CN109956896B

CN109956896B - Spiro [ cyclopropane-1, 3-indoline ] framework, crystal thereof, preparation method and application thereof

Info

Publication number: CN109956896B
Application number: CN201711338045.XA
Authority: CN
Inventors: 杨开川; 李俊龙; 李青竹; 冷海军; 沈旭东; 刘悦; 朱红萍; 赖含雨; 张翔; 戴青松; 刘宇; 曾荣; 张鹰
Original assignee: Sichuan Industrial Institute of Antibiotics
Current assignee: Sichuan Industrial Institute of Antibiotics
Priority date: 2017-12-14
Filing date: 2017-12-14
Publication date: 2021-03-12
Anticipated expiration: 2037-12-14
Also published as: CN109956896A

Abstract

The invention discloses spiro [ cyclopropane-1, 3-indoline]The crystal form of the compound is a trigonal system, and the unit cell parameter is

A is 73.580(4) °, β is 83.539(4) °, γ is 85.980(4) °, space group is P-1, Z is 2, unit cell volume is

The invention also discloses a method for preparing the crystal form, and discloses application of the compound, or the crystal form, or the solvate, or the pharmaceutically acceptable salt thereof in preparing antibacterial drugs. The invention has high yield and purity of the crystal form, is easy to obtain the crystal form of the high-purity compound, has simple and convenient operation and low cost, and is very beneficial to controlling the quality of the compound shown in the formula I; meanwhile, the crystal form hygroscopicity of the compound is not obviously increased, the stability is good, and great convenience is provided for later-stage product transportation, storage or preparation processes.

Description

Spiro [ cyclopropane-1, 3-indoline ] framework, crystal thereof, preparation method and application thereof

Technical Field

The invention relates to a spiro [ cyclopropane-1, 3-indoline ] framework, a crystal thereof, a preparation method and application thereof.

Background

The spiro [ cyclopropane-1, 3-indoline ] skeleton is widely existed in natural products and synthetic drugs, and related researches show that the compound containing the skeleton has various important biological activities and pharmaceutical activities and has wide application prospects.

Two or more different crystalline states are common for the same compound, while different crystalline forms typically exhibit different bioavailability, dissolution rates, stability, melting points, color, filterability, density, and flowability, among others. Therefore, the development of a crystal form with better solubility and stability is very important.

Disclosure of Invention

In order to solve the problems, the invention provides a spiro [ cyclopropane-1, 3-indoline ] framework, a crystal thereof, a preparation method and application thereof.

The invention provides a spiro [ cyclopropane-1, 3-indoline ] framework, which has a structural formula shown as a formula I:

the invention provides a method for preparing a compound shown as a formula I, which comprises the following steps:

under the condition of argon, adding a compound shown as a formula II, potassium carbonate and alpha-chloropropylaldehyde into tetrahydrofuran, and reacting completely to obtain a reaction solution; removing solvent, performing column chromatography to obtain eluent, and removing solvent to obtain compound shown in formula I.

Wherein the reaction temperature is 60 ℃; the reaction time is 1 h; the feeding ratio of the compound shown in the formula II, potassium carbonate, alpha-chloropropylaldehyde and tetrahydrofuran is 1:1.5:2.5:10 mmol/mmol/mmol/mL; the eluent of the column chromatography is petroleum ether: ethyl acetate 10: 1; the eluent is prepared by thin-layer chromatography, and the collecting and developing agent is petroleum ether: and (3) an eluent with ethyl acetate being 5:1 and Rf being 0.3-0.4.

Wherein the compound shown in the formula II is prepared by the following method:

(a) preparing a compound represented by the formula III: adding a compound shown as a formula IV and a compound shown as a formula V into methanol, and filtering and drying after complete reaction to obtain a compound shown as a formula III;

(b) preparing a compound represented by the formula II: and (3) sequentially adding the compound shown in the formula III, benzaldehyde and titanium tetrachloride into dichloromethane, and after complete reaction, filtering and drying to obtain the compound shown in the formula II.

In the step (a), the reaction temperature is 60 ℃; the reaction time is 4 h; the feeding ratio of the compound shown in the formula IV to the compound shown in the formula V to methanol is 2:1:1 mmol/mmol/mL.

In the step (b), the reaction temperature is 25 +/-5 ℃; the reaction time is 4-16 h; the feeding ratio of the compound shown in the formula III, benzaldehyde, titanium tetrachloride and dichloromethane is 1:3:2: 2.5-3 mmol/mmol/mL.

The invention provides a crystal form of a compound shown as a formula I, wherein the crystal form is a trigonal crystal system, and unit cell parameters are

α＝73.580(4)°，β＝83.539(4)°，γ＝85.980(4)°。

Wherein the space group of the crystal form is P-1, Z is 2, and the unit cell volume is

Wherein the ee value of the crystal form is more than 99%.

Wherein the melting point of the crystal form is 156-160 ℃.

The invention provides a method for preparing the crystal form, which comprises the following steps:

(1) obtaining the compound shown in the formula I according to the preparation method;

(2) and (2) crystallizing the compound shown in the formula I obtained in the step (1) in an ethyl acetate/petroleum ether mixed solvent to obtain the crystal form of the compound shown in the formula I.

In the step (2), the volume ratio of ethyl acetate to petroleum ether is 10-20: 80-90, preferably 15: 85.

The application of the compound shown in the formula I, or crystal form, solvate or pharmaceutically acceptable salt thereof in preparing antibacterial drugs.

The invention provides a pharmaceutical composition, which is prepared into a commonly used pharmaceutical preparation by taking a compound shown as the formula I, or a crystal form, or a solvate, or a pharmaceutically acceptable salt thereof as an active ingredient and adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients.

The room temperature of the invention is 25 +/-5 ℃.

The invention has the beneficial effects that: the invention provides a compound shown in formula I and a preparation method thereof, and also provides a crystal form and a preparation method thereof, the crystal form has high yield and purity, the crystal form of the compound with high purity can be easily obtained, the operation is simple and convenient, the cost is low, and the quality control of the compound shown in formula I is very facilitated; meanwhile, the compound or the crystal form thereof has antibacterial activity, and the crystal form of the compound has unobvious moisture absorption increase and good stability, so that great convenience is provided for later-stage product transportation, storage or preparation processes.

Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.

The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Drawings

FIG. 1 is a perspective view of a crystalline form of a compound of formula I according to the present invention.

Detailed Description

The raw materials and equipment used in the embodiment of the present invention are known products and obtained by purchasing commercially available products.

The raw materials used in the invention can be purchased from the market, wherein the p-toluenesulfonyl azide can be purchased or obtained by a related synthesis method, and the invention is obtained by the following synthesis method:

at room temperature, 13.8g of p-toluenesulfonyl chloride (73mmol), 50mL of water and 5.7g of sodium azide (87mmol) are sequentially added into a 500mL flask, the mixture is stirred for reaction for 2 hours, after the reaction is monitored by a TLC method to be complete, 50mL of dichloromethane is used for extraction for 3 times, organic phases are combined, anhydrous sodium sulfate is used for drying, and the p-toluenesulfonyl azide product is obtained by concentration, wherein the yield is 88%.

EXAMPLE 1 preparation of Compounds of formula I

(1) Preparation of the Compound of formula II

a) Preparation of Compounds of formula III

At 60 ℃, 50mL of methanol, 11.7g of indole (IV, 100mmol) and 9.9g of p-toluenesulfonyl azide (V, 50mmol) are sequentially added into a 100mL flask, the mixture reacts for 4 hours, the reaction is monitored by a TLC method until the reaction is completed, a solid is separated out, the solid is filtered and dried in vacuum, 13.0g of pink solid product shown in a formula III is obtained, and the yield is 90%.

b) Preparation of a Compound of formula II

In a 100mL flask at room temperature, 50mL of dichloromethane, 5.0g of the substrate of formula III (17.5mmol) and 6.0g of benzaldehyde (52.5mmol) were added sequentially, and 3.5mL of titanium tetrachloride (35.0mmol) were added to promote the reaction to occur for 4-16 hours, followed by TLC until the reaction was complete, precipitation of a solid, filtration to give a solid product, and vacuum drying to give 5.5g of a yellow powder of formula II in 75% yield.

(2) Preparation of the Compound of formula I

Under argon atmosphere, 40.0mg of indole electron-deficient olefin substrate of formula II (0.1mmol), 21.0mg of potassium carbonate (0.15mmol), 1mL of tetrahydrofuran and 42.1mg of alpha-chloropropylaldehyde (0.25mmol) are added sequentially to a reaction tube, the tube is sealed and stirred at 60 ℃ for 1h, the reaction is monitored by TLC, after the reaction is completed, the reaction solvent is removed under reduced pressure, and the residue is loaded onto a silica gel column and treated with petroleum ether: eluting with ethyl acetate 10:1 as eluent, and performing thin layer chromatography with petroleum ether as developing agent: and (3) combining the eluent with Rf of 0.3-0.4, and removing the solvent to obtain a purified product, namely the final product of the formula I, wherein the ethyl acetate is 5: 1.

The structure of the final product obtained by preparation is characterized to obtain HRMS (ESI) of the final product,¹H NMR and¹³c NMR data confirmed that the final product was indeed the compound of formula I.

HRMS(ESI):m/z calculated for C₃₁H₂₆N₂O₃S+Na⁺:529.1562,found:529.1567。

¹H NMR(600MHz,Chloroform-d)δ10.26(s,1H),9.82(s,1H),7.45–7.41(m,2H),7.32–7.26(m,6H),7.08–7.04(m,3H),6.87–6.84(m,2H),6.72–6.64(m,1H),3.83(s,1H),3.74(d,J＝15.0Hz,1H),2.93(d,J＝15.0Hz,1H),2.38(s,3H)。

¹³C NMR(150MHz,Chloroform-d)δ199.6,143.1,141.7,138.7,130.6,130.1,129.7,129.3,129.2,128.9,128.8,128.4,128.2,127.9,126.9,126.4,126.3,123.3,122.6,111.6,51.4,47.4,46.6,31.6,21.6.

Example 2 preparation of form I of the invention

30.0mg of the compound represented by the formula 1 prepared in example 1 was taken in petroleum ether: ethyl acetate 85: 15(v/v), slowly volatilizing at normal temperature to crystallize to obtain 18.0mg of a single crystal of the compound shown in the formula I, wherein the yield is 60%, the ee value is more than 99%, the crystal form is diffracted by the single crystal, the crystal structure data is shown in table 1, and the projection view of the three-dimensional structure is shown in figure 1.

TABLE 1 Crystal Structure data in Single Crystal diffraction

The advantageous effects of the present invention are described below by way of experimental examples.

Experimental example 1 antibacterial Activity study

The antibacterial activity of the compound was determined by an equal dilution method. Firstly, accurately weighing the same amount of crystal form I and levofloxacin, respectively dissolving the crystal form I and the levofloxacin in 2ml of DMSO, respectively making 10 concentration gradients according to an equal-time dilution method, adding 1ml of drug-containing solution into an MH culture dish for each gradient, and uniformly mixing the drug-containing solution with 14ml of MH solid culture medium to prepare culture dishes with different drug contents. Then using a 27-hole punch to obtain a bacteria content of 10⁶Inoculating the bacterial liquid on a culture dish, putting the culture dish into a constant-temperature incubator at 37 ℃, culturing for 18-24h, and observing whether bacteria grow on the inoculated part to judge the bacteriostatic effect. The results are shown in table 2:

TABLE 2 determination of the MIC (mg/ml) of the antibacterial Activity of form I

Bacterial strains	Crystal form I	Levofloxacin
			Pseudomonas aeruginosa ATCC27853	2.17	0.0001
Proteus mirabilis ATCC29315	3.73	0.0001
			Escherichia coli (ATCC25922)	1.82	0.0001
Enterococcus faecalis ATCC29212	2.55	0.0001
			Salmonella enteritidis (CICC21527)	2.67	0.0001
Aspergillus flavus (CICC 2062)	1.96	0.0001

Note: the strains used above are all pharmacological laboratory preservation strains.

As can be seen from the above tests, the compound of the crystal form I provided by the invention has antibacterial activity.

Test example 2 investigation of stability and hygroscopicity of the crystalline form of the present invention

1. Stability of

Putting the crystal form I compound into a stability test box for accelerated test, wherein the test conditions are as follows: the temperature is 40 +/-2 ℃; humidity, RH 75% + -5%, time 3 months.

The results show that the compound of the crystal form I is not obviously changed by adopting TLC and HPLC, which indicates that the crystal form of the invention has good stability.

2. Moisture absorption property

The following determination results are obtained by adopting the guiding principle of the moisture absorption test of the second part, appendix XIX J of the pharmacopoeia 2010 edition of the people's republic of China:

TABLE 3 moisture absorption examination

Use time (sky)	0	5	10	15
					Compound induced wet weight gain	1.4％	1.5％	1.5％	1.5％

The results in the table show that the crystal form I is not obvious in moisture absorption and weight increment after being placed in a humid environment for 15 days, and the crystal form I can effectively avoid compound moisture absorption and deliquescence.

The spiro [ cyclopropane-1, 3-indoline ] skeleton compound and the crystal form thereof prepared by the invention have antibacterial activity, unobvious increase of hygroscopicity and good stability, and provide great convenience for later-stage product transportation, storage or preparation process.

In conclusion, the invention provides the compound shown in the formula I and the preparation method thereof, and simultaneously provides the crystal form of the compound, the crystal form has high yield and purity, the compound with high purity can be easily obtained, the difficulty of the purification process of the compound shown in the formula I is reduced, the operation is simple and convenient, the cost is low, and the quality control of the compound shown in the formula I is very facilitated; meanwhile, the compound or the crystal form thereof has antibacterial activity, and the crystal form of the compound has unobvious moisture absorption increase and good stability, so that great convenience is provided for later-stage product transportation, storage or preparation processes.

Claims

1. A spiro [ cyclopropane-1, 3-indoline ] skeleton which is characterized in that: it has a structural formula shown in formula I:

2. a process for preparing a compound of formula i, which comprises: it comprises the following steps:

3. The method of claim 2, wherein: the temperature of the reaction is 60 ℃; the reaction time is 1 h; the feeding ratio of the compound shown in the formula II, potassium carbonate, alpha-chloropropylaldehyde and tetrahydrofuran is 1:1.5:2.5:10 mmol/mmol/mmol/mL; the eluent of the column chromatography is petroleum ether: ethyl acetate 10: 1; the eluent is prepared by thin-layer chromatography, and the collecting and developing agent is petroleum ether: and (3) an eluent with ethyl acetate being 5:1 and Rf being 0.3-0.4.

4. The method of claim 2, wherein: the compound shown in the formula II is prepared by the following method:

5. The method of claim 4, wherein: in the step (a), the reaction temperature is 60 ℃; the reaction time is 4 h; the feeding ratio of the compound shown in the formula IV to the compound shown in the formula V to methanol is 2:1:1 mmol/mmol/mL; in the step (b), the reaction temperature is 25 +/-5 ℃; the reaction time is 4-16 h; the feeding ratio of the compound shown in the formula III, benzaldehyde, titanium tetrachloride and dichloromethane is 1:3:2: 2.5-3 mmol/mmol/mL.

6. A crystalline form of a compound of formula i, characterized in that: the crystal form is trigonal, and the cell parameter is

α＝73.580(4)°，β＝83.539(4)°，γ＝85.980(4)°；

7. The crystalline form of claim 6, characterized in that: the space group of the crystal form is P-1, Z is 2, and the unit cell volume is

Ee value of the crystal form>99 percent; the melting point of the crystal form is 156-160 ℃.

8. A process for preparing the crystalline form of claim 6 or 7, characterized in that: it comprises the following steps:

(1) the preparation method according to any one of claims 2 to 5, wherein the compound shown as the formula I is obtained;

9. The method of claim 8, wherein: in the step (2), the volume ratio of ethyl acetate to petroleum ether is 10-20: 80-90.

10. The method of claim 9, wherein: in the step (2), the volume ratio of the ethyl acetate to the petroleum ether is 15: 85.

11. The use of a compound of formula i, or a crystalline form thereof, or a pharmaceutically acceptable salt thereof, as claimed in claim 1, in the manufacture of an antibacterial medicament.

12. A pharmaceutical composition characterized by: the compound of formula I as shown in claim 1, or a crystal form thereof, or a pharmaceutically acceptable salt thereof is used as an active ingredient, and pharmaceutically acceptable auxiliary materials or auxiliary ingredients are added to prepare a pharmaceutically common preparation.