CN103130814B - Two kinds of 9-nitrocamptothecin new crystal and preparation method thereof - Google Patents

Abstract

The invention provides a new crystal form I of 9-nitrocamptothecin and a preparation method thereof. The heat and humidity stability of the crystal form I is better than that of the existing 9-nitrocamptothecin crystal form. The invention also provides 9-nitrocamptothecin crystal form II and a preparation method thereof. The solubility of the crystal form II is greater than that of the existing 9-nitrocamptothecin crystal form. The preparation method of the crystal form I and the crystal form II of 9-nitrocamptothecin of the present invention is simple and easy to operate, and is suitable for technological production.

Description

Two new crystal forms of 9-nitrocamptothecin and preparation method thereof

technical field

The present invention relates to two new crystal forms of 9-nitrocamptothecin (also known as Rubicon): crystal form I and crystal form II, and a preparation method thereof.

Background technique

9-Nitrocamptothecin, chemical name: 4(S)-ethyl-4-hydroxy-10-nitro-1H-pyrano[3',4':6,7]indeno[1,2 -b] Quinoline-3,14(4H,12H)-dione is a new type of semi-synthetic camptothecin derivative, which belongs to the topoisomerase inhibitor newly developed in recent years. 9-Nitrocamptothecin is an oral preparation. The new drug developed by SuperGen is used to treat pancreatic cancer and other solid tumors. The biggest advantage is that it can be taken during non-hospitalization, so patients can be treated at home.

Clinical pharmacological experiments have proved that 9-nitrocamptothecin has less toxic and side effects than other camptothecin drugs that have been put into clinical use, and its unique advantage is that it is effective orally. 9-Nitrocamptothecin has strong tumor suppressor activity on various cancer cells, and is widely distributed in pancreas, ovary, prostate and other parts, and may become a specific drug for pancreatic cancer, ovarian cancer and prostate cancer, especially for The treatment of pancreatic cancer has special effects. The American Cancer Research Organization believes that 9-nitrocamptothecin is very likely to become the first-line drug for the treatment of advanced pancreatic cancer and has great clinical value.

There are currently seven crystal forms of 9-nitrocamptothecin disclosed in WO03/072027 that have been reported in patents. The patent also discloses that the solvents for recrystallization of the seven crystal forms can be: acetone, dichloromethane, tetrahydrofuran, and acetonitrile, etc., and the crystal form obtained by using acetone, dichloromethane, tetrahydrofuran, and acetonitrile for the powder cake in anhydrous DMF and obtained by recrystallization from aqueous DMF. CN1611505 discloses a crystal form obtained by cooling in methanol.

Contents of the invention

The technical problem to be solved by the present invention is to provide two novel crystal forms of 9-nitrocamptothecin, crystal form I and crystal form II. The heat and humidity stability of the crystal form I of the present invention is better than that of the existing 9-nitrocamptothecin crystal form, and the solubility of the crystal form II is greater than that of the existing 9-nitrocamptothecin crystal form. The present invention also provides preparation methods of 9-nitrocamptothecin crystal form I and crystal form II respectively.

The present invention provides a crystal form I of 9-nitrocamptothecin. In the powder X-ray diffraction spectrum using a radiation source of Cu-Kα, the crystal form I has diffraction angles 2θ=6.100, 8.392, 10.932, and 12.197 、12.591、13.381、13.932、15.055、16.535、16.873、18.332、18.885、19.674、21.292、21.607、21.964、22.753、23.958、24.433、25.041、25.360、25.674、26.957、27.962、28.771、29.719、30.352、31.596、32.603 There are characteristic peaks at , 35.937, 37.001, 38.463 and 39.960 degrees, and the 2θ error range is ±0.2.

Wherein, the melting point of the crystal form I of 9-nitrocamptothecin is generally 263±1°C.

The present invention also provides the preparation method of the crystalline form I of described 9-nitrocamptothecin, which comprises the following steps: (1) dissolving 9-nitrocamptothecin in an organic solvent; (2) using evaporation method to remove the solvent to obtain the crystal form I of 9-nitrocamptothecin; wherein the organic solvent is a single solvent selected from the following solvents or a mixed solvent consisting of the following solvents: methanol, ethyl formate, acetic acid Ethyl ester, isobutyl acetate, carbon disulfide, tetrahydrofuran, 2-butanone and n-propanol.

In step (1), the 9-nitrocamptothecin may be various forms of 9-nitrocamptothecin crystals or amorphous 9-nitrocamptothecin in the art.

In step (1), there is no special requirement on the amount of the organic solvent, as long as it can completely dissolve 9-nitrocamptothecin.

In step (1), when the organic solvent is a mixed solvent, the mixed solvent is preferably composed of two solvents, and the volume ratio of the two solvents is preferably 5:1-7:1.

In step (1), the single solvent is preferably tetrahydrofuran, n-propanol or 2-butanone. Described mixed solvent is preferably ethyl acetate and methanol, isobutyl acetate and methanol, ethyl formate and methanol, or carbon disulfide and methanol, wherein ethyl acetate, isobutyl acetate, ethyl formate or carbon disulfide and methanol The volume ratio is preferably 5:1-7:1.

In step (2), the evaporation temperature of the evaporation method is preferably 25-60°C, more preferably 25-40°C.

In step (2), the evaporation pressure of the evaporation method is preferably 0.05-0.1MP.

In step (2), vacuum drying is preferably performed after the solvent is removed by evaporation, and the pressure of the vacuum drying is preferably 65-165 mBar.

The present invention also provides a crystal form II of 9-nitrocamptothecin. In the powder X-ray diffraction spectrum using Cu-Kα as the radiation source, the crystal form II has diffraction angles 2θ=7.834, 8.863, 12.058, There are characteristic peaks at 14.248, 14.623, 16.025, 18.035, 19.755, 21.156, 22.004, 23.524, 25.773, 26.565, 27.054, 28.911 and 32.700 degrees, and the 2θ error range is ±0.2.

Wherein, the melting point of the crystal form II of 9-nitrocamptothecin is 156±1°C.

The present invention also provides a preparation method for the crystal form II of 9-nitrocamptothecin, which includes the following steps: (1) mixing 9-nitrocamptothecin with an organic solvent, heating and stirring to make 9-nitrocamptothecin (2) cooling down to obtain the crystal form II of 9-nitrocamptothecin; wherein the organic solvent is selected from acetone or 2-butanone, etc.

In step (1), the 9-nitrocamptothecin may be various forms of 9-nitrocamptothecin crystals or amorphous 9-nitrocamptothecin in the art.

In step (1), there is no special requirement on the amount of the organic solvent, as long as it can completely dissolve 9-nitrocamptothecin.

In step (1), the temperature at the end of the heating is preferably 45-55°C.

In step (1), the heating process is preferably carried out under stirring conditions.

In step (2), the cooling temperature is preferably below 28°C.

In step (2), after the cooling, the solid-liquid separation is preferably carried out by conventional methods in the art, followed by drying. The solid-liquid separation adopts conventional solid-liquid separation methods, such as evaporating the solvent or filtering at normal temperature and pressure. The filtration can adopt various conventional filtration methods in the art, such as vacuum filtration.

In the present invention, the above-mentioned preferred conditions can be combined arbitrarily on the basis of conforming to common knowledge in the field, so as to obtain various preferred embodiments of the present invention.

The starting materials and reagents of the present invention are all commercially available.

The positive progress effect of the present invention is:

1. The present invention provides a crystal form I of 9-nitrocamptothecin, which is more stable to heat and humidity than the existing 9-nitrocamptothecin crystal form.

2. The present invention also provides a crystal form II of 9-nitrocamptothecin. The solubility of the crystal form II is greater than that of the existing 9-nitrocamptothecin crystal form.

3. The preparation method of crystal form I and crystal form II of 9-nitrocamptothecin in the present invention is simple and easy to operate, and is suitable for industrial production.

Description of drawings

Fig. 1 is a DSC-TGA analysis chart of the crystal form I of 9-nitrocamptothecin.

Fig. 2 is the powder XRD pattern of the crystal form I of 9-nitrocamptothecin.

Fig. 3 is a DSC-TGA analysis diagram of the crystal form II of 9-nitrocamptothecin.

Fig. 4 is the powder XRD pattern of the crystal form II of 9-nitrocamptothecin.

Fig. 5 is the moisture stability graph of 9-nitrocamptothecin crystal form I, crystal form II and bulk drug.

The above description and the following examples are only exemplary and explanatory, and their protection should not be limited thereto.

detailed description

The present invention will be described in further detail below in conjunction with the examples, but the present invention is not limited thereto.

The preparation of embodiment 1-169-nitrocamptothecin crystal form I

Example 1

Put 1 g of 9-nitrocamptothecin into a 1-liter reaction bottle, add THF 200 ml, stir to dissolve, evaporate the solvent under normal temperature and pressure to obtain crystal form I, and store the obtained crystal after vacuum drying.

Example 2

Put 1 g of 9-nitrocamptothecin into a 1-liter reaction bottle, add 700 ml of ethyl acetate and 100 ml of methanol, stir to dissolve, evaporate the solvent under normal temperature and pressure to obtain crystal form I, and store the obtained crystals after vacuum drying.

Example 3

Put 1 g of 9-nitrocamptothecin into a 1-liter reaction bottle, add 700 ml of isobutyl acetate and 100 ml of methanol, stir to dissolve, evaporate the solvent at normal temperature and pressure to obtain crystal form I, and store the obtained crystal after vacuum drying .

Example 4

Put 1 g of 9-nitrocamptothecin into a 1-liter reaction bottle, add 700 ml of ethyl formate and 100 ml of methanol, stir to dissolve, evaporate the solvent under normal temperature and pressure to obtain crystal form I, and store the obtained crystals after vacuum drying.

Example 5

Put 1 g of 9-nitrocamptothecin into a 1-liter reaction bottle, add 700 ml of carbon disulfide and 100 ml of methanol, stir to dissolve, evaporate the solvent under normal temperature and pressure to obtain crystal form I, and store the obtained crystals after vacuum drying.

Example 6

Put 1 g of 9-nitrocamptothecin into a 1-liter reaction bottle, add 700 ml of n-propanol, stir for 8 hours, evaporate the solvent under normal temperature and pressure to obtain crystal form I, and store the obtained crystals after vacuum drying.

Example 7

Put 1 g of 9-nitrocamptothecin in a 1-liter reaction bottle, add 500 ml of 2-butanone, stir to dissolve, evaporate the solvent under normal temperature and pressure to obtain crystal form I, and store the obtained crystals after vacuum drying.

Example 8

Put 1 g of 9-nitrocamptothecin into a 1-liter reaction bottle, add 200 ml of THF, stir to dissolve, evaporate the solvent at 40° C. under normal pressure to obtain crystal form I, and store the obtained crystals after vacuum drying.

Example 9

Put 1g of 9-nitrocamptothecin into a 1-liter reaction bottle, add 700ml of ethyl acetate and 100ml of methanol, stir to dissolve, evaporate the solvent at normal pressure at 40°C to obtain crystal form I, and store the obtained crystals after vacuum drying .

Example 10

Put 1g of 9-nitrocamptothecin in a 1-liter reaction bottle, add 700ml of isobutyl acetate and 100ml of methanol, stir to dissolve, evaporate the solvent at normal pressure at 40°C to obtain crystal form I, and vacuum-dry the obtained crystal save.

Example 11

Put 1g of 9-nitrocamptothecin in a 1-liter reaction bottle, add 700ml of ethyl formate and 100ml of methanol, stir to dissolve, evaporate the solvent at normal pressure at 40°C to obtain crystal form I, and store the obtained crystals after vacuum drying .

Example 12

Put 1 g of 9-nitrocamptothecin into a 1-liter reaction bottle, add 700 ml of carbon disulfide and 100 ml of methanol, stir to dissolve, evaporate the solvent at normal pressure at 40° C. to obtain crystal form I, and store the obtained crystals after vacuum drying.

Example 13

Put 1 g of 9-nitrocamptothecin into a 1-liter reaction bottle, add 700 ml of n-propanol, stir for 8 hours, and evaporate the solvent at 60 °C under normal pressure to crystallize (what kind of solvent is used for recrystallization?) to obtain crystal form I. The crystals were dried in vacuo and stored.

Example 14

Put 1 g of 9-nitrocamptothecin into a 1-liter reaction bottle, add 500 ml of 2-butanone, stir to dissolve, evaporate the solvent at 40° C. under normal pressure to obtain crystal form I, and store the obtained crystals after vacuum drying.

Example 15

Put 1g of 9-nitrocamptothecin into a 1-liter reaction bottle, add 700ml of methanol and stir at 60°C to dissolve it, keep stirring at 60°C for 3 hours, cool down to 40°C and evaporate the solvent to crystallize to obtain crystal form I. Store after vacuum drying.

Example 16

Put 1 g of 9-nitrocamptothecin into a 1-liter reaction bottle, add 800 ml of ethyl acetate and stir to dissolve it, evaporate the solvent at room temperature to crystallize to obtain crystal form I, and store the obtained crystal after vacuum drying.

The preparation of the crystal form II of embodiment 17-199-nitrocamptothecin

Example 17

Put 0.5g of 9-nitrocamptothecin into a 1000ml crystallizer, add 250ml of acetone, heat up to 55°C and stir for 3h, slowly cool down to room temperature, and evaporate the solvent under normal temperature and pressure to obtain crystal form II. Store after drying.

Example 18

Weigh 0.5mg of 9-nitrocamptothecin into 1000ml of crystallization, add 350ml of butanone, stir at 45°C for 3h, slowly cool down to 5°C and stop stirring for 0.5h to grow crystals, filter, drain, and vacuum at room temperature Oven-dried for 48 hours to obtain the crystal form II.

Example 19

Add 0.5 g of 9-nitrocamptothecin to 1000 ml of crystallization, add 300 ml of acetone, heat up to 55°C and stir for 3 hours, slowly cool down to 5°C, and evaporate the solvent under normal temperature and pressure to obtain crystal form II.

Effect Example 19-Crystal Form I of Nitrocamptothecin

The crystal form I of 9-nitrocamptothecin obtained in Examples 1-16 was subjected to powder X-ray diffraction. The radiation source was Cu-Kα. The spectrum is shown in FIG. 2 .

Table 19 - PXRD characteristic peaks of the crystal form I of nitrocamptothecin

Effect Example 29-Crystal Form II of Nitrocamptothecin

The crystal form II of 9-nitrocamptothecin obtained in Examples 17-19 was subjected to powder X-ray diffraction. The radiation source was Cu-Kα. The spectrum is shown in FIG. 4 .

Table 29 - PXRD characteristic peaks of the crystalline form II of nitrocamptothecin

Effect Example 3

The crystal form I of 9-nitrocamptothecin in Example 1 was detected by differential scanning calorimetry (DSC). Starting from 20°C, the temperature was raised to 350°C at a rate of 10°C/min. There is an endothermic peak at 1°C, see Figure 1 for details. The TGA diagram of the crystal form I is also shown in FIG. 1 . The DSC and TGA figures of other embodiments are the same as this embodiment.

The crystal form II of 9-nitrocamptothecin in Example 17 was detected by differential scanning calorimetry (DSC). Starting from 20°C, the temperature was raised to 350°C at a rate of 10°C/min. There is an endothermic peak at ±1°C, see Figure 3 for details. The TGA diagram of the crystal form II is also shown in FIG. 3 . The DSC and TGA figures of other crystal form II examples are the same as this example.

Effect Example 4

1. Solubility

What the test of solubility adopts is to apply HPLC method, measures with external standard method, and the condition of HPLC is as follows:

Column model: Synergi4μhydro-RP80A250mm*4.60mm

Mobile phase: methanol: water = 0.6: 0.4

Column temperature: 30°C

Wavelength: 287nm

The test results are as follows:

sample name Form I Form II API Solubility μg/ml 0.946 2.329 1.234

2. Humidity stability

The crystalline form I of the present invention is the most stable crystalline form after testing, and it is found that both the bulk drug and the newly discovered crystalline form II tend to be converted into crystalline form I, while the crystalline form I does not undergo conversion under experimental conditions. Experimental conditions: Take appropriate amount of crystal form I, crystal form II and bulk drug under the condition of 97% humidity, and regularly sample and test respectively.

Claims (4)

1. A preparation method for the crystal form I of 9-nitrocamptothecin, characterized in that: the crystal form I uses the radiation source as Cu-Kα in the powder X-ray diffraction spectrum, at the diffraction angle 2θ= 6.100、8.392、10.932、12.197、12.591、13.381、13.932、15.055、16.535、16.873、18.332、18.885、19.674、21.292、21.607、21.964、22.753、23.958、24.433、25.041、25.360、25.674、26.957、27.962、28.771、 There are characteristic peaks at 29.719, 30.352, 31.596, 32.603, 35.937, 37.001, 38.463 and 39.960 degrees, and the 2θ error range is ±0.2; The preparation method comprises the following steps: (1) dissolving 9-nitrocamptothecin in an organic solvent; (2) removing the solvent by evaporation to obtain crystal form I of 9-nitrocamptothecin; Wherein, in step (1), the organic solvent is a single solvent or a mixed solvent: the single solvent is tetrahydrofuran, n-propanol or 2-butanone; the mixed solvent is ethyl acetate and methanol, acetic acid Isobutyl and methanol, ethyl formate and methanol, or carbon disulfide and methanol, wherein the volume ratio of ethyl acetate, isobutyl acetate, ethyl formate or carbon disulfide to methanol is 5:1-7:1; In step (2), the evaporation temperature of the evaporation method is 25-60°C; the evaporation pressure of the evaporation method is 0.05-0.1MP. 2. The preparation method according to claim 1, characterized in that: the melting point of the crystal form I is 263±1°C. 3. The preparation method according to claim 1 or 2, characterized in that: in step (2), vacuum drying is carried out after the solvent is removed by evaporation, and the pressure of the vacuum drying is 65-165 mBar. 4. The preparation method according to claim 1, characterized in that: in step (2), the evaporation temperature of the evaporation method is 25-40°C.