has antitumor activity against malignant melanoma and colon cancer. Can be used for treating lung cancer and ovarian cancer in combination with other anticancer drugs. Therefore, the compound plays an increasingly important role in human antitumor clinical application, and the clinical demand is increasing year by year.

Patent JP2007261976A provides a synthetic route for epirubicin hydrochloride, which is as follows:

The method comprises the steps of taking 4' -epidaunorubicin as a raw material, reacting under the action of methyl formate and bromine to obtain a brominated product, hydrolyzing with epoxypropane and hydrogen bromide in sequence to prepare a bromoketone intermediate, and hydrolyzing with sodium bicarbonate, sodium hydroxide and sodium chloride aqueous solution in sequence to obtain epirubicin hydrochloride. Organic solvents such as propylene oxide and the like used in the method cause great harm to physical and psychological health of producers, and the generated waste liquid is difficult to treat and has great environmental pressure; meanwhile, the final hydrolysis stage adopts multi-step hydrolysis conditions with different pH values, and is difficult to accurately control, so that extremely high requirements are put on the technology of operators, and the process has the disadvantages of high production cost, serious environmental pollution and difficult industrial mass production. The process has strict requirements on the water content of the raw material 4 '-epidaunorubicin and the humidity during reaction, the defect of overlarge residual amount of the 4' -epidaunorubicin can be caused by slight change of the water content in the actual application process, the quality of the final product is difficult to guarantee, and the material loss rate is extremely high.

Patent US20070142309 provides synthetic routes of various epirubicin hydrochloride, the synthetic principle is different, but basically all adopt the treatment method of reductive amination with sodium triacetoxyborohydride, then the mode of bromination and sodium formate hydrolysis of the intermediate produced obtains final product epirubicin hydrochloride, the cost of the reagent is higher than that of the traditional organic synthetic reagent, and the applied reagent is difficult to preserve in workshop environment, easy to generate safety production accident, and not suitable for industrial mass production practice.

Therefore, the epirubicin hydrochloride preparation methods described above all have problems, such as low yield, large impurities, high technical requirements, serious environmental pollution and high production cost, and the intermediates generated by the epirubicin hydrochloride preparation methods in the prior art have extremely high requirements for moisture, are easy to decompose, and are not suitable for industrial mass production.

disclosure of Invention

In view of the defects of the prior art, the invention provides a novel intermediate compound III and a novel route for synthesizing epirubicin hydrochloride by using the intermediate. The preparation method is simple, cheap and efficient, in order to better reduce the production cost and reduce the pollution to the environment, the preparation method basically uses an economic and less-pollution organic reagent, and the intermediate generated by the preparation method has low requirement on moisture, high stability and high quality yield.

The invention is realized by the following technical scheme:

An epirubicin hydrochloride compound is shown in a formula III, and the structural formula is as follows:

A process for the preparation of an intermediate compound iii comprising the steps of: adding methanol, an acidic catalyst and an esterification reagent B to enable daunorubicin hydrochloride II to generate an unseparated intermediate compound with a ketal structure, protecting active amino of an aminosugar part in the intermediate structure with trifluoroacetic anhydride to generate an intermediate compound III,

The synthesis of the compound III from the compound II comprises the following steps: adding methanol, an acidic catalyst, an esterification reagent B and daunorubicin hydrochloride II into an organic solvent A, reacting for 2-3 hours to generate an unseparated intermediate compound II-1 with a ketal structure, then adding trifluoroacetic anhydride into a reaction system, and stirring and reacting for 1-1.5 hours to generate an intermediate compound III with an amino trifluoroacetate structure.

further preferably, the steps are: adding an organic solvent A and daunorubicin hydrochloride II into a three-neck glass reaction bottle. Starting stirring, controlling the temperature to be 5-10 ℃, checking the dissolution condition of the material after 10-15 minutes, and continuing stirring if the material is not completely dissolved; if the solution is completely dissolved, cooling the feed liquid to 0-5 ℃, adding a methanol solution of an acid catalyst, stirring for 1 hour, and then adding an esterification reagent B; stirring for 2-3 hours after the addition is finished; and after stirring, controlling the temperature of the reaction liquid to be 3-10 ℃, adding trifluoroacetic anhydride into the bottle, and stirring for 1-1.5 hours after the addition is finished. To generate an intermediate compound III with an amino trifluoroacetate structure.

The dosage of the added acidic catalyst methanol solution is as follows: in terms of weight g/volume mL/weight g (the following sections are expressed as (W/V/W, g/mL/g)), daunorubicin hydrochloride: methanol: the acidic catalyst is 1: 177-200: 1-3, and the acidic catalyst is preferably camphorsulfonic acid or p-methylbenzenesulfonic acid. The temperature for the reaction with the esterification reagent B is preferably 0-10 ℃.

The organic solvent A is preferably one of aromatic hydrocarbon, aliphatic hydrocarbon, alicyclic hydrocarbon, halogenated hydrocarbon, alcohol, alkane, ether, amide, glycol derivative and ester solvent; more preferably one or more selected from benzene, toluene, cyclohexane, methanol, ethanol, tert-butanol, dichloromethane, 1, 4-dioxane, diethyl ether, acetone, trichloroethylene, tetrahydrofuran, methyl tert-butyl ether, ethyl acetate and DMF; further preferred is methanol or methylene chloride or 1, 4-dioxane.

The esterifying reagent B is preferably one or more of ethyl formate, methyl acetate, ethyl acetate, triethyl orthoformate, ethyl chloroformate and propyl formate; further preferred is triethyl orthoformate or trimethyl orthoformate.

The dosage of the organic solvent A is as follows: daunorubicin hydrochloride: the organic solvent A is 1: 50-70, preferably 1: 60.

The dosage of the organic solvent B is as follows: daunorubicin hydrochloride: the ratio of the esterification reagent B to the esterification reagent B is 1: 1-4, preferably 1:2(W/V, g/mL).

the dosage of the trifluoroacetic anhydride is as follows: daunorubicin hydrochloride: trifluoroacetic anhydride is 1:1 to 4, preferably 1:2(W/V, g/mL).

After the reaction, methanol (daunorubicin hydrochloride: methanol 1: 20-40 (W/V, g/mL)) is added to the reaction solution, and after stirring for 10-15 minutes, 8% sodium bicarbonate solution is added to adjust the pH to 7.0-8.0. Controlling the temperature to be 15-25 ℃, and carrying out heat preservation stirring reaction for 3-5 hours. And (3) TLC detection: the chloroform/isopropanol (volume ratio 96/4) is used as developing solvent, and the raw material spots are detected to be basically disappeared. The intermediate compound III Rf is approximately equal to 0.3, and the daunorubicin hydrochloride Rf is approximately equal to 0. Standing and separating liquid, and collecting a lower organic phase. Concentrating under vacuum to a certain volume (daunorubicin hydrochloride: the volume of the concentrated solution is 1:10(W/V, g/mL)), adding n-hexane under stirring for crystallization, and performing suction filtration and drying to obtain a solid intermediate compound III. Preferably, the adding amount of the n-hexane is as follows: daunorubicin hydrochloride: n-hexane 1:10(W/V, g/mL).

The application also provides a method for preparing epirubicin hydrochloride by using the intermediate III, which comprises the following steps: step 1, a step of preparing intermediate iii as described above; step 2, further dehydrating, oxidizing the alcohol hydroxyl group with the amino sugar structure into carbonyl, and generating an unseparated intermediate compound IV; step 3, reducing carbonyl in the amino sugar into hydroxyl by using a selective reducing agent D, and converting the hydroxyl into an intermediate V with a 4-OH configuration opposite to that of the daunorubicin amino sugar structure; step 4, removing the protection of the amino group in the intermediate V structure by using strong base to generate an intermediate VI; and 5, carrying out bromination and acid hydrolysis reaction in sequence in an acidic environment to generate a corresponding bromoketone intermediate, hydrolyzing in a sodium formate solution, and substituting bromine into alcoholic hydroxyl to finally obtain the target product epirubicin hydrochloride I. The specific synthetic route is as follows:

Step 2, synthesizing a compound IV by using a compound III specifically comprises the following steps: and (2) adding trifluoroacetic anhydride into an organic solvent A, stirring and reacting for 1-1.5 hours after the addition is finished, then adding an intermediate compound III to react with 1, 5-diazabicyclo (4,3,0) non-5-ene for 0.5 hour, oxidizing alcoholic hydroxyl with an amino sugar structure to carbonyl, then adding an organic acid C into a reaction system, and stirring and reacting for 0.5 hour to generate an intermediate compound IV.

The temperature for the reaction with 1, 5-diazabicyclo (4,3,0) non-5-ene described in step 2 is preferably-75 ℃ to 0 ℃, most preferably-70 ℃ to-40 ℃.

The organic acid C in the step 2 is preferably selected from one or more of formic acid, acetic acid, propionic acid, butyric acid, caprylic acid, adipic acid, oxalic acid, malonic acid, succinic acid, maleic acid, tartaric acid, benzoic acid, phenylacetic acid, phthalic acid, terephthalic acid, valeric acid, caproic acid, capric acid, stearic acid, palmitic acid and acrylic acid; further preferably one of formic acid, acetic acid, propionic acid, butyric acid, caprylic acid, adipic acid, oxalic acid, malonic acid, succinic acid and maleic acid; most preferred are glacial acetic acid and malonic acid.

The organic solvent A in the step 2 is selected from one or more of aromatic hydrocarbons, aliphatic hydrocarbons, alicyclic hydrocarbons, halogenated hydrocarbons, alcohols, alkanes, ethers, amides, glycol derivatives, ester solvents and phenol; more preferably one or more of benzene, toluene, cyclohexane, tert-butanol, dichloromethane, 1, 4-dioxane, diethyl ether, acetone, trichloroethylene, tetrahydrofuran, methyl tert-butyl ether, ethyl acetate and DMF; more preferably, methylene chloride is used.

And 2, the dosage of the organic solvent A is as follows: intermediate III: the organic solvent A is 1: 20-40, preferably 1:40(W/V, g/mL).

step 3, the synthesis of the compound V from the compound IV specifically comprises the following steps: and (3) reacting the intermediate IV with a reducing agent D in an organic solvent A for 0.5-1 hour, reducing carbonyl in amino sugar into hydroxyl, and converting the hydroxyl into an intermediate V with a structure opposite to that of 4-OH of the daunorubicin amino sugar structure.

The temperature for the reaction with the reducing agent D in the step 3 is preferably-75 ℃ to 0 ℃, and most preferably-60 ℃ to-30 ℃.

the reducing agent D in the step 3 is preferably one or more of sodium hydride, lithium aluminum hydride, sodium borohydride, lithium hydride, bis (methoxyethoxy) aluminum hydride, lithium triethylborohydride and sodium cyanoborohydride; the selective reducing agent is preferably used for minimizing isomer impurities of the intermediate V, and the purity of the intermediate V is more than 85%.

The organic solvent A in the step 3 is preferably one or more of aromatic hydrocarbon, aliphatic hydrocarbon, alicyclic hydrocarbon, halogenated hydrocarbon, alcohol, alkane, ether, amide, glycol derivative, ester solvent and phenol; more preferably one or more of benzene, toluene, cyclohexane, tert-butanol, dichloromethane, 1, 4-dioxane, diethyl ether, acetone, trichloroethylene, tetrahydrofuran, methyl tert-butyl ether, ethyl acetate and DMF; further preferred is dichloromethane or acetone.

and 4, step 4: the preparation method of the intermediate compound VI comprises the following specific steps: reacting the intermediate compound V with alkali E for 0.5 hour at a certain temperature in purified water to obtain an intermediate compound VI; the temperature is preferably 0 ℃ to 20 ℃, and most preferably 0 ℃ to 10 ℃.

Step 4 the base E is preferably one selected from the group consisting of sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium carbonate and sodium bicarbonate, most preferably sodium hydroxide.

and 5: the preparation method of the intermediate compound I comprises the following specific steps: the method comprises the following steps of firstly carrying out bromination reaction in an acidic environment to convert the intermediate into a bromo-intermediate VI-1 with a ketal structure, carrying out acid hydrolysis reaction on the intermediate with the structure in the acidic environment to remove the protection of the ketal structure on carbonyl to generate a corresponding bromo-ketone intermediate VI-2, then carrying out hydrolysis in a sodium formate solution to replace bromine into alcoholic hydroxyl, and salifying in a hydrochloric acid-methanol solution to finally obtain a target product epirubicin hydrochloride I.

And 5: the specific steps for the preparation of intermediate compound i are further detailed as follows:

in an organic solvent A, reacting the intermediate compound VI with an organic solvent A solution of hydrogen bromide for 2-3 hours, and dropwise adding an organic solvent A solution of bromine into a reaction solution in the reaction process; the temperature for the reaction with the solution of the organic solvent A of hydrogen bromide is preferably 0 ℃ to 30 ℃, and most preferably 10 ℃ to 20 ℃.

Adding sufficient reducing agent F to neutralize excessive bromine, quenching, and adding a proper amount of saturated alkali solution G to adjust the pH of the reaction solution to 4.5-5.0, wherein the most preferable pH is 4.7.

Adding an inorganic acid H aqueous solution, adjusting the pH value to 1.3-1.5, optimally selecting the pH value to 1.5, and stirring and reacting for 2-2.5 hours at the temperature of 25-35 ℃.

and adding a sodium formate aqueous solution, adjusting the pH value to 3.0-3.5, optimally selecting 3.2, controlling the temperature to 25-35 ℃, and hydrolyzing the sodium formate for 1-3 hours to finally obtain a target compound epirubicin hydrochloride I crude product solution. The mass concentration (g/g) of the sodium formate aqueous solution is preferably 20-30%.

The organic solvent A in the step 5 is preferably one or more of aromatic hydrocarbon, aliphatic hydrocarbon, alicyclic hydrocarbon, halogenated hydrocarbon, alcohol, alkane, ether, amide, glycol derivative, ester solvent and phenol; more preferably one or more of benzene, toluene, cyclohexane, tert-butanol, dichloromethane, 1, 4-dioxane, diethyl ether, acetone, trichloroethylene, tetrahydrofuran, methyl tert-butyl ether, ethyl acetate and DMF; further preferably methanol, or 1, 4-dioxane, or acetone, or ethyl acetate, or a mixed solvent of acetone and ethyl acetate (the volume ratio of the two is 1: 3).

the reducing agent F is preferably one selected from sodium sulfite, sodium bisulfite, ferrous sulfate, sodium nitrite and oxalic acid, and most preferably sodium sulfite.

The base G is preferably selected from one of sodium hydroxide, potassium hydroxide, calcium hydroxide, aqueous ammonia and sodium bicarbonate, most preferably aqueous ammonia.

The inorganic acid H is preferably selected from one of phosphoric acid, sulfuric acid, perchloric acid, boric acid, nitric acid, hydrochloric acid and hydrobromic acid, and most preferably hydrochloric acid; the concentration of the inorganic acid is preferably 6-8%.

Wherein the following compounds are novel substances:

The above steps are described in further detail in the following sections:

Step 1, preparation of intermediate compound iii:

Adding an organic solvent A and daunorubicin hydrochloride II into a three-neck glass reaction bottle. Starting stirring, controlling the temperature to be 5-10 ℃, checking the dissolution condition of the material after 10-15 minutes, and continuing stirring if the material is not completely dissolved; if the solution is completely dissolved, cooling the feed liquid to 0-5 ℃, adding a methanol solution of an acid catalyst, stirring for 1 hour, and then adding an esterification reagent B; stirring for 2-3 hours after the addition is finished; and after stirring, controlling the temperature of the reaction liquid to be 3-10 ℃, adding trifluoroacetic anhydride into the bottle, and stirring for 1-1.5 hours after the addition is finished.

The dosage of the organic solvent A is as follows: in terms of weight g/volume mL (the following sections are expressed as (W/V, g/mL)), daunorubicin hydrochloride: the organic solvent A is 1: 50-70, preferably 1: 60; the organic solvent A is preferably one or more of methanol, dichloromethane and 1, 4-dioxane.

The dosage of the added acidic catalyst methanol solution is as follows: daunorubicin hydrochloride: methanol: the acid catalyst is 1:177:1(W/V/W, g/mL/g), and the acid catalyst is camphorsulfonic acid.

The dosage of the esterification reagent B is as follows: daunorubicin hydrochloride: the ratio of the esterification reagent B to the esterification reagent B is 1: 1-4, preferably 1:2(W/V, g/mL); the esterification reagent B is preferably triethyl orthoformate or trimethyl orthoformate.

The dosage of the trifluoroacetic anhydride is as follows: daunorubicin hydrochloride: trifluoroacetic anhydride is 1:1 to 4, preferably 1:2(W/V, g/mL).

After the reaction, methanol (daunorubicin hydrochloride: methanol 1: 20-40 (W/V, g/mL)) is added to the reaction solution, and after stirring for 10-15 minutes, 8% sodium bicarbonate solution is added to adjust the pH to 7.0-8.0. Controlling the temperature to be 15-25 ℃, and carrying out heat preservation stirring reaction for 3-5 hours. And (3) TLC detection: the chloroform/isopropanol (volume ratio 96/4) is used as developing solvent, and the raw material spots are detected to be basically disappeared. The intermediate compound III Rf is approximately equal to 0.3, and the daunorubicin hydrochloride Rf is approximately equal to 0. Standing and separating liquid, and collecting a lower organic phase. Concentrating under vacuum to a certain volume (daunorubicin hydrochloride: the volume of the concentrated solution is 1:10(W/V, g/mL)), adding n-hexane under stirring for crystallization, and performing suction filtration and drying to obtain a solid intermediate compound III. Preferably, the adding amount of the n-hexane is as follows: daunorubicin hydrochloride: n-hexane 1:10(W/V, g/mL).

Step 2, preparing an intermediate compound IV:

The organic solvent A was added to a three-necked glass reaction flask. Starting stirring, controlling the temperature to be-70 to-40 ℃, slowly adding an organic solvent A solution of trifluoroacetic anhydride, and stirring and reacting for 1-1.5 hours at-70 to-40 ℃.

the organic solvent A is preferably dichloromethane.

the dosage of the organic solvent A is as follows: intermediate III: the organic solvent A is 1: 20-40, preferably 1:40(W/V, g/mL).

The dosage of the trifluoroacetic anhydride organic solvent A solution is as follows: intermediate III: trifluoroacetic anhydride: 1-dichloromethane: 0.5-0.8: 2.4 to 3(W/W/V, g/g/mL).

After the reaction is finished, slowly dropwise adding an organic solvent A solution of an epirubicin hydrochloride intermediate III into the reaction solution; after the completion of the addition, the mixture was stirred for 20 minutes, and then an organic solvent A solution of 1, 5-diazabicyclo (4,3,0) non-5-ene was added dropwise thereto, and after the completion of the addition, the mixture was stirred for 10 minutes, and then an organic solvent A solution of an organic acid C was added thereto, and the mixture was stirred and reacted for 0.5 hour. The organic solvent A is preferably dichloromethane. The organic acid C is preferably one of formic acid, acetic acid, propionic acid, butyric acid, caprylic acid, adipic acid, oxalic acid, malonic acid, succinic acid and maleic acid; most preferred are glacial acetic acid and malonic acid.

The solution A of the intermediate III in the organic solvent is as follows: intermediate III: the organic solvent A is 1: 5-8 (W/V, g/mL).

The dosage of the organic solvent A solution of the 1, 5-diazabicyclo (4,3,0) non-5-ene is as follows: intermediate III: 1, 5-diazabicyclo (4,3,0) non-5-ene: 1-dichloromethane: 0.84-0.9: 5 to 8(W/W/V, g/g/mL).

The dosage of the organic solvent A solution of the organic acid C is as follows: intermediate III: an organic acid C: organic solvent a ═ 1: 0.85-0.9: 10 to 14(W/V/V, g/mL/mL).

And (3) TLC detection: using trichloromethane/isopropanol (volume ratio 96/4) as developing agent, detecting that intermediate III spot should basically disappear, wherein intermediate III Rf is about 0.3, and intermediate IV Rf is about 0.8. Heating the reaction solution to 0-10 ℃, washing the organic phase with purified water, 0.1mol/L hydrochloric acid solution and 2% sodium bicarbonate solution in turn, concentrating the organic phase to a certain volume in a vacuum state (an intermediate III: the volume of the concentrated solution is 1:10(W/V, g/mL)), adding n-hexane for crystallization in a stirring state, and performing suction filtration and drying to obtain a solid intermediate compound IV. Preferably, the adding amount of the n-hexane is as follows: intermediate III: n-hexane 1:10(W/V, g/mL).

Step 3, preparation of intermediate compound v:

Adding the organic solvent A and the intermediate IV into a three-neck glass reaction bottle. Starting stirring, controlling the temperature to be-60 to-30 ℃, dropwise adding the ethanol solution of the reducing agent D, and stirring and reacting for 0.5 to 1 hour after the addition.

The organic solvent A is preferably dichloromethane or acetone.

The dosage of the organic solvent A is as follows: an intermediate IV: the organic solvent A is 1: 40-50 (W/V, g/mL).

The reducing agent D is preferably sodium hydride or lithium aluminum hydride or sodium borohydride; further preferred is sodium borohydride. The selective reducing agent is preferred to minimize the isomer impurities of intermediate v.

The dosage of the ethanol solution of the reducing agent D is as follows: an intermediate IV: reducing agent D: the absolute ethyl alcohol is 1: 0.02-0.04: 80-100 (W/W/V, g/g/mL).

After the reaction is finished, adding acetone into the reaction solution, and continuously stirring for 20-40 minutes, wherein the dosage of the acetone is as follows: an intermediate IV: acetone is 1: 1-3 (W/V, g/mL). Heating the reaction solution to 0-10 ℃, and detecting by TLC: with trichloromethane/isopropanol (volume ratio 96/4) as a developing solvent, the intermediate IV spots to be detected basically disappear, the intermediate III Rf is approximately equal to 0.3, the intermediate IV Rf is approximately equal to 0.8, and the intermediate V Rf is approximately equal to 0.25. Adding dichloromethane and purified water into the reaction solution (intermediate IV: purified water: dichloromethane is 1: 40-60: 30-40 (W/V/V, g/mL/mL)), stirring, standing for liquid separation, extracting the water phase with dichloromethane solution for 3 times (intermediate IV: dichloromethane is 1: 30-40 (W/V, g/mL)), combining the organic phases, concentrating under vacuum to a certain volume (intermediate IV: concentrated solution is 1:10(W/V, g/mL)), adding n-hexane under stirring for crystallization, and performing suction filtration and drying to obtain a solid intermediate compound V. Preferably, the adding amount of the n-hexane is as follows: an intermediate IV: n-hexane 1:10(W/V, g/mL).

Step 4, preparation of intermediate compound VI:

Purified water, intermediate v, was added to a three-necked glass reaction flask. Starting stirring, controlling the temperature to be 0-10 ℃, dropwise adding the alkali E, and stirring and reacting for 0.5 hour after the addition is finished.

The amount of the purified water is as follows: an intermediate V: purified water is 1: 40-60 (W/V, g/mL).

Preferably, the base E is a sodium hydroxide solution (sodium hydroxide: purified water 0.25 to 0.4:10 to 14(W/V, g/g/mL)) in an amount of: an intermediate V: and (3) sodium hydroxide is 1: 0.25-0.4 (W/W, g/g). The sodium hydroxide addition can also be expressed as: an intermediate V: sodium hydroxide: purified water is 1: 0.25-0.4: 10-14 (W/W/V, g/g/mL).

After the preparation, TLC detection: using trichloromethane/isopropanol (volume ratio: 96/4) as developing solvent, detecting that intermediate V spot should basically disappear, intermediate V Rf is about 0.25, and intermediate VI Rf is about 0. Adding a dichloromethane solution of methanol (intermediate V: methanol: dichloromethane: 1: 40-60: 8-10 (W/V/V, g/mL/mL)), stirring, and dropwise adding 0.5mol/L sodium bicarbonate solution to adjust the pH value to 7.6-7.7 and 7.7-7.8. Finally, 0.5mol/L sodium carbonate solution is used for adjusting the pH value to 8.0-8.2. Standing, separating, collecting organic phase, concentrating under vacuum to a certain volume (intermediate V: the volume of concentrated solution is 1:10(W/V, g/mL)), adding n-hexane under stirring for crystallization, and filtering and drying to obtain solid intermediate compound VI. Preferably, the adding amount of the n-hexane is as follows: an intermediate V: n-hexane 1:10(W/V, g/mL).

Step 5, preparing epirubicin hydrochloride I:

Adding an organic solvent A and an intermediate VI into a three-neck glass reaction bottle, starting stirring, controlling the temperature to be 15 ℃, dropwise adding a 4% hydrogen bromide organic solvent A solution into the reaction solution, stirring for 20-30 minutes after dropwise adding, dropwise adding a bromine organic solvent A solution into the reaction solution, keeping the temperature and stirring for reacting for 2-2.5 hours after dropwise adding. After the reaction is finished, adding a reducing agent F solution into the reaction solution, stirring for 10-15 minutes, adjusting the pH value to 4.5-5.0 by using an alkali G, stirring for reacting for 10-15 minutes, adjusting the pH value to 1.3-1.5 by using a 7% inorganic acid H solution, and carrying out heat preservation stirring reaction for 2-2.5 hours at the temperature of 25-35 ℃. And after the reaction is finished, adding 25% of sodium formate water solution, adjusting the pH value to 3.0-3.5, controlling the temperature to 25-35 ℃ and hydrolyzing the sodium formate for 1-3 hours to finally obtain the target compound epirubicin hydrochloride I crude product solution.

The dosage of the organic solvent A added into the three-neck glass reaction bottle is as follows: intermediate VI: the organic solvent A is 1: 40-60 (W/V, g/mL), and the organic solvent A is preferably methanol or 1, 4-dioxane.

The dosage of the organic solvent A solution of the 4 percent hydrogen bromide is as follows: intermediate VI: 4% hydrogen bromide in organic solvent a ═ 1: 4-8 (W/V, g/mL), and the organic solvent A is preferably methanol or 1, 4-dioxane.

the dosage of the bromine organic solvent A solution is as follows: intermediate VI: bromine: 1, 4-dioxane is 1: 0.2-0.5: 5-8 (W/W/V, g/g/mL), and the organic solvent A is preferably methanol or 1, 4-dioxane.

The dosage of the reducing agent F solution is as follows: intermediate VI: reducing agent F: purified water is 1:0.05 to 0.07:0.5 to 0.7(W/W/V, g/g/mL). Wherein the concentration of the reducing agent F solution is as follows: reducing agent F: purified water is 0.05 to 0.07:0.5 to 0.7(W/V, g/mL). The reducing agent F solution is preferably a sodium sulfite solution.

The alkali G is selected from one of sodium hydroxide, potassium hydroxide, calcium hydroxide, ammonia water and sodium bicarbonate, and most preferably, the ammonia water.

the inorganic acid H is selected from one of phosphoric acid, sulfuric acid, perchloric acid, boric acid, nitric acid, hydrochloric acid and hydrobromic acid, and hydrochloric acid is most preferable; the concentration of the inorganic acid is preferably 6-8%.

after the preparation, dichloromethane and purified water are added into the reaction liquid (intermediate VI: purified water: dichloromethane is 1: 40-60: 30-40 (W/V/V, g/mL/mL)), the mixture is stirred and kept stand for liquid separation, the water phase is extracted three times by using a dichloromethane solution (intermediate VI: dichloromethane is 1: 30-40 (W/V, g/mL)), the organic phases are combined and concentrated to a certain volume under a vacuum state (intermediate VI: the volume of the concentrated solution is 1:10(W/V, g/mL)), n-hexane is added under a stirring state for crystallization, and solid epirubicin hydrochloride is obtained after suction filtration and drying. Preferably, the addition amount of the n-hexane is as follows: intermediate VI: n-hexane 1:10(W/V, g/mL).

The invention has the advantages that:

Various reagents adopted are cheap and easy to obtain, and mature treatment methods are provided for generated pollutants, so that the generated environmental pollution is small. The whole method is simple and easy to operate, has no strict technical requirements on practitioners, has stable reaction of each reaction step, is easy to control, has loose requirements on water content of the generated intermediate, produces few impurities, and has high purity, stable quality and high yield.

The intermediate generated in the prior art is easy to interconvert ketone type and enol type, the transition state intermediate with enol type structure is extremely unstable and is easy to be affected by moisture in air (for example, in summer with high air humidity, the conversion rate of the intermediate I to the intermediate II in the original process is extremely unstable, the humidity and the yield are in an obvious negative correlation state, namely the higher the humidity is, the lower the conversion rate is, the yield in summer is 30% -40%, the conversion rate in other steps is correspondingly reduced under the high humidity condition), the reaction is hindered due to the influence of temperature, and even the conversion is changed into other impurities, so that the product yield and the product purity are directly affected. According to the invention, triethyl orthoformate is directly reacted with the raw material to protect the ketone group, so that tautomerism between the ketone group and an enol structure is inhibited, and smooth reaction is ensured.

The route for synthesizing epirubicin hydrochloride provided by the prior art takes 4' -epidaunorubicin as a raw material, the pH is adjusted by 5% sodium formate solution after bromination is finished, and excess bromine is neutralized by sodium bisulfite solution. In the process, the excessive bromine is neutralized by sodium bisulfite with the prescription amount directly, and then the pH value is adjusted by saturated sodium bicarbonate solution, so that the obtained reaction liquid intermediate can exist stably, the impurities can not be generated under the same concentration condition, and the yield (the mass yield, namely the yield of the conversion step from the compound VI to the VI-1) of about 40 percent of the original process can be increased to more than 95 percent; on the basis, the mass yield of the crude product before column chromatography separation is improved to more than 85 percent from the original 40 to 60 percent, and the concentration time can be greatly shortened after the process is improved, thereby directly saving the consumption of manpower, energy and material resources.

In conclusion, the method is efficient, economical and simple in production operation, and completely meets the requirements of industrial mass production.

Detailed Description

The present invention will be further explained with reference to specific examples, which are not intended to limit the invention itself in any manner.

example one

Preparation of intermediate compound iii:

to a 1000mL three-necked glass reaction bottle were added dichloromethane and daunorubicin hydrochloride (daunorubicin hydrochloride: dichloromethane ═ 1:60(W/V, g/mL)). Starting stirring, controlling the temperature to be 5 ℃, checking the dissolution condition of the materials after 10-15 minutes, and continuing stirring if the materials are not completely dissolved; if the solution is completely dissolved, the solution is cooled to 0 ℃, a methanol solution of camphorsulfonic acid (daunorubicin hydrochloride: methanol: camphorsulfonic acid ═ 1:177:1(W/V/W, g/mL/g)) is added thereto, and after stirring for 1 hour, trimethyl orthoformate (daunorubicin hydrochloride: trimethyl orthoformate ═ 1:2(W/V, g/mL)) is added; stirring for 2 hours after the addition is finished; after completion of the stirring, the reaction solution was warmed to 5 ℃ and trifluoroacetic anhydride (daunorubicin hydrochloride: trifluoroacetic anhydride ═ 1:2(W/V, g/mL) was added to the flask, and the mixture was stirred for 1 hour.

After the reaction, methanol (daunorubicin hydrochloride: methanol 1:30(W/V, g/mL)) was added to the reaction mixture, and after stirring for 10 minutes, 8% sodium bicarbonate solution was added to adjust the pH to 7.0 to 8.0. Controlling the temperature to be 15-25 ℃, and carrying out heat preservation stirring reaction for 3-5 hours. And (3) TLC detection: the chloroform/isopropanol (volume ratio 96/4) is used as developing solvent, and the raw material spots are detected to be basically disappeared. The intermediate compound III Rf is approximately equal to 0.3, and the daunorubicin hydrochloride Rf is approximately equal to 0. Standing and separating liquid, and collecting a lower organic phase. Concentrating under vacuum to a certain volume (daunorubicin hydrochloride: the volume of the concentrated solution is 1:10(W/V, g/mL)), adding n-hexane (daunorubicin hydrochloride: the volume of n-hexane is 1:10(W/V, g/mL)) under stirring, crystallizing, and performing suction filtration and drying to obtain a solid intermediate compound III with the mass yield of 95.6%. The purity is 95.5%.

Example two

Preparation of intermediate compound iii:

To a 1000mL three-necked glass reaction flask were added 1, 4-dioxane and daunorubicin hydrochloride (daunorubicin hydrochloride: 1, 4-dioxane ═ 1:70(W/V, g/mL)). Starting stirring, controlling the temperature to be 10 ℃ for 10-15 minutes, checking the dissolution condition of the materials, and continuing stirring if the materials are not completely dissolved; if the solution is completely dissolved, the temperature of the feed solution is reduced to 5 ℃, a methanol solution of camphorsulfonic acid (daunorubicin hydrochloride: methanol: camphorsulfonic acid ═ 1:177:1(W/V/W, g/mL/g)) is added thereto, and after stirring for 1 hour, trimethyl orthoformate (daunorubicin hydrochloride: trimethyl orthoformate ═ 1:1(W/V, g/mL)) is added; stirring for 3 hours after the addition is finished; after completion of the stirring, the reaction solution was warmed to 10 ℃ and trifluoroacetic anhydride (daunorubicin hydrochloride: trifluoroacetic anhydride 1:4(W/V, g/mL)) was added to the flask, and the mixture was stirred for 1.5 hours.

After the reaction, methanol (daunorubicin hydrochloride: methanol 1:40(W/V, g/mL)) was added to the reaction mixture, and after stirring for 10 minutes, 8% sodium bicarbonate solution was added to adjust the pH to 7.0 to 8.0. Controlling the temperature to be 15-25 ℃, and carrying out heat preservation stirring reaction for 3-5 hours. And (3) TLC detection: the chloroform/isopropanol (volume ratio 96/4) is used as developing solvent, and the raw material spots are detected to be basically disappeared. The intermediate compound III Rf is approximately equal to 0.3, and the daunorubicin hydrochloride Rf is approximately equal to 0. Standing and separating liquid, and collecting a lower organic phase. Concentrating under vacuum to a certain volume (daunorubicin hydrochloride: the volume of the concentrated solution is 1:10(W/V, g/mL)), adding n-hexane (daunorubicin hydrochloride: the volume of n-hexane is 1:10(W/V, g/mL)) under stirring, crystallizing, and performing suction filtration and drying to obtain a solid intermediate compound III with the mass yield of 93.7%. The purity is 95.4%.

EXAMPLE III

preparation of intermediate compound iii:

To a 1000mL three-necked glass reaction flask were added tetrahydrofuran and daunorubicin hydrochloride (daunorubicin hydrochloride: tetrahydrofuran ═ 1:50(W/V, g/mL)). Starting stirring, controlling the temperature to be 8 ℃, checking the dissolution condition of the materials after 10-15 minutes, and continuing stirring if the materials are not completely dissolved; if the solution is completely dissolved, the temperature of the feed solution is reduced to 3 ℃, a methanol solution of camphorsulfonic acid (daunorubicin hydrochloride: methanol: camphorsulfonic acid ═ 1:177:1(W/V/W, g/mL/g)) is added thereto, and after stirring for 1 hour, trimethyl orthoformate (daunorubicin hydrochloride: trimethyl orthoformate ═ 1:5(W/V, g/mL)) is added; stirring for 3 hours after the addition is finished; after completion of the stirring, the reaction solution was warmed to 3 ℃ and trifluoroacetic anhydride (daunorubicin hydrochloride: trifluoroacetic anhydride 1:5(W/V, g/mL)) was added to the flask, and the mixture was stirred for 1.5 hours.

After the reaction, methanol (daunorubicin hydrochloride: methanol 1:20(W/V, g/mL)) was added to the reaction mixture, and after stirring for 15 minutes, 8% sodium bicarbonate solution was added to adjust the pH to 7.0 to 8.0. Controlling the temperature to be 15-25 ℃, and carrying out heat preservation stirring reaction for 3-5 hours. And (3) TLC detection: the chloroform/isopropanol (volume ratio 96/4) is used as developing solvent, and the raw material spots are detected to be basically disappeared. The intermediate compound III Rf is approximately equal to 0.3, and the daunorubicin hydrochloride Rf is approximately equal to 0. Standing and separating liquid, and collecting a lower organic phase. Concentrating under vacuum to a certain volume (daunorubicin hydrochloride: the volume of the concentrated solution is 1:10(W/V, g/mL)), adding n-hexane (daunorubicin hydrochloride: the volume of n-hexane is 1:10(W/V, g/mL)) under stirring, crystallizing, and performing suction filtration and drying to obtain a solid intermediate compound III with the mass yield of 92.4%. The purity is 95.0%.

Example four

preparation of intermediate compound iii:

To a 1000mL three-necked glass reaction flask were added n-heptane and daunorubicin hydrochloride (daunorubicin hydrochloride: n-heptane ═ 1:40(W/V, g/mL)). Starting stirring, controlling the temperature at 15 ℃, checking the dissolution condition of the materials after 10-15 minutes, and continuing stirring if the materials are not completely dissolved; if the solution is completely dissolved, the solution is cooled to 11 ℃, a methanol solution of camphorsulfonic acid (daunorubicin hydrochloride: methanol: camphorsulfonic acid ═ 1:200:3(W/V/W, g/mL/g)) is added thereto, and after stirring for 1 hour, triethyl orthoformate (daunorubicin hydrochloride: triethyl orthoformate ═ 1:7(W/V, g/mL)) is added; stirring for 3 hours after the addition is finished; after completion of the stirring, the reaction solution was warmed to 15 ℃ and trifluoroacetic anhydride (daunorubicin hydrochloride: trifluoroacetic anhydride 1:7(W/V, g/mL)) was added to the flask, and the mixture was stirred for 1.5 hours.

After completion of the reaction, methanol (daunorubicin hydrochloride: methanol 1:50(W/V, g/mL)) was added to the reaction mixture, and after stirring for 15 minutes, 8% sodium hydrogencarbonate solution was added to adjust the pH to 8.5. Controlling the temperature to be 15-25 ℃, and carrying out heat preservation stirring reaction for 3-5 hours. And (3) TLC detection: the chloroform/isopropanol (volume ratio 96/4) is used as developing solvent, and the raw material spots are detected to be basically disappeared. The intermediate compound III Rf is approximately equal to 0.3, and the daunorubicin hydrochloride Rf is approximately equal to 0. Standing and separating liquid, and collecting a lower organic phase. Concentrating under vacuum to a certain volume (daunorubicin hydrochloride: the volume of the concentrated solution is 1:15(W/V, g/mL)), adding n-hexane (daunorubicin hydrochloride: the volume of n-hexane is 1:8(W/V, g/mL)) under stirring, crystallizing, and performing suction filtration and drying to obtain a solid intermediate compound III with the mass yield of 90.0%. The purity was 93.7%.

EXAMPLE five

preparation of intermediate compound iv:

To a 1000mL three-necked glass reaction vessel was added methylene chloride (intermediate III: methylene chloride: 1:40(W/V, g/mL)). Stirring is started, the temperature is controlled to be-40 ℃, a dichloromethane solution of trifluoroacetic anhydride is slowly added (intermediate III: trifluoroacetic anhydride: dichloromethane ═ 1:0.8: 2.4(W/W/V, g/g/mL)) and after stirring reaction is carried out for 1 hour at-40 ℃, a dichloromethane solution of epirubicin hydrochloride intermediate III (intermediate III: dichloromethane ═ 1:5(W/V, g/mL)) is slowly dropped into the reaction liquid; after completion of the addition, the mixture was stirred for 20 minutes, a dichloromethane solution of 1, 5-diazabicyclo (4,3,0) non-5-ene (intermediate iii: 1, 5-diazabicyclo (4,3,0) non-5-ene: dichloromethane ═ 1: 0.84: 5(W/V, g/g/mL)) was added dropwise thereto, and after completion of the addition and stirring for 10 minutes, a dichloromethane solution of glacial acetic acid (intermediate iii: glacial acetic acid: dichloromethane ═ 1: 0.85: 10(W/V, g/mL)) was added thereto, and the reaction was stirred for 0.5 hour.

and (3) TLC detection: using trichloromethane/isopropanol (volume ratio 96/4) as developing agent, detecting that intermediate III spot should basically disappear, wherein intermediate III Rf is about 0.3, and intermediate IV Rf is about 0.8. Heating the reaction solution to 0-10 ℃, washing the organic phase with purified water, 0.1mol/L hydrochloric acid solution and 2% sodium bicarbonate solution in turn, concentrating the organic phase to a certain volume under a vacuum state (intermediate III: the volume of the concentrated solution is 1:10(W/V, g/mL)), adding n-hexane (intermediate III: the volume of the n-hexane is 1:10(W/V, g/mL)) under a stirring state for crystallization, and performing suction filtration and drying to obtain a solid intermediate compound IV with the mass yield of 93.2%. The purity is 86.2 percent, and the content of related substances (intermediate compounds III) is less than 8.0 percent.

EXAMPLE six

preparation of intermediate compound iv:

To a 1000mL three-necked glass reaction flask was added ethyl acetate (intermediate III: ethyl acetate: 1:20(W/V/V, g/mL/mL)). Starting stirring, controlling the temperature to be 70 ℃ below zero, slowly adding an ethyl acetate solution of trifluoroacetic anhydride (intermediate III: trifluoroacetic anhydride: ethyl acetate ═ 1:0.5: 3(W/W/V, g/g/mL)), stirring at 70 ℃ below zero for 1 hour, and then slowly dropwise adding an ethyl acetate solution of epirubicin hydrochloride intermediate III (intermediate III: ethyl acetate ═ 1:8(W/V, g/mL)); after the completion of the addition, the mixture was stirred for 20 minutes, and then an ethyl acetate solution of 1, 5-diazabicyclo (4,3,0) non-5-ene (intermediate iii: 1, 5-diazabicyclo (4,3,0) non-5-ene: ethyl acetate: 1: 0.9: 8(W/V, g/g/mL)) was added dropwise thereto, and after the completion of the addition, an ethyl acetate solution of malonic acid (intermediate iii: malonic acid: ethyl acetate: 1: 0.9: 14(W/V, g/mL)) was added thereto and the reaction was stirred for 0.5 hour.

And (3) TLC detection: using trichloromethane/isopropanol (volume ratio 96/4) as developing agent, detecting that intermediate III spot should basically disappear, wherein intermediate III Rf is about 0.3, and intermediate IV Rf is about 0.8. Heating the reaction solution to 0-10 ℃, washing the organic phase with purified water, 0.1mol/L hydrochloric acid solution and 2% sodium bicarbonate solution in turn, concentrating the organic phase to a certain volume under a vacuum state (intermediate III: the volume of the concentrated solution is 1:10(W/V, g/mL)), adding n-hexane (intermediate III: the volume of the n-hexane is 1:10(W/V, g/mL)) under a stirring state for crystallization, and performing suction filtration and drying to obtain a solid intermediate compound IV with the mass yield of 93.1%. The purity is 85.5%, and the content of related substances (intermediate compound III) is less than 8.0%.

EXAMPLE seven

Preparation of intermediate compound iv:

To a 1000mL three-necked glass reaction flask was added DMF (intermediate iii: DMF ═ 1:50(W/V, g/mL)). Stirring is started, the temperature is controlled to be-30 ℃, a DMF solution of trifluoroacetic anhydride (intermediate III: trifluoroacetic anhydride: DMF 1:0.4: 4(W/W/V, g/g/mL)) is slowly added, and after stirring reaction is carried out for 1.5 hours at-30 ℃, a DMF solution of epirubicin hydrochloride intermediate III (intermediate III: DMF 1:10(W/V, g/mL)) is slowly dropped into the reaction liquid; after completion of the addition, the mixture was stirred for 20 minutes, a DMF solution of 1, 5-diazabicyclo (4,3,0) non-5-ene (intermediate III: 1, 5-diazabicyclo (4,3,0) non-5-ene: DMF 1: 0.95: 10(W/W/V, g/g/mL)) was added dropwise thereto, and after completion of the addition and stirring for 10 minutes, a DMF solution of formic acid (intermediate III: malonic acid: DMF 1: 0.9: 16(W/V/V, g/mL/mL)) was added thereto, and the reaction was stirred for 0.5 hour.

And (3) TLC detection: using trichloromethane/isopropanol (volume ratio 96/4) as developing agent, detecting that intermediate III spot should basically disappear, wherein intermediate III Rf is about 0.3, and intermediate IV Rf is about 0.8. Heating the reaction solution to 0-10 ℃, washing the organic phase with purified water, 0.1mol/L hydrochloric acid solution and 2% sodium bicarbonate solution in turn, concentrating the organic phase to a certain volume under a vacuum state (intermediate III: the volume of the concentrated solution is 1:15(W/V, g/mL)), adding n-hexane (intermediate III: the volume of the n-hexane is 1:7(W/V, g/mL)) under a stirring state for crystallization, and performing suction filtration and drying to obtain a solid intermediate compound IV with the mass yield of 91.7%. Purity 84.1%, content of related substance (intermediate compound III) is less than 8.0%.

Example eight

Preparation of intermediate compound v:

To a 1000mL three-necked glass reaction flask were added dichloromethane and intermediate iv (intermediate iv: dichloromethane ═ 1:40(W/V, g/mL)). Stirring was started, the temperature was controlled at-60 ℃, an ethanol solution of sodium borohydride (intermediate iv: sodium borohydride: absolute ethanol: 1:0.02:100 (W/V, g/g/mL)) was added dropwise, and after completion of the addition, the reaction mixture was stirred for 0.5 hour, acetone (intermediate iv: acetone: 1(W/V, g/mL)) was added to the reaction mixture, and stirring was continued for 20 minutes.

heating the reaction solution to 0-10 ℃, and detecting by TLC: with trichloromethane/isopropanol (volume ratio 96/4) as a developing solvent, the intermediate IV spots to be detected basically disappear, the intermediate III Rf is approximately equal to 0.3, the intermediate IV Rf is approximately equal to 0.8, and the intermediate V Rf is approximately equal to 0.25. Adding dichloromethane and purified water into the reaction solution (intermediate IV: purified water: dichloromethane is 1: 60: 30(W/V/V, g/mL/mL)), stirring, standing for liquid separation, extracting the water phase with dichloromethane solution three times (intermediate IV: dichloromethane is 1:30(W/V, g/mL)), combining the organic phases, concentrating under vacuum to a certain volume (intermediate IV: concentrated solution is 1:10(W/V, g/mL)), detecting the intermediate V compound, wherein the purity is 87.3%, adding n-hexane (intermediate IV: n-hexane is 1:10(W/V, g/mL)) under stirring for crystallization, and performing suction filtration and drying to obtain the solid intermediate compound V, wherein the mass yield is 93.9%. The purity is 99.7%, and the content of related substance intermediate compound I is less than 0.6%.

Example nine

Preparation of intermediate compound v:

to a 1000mL three-necked glass reaction flask was added acetone and intermediate iv (intermediate iv: acetone ═ 1:50(W/V, g/mL)). Stirring was started, the temperature was controlled at-30 ℃, an ethanol solution of lithium aluminum hydride (intermediate iv: lithium aluminum hydride: absolute ethanol: 1:0.04:80 (W/V, g/g/mL)) was added dropwise, and after completion of the addition, the reaction mixture was stirred for 1 hour, acetone (intermediate iv: acetone: 1:2(W/V, g/mL)) was added to the reaction mixture, and stirring was continued for 20 minutes.

Heating the reaction solution to 0-10 ℃, and detecting by TLC: with trichloromethane/isopropanol (volume ratio 96/4) as a developing solvent, the intermediate IV spots to be detected basically disappear, the intermediate III Rf is approximately equal to 0.3, the intermediate IV Rf is approximately equal to 0.8, and the intermediate V Rf is approximately equal to 0.25. Adding dichloromethane and purified water into the reaction solution (intermediate IV: purified water: dichloromethane is 1: 40: 40(W/V, g/mL/mL)), stirring, standing for liquid separation, extracting the water phase with dichloromethane solution three times (intermediate IV: dichloromethane is 1:40(W/V, g/mL)), combining the organic phases, concentrating under vacuum to a certain volume (intermediate IV: concentrated solution is 1:10(W/V, g/mL)), detecting the intermediate V compound, detecting the purity of 88.1%, adding n-hexane (intermediate IV: n-hexane is 1:10(W/V, g/mL)) under stirring for crystallization, and performing suction filtration and drying to obtain a solid intermediate compound V with the mass yield of 92.7%. The purity is 99.6%, and the content of related substance intermediate compound I is less than 0.6%.

example ten

Preparation of intermediate compound v:

to a 1000mL three-necked glass reaction flask, tetrahydrofuran and intermediate iv (intermediate iv: tetrahydrofuran ═ 1:60(W/V, g/mL)) were added. Stirring was started, the temperature was controlled at-75 ℃, an ethanol solution of sodium cyanoborohydride (intermediate iv: sodium cyanoborohydride: anhydrous ethanol: 1:0.06:70 (W/V, g/g/mL)) was added dropwise, and after completion of the addition, the reaction mixture was stirred for 1 hour, acetone (intermediate iv: acetone: 1:4(W/V, g/mL)) was added to the reaction mixture, and stirring was continued for 20 minutes.

Heating the reaction solution to 0-10 ℃, and detecting by TLC: with trichloromethane/isopropanol (volume ratio 96/4) as a developing solvent, the intermediate IV spots to be detected basically disappear, the intermediate III Rf is approximately equal to 0.3, the intermediate IV Rf is approximately equal to 0.8, and the intermediate V Rf is approximately equal to 0.25. Adding dichloromethane and purified water into the reaction solution (intermediate IV: purified water: dichloromethane is 1: 30: 50(W/V, g/mL/mL)), stirring, standing for liquid separation, extracting the water phase with dichloromethane solution three times (intermediate IV: dichloromethane is 1:50(W/V, g/mL)), combining the organic phases, concentrating under vacuum to a certain volume (intermediate IV: concentrated solution is 1:15(W/V, g/mL)), detecting the intermediate V compound, detecting the purity of 88.3%, adding n-hexane (intermediate IV: n-hexane is 1:12(W/V, g/mL)) under stirring for crystallization, and performing suction filtration and drying to obtain a solid intermediate compound V with the mass yield of 92.5%. The purity is 99.5%, and the content of related substance intermediate compound I is less than 0.6%.

EXAMPLE eleven

preparation of intermediate compound v:

To a 1000mL three-necked glass reaction flask, cyclohexane and intermediate iv (intermediate iv: cyclohexane 1:40(W/V, g/mL)) were added. Stirring was started, the temperature was controlled at 5 ℃, an ethanol solution of lithium triethylborohydride (intermediate iv: lithium triethylborohydride: absolute ethanol: 1:0.02:100 (W/V, g/g/mL)) was added dropwise, and after completion of the addition, the reaction mixture was stirred for 1.5 hours, acetone (intermediate iv: acetone: 1:4(W/V, g/mL)) was added to the reaction mixture, and stirring was continued for 20 minutes.

Heating the reaction solution to 0-10 ℃, and detecting by TLC: with trichloromethane/isopropanol (volume ratio 96/4) as a developing solvent, the intermediate IV spots to be detected basically disappear, the intermediate III Rf is approximately equal to 0.3, the intermediate IV Rf is approximately equal to 0.8, and the intermediate V Rf is approximately equal to 0.25. Adding dichloromethane and purified water into the reaction solution (intermediate IV: purified water: dichloromethane is 1: 50: 50(W/V, g/mL/mL)), stirring, standing for liquid separation, extracting the water phase with dichloromethane solution three times (intermediate IV: dichloromethane is 1:50(W/V, g/mL)), combining the organic phases, concentrating under vacuum to a certain volume (intermediate IV: concentrated solution is 1:8(W/V, g/mL)), detecting the intermediate V compound, wherein the purity is 87.8%, adding n-hexane (intermediate IV: n-hexane is 1:8(W/V, g/mL)) under stirring for crystallization, and performing suction filtration and drying to obtain the solid intermediate compound V, wherein the mass yield is 89.1%. The purity is 98.4%, and the content of related substance intermediate compound I is less than 0.6%.

Example twelve

Preparation of intermediate compound vi:

To a 1000mL three-necked glass reaction flask was added purified water, intermediate V (intermediate V: purified water ═ 1:60(W/V, g/mL)). Stirring was started, the temperature was controlled at 0 ℃, and a sodium hydroxide solution was added dropwise (intermediate V: sodium hydroxide: purified water ═ 1:0.25:10 (W/V, g/g/mL)), and after completion of the addition, the reaction was stirred for 0.5 hour, and TLC detection: using trichloromethane/isopropanol (volume ratio: 96/4) as developing solvent, detecting that intermediate V spot should basically disappear, intermediate V Rf is about 0.25, and intermediate VI Rf is about 0.

A dichloromethane solution of methanol (intermediate V: methanol: dichloromethane: 1: 60: 8(W/V, g/mL)) was added to the reaction solution, stirring was started, and a 0.5mol/L sodium bicarbonate solution was added dropwise to adjust the pH to 7.6 to 7.8. Finally, 0.5mol/L sodium carbonate solution is used for adjusting the pH value to 8.0-8.2. Standing, separating, collecting an organic phase, concentrating under vacuum to a certain volume (intermediate V: the volume of the concentrated solution is 1:10(W/V, g/mL)), adding n-hexane (intermediate V: the volume of the n-hexane is 1:10(W/V, g/mL)) under stirring, crystallizing, and performing suction filtration and drying to obtain a solid intermediate compound VI with the mass yield of 94.6%. The purity is 98.9%, the maximum impurity content is less than 0.6%, and the total impurity content is less than 3.0%.

EXAMPLE thirteen

Preparation of intermediate compound vi:

To a 1000mL three-necked glass reaction flask was added purified water and intermediate V (intermediate V: purified water ═ 1:40(W/V, g/mL)). Stirring was started, the temperature was controlled at 10 ℃, and sodium hydroxide solution was added dropwise (intermediate V: sodium hydroxide: purified water ═ 1:0.4:14 (W/V, g/g/mL)), and after completion of the addition, the reaction was stirred for 0.5 hour, and TLC detection: using trichloromethane/isopropanol (volume ratio: 96/4) as developing solvent, detecting that intermediate V spot should basically disappear, intermediate V Rf is about 0.25, and intermediate VI Rf is about 0.

a dichloromethane solution of methanol (intermediate V: methanol: dichloromethane: 1: 40: 10(W/V, g/mL)) was added to the reaction solution, stirring was started, and a 0.5mol/L sodium bicarbonate solution was added dropwise to adjust the pH to 7.6 to 7.8. Finally, 0.5mol/L sodium carbonate solution is used for adjusting the pH value to 8.0-8.2. Standing, separating, collecting an organic phase, concentrating under vacuum to a certain volume (intermediate V: the volume of the concentrated solution is 1:10(W/V, g/mL)), adding n-hexane (intermediate V: the volume of the n-hexane is 1:10(W/V, g/mL)) under stirring, crystallizing, and performing suction filtration and drying to obtain a solid intermediate compound VI with the mass yield of 93.1%. The purity is 98.8%, the maximum impurity content is less than 0.6%, and the total impurity content is less than 3.0%.

Example fourteen

Preparation of intermediate compound vi:

to a 1000mL three-necked glass reaction flask was added purified water and intermediate V (intermediate V: purified water ═ 1:70(W/V, g/mL)). Stirring was started, the temperature was controlled at 20 ℃, a sodium bicarbonate solution was added dropwise (intermediate V: sodium bicarbonate: purified water: 1:1.5:10 (W/V, g/g/mL)), and the addition was completed, the reaction was stirred for 1 hour, and TLC detection: using trichloromethane/isopropanol (volume ratio: 96/4) as developing solvent, detecting that intermediate V spot should basically disappear, intermediate V Rf is about 0.25, and intermediate VI Rf is about 0.

A dichloromethane solution of methanol (intermediate V: methanol: dichloromethane: 1: 35: 15(W/V, g/mL)) was added to the reaction solution, stirring was started, and a 0.5mol/L sodium bicarbonate solution was added dropwise to adjust the pH to 7.6 to 7.8. Finally, 0.5mol/L sodium carbonate solution is used for adjusting the pH value to 8.0-8.2. Standing, separating, collecting an organic phase, concentrating under vacuum to a certain volume (intermediate V: the volume of the concentrated solution is 1:10(W/V, g/mL)), adding n-hexane (intermediate V: the volume of the n-hexane is 1:10(W/V, g/mL)) under stirring, crystallizing, and performing suction filtration and drying to obtain a solid intermediate compound VI with the mass yield of 92.1%. The purity is 98.2%, the maximum impurity content is less than 0.6%, and the total impurity content is less than 3.0%.

Example fifteen

Preparation of intermediate compound vi:

To a 1000mL three-necked glass reaction flask was added purified water and intermediate V (intermediate V: purified water ═ 1:30(W/V, g/mL)). Stirring is started, 25 ℃ is controlled, calcium hydroxide solution is added dropwise, (intermediate V: calcium hydroxide: purified water is 1:1.1:10(W/W/V, g/g/mL)), the addition is finished, the reaction is stirred for 1.5 hours, and TLC detection: using trichloromethane/isopropanol (volume ratio: 96/4) as developing solvent, detecting that intermediate V spot should basically disappear, intermediate V Rf is about 0.25, and intermediate VI Rf is about 0.

A dichloromethane solution of methanol (intermediate V: methanol: dichloromethane: 1: 65: 15(W/V, g/mL)) was added to the reaction solution, stirring was started, and a 0.5mol/L sodium bicarbonate solution was added dropwise to adjust the pH to 7.6 to 7.8. Finally, 0.5mol/L sodium carbonate solution is used for adjusting the pH value to 8.0-8.2. Standing, separating, collecting an organic phase, concentrating under vacuum to a certain volume (intermediate V: the volume of the concentrated solution is 1:12(W/V, g/mL)), adding n-hexane (intermediate V: the volume of the n-hexane is 1:8(W/V, g/mL)) under stirring, crystallizing, and performing suction filtration and drying to obtain a solid intermediate compound VI with the mass yield of 91.2%. The purity is 97.8%, the maximum impurity content is less than 0.6%, and the total impurity content is less than 3.0%.

Example sixteen

Preparation of epirubicin hydrochloride I:

To a 1000mL three-necked glass reaction flask, methanol and intermediate vi (intermediate vi: methanol ═ 1:40(W/V, g/mL)) were added. Stirring was started, the temperature was controlled at 15 ℃, a 4% methanol solution of hydrogen bromide (intermediate vi: 4% methanol solution of hydrogen bromide: 1:4(W/V, g/mL)) was added dropwise to the reaction solution, and after stirring was completed for 20 minutes, a methanol solution of bromine (intermediate vi: bromine: methanol: 1:0.5: 5(W/V, g/g/mL)) was added dropwise to the reaction solution, and after completion of the addition, the reaction was stirred for 2 hours under heat preservation. After the reaction, a sodium sulfite solution (intermediate vi: sodium bisulfite: purified water: 1:0.05:0.5 (W/V, g/g/mL)) was added to the reaction mixture, stirred for 10 minutes, then adjusted to pH 4.5 to 5.0 with ammonia water, stirred for 10 minutes, adjusted to pH 1.3 to 1.5 with 7% hydrochloric acid solution, and then stirred and reacted at 25 ℃ to 35 ℃ for 2 hours with heat preservation. And after the reaction is finished, adding 25% sodium formate aqueous solution, adjusting the pH to 3.0-3.5, controlling the temperature to 25-35 ℃ and hydrolyzing the sodium formate for 1 hour to finally obtain the target compound epirubicin hydrochloride I crude product solution.

Dichloromethane and purified water are added into the reaction liquid (intermediate VI: purified water: dichloromethane is 1: 60: 30(W/V/V, g/mL)), the mixture is stirred and kept stand for liquid separation, the water phase is extracted three times by using dichloromethane solution (intermediate VI: dichloromethane is 1:30(W/V, g/mL)), the organic phases are combined and concentrated to a certain volume under vacuum (intermediate VI: concentrated solution is 1:10(W/V, g/mL)), normal hexane (intermediate VI: normal hexane is 1:10(W/V, g/mL)) is added under stirring for crystallization, solid epirubicin hydrochloride I is obtained after suction filtration and drying, and the mass yield is 94.1%. The product is red powder with the purity of 98.7 percent, and meets the requirements of pharmacopoeia: if an impurity peak exists in the chromatogram of the test solution, the area of the doxorubicin (relative retention time is about 0.3) is calculated according to the corrected peak area (multiplied by a correction factor of 0.7), and is not more than the main peak area (1.0) of the control solution, the area of the doxorubicin (relative retention time is about 0.8) is not more than the main peak area (1.0) of the control solution, the area of the other single impurity peak is not more than 0.5 times (0.5) of the main peak area of the control solution, and the sum of the areas of the impurities is not more than 2 times (2.0) of the main peak area of the control solution.

Example seventeen

Preparation of epirubicin hydrochloride I:

To a 1000mL three-necked glass reaction flask, 1, 4-dioxane and intermediate vi (intermediate vi: 1, 4-dioxane: 1:60(W/V, g/mL)) were added. Stirring was started, the temperature was controlled at 0 ℃, a 4% 1, 4-dioxane solution of hydrogen bromide (intermediate vi: 4% 1, 4-dioxane solution of hydrogen bromide: 1:8(W/V, g/mL)) was added dropwise to the reaction solution, after completion of the addition, a 1, 4-dioxane solution of bromine (intermediate vi: 1, 4-dioxane: 1:0.2: 8(W/V, g/mL)) was added dropwise to the reaction solution after stirring for 20 minutes, and after completion of the addition, the reaction was stirred for 2.5 hours under heat preservation. After the reaction, a ferrous sulfate solution (intermediate vi: ferrous sulfate: purified water: 1:0.07:0.7 (W/V, g/g/mL)) was added to the reaction mixture, and stirred for 15 minutes, then the PH was adjusted to 4.5 to 5.0 with a potassium hydroxide solution (intermediate V: potassium hydroxide: purified water: 1:0.25:10 (W/V, g/g/mL)), stirred for 10 minutes, adjusted to 1.3 to 1.5 with a 7% phosphoric acid solution, and stirred for 2.5 hours at 25 to 35 ℃. And after the reaction is finished, adding 20% sodium formate aqueous solution, adjusting the pH value to 3.0-3.5, controlling the temperature to 25-35 ℃ and hydrolyzing the sodium formate for 3 hours to finally obtain the target compound epirubicin hydrochloride I crude product solution.

Dichloromethane and purified water are added into the reaction liquid (intermediate VI: purified water: dichloromethane is 1: 40: 40(W/V, g/mL)), the mixture is stirred and kept stand for liquid separation, the water phase is extracted three times by using dichloromethane solution (intermediate VI: dichloromethane is 1:40(W/V, g/mL)), the organic phases are combined and concentrated to a certain volume under vacuum (intermediate VI: concentrated solution is 1:10(W/V, g/mL)), normal hexane is added under stirring (intermediate VI: normal hexane is 1:10(W/V, g/mL)) for crystallization, solid epirubicin hydrochloride is obtained after suction filtration and drying, and the mass yield is 92.2%. The product is red powder, the purity is 98.1 percent, and the product meets the requirements of pharmacopoeia: if an impurity peak exists in the chromatogram of the test solution, the area of the doxorubicin (relative retention time is about 0.3) is calculated according to the corrected peak area (multiplied by a correction factor of 0.7), and is not more than the main peak area (1.0) of the control solution, the area of the doxorubicin (relative retention time is about 0.8) is not more than the main peak area (1.0) of the control solution, the area of the other single impurity peak is not more than 0.5 times (0.5) of the main peak area of the control solution, and the sum of the areas of the impurities is not more than 2 times (2.0) of the main peak area of the control solution.

EXAMPLE eighteen

Preparation of epirubicin hydrochloride I:

To a 1000mL three-necked glass reaction flask was added chlorotrifluoroethylene, intermediate vi (intermediate vi: trichloroethylene ═ 1:40(W/V, g/mL)). Stirring was started, the temperature was controlled at 30 ℃, a 4% hydrogen bromide trichloroethylene solution (intermediate vi: 4% hydrogen bromide trichloroethylene solution 1:4(W/V, g/mL)) was added dropwise to the reaction mixture, after completion of the addition, a bromine trichloroethylene solution (intermediate vi: bromine: trichloroethylene 1:0.5: 5(W/V, g/g/mL)) was added dropwise to the reaction mixture, and after completion of the addition, the reaction mixture was stirred while maintaining the temperature for 3 hours. After the reaction, a ferrous sulfate solution (intermediate vi: ferrous sulfate: purified water: 1:0.05:0.5 (W/V, g/g/mL)) was added to the reaction mixture, and stirred for 10 minutes, then the PH was adjusted to 4.5 to 5.0 with a potassium hydroxide solution (intermediate V: potassium hydroxide: purified water: 1:0.25:10 (W/V, g/g/mL)), and stirred for 10 minutes, then adjusted to 1.3 to 1.5 with a 7% phosphoric acid solution, and stirred and reacted at 25 to 35 ℃ for 2 hours. And after the reaction is finished, adding 30% sodium formate aqueous solution, adjusting the pH to 3.0-3.5, controlling the temperature to 25-35 ℃ and hydrolyzing the sodium formate for 3 hours to finally obtain the target compound epirubicin hydrochloride I crude product solution.

Dichloromethane and purified water are added into the reaction liquid (intermediate VI: purified water: dichloromethane is 1: 65: 45(W/V/V, g/mL)), the mixture is stirred and kept stand for liquid separation, the water phase is extracted three times by using dichloromethane solution (intermediate VI: dichloromethane is 1: 45(W/V, g/mL)), the organic phases are combined and concentrated to a certain volume under vacuum (intermediate VI: concentrated solution is 1:8(W/V, g/mL)), normal hexane (intermediate VI: normal hexane is 1:12(W/V, g/mL)) is added under stirring for crystallization, solid epirubicin hydrochloride I is obtained after suction filtration and drying, and the mass yield is 91.2%. The product is red powder, the purity is 97.8 percent, and the product meets the requirements of pharmacopoeia: if an impurity peak exists in the chromatogram of the test solution, the area of the doxorubicin (relative retention time is about 0.3) is calculated according to the corrected peak area (multiplied by a correction factor of 0.7), and is not more than the main peak area (1.0) of the control solution, the area of the doxorubicin (relative retention time is about 0.8) is not more than the main peak area (1.0) of the control solution, the area of the other single impurity peak is not more than 0.5 times (0.5) of the main peak area of the control solution, and the sum of the areas of the impurities is not more than 2 times (2.0) of the main peak area of the control solution.

Example nineteen

Preparation of epirubicin hydrochloride I:

to a 1000mL three-necked glass reaction flask, heptane, intermediate vi (intermediate vi: heptane ═ 1:70(W/V, g/mL)) was added. Stirring was started, the temperature was controlled at 35 ℃, a 4% heptane solution of hydrogen bromide (intermediate vi: 4% heptane solution of hydrogen bromide: 1:2(W/V, g/mL)) was added dropwise to the reaction solution, and after stirring was completed for 20 minutes, a heptane solution of bromine (intermediate vi: bromine: heptane: 1:0.7:10 (W/V, g/g/mL)) was added dropwise to the reaction solution, and after completion of the addition, the reaction was stirred at constant temperature for 1.5 hours. After the reaction, a ferrous sulfate solution (intermediate vi: ferrous sulfate: purified water 1:0.09:0.9 (W/V, g/g/mL)) was added to the reaction mixture, and the mixture was stirred for 10 minutes, then the PH was adjusted to 4.0 to 4.5 with a potassium hydroxide solution (intermediate V: potassium hydroxide: purified water 1:0.2: 6(W/V, g/g/mL)), stirred for 15 minutes, adjusted to 1.1 to 1.2 with a 12% phosphoric acid solution, and then stirred and reacted at 25 to 35 ℃ for 2 hours with heat preservation. And after the reaction is finished, adding 35% sodium formate aqueous solution, adjusting the pH value to 2.5-3.0, controlling the temperature to 25-35 ℃ and hydrolyzing the sodium formate for 1 hour to finally obtain the target compound epirubicin hydrochloride I crude product solution.

Dichloromethane and purified water are added into the reaction liquid (intermediate VI: purified water: dichloromethane is 1: 35: 50(W/V/V, g/mL/mL)), the mixture is stirred and kept stand for liquid separation, the water phase is extracted three times by using dichloromethane solution (intermediate VI: dichloromethane is 1:50(W/V, g/mL)), the organic phases are combined and concentrated to a certain volume under vacuum (intermediate VI: concentrated solution is 1:15(W/V, g/mL)), normal hexane (intermediate VI: normal hexane is 1:15(W/V, g/mL)) is added under stirring for crystallization, solid epirubicin hydrochloride I is obtained after suction filtration and drying, and the mass yield is 90.0%. The product is red powder, the purity is 97.2 percent, and the product meets the requirements of pharmacopoeia: if an impurity peak exists in the chromatogram of the test solution, the area of the doxorubicin (relative retention time is about 0.3) is calculated according to the corrected peak area (multiplied by a correction factor of 0.7), and is not more than the main peak area (1.0) of the control solution, the area of the doxorubicin (relative retention time is about 0.8) is not more than the main peak area (1.0) of the control solution, the area of the other single impurity peak is not more than 0.5 times (0.5) of the main peak area of the control solution, and the sum of the areas of the impurities is not more than 2 times (2.0) of the main peak area of the control solution.

Example twenty

Preparation of epirubicin hydrochloride I:

To a 1000mL three-necked glass reaction flask, heptane, intermediate vi (intermediate vi: heptane ═ 1:70(W/V, g/mL)) was added. Stirring was started, the temperature was controlled at 35 ℃, a 4% heptane solution of hydrogen bromide (intermediate vi: 4% heptane solution of hydrogen bromide: 1:2(W/V, g/mL)) was added dropwise to the reaction solution, and after stirring was completed for 20 minutes, a heptane solution of bromine (intermediate vi: bromine: heptane: 1:0.7:10 (W/V, g/g/mL)) was added dropwise to the reaction solution, and after completion of the addition, the reaction was stirred at constant temperature for 1.5 hours. After the reaction, a ferrous sulfate solution (intermediate vi: ferrous sulfate: purified water 1:0.09:0.9 (W/V, g/g/mL)) was added to the reaction mixture, and the mixture was stirred for 10 minutes, then the PH was adjusted to 4.0 to 4.5 with a potassium hydroxide solution (intermediate V: potassium hydroxide: purified water 1:0.2: 6(W/V, g/g/mL)), stirred for 15 minutes, adjusted to 1.1 to 1.2 with a 12% phosphoric acid solution, and then stirred and reacted at 25 to 35 ℃ for 2 hours with heat preservation. And after the reaction is finished, adding 35% sodium formate aqueous solution, adjusting the pH value to 2.5-3.0, controlling the temperature to 25-35 ℃ and hydrolyzing the sodium formate for 1 hour to finally obtain the target compound epirubicin hydrochloride I crude product solution.

And after the reaction is finished, adding 250mL of purified water into the reaction solution, adjusting the pH value to 3 with dilute hydrochloric acid, concentrating, performing suction filtration on the concentrated solution, separating with cation exchange resin, collecting the main component, and concentrating to dryness to finally obtain an epirubicin hydrochloride I solid with the mass yield of 85.9%. The product is red powder with the purity of 96.1 percent, and meets the requirements of pharmacopoeia: if an impurity peak exists in the chromatogram of the test solution, the area of the doxorubicin (relative retention time is about 0.3) is calculated according to the corrected peak area (multiplied by a correction factor of 0.7), and is not more than the main peak area (1.0) of the control solution, the area of the doxorubicin (relative retention time is about 0.8) is not more than the main peak area (1.0) of the control solution, the area of the other single impurity peak is not more than 0.5 times (0.5) of the main peak area of the control solution, and the sum of the areas of the impurities is not more than 2 times (2.0) of the main peak area of the control solution.

Example twenty one

Weighing 6.7g of 4' -epidaunorubicin, adding 67mL of methanol to dissolve at room temperature, adding 67mL of 1, 4-dioxane, then adding 6mL of methyl formate and 0.73mL of bromine, reacting for 1h, then adding 2.16mL of acidified propylene oxide, continuously stirring for 30min, concentrating the reaction solution to 65mL, cooling, concentrating, then adding 740mL of isopropanol to crystallize, then adding 150mL of isopropanol to precipitate to wash the crystal, fully drying the crystal, dissolving with 142mL of water, adding 4.2mL of 146mL of hydrogen bromide in acetone to acidify, reacting for 18h at room temperature, adding a solution of 10g of sodium formate and 42mL of water to react for 24h, adjusting the pH to 5 with dilute hydrochloric acid, and continuously stirring for 24 h. And after the reaction is finished, adding 250mL of purified water into the reaction solution, adjusting the pH value to 3 with dilute hydrochloric acid, concentrating, performing suction filtration on the concentrated solution, separating with cation exchange resin, collecting the main component, and concentrating to dryness to finally obtain the epirubicin hydrochloride I solid, wherein the mass yield is 59.7%, and the purity is 90.1%.

Example twenty two

Weighing 6.7g of 4' -epidaunorubicin, adding 67mL of methanol to dissolve at room temperature, adding 67mL of 1, 4-dioxane, then adding 6mL of methyl formate and 0.73mL of bromine, reacting for 1h, then adding 2.16mL of acidified propylene oxide, continuously stirring for 30min, concentrating the reaction solution to 65mL, cooling, concentrating, then adding 740mL of isopropanol to crystallize, then adding 150mL of isopropanol to precipitate to wash the crystal, fully drying the crystal, dissolving with 142mL of water, adding 4.2mL of 146mL of hydrogen bromide in acetone to acidify, reacting for 18h at room temperature, adding a solution of 10g of sodium formate and 42mL of water to react for 24h, adjusting the pH to 5 with dilute hydrochloric acid, and continuously stirring for 24 h.

After the reaction, dichloromethane and purified water were added to the reaction solution (intermediate vi: purified water: dichloromethane 1: 35: 50(W/V, g/mL)), followed by stirring, standing for liquid separation, extracting the aqueous phase three times with a dichloromethane solution (intermediate vi: dichloromethane 1:50(W/V, g/mL)), combining the organic phases, concentrating under vacuum to a certain volume (intermediate vi: concentrated solution volume: 1:15(W/V, g/mL)), adding n-hexane (intermediate vi: n-hexane 1:15(W/V, g/mL)) under stirring for crystallization, and performing suction filtration and drying to obtain an epirubicin hydrochloride solid with a mass yield of 66.4%. The product is red powder with purity of 90.2%, and meets the requirements of pharmacopoeia: if an impurity peak exists in the chromatogram of the test solution, the area of the doxorubicin (relative retention time is about 0.3) is calculated according to the corrected peak area (multiplied by a correction factor of 0.7), and is not more than the main peak area (1.0) of the control solution, the area of the doxorubicin (relative retention time is about 0.8) is not more than the main peak area (1.0) of the control solution, the area of the other single impurity peak is not more than 0.5 times (0.5) of the main peak area of the control solution, and the sum of the areas of the impurities is not more than 2 times (2.0) of the main peak area of the control solution.

example twenty three

synthetic route

The preparation method comprises the following steps:

preparation of intermediate compound iii:

Daunorubicin hydrochloride (5g, 8.63mmol) was suspended in dichloromethane (200 mL; intermediate ii: dichloromethane ═ 1:40(W/V, g/mL)), cooled to 0 ℃ under nitrogen protection, and a solution of trifluoroacetic anhydride (7 mL; intermediate ii: trifluoroacetic anhydride ═ 1:1.4(W/V, g/mL)) in dichloromethane (15 mL; intermediate ii: dichloromethane ═ 1:3(W/V, g/mL)) was added dropwise over 60min with vigorous stirring, the mixture was allowed to react at the same temperature for 0.5h, then 250mL of purified water (intermediate ii: purified water ═ 1:50(W/V, g/mL)) was added thereto, the mixture was shaken well, the organic phase was collected by standing and separation, 200mL of a saturated sodium bicarbonate solution was added thereto (intermediate ii: saturated sodium bicarbonate solution ═ 1:40(W/V, g/mL)), the mixture is fully shaken up and then is put to the room temperature, the mixture is vigorously stirred for 15 to 24 hours, an organic phase is obtained after the hydrolysis is finished and is separated, and the mixture is decompressed and dried at the temperature of 35 ℃ to constant weight, thus obtaining an intermediate III (4.1g, the mass yield is 82.0 percent) and the purity is 85.5 percent.

preparation of intermediate compound iv:

In a 500mL three-necked glass reaction flask, 5mL of DMSO was dissolved in 100mL of dichloromethane (intermediate iii: DMSO: dichloromethane ═ 1:1:20(W/V, g/mL)), the solution was sufficiently stirred and cooled to-60 ℃, after which 1mL of oxalyl chloride and 5mL of dichloromethane solution (intermediate iii: oxalyl chloride: dichloromethane ═ 1:0.2:1(W/V, g/mL)) were added, followed by stirring and reaction at-60 ℃ for 40 minutes with heat preservation. 5g of intermediate III was dissolved in 50mL of dichloromethane (intermediate III: dichloromethane ═ 1:10(W/V, g/mL)), and this solution was added to the above-mentioned cooled solution over 20 minutes, while maintaining the temperature at-60. + -. 5 ℃ for 1 hour, and 7mL of triethylamine (intermediate III: triethylamine ═ 1:1.4(W/V, g/mL)) was added over 10 minutes after the end of the reaction, while maintaining the temperature at ≤ 60 ℃. 5mL of glacial acetic acid is dissolved in 10mL of dichloromethane (intermediate III: glacial acetic acid: dichloromethane: 1:2(W/V/V, g/mL/mL)), the solution is added into the reaction solution and stirred for 2 minutes, 300mL of purified water (intermediate III: purified water: 1:60(W/V, g/mL)) is added into the reaction solution, the mixture is fully stirred and then kept stand to separate an organic phase, the process is repeated for 3 times, the organic phases obtained are combined for three times, and the mixture is concentrated to dryness in a vacuum state to finally obtain 4.4g of intermediate IV, wherein the mass yield is 88.0% and the purity is 82.2%.

Preparation of intermediate compound v:

In a 1000mL three-necked glass reaction flask, 4.7g of intermediate iv was dissolved in 180mL of tetrahydrofuran (intermediate iv: tetrahydrofuran 1:38.3(W/V, g/mL)), and after stirring was started, 2.1g of sodium triethylborohydride (intermediate iv: sodium triethylborohydride 1:1.5(W/W, g/g)) was added over 40 minutes, and after completion of the addition, the reaction was stirred at 20 ± 2 ℃ for 1 hour, and after completion of the reaction, the reaction mixture was added to a mixture of 150mL of dichloromethane, 300mL of purified water and 2mL of 1mol/L hydrochloric acid (intermediate iv: dichloromethane: purified water: 1mol/L hydrochloric acid 1:32:63.8:0.43(W/V, g/mL)), and after sufficient stirring, the mixture was allowed to stand to obtain an organic phase, and 300mL of purified water was added (intermediate iv: purified water 1:63.8(W/V, g/mL)) was washed 2 times and the organic phase was concentrated under vacuum to dryness to give intermediate V4.6 g, in 92.0% mass yield and 77.3% purity. The crude product is further purified by preparative high performance liquid chromatography, and main peak fractions are collected and concentrated to dryness to obtain intermediate V3.0 g, with a mass yield of 65.2% and a purity of 94%.

Preparation of intermediate compound vi:

After 3.0g of intermediate V was sufficiently dissolved in 200mL of purified water at 30 ℃ (intermediate V: purified water 1:66.7(W/V, g/mL)) in a 500mL three-necked glass reaction flask, 10mL of 1.0mol/L NaOH solution (intermediate V: 1.0mol/L NaOH solution 1:3.3(W/V, g/mL)) was added to the solution, the reaction was incubated at the same temperature for 30 minutes, and then pH was adjusted to 7.0 with 1mol/L hydrochloric acid solution, the reaction solution was separated by preparative high performance liquid chromatography, and the major peak fractions were collected and concentrated to dryness to obtain 2.1g of intermediate vi in a mass yield of 70.0% and a purity of 94.8%.

Preparation of epirubicin hydrochloride I:

After 2.1g of intermediate vi was dissolved in 70mL of DMF (intermediate vi: DMF 1:33.3(W/V, g/mL)) in a 1000mL three-necked glass reaction flask, 2.8g of bis (N, N-dimethylacetamide) dibromohydrobromide (intermediate vi: bis (N, N-dimethylacetamide) dibromohydrobromide 1:1.3(W/W, g/g)) was added to the solution, the reaction was incubated at 40 ℃ for 2 hours, 350mL of acetonitrile was added to the reaction solution after the reaction was completed (intermediate vi: acetonitrile 1:167(W/V, g/mL)), the precipitate was suction-filtered and washed with acetonitrile repeatedly, and the precipitate was dried and dissolved in a mixture of 80mL of acetone, 80mL of a 0.25mol/L aqueous solution of sodium bromide and 8g of sodium formate (intermediate vi: acetone: 0.25mol/L aqueous solution of hydrogen bromide: 1:38: sodium formate: 3.8 (W/V) in a 1000mL three-necked glass reaction flask, and then the mixture of sodium formate was dissolved in a mixture of 80mL of water (intermediate vi: acetone: 0.25mol/L aqueous solution of sodium bromide) and 8g of sodium formate (W/ V/W, g/mL/mL/g)) was stirred at 35 ℃ for 30 hours, and after the reaction was completed, the yield was 40.9% and the purity was 60.2%.

separating the reaction liquid by preparative high performance liquid chromatography, collecting main peak fractions, concentrating under reduced pressure to dryness, and adding acetone for recrystallization to obtain 1.3g epirubicin hydrochloride with a mass yield of 61.9%, a final mass yield of 25.3% and a purity of 99.8%.

Example twenty-four

the preparation of intermediates II to VI is the same as that of example twenty-three, changing the last step of the preparation of epirubicin hydrochloride I.

Preparation of epirubicin hydrochloride I:

In a 1000mL three-necked glass reaction flask, 35.2g of intermediate vi (49.6mmol) was dissolved in 321mL of anhydrous dichloromethane (intermediate vi: dichloromethane ═ 1:9.1(W/V, g/mL)), and 19.7g of anhydrous pyridine (249.4 mmol; intermediate vi: anhydrous pyridine ═ 1:7(W/V, g/mL)) was added. The solution was cooled to-5 to 0 ℃ and a solution of 28.1g of trifluoromethanesulfonic anhydride (99.6mmol) in anhydrous dichloromethane (30mL) (intermediate VI: trifluoromethanesulfonic anhydride: anhydrous dichloromethane ═ 1:0.8:0.85(W/W/V, g/g/mL)) was added dropwise over 30min, and the reaction was continued at the same temperature for 1 hour after completion of the dropwise addition. A prepared triethylamine formate solution in methylene chloride (triethylamine 62.7g, anhydrous formic acid 28.7g, anhydrous methylene chloride 209 mL; intermediate VI triethylamine: anhydrous formic acid: anhydrous methylene chloride 1:1.8:0.82:6.0(W/W/W/V, g/g/mL)) was added, the mixture was stirred at 20 to 25 ℃ for 16 hours, 562mL of methanol (intermediate VI: methanol 1:16(W/V, g/mL)) was added, the mixture was cooled to-10 to-5 ℃, and 950g of a 1.6mol/L sodium hydroxide solution (intermediate VI: 1.6mol/L sodium hydroxide solution 1:27(W/W, g/g)) was added and the mixture was reacted at the same temperature for 6 hours. Adding 621mL of 3mol/L hydrochloric acid (intermediate VI: 3mol/L hydrochloric acid is 1:17.6(W/V, g/mL)), heating to 10-15 ℃, stirring for reaction for 7h, standing for layering after the reaction is finished, washing an aqueous phase twice with 562mL of dichloromethane (intermediate VI: dichloromethane is 1:16(W/V, g/mL)), merging organic phases, washing twice with 562mL of water (intermediate VI: water is 1:16(W/V, g/mL)), merging aqueous phases, and adsorbing and purifying by NM100 macroporous adsorption resin [ eluent is acetonitrile: and obtaining 16.8g of crude epirubicin hydrochloride by using a pH value of 4.5 aqueous solution (6: 1), wherein the mass yield is 47.7 percent and the purity is 90.1 percent. The crude product is chromatographed by NM100 macroporous adsorption resin column [ mobile phase is acetonitrile: and (3) re-purifying the solution with pH 4.5 (1: 7.3) to obtain 15.1g of epirubicin hydrochloride pure product, wherein the yield of the epirubicin hydrochloride pure product is 89.9 percent, the final mass yield is 42.9 percent, and the purity is 94.7 percent.

Claims (31)

1. An epirubicin hydrochloride compound is shown in a formula III, and the structural formula is as follows:

。

2. A process for the preparation of a compound according to claim 1, comprising the steps of: adding methanol, an acidic catalyst and an esterification reagent B to enable daunorubicin hydrochloride II to generate an unseparated intermediate compound II-1 with a ketal structure, and protecting active amino groups of amino sugar parts in the intermediate structure by trifluoroacetic anhydride to generate an intermediate compound III.

3. The process according to claim 2, wherein the synthesis of compound III from compound II comprises the following steps: adding methanol, an acidic catalyst, an esterification reagent B and daunorubicin hydrochloride II into an organic solvent A, reacting for 2 hours to generate an unseparated intermediate compound II-1 with a ketal structure, then adding trifluoroacetic anhydride into a reaction system, and stirring for reacting for 1 hour to generate an intermediate compound III with an amino trifluoroacetate structure.

4. The method according to claim 2 or 3, wherein the temperature for the reaction with the esterifying reagent B is 0 to 10 ℃.

5. The process according to claim 2 or 3, wherein the reaction temperature with the esterifying reagent B is 5 ℃.

6. The method according to claim 3, wherein the organic solvent A is selected from one of aromatic hydrocarbons, aliphatic hydrocarbons, halogenated hydrocarbons, alcohols, ethers, amides, ester solvents, and phenols.

7. The method according to claim 3, wherein the organic solvent A is selected from one or more of benzene, toluene, cyclohexane, methanol, ethanol, tert-butanol, dichloromethane, 1, 4-dioxane, diethyl ether, acetone, trichloroethylene, tetrahydrofuran, methyl tert-butyl ether, ethyl acetate, and DMF.

8. The preparation method according to claim 3, wherein the organic solvent A is dichloromethane.

9. A process according to claim 3, wherein said esterifying reagent B is triethyl orthoformate or trimethyl orthoformate.

10. The preparation method according to claim 3, wherein the organic solvent A is used in an amount of: daunorubicin hydrochloride in g/mL by weight: the organic solvent A =1: 50-70.

11. The preparation method according to claim 3, wherein the organic solvent A is used in an amount of: daunorubicin hydrochloride in g/mL by weight: organic solvent a =1: 60.

12. The process according to claim 3, wherein the esterification reagent B is used in an amount of: daunorubicin hydrochloride in g/mL by weight: esterification reagent B =1: 1 to 4.

13. The process according to claim 3, wherein the esterification reagent B is used in an amount of: daunorubicin hydrochloride in g/mL by weight: esterification reagent B =1: 2.

14. The preparation method according to claim 3, wherein the trifluoroacetic anhydride is used in an amount of: daunorubicin hydrochloride in g/mL by weight: trifluoroacetic anhydride is 1: 1-4.

15. The preparation method according to claim 3, wherein the trifluoroacetic anhydride is used in an amount of: daunorubicin hydrochloride in g/mL by weight: trifluoroacetic anhydride ═ 1: 2.

16. the preparation method according to claim 2, wherein the synthetic route is as follows:

。

17. A process for the preparation of epirubicin hydrochloride, compound III according to claim 1, characterized in that it comprises the following steps: dehydrating the intermediate compound III, and oxidizing the alcohol hydroxyl group with the amino sugar structure into carbonyl to generate an unseparated intermediate compound IV; reducing carbonyl in the amino sugar of the intermediate compound IV into hydroxyl by using a selective reducing agent D, and converting the hydroxyl into an intermediate V with a 4-OH configuration opposite to that of the daunorubicin amino sugar structure; removing the protection of amino group in the intermediate V structure by using strong base E to generate an intermediate VI; the intermediate compound VI is subjected to bromination and acid hydrolysis reaction in sequence in an acidic environment to generate a corresponding bromoketone intermediate, the bromoketone intermediate is hydrolyzed in a sodium formate solution, bromine is substituted by alcoholic hydroxyl to finally obtain a target product epirubicin hydrochloride I,

。

18. The process according to claim 17, wherein the synthesis of compound iv from compound iii comprises the following steps: trifluoroacetic anhydride is added into an organic solvent A, after the addition is finished, the mixture is stirred and reacts for 1 hour, then an intermediate compound III is added, the mixture reacts with 1, 5-diazabicyclo (4,3,0) non-5-ene for 0.5 hour, the alcoholic hydroxyl group with the amino sugar structure is oxidized into carbonyl, then an organic acid C is added into a reaction system, and the mixture is stirred and reacts for 0.5 hour to generate an intermediate compound IV.

19. The method according to claim 18, wherein the organic acid C is one or more selected from formic acid, acetic acid, propionic acid, butyric acid, caprylic acid, adipic acid, oxalic acid, malonic acid, succinic acid, maleic acid, tartaric acid, benzoic acid, phenylacetic acid, phthalic acid, terephthalic acid, valeric acid, caproic acid, capric acid, stearic acid, palmitic acid, and acrylic acid.

20. the process of claim 17, wherein the synthesis of compound V from compound IV comprises the steps of: and (3) reacting the intermediate IV with a reducing agent D in an organic solvent A for 0.5 hour to reduce carbonyl in the amino sugar into hydroxyl, and converting the hydroxyl into an intermediate V with a 4-OH configuration opposite to that of the daunorubicin amino sugar structure.

21. The preparation method according to claim 20, wherein the reducing agent D is selected from one or more of sodium hydride, lithium aluminum tetrahydroborate, sodium borohydride, lithium hydride, bis (methoxyethoxyaluminum) hydride, lithium triethylborohydride and sodium cyanoborohydride.

22. The process according to claim 17, wherein the synthesis of compound VI from compound V comprises the following steps: the intermediate compound V is reacted with a base E in purified water for 0.5 hour to deprotect the amino group in the structure of intermediate V with a strong base to produce intermediate VI.

23. The method according to claim 22, wherein the base E is one selected from the group consisting of sodium hydroxide, potassium hydroxide, calcium hydroxide, aqueous ammonia, and sodium hydrogencarbonate.

24. The method according to claim 17, wherein the synthesis of compound I from compound VI comprises the following steps: in an organic solvent A, reacting an intermediate compound VI with an organic solvent A solution of 4% hydrogen bromide for 2 hours, and dropwise adding a bromine organic solvent A solution in the reaction process to convert the intermediate compound VI into a bromo intermediate VI-1 with a ketal structure; adding sufficient reducing agent F to neutralize excessive bromine for quenching reaction, adding a proper amount of saturated alkali solution G to adjust the pH of the reaction solution to 4.5-5.0, adding an inorganic acid H aqueous solution, adjusting the pH to = 1.3-1.5, carrying out heat preservation stirring reaction at 25-35 ℃ for 2 hours, carrying out acid hydrolysis reaction in an acidic environment, and removing the protection of a ketal structure on a carbonyl group to generate a corresponding bromoketone intermediate VI-2; and adding a sodium formate aqueous solution, adjusting the pH value to 3.0-3.5, controlling the temperature to 25-35 ℃ to hydrolyze the sodium formate for 1-3 hours, substituting bromine into alcoholic hydroxyl, and salifying in a hydrochloric acid-methanol solution to finally obtain a target compound epirubicin hydrochloride I crude product solution.

25. The method according to claim 24, wherein the reducing agent F is one selected from sodium sulfite, sodium bisulfite, ferrous sulfate, sodium nitrite, and oxalic acid.

26. The method of claim 24, wherein the reducing agent F is selected from sodium sulfite.

27. The method according to claim 24, wherein the base G is one selected from the group consisting of sodium hydroxide, potassium hydroxide, calcium hydroxide, aqueous ammonia, and sodium hydrogencarbonate.

28. The method as claimed in claim 24, wherein the base G is selected from ammonia.

29. the method according to claim 24, wherein the inorganic acid H is one or more selected from phosphoric acid, sulfuric acid, perchloric acid, boric acid, nitric acid, hydrochloric acid, and hydrobromic acid.

30. The method according to any one of claims 18,20 or 24, wherein the organic solvent a is selected from one of aromatic hydrocarbons, aliphatic hydrocarbons, halogenated hydrocarbons, alcohols, ethers, amides, and ester solvents.

31. The method according to any one of claims 18,20, or 24, wherein the organic solvent a is selected from one or more of benzene, toluene, cyclohexane, methanol, ethanol, t-butanol, dichloromethane, 1, 4-dioxane solution, diethyl ether, acetone, trichloroethylene, tetrahydrofuran, methyl t-butyl ether, ethyl acetate, and DMF.