Camptothecin derivative compounds and their applications

Abstract

The invention discloses a camptothecin derivative compound and its application, and belongs to the technical field of pharmaceutical chemistry. Dissolve exotecan mesylate in a solvent, react with an amino alcohol compound, filter, wash with water, and dry to obtain a camptothecin derivative compound. The invention performs a series of derivatizations on exotecan on the basis of maintaining the pharmacodynamic skeleton of exotecan to form a series of exotecan derivatives, which have the characteristics of high anti-tumor activity, low toxic side effects, and easy solubility in water, and have potential application value as anti-tumor single drugs or antibody-coupled drugs.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical chemistry, and specifically relates to camptothecin derivative compounds and applications thereof. Background Art

[0002] In the field of cancer treatment, traditional chemotherapy drugs, due to their non-selectivity and high systemic toxicity, often inhibit tumor cell proliferation while also causing damage to normal cells, leading to a variety of adverse events in patients. In order to improve the therapeutic effect and reduce side effects, the research and development of targeted drugs and antibody-drug conjugates (ADCs) have become a research hotspot. ADCs can deliver drugs directly to tumor cells by combining cytotoxic drugs with specific antibodies, thereby improving the therapeutic effect and reducing damage to normal cells.

[0003] In the existing technology, the design and application of ADCs face a series of challenges. First, the selection of cytotoxic drugs is crucial. They need to have sufficient water solubility, serum stability, conjugated functional groups, and insensitivity to lysosomal enzyme degradation. In addition, the drug's lipid-water partition coefficient (LogP) and charge state also affect its ability to penetrate the cell membrane, thereby affecting the efficacy of ADC. Exatecan is a fully synthetic derivative of the natural product camptothecin. It has extremely strong inhibitory activity on the topoisomerase Top1, can promote cell apoptosis, and has a broad spectrum of anti-tumor activity, but its toxic side effects limit its clinical application. Therefore, the development of new exatecan derivatives, aimed at improving its water solubility, reducing toxic side effects, and maintaining or enhancing its anti-tumor activity, is a technical problem that needs to be solved urgently in the existing technology. Summary of the invention

[0004] In view of the above problems existing in the prior art, the present application provides a camptothecin derivative compound and its application, which has the characteristics of high anti-tumor activity, low toxicity and side effects, and easy solubility in water.

[0005] To solve the above problems, the present invention provides the following technical solutions:

[0006] In a first aspect, the present application provides a camptothecin derivative compound as shown in formula (1),

[0007] ;

[0008] or a pharmaceutically acceptable salt, hydrate, solvate, optical isomer or crystal form thereof, wherein:

[0009] X is oxygen, nitrogen, a nitrogen atom with an alkyl group, a sulfur atom, a carbon atom, and -SO2-;

[0010] n is 1 or 2;

[0011] R1 and R2 are each independently selected from -(CH2) m -; m is 0, 1, 2 or -(CH2) y -CO-; y is 0, 1 or 2.

[0012] In one embodiment of the present application, the carbon at the position marked with * is a carbon atom having a chiral center, and its chiral configuration is R or S alone.

[0013] In one embodiment of the present application, when X is a nitrogen atom carrying an alkyl group, the alkyl group is selected from a C1-C3 alkyl group.

[0014] In one embodiment of the present application, the camptothecin derivative compound has one of the following chemical formulas:

[0015] , , ,

[0016] , , ,

[0017] , , ,

[0018] , , ,

[0019] , , ,

[0020] , , ,

[0021] , , ,

[0022] , , .

[0023] In a second aspect, the present application provides a method for preparing a camptothecin derivative compound, comprising the following steps:

[0024] Exitecan mesylate is dissolved in a solvent, firstly reacted with chloroformate for condensation reaction, then reacted with amino alcohol compound for amine ester exchange, and then filtered, washed, dried and / or post-treated to obtain camptothecin derivative compounds.

[0025] In one embodiment of the present application, the preparation method of camptothecin derivative compounds specifically comprises the following steps:

[0026] Dissolve exitecan mesylate in a solvent, add triethylamine under stirring, then drop phenyl chloroformate, stir at room temperature, add triethylamine and amino alcohol compounds, react, and obtain camptothecin derivative compounds through filtering, washing, drying and / or post-treatment.

[0027] In one embodiment of the present application, the solvent is selected from one of dimethyl sulfoxide, toluene, xylene, and N,N-dimethylformamide.

[0028] In one embodiment of the present application, the solvent is selected from dimethyl sulfoxide.

[0029] In one embodiment of the present application, the amino alcohol compound can be selected from but is not limited to D-prolinol, L-prolinol, (S)-2-piperidinemethanol, (R)-2-piperidinemethanol, (R)-3-hydroxymethylmorpholine, (S)-3-hydroxymethylmorpholine, (R)-2-(pyrrolidin-2-yl)ethan-1-ol, (S)-3-(hydroxymethyl)piperazine-2-one, (R)-2-hydroxymethylazetidine, and (S)-1-Boc-3-hydroxymethylpiperazine.

[0030] In one embodiment of the present application, the molar ratio of exitecan mesylate to the amino alcohol compound is 1:1.5-2.5.

[0031] In one embodiment of the present application, the molar ratio of exitecan mesylate to the amino alcohol compound is 1:2.0.

[0032] In one embodiment of the present application, the molar ratio of exitecan mesylate to phenyl chloroformate is 1:1.0-2.0.

[0033] In one embodiment of the present application, the molar ratio of exitecan mesylate to phenyl chloroformate is 1:1.5.

[0034] In one embodiment of the present application, the molar ratio of exitecan mesylate to triethylamine is 1:4-8.

[0035] In one embodiment of the present application, the molar ratio of exitecan mesylate to triethylamine is 1:5-7.

[0036] In one embodiment of the present application, the molar ratio of exitecan mesylate to triethylamine is 1:6.

[0037] In one embodiment of the present application, the stirring temperature at room temperature is 20-30° C., and the stirring time at room temperature is 6-8 h.

[0038] In one embodiment of the present application, the reaction temperature is 20-30° C., and the reaction time is 1-24 h.

[0039] In one embodiment of the present application, the preparation of compound HG-YX-7 requires a post-processing step, and the post-processing step is specifically as follows:

[0040] The dried product was dissolved in trifluoroacetic acid, heated for reaction, concentrated under reduced pressure, extracted with an organic solvent, the pH value was adjusted to neutral, and the organic solvent was dried and concentrated to dryness to obtain compound HG-YX-7.

[0041] In one embodiment of the present application, the organic solvent is dichloromethane.

[0042] In a third aspect, the present application provides the use of camptothecin derivative compounds, or pharmaceutically acceptable salts, hydrates, solvates, optical isomers or crystal forms thereof as small molecule toxins for use in the preparation of antibody-drug conjugates.

[0043] In a fourth aspect, the present application provides a pharmaceutical composition comprising a pharmaceutically acceptable excipient or carrier and a camptothecin derivative compound, or a pharmaceutically acceptable salt, hydrate, solvate, optical isomer or crystal form thereof.

[0044] Compared with the prior art, the present invention has the following beneficial effects:

[0045] The present invention performs a series of derivatizations on isotecan on the basis of maintaining the pharmacodynamic skeleton of isotecan, thereby forming a series of isotecan derivatives. These compounds have the characteristics of high anti-tumor activity, low toxic side effects, and easy solubility in water, and have potential application value as anti-tumor single drugs or antibody-coupled drugs. DETAILED DESCRIPTION

[0046] The following is a clear and complete description of the technical solutions in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all of the embodiments.

[0047] The "range" disclosed in this application is defined in the form of a lower limit and an upper limit, and a given range is defined by selecting a lower limit and an upper limit, and the selected lower limit and upper limit define the boundaries of the particular range. The range defined in this way can be inclusive or exclusive of the end values, and can be arbitrarily combined, that is, any lower limit can be combined with any upper limit to form a range.

[0048] Unless otherwise specified, when the present invention relates to the percentage between liquids, the percentage is volume / volume percentage; when the present invention relates to the percentage between liquids and solids, the percentage is volume / weight percentage; when the present invention relates to the percentage between solids and liquids, the percentage is weight / volume percentage; the rest are weight / weight percentages.

[0049] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0050] Example 1: Preparation of compound (HG-YX-1)

[0051] ;

[0052] Dissolve exitecan mesylate (100 mg) in dimethyl sulfoxide (DMSO) (5 mL). Add triethylamine (58 mg) under stirring at room temperature and stir for 5 minutes. Then add phenyl chloroformate (45.02 mg) dropwise. Stir the reaction solution at room temperature for 7 hours. Add triethylamine (56 mg) and D-prolinol (38.0 mg). The reaction solution reacts at room temperature overnight.

[0053] Add water (15 mL), stir, and filter the precipitate. The filter cake was rinsed with water (10 mL) and dried to obtain compound (HG-YX-1) (62.0 mg) with a yield of 58.6%. MS m / z: 563.226 ([M+H] + ). 1 HNMR (400MHz, DMSO): 7.77-7.80, 7.31, 6.76-6.78, 6.53, 5.16-5.50, 3.21-3.34, 2.54, 2.40, 2.24-2.29, 1.87-1.92, 0.87.

[0054] Example 2: Preparation of compound (HG-YX-2)

[0055] The difference between Example 2 and Example 1 is that L-prolinol is used instead of D-prolinol to obtain compound (HG-YX-2) (48.2 mg) with a yield of 35.9%. MS m / z: 563.215 ([M+H] + ). 1 HNMR (400MHz, DMSO): 7.75-7.79, 7.31, 6.75-6.93, 6.53, 5.04-5.45, 3.13-3.20, 2.52, 2.38, 2.17-2.38, 1.80-1.86, 0.87.

[0056] Example 3: Preparation of compound (HG-YX-5)

[0057] The difference between Example 3 and Example 1 is that (R)-3-hydroxymethylmorpholine is used instead of D-prolinol to obtain compound (HG-YX-5) (56.8 mg) with a yield of 52.2%. MS m / z: 579.274 ([M+H] + ). 1 HNMR (400MHz, DMSO): 7.77, 7.31, 6.95, 6.52, 5.43, 5.23, 3.60-3.82, 3.42-3.52, 2.98-3.15, 2.39, 2.17, 1.80-1.92, 0.88.

[0058] Example 4: Preparation of compound (HG-YX-6)

[0059] The difference between Example 4 and Example 1 is that (S)-3-hydroxymethylmorpholine is used instead of D-prolinol to obtain compound (HG-YX-6) (55.6 mg) with a yield of 51.1%. MS m / z: 579.312 ([M+H] + ). 1 HNMR (400MHz, DMSO): 7.77, 7.30, 6.94, 6.43-5.49, 5.17, 3.86-3.94, 3.67-3.84, 3.39-3.60, 2.90-3.14, 2.38, 2.17, 1.81-1.95, 0.88.

[0060] Example 5: Preparation of Compound (HG-YX-11)

[0061] The difference between Example 5 and Example 1 is that (R)-2-(pyrrolidin-2-yl)ethan-1-ol was used instead of D-prolinol to obtain compound (HG-YX-11) (66.8 mg) with a yield of 61.6%. MS m / z: 577.312 ([M+H] + ). 1 HNMR (400MHz, DMSO): 7.76, 7.31, 6.77, 6.50, 5.41, 5.18, 3.89, 3.10, 2.38, 2.09-2.27, 1.81-1.95, 1.49-1.57, 0.88.

[0062] Example 6: Preparation of Compound (HG-YX-21)

[0063] The difference between Example 6 and Example 1 is that (S)-2-hydroxymethylazetidine was used instead of D-prolinol to obtain compound (HG-YX-21) (55.9 mg) with a yield of 54.2%. MS m / z: 559.264 ([M+H]+ ). 1 HNMR (400MHz, DMSO): 7.77, 7.30, 7.08, 6.51, 5.44, 5.09-5.36, 4.26, 3.80, 3.65, 3.52, 3.13, 2.37, 2.13, 1.81-1.95, 0.87.

[0064] Example 7: Preparation of Compound (HG-YX-22)

[0065] The difference between Example 7 and Example 1 is that (R)-2-hydroxymethylazetidine is used instead of D-prolinol to obtain compound (HG-YX-22) (50.6 mg) with a yield of 49.0%. MS m / z: 549.271 ([M+H] + ). 1 HNMR (400MHz, DMSO): 7.77, 7.32, 7.10, 6.50, 5.44, 5.10-5.36, 4.20, 3.82, 3.67, 3.5, 8, 3.14, 2.38, 2.14, 1.81-1.95, 0.88.

[0066] Example 8: Preparation of Compound (HG-YX-7)

[0067] ;

[0068] Dissolve exitecan mesylate (100 mg) in DMSO (5 mL). Add triethylamine (58 mg) under stirring at room temperature and stir for 5 minutes. Then add phenyl chloroformate (45.02 mg) dropwise. Stir the reaction solution at room temperature for 7 hours. Add triethylamine (56 mg) and (S)-1-Boc-3-hydroxymethylpiperazine (82.3 mg). The reaction solution reacts at room temperature overnight.

[0069] Water (15 mL) was added, stirred, and the precipitate was filtered. The filter cake was rinsed with water (10 mL) and dried to obtain compound (7-1) (58.2 mg) with a yield of 45.7%.

[0070] Compound (7-1) (50.0 mg) was dissolved in trifluoroacetic acid (10.0 mL), heated to 70°C, and reacted for 2 hours. The mixture was concentrated to dryness under reduced pressure, and dichloromethane (30.0 mL) was added. The mixture was washed with saturated sodium carbonate until neutral. The organic solvent was dried and concentrated to dryness to obtain compound (HG-YX-7) (36.0 mg) with a yield of 84.6%. MS m / z: 578.317 ([M+H] + ). 1HNMR (400MHz, DMSO): 7.79, 7.31, 6.89, 6.52, 5.44, 5.23, 3.42-3.52, 2.98-3.15, 2.62-2.83, 2.3, 6, 2.16, 1.83-1.92, 0.88.

[0071] The compounds prepared in the examples (compound (HG-YX-1), compound (HG-YX-2), compound (HG-YX-5), compound (HG-YX-6), compound (HG-YX-11), compound (HG-YX-21) and compound (HG-YX-22)) were tested for cell proliferation inhibition:

[0072] Take the required number of NCI-N87 cells (1500 cells / well / 40µL) (ATCC, CRL-5822) and inoculate them in a 384-well plate (Corning, 3764) and incubate them in a 37°C incubator overnight. The DMSO-diluted compounds were added to the cell culture plate through IDOT (DISPENDIX) and the compounds were allowed to act for 6 days. The CellTiter-Glo reagent (CTG, Vazyme, DD1101-03) was equilibrated with the cell culture plate at room temperature for 30 min, 20 µL CTG was added and shaken for 2 min, and incubated at room temperature in the dark for 30 min. The luminescent signal value was read using BMG (PHERAstar FSX).

[0073] The inhibition rate of the compound on the cells was standardized between the High control and the Low control. The wells containing cell culture medium were used as the Low control. The wells containing cells and the same proportion of DMSO were used as the High control. Inhibition%=(Ave_H-Sample) / (Ave_H-Ave_L), where Inhibition% is the inhibition rate, Ave_H represents the average signal value of the High control group, Ave_L represents the average signal value of the Low control group, and Sample is the signal value of the experimental group. IC was calculated by fitting a four-parameter logistic model or Excel 50 Value, IC 50 The value is the concentration that inhibits 50% of the curve.

[0074] The results are shown in Table 1. Compared with Dxd, the compound (HG-YX-1) is more sensitive to the cytotoxicity of NCI-N87 cells.

[0075] Table 1 Toxicity of each compound to NCI-N87 cells

[0076]

[0077] The present application is described in detail above in conjunction with specific implementation methods and exemplary embodiments, but these descriptions cannot be understood as limiting the present application. Those skilled in the art understand that, without departing from the spirit and scope of the present application, the technical solution of the present application and its implementation methods may be subjected to a variety of equivalent substitutions, modifications or improvements, all of which fall within the scope of the present application.

Claims

1. A camptothecin derivative compound, characterized in that: The structural formula of the camptothecin derivative compound is shown in formula (1): ； in: X is oxygen, nitrogen, a nitrogen atom with a C1-C3 alkyl group, a sulfur atom, a carbon atom, and -SO2-; n is 1 or 2; R1 and R2 are each independently selected from -(CH2) m -; m is 0, 1, 2 or -(CH2) y -CO-; y is 0, 1 or 2; It has one of the following chemical formulas: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 。 2. Use of the camptothecin derivative compound according to claim 1 as a small molecule toxin in the preparation of antibody-drug conjugates.

3. A pharmaceutical composition, characterized in that The composition comprises a pharmaceutically acceptable excipient or carrier and the camptothecin derivative compound as claimed in claim 1.

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