A glycosylated camptothecin compound Gly-CPTR and its pharmaceutical use, drug or prodrug and preparation method

Abstract

The present invention "a glycosylated camptothecin compound Gly-CPTR and its pharmaceutical use, drug or prodrug and preparation method" belongs to the field of chemical drugs. The glycosylated camptothecin compound Gly-CPTR is a compound in which a glycosyl molecule is connected to a camptothecin derivative CPTR through an ether bond. The present invention also provides the use of Gly-CPTR in which a glycosyl molecule is connected to CPTR through an ether bond in the preparation of an anti-colorectal cancer drug or prodrug, as well as an anti-colorectal cancer drug or prodrug based on the Gly-CPTR and a preparation method thereof. The Gly-CPTR and its drug or prodrug of the present invention can be administered orally and targeted to the large intestine, and have significant tumor inhibition ability against in situ colorectal cancer in mice, and have high safety.

Description

Technical Field

[0001] The present invention relates to the field of chemical medicines, and in particular to a glycosylated camptothecin compound Gly-CPTR and its pharmaceutical use, medicine or prodrug and preparation method. Background Art

[0002] The drugs currently used in clinical chemotherapy of colorectal cancer mainly include fluorouracil, oxaliplatin, capecitabine, and irinotecan. Except for capecitabine, other drugs are prone to produce serious systemic toxic side effects and need to be administered by drip or push injection, which brings great inconvenience to clinical medication and also brings high risks and pain to patients.

[0003] Irinotecan is a prodrug of camptothecins, such as Figure 1 As shown, it is converted into its active form SN38 (7-ethyl-10-hydroxycamptothecin) under the action of carboxylesterase (CES) in the body, and is an important drug for the clinical treatment of metastatic colorectal cancer. Although irinotecan has a wide range of anti-tumor activity, it has problems of poor water solubility and high systemic toxicity. The main side effects include more severe delayed diarrhea, neutropenia, nausea and vomiting, acute cholinergic syndrome, etc. Therefore, it can only be administered by intravenous drip to achieve the anti-cancer active dose while minimizing the side effects.

[0004] The clinical intravenous drip preparation is an injection solution dissolved in water after acidification, which leads to unstable preparation quality and reduced anticancer activity, and requires repeated administration or extended treatment, which leads to increased dose-limiting toxicity. In addition, hydroxycamptothecin is mainly distributed in the gallbladder after intravenous administration, while the distribution in the rectum, a common tissue and organ for cancer, is not ideal.

[0005] The anti-tumor activity of SN38 is 100 to 1000 times that of its prodrug irinotecan, but its water solubility is extremely poor and difficult to make into a drug, and it has high systemic toxicity. How to improve the water solubility of SN38 and release it in the targeted colon cancer area has become an urgent problem to be solved in the current clinical treatment of colorectal cancer. Therefore, the development of chemotherapy drugs with targeted effects and oral administration is an important way to optimize the clinical treatment of colorectal cancer.

[0006] The lesion site of colorectal cancer, the colorectum, has a rich and diverse intestinal flora, which secretes a large number of biological enzymes, of which glycosidase is a large category. The existing technology has reported the modification of sugar chains of camptothecin drugs, which physically shields camptothecin drugs in structure. This type of [sugar chain-drug] prodrug molecules are inactive before reaching the colorectal site, greatly reducing their toxicity. After reaching the colorectum, the sugar chains are cut off under the action of the intestinal flora enzyme-glycosidase, releasing the active molecules of camptothecin, achieving the effect of anti-colorectal cancer.

[0007] However, the sugar chain-modified prodrugs of camptothecin drugs reported so far usually use a linker containing a benzene ring to connect the sugar chain to the camptothecin drugs, and the sugar chain molecule generally uses glucuronic acid. This type of prodrug has a complex structure and high preparation cost. Its release mechanism in the body is to release active camptothecin drugs after entering the cells. The metabolites of the benzene ring part after release may have certain toxic side effects, and the efficacy depends to a large extent on the membrane permeability of this type of prodrug.

[0008] There is an urgent need in the art to develop other camptothecin prodrug compounds with better cell activity, water solubility and toxicity after drug release, simpler structure and the ability to be released outside the cell. Summary of the invention

[0009] Based on the above-mentioned deficiencies and needs of the prior art in the art, the purpose of the present invention is to provide a camptothecin prodrug compound with a new structure, which has a simpler structure, a simpler drug release mechanism, can release the drug extracellularly, and can also achieve better cell activity and water solubility and completely eliminate toxicity after drug release.

[0010] The glycosylated camptothecin compound Gly-CPTR prepared by the present invention has low toxicity, greatly enhanced water solubility, is not degraded in the stomach and small intestine, can release the drug in the targeted colon cancer site, and is an ideal oral anti-colorectal cancer drug.

[0011] The technical solution of the present invention is as follows:

[0012] A glycosylated camptothecin compound Gly-CPTR, characterized in that it is a compound in which a glycosyl molecule is connected to a camptothecin derivative CPTR via an ether bond.

[0013] The advantage of using ether bonds to connect the prodrugs of the present invention is that ether bonds are the simplest form, and under the action of intestinal microbial enzymes, ether bonds can be directly broken to release sugars and camptothecin drugs; if other bonds, such as ester bonds, are used, they may be degraded by intestinal esterases in the small intestine, and the purpose of safely targeting and delivering camptothecin drugs to the colorectum cannot be achieved. If the existing benzene ring-containing linker is used, the metabolites of the benzene ring part of the prodrug after release in vivo have greater cytotoxicity (as shown in Table 1 below), and the drug release efficiency and anti-tumor activity are poor (as shown in Tables 2, Figure 3 ).

[0014] The camptothecin derivative CPTR has the structure of the following formula I:

[0015] Formula I:

[0016]

[0017] In Formula I, R 1 Selected from: H, NO 2 , CH2 CH=CH 2 , CH 2 NMe 2 ; R 2 Selected from: H or CH 2 CH 3 ;

[0018] Preferably, the saccharide group is selected from the group consisting of glucosyl, galactosyl, fucosyl, lactosyl, xylosyl, mannosyl, cellobiosyl, maltobiosyl or maltotriosyl.

[0019] The modified group used in the prodrug of the present invention is a sugar group instead of a uronic acid group, and the positive effects are: the sugar group is diversified and the preparation cost is lower. The modification of camptothecin with uronic acid requires a multi-step chemical reaction operation. In addition, the camptothecin drugs modified with uronic acid are mainly concentrated inside the cancer cells and released by uronidase degradation. The main purpose of the present invention is to transport the active drug to the colorectal part, release the active camptothecin molecules in the colorectal part, and these active molecules release anti-cancer effects after entering the cells.

[0020] The glycosylated camptothecin compound Gly-CPTR has a structure shown in the following formula II:

[0021] Formula II:

[0022]

[0023] In Formula II, R 1 Selected from: H, NO 2 , CH 2 CH=CH 2 , CH 2 NMe 2 ; R 2 Selected from: H or CH 2 CH 3 .

[0024] The glycosylated camptothecin compound Gly-CPTR is selected from the compounds shown in the following structural formula:

[0025]

[0026] Preferably, the glycosylated camptothecin compound Gly-CPTR is a prodrug of a drug for treating colorectal cancer;

[0027] Preferably, the drug for treating colorectal cancer uses a camptothecin derivative as a pharmacologically active ingredient;

[0028] Preferably, the camptothecin derivative CPTR is selected from: 7-ethyl-10-hydroxycamptothecin, 10-hydroxycamptothecin, 9-methylene-N,N-dimethyl-10-hydroxycamptothecin, 9-nitro-10-hydroxycamptothecin, 9-allyl-10-hydroxycamptothecin;

[0029] Preferably, the prodrug further comprises a pharmaceutically acceptable excipient.

[0030] Use of Gly-CPTR in which a glycosyl molecule is connected to a camptothecin drug via an ether bond in the preparation of an anti-colorectal cancer drug or prodrug.

[0031] Gly-CPTR has the structure shown in Formula II below:

[0032]

[0033] In Formula II, R 1 Selected from: H, NO 2 , CH 2 CH=CH 2 , CH 2 NMe 2 ; R 2 Selected from: H or CH 2 CH 3 ;

[0034] Preferably, the glycosyl is selected from the group consisting of glucosyl, galactosyl, fucosyl, lactosyl, mannosyl, xylosyl, mannosyl, cellobiosyl-, maltobiosyl, maltotriosyl;

[0035] Preferably, the anti-colorectal cancer drug or prodrug is selected from the compounds shown in the following structural formula:

[0036]

[0037] The anti-colorectal cancer drug uses the camptothecin derivative CPTR as the drug's pharmacologically active ingredient;

[0038] Preferably, the camptothecin derivative CPTR is selected from: 7-ethyl-10-hydroxycamptothecin, 10-hydroxycamptothecin, 9-methylene-N,N-dimethyl-10-hydroxycamptothecin, 9-nitro-10-hydroxycamptothecin, 9-allyl-10-hydroxycamptothecin;

[0039] Preferably, the drug or prodrug further comprises a pharmaceutically acceptable excipient;

[0040] Preferably, the anti-colorectal cancer drug or prodrug is an oral drug.

[0041] An anti-colorectal cancer drug or prodrug, comprising an active ingredient; characterized in that the active ingredient comprises: the glycosylated camptothecin compound Gly-CPTR.

[0042] The anti-colorectal cancer drug or prodrug further includes: pharmaceutically acceptable excipients.

[0043] A method for preparing an anti-colorectal cancer drug or prodrug, characterized in that it is prepared by using a fully acyl halide sugar and CPTR through a Koenigs-Knorr glycosidation reaction.

[0044] The peracyl halide sugar is selected from the group consisting of peracetyl bromide sugar, peracetyl chloride sugar, peracetyl iodide sugar, perbenzoyl bromide sugar, perbenzoyl chloride sugar, and perbenzoyl iodide sugar;

[0045] Preferably, the peracetyl brominated sugar is selected from the compounds of the following structural formula:

[0046]

[0047] Preferably, the Koenigs-Knorr glycosidation reaction refers to: the fully acyl halide sugar and CPTR are dissolved in an organic solvent and then a catalyst is added to react;

[0048] Preferably, the catalyst is selected from: silver oxide or anhydrous potassium carbonate or silver trifluoromethanesulfonate;

[0049] Preferably, the organic solvent is selected from: dry acetonitrile, dichloromethane, tetrahydrofuran, N,N-dimethylformamide, acetone, N,N-dimethyl sulfoxide;

[0050] Preferably, the reaction temperature is 25-45°C;

[0051] Preferably, the reaction time is 3 hours;

[0052] Preferably, the usage ratio of the full acyl halide sugar, CPTR, and catalyst is: 1.0-2.0eq: 1.0-2.0eq: 1.0-2.0eq;

[0053] Preferably, after the reaction is completed, the liquid obtained by filtration is concentrated to obtain a crude product;

[0054] Preferably, the crude product is purified by acidic resin and silica gel column in sequence to obtain the final product;

[0055] Preferably, the acid resin purification refers to the crude product being dissolved in a mixed solution of methanol and dichloromethane and reacting at room temperature for 2 hours before adding the acid resin;

[0056] Preferably, the methanol-dichloromethane mixed solution refers to a solution obtained by mixing methanol and dichloromethane in a volume ratio of 1:1 to 1:4;

[0057] Preferably, the pH of the solution dissolved in methanol-dichloromethane is adjusted to 9-11;

[0058] Preferably, the pH adjustment refers to adding 1M sodium methoxide dropwise for adjustment;

[0059] Preferably, the acidic resin refers to acidic resin Resin IR120;

[0060] Preferably, the acidic resin is added and stirred until the pH of the system is 7.0;

[0061] Preferably, the silica gel column purification refers to: after the acid resin purification, the product is filtered and the filtrate obtained is passed through a silica gel column, and the mobile phase is ethyl acetate: methanol: water = 7:2:0 to 7:2:1.

[0062] The beneficial effects of the present invention are:

[0063] 1. The present invention modifies CPTR with sugar chains by chemically synthesizing all-acyl halide sugars and camptothecin derivatives CPTR to form glycosylated camptothecin drugs Gly-CPTR, thereby greatly reducing the toxicity of SN38 and improving the water solubility of Gly-CPTR. Under the action of glycosidase, the drug is released in a targeted manner to the colon cancer site, which greatly improves the targeted therapeutic effect of Gly-CPTR on colon cancer and has great application value in clinical treatment.

[0064] 2. The present invention not only produces the glycosylated camptothecin drug Gly-CPTR by chemical synthesis, but also confirms the targeting ability and tumor inhibition ability of Gly-CPTR on colorectal cancer through a large number of biological activity experiments. It can directly reach the colorectal area under oral administration and release active CPTR in the colorectal area. It has very good targeted drug release efficacy and can effectively inhibit the growth of tumors in situ colorectal mice. Compared with irinotecan, the prodrug of SN38, Gly-CPTR does not cause toxic side effects such as severe weight loss (caused by delayed diarrhea reaction) and a significant reduction in the number and ratio of neutrophils. It has stronger drug safety and is a very ideal oral anti-colorectal cancer drug with good market application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0065] In order to more clearly illustrate the technical solutions in the experimental examples of the present invention, the drawings required for use in the description of the experimental examples will be briefly introduced below. Obviously, the drawings described below are only some experimental examples of the present invention. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without paying creative work.

[0066] Figure 1Schematic diagram of the release of active drug SN38 of irinotecan in vivo in the prior art.

[0067] Figure 2 This is the structure of compound 14 used for comparison in Experimental Examples 2-4 of the present invention.

[0068] Figure 3 This is the tumor inhibition ability of Gly-CPTR in Experimental Example 4 of the present invention on the orthotopic colorectal cancer nude mouse model.

[0069] Figure 4 It is a line graph of the acute toxicity (body weight change) of Gly-CPTR and irinotecan in mice in Experimental Example 5 of the present invention. DETAILED DESCRIPTION

[0070] The technical scheme of the present invention will be clearly and completely described below in conjunction with specific embodiments and experimental examples. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0071] Sources of biological materials

[0072] The SD rats used in Experimental Example 3 of the present invention, the SPF male nude mice used in Experimental Example 3, and the Balb / C mice used in Experimental Example 5 can all be commercially available.

[0073] Sources of experimental materials and reagents

[0074] The experimental methods used in the experimental examples of the present invention are all conventional methods.

[0075] The materials and reagents used in the experimental examples of the present invention can be obtained from commercial sources.

[0076] Group 1 Example: Glycosylated camptothecin compound Gly-CPTR of the present invention

[0077] This group of embodiments provides a glycosylated camptothecin compound Gly-CPTR. All embodiments in this group have the following common features: the glycosylated camptothecin compound Gly-CPTR is a compound in which a glycosyl molecule is connected to a camptothecin derivative CPTR via an ether bond.

[0078] Based on the teachings of the present invention, those skilled in the art can select appropriate chemical reaction conditions in combination with common technical means in the field of chemistry to connect the glycosyl molecule and the camptothecin derivative through an ether bond. Any act of connecting the glycosyl molecule and the camptothecin derivative in the form of an ether bond and / or synthesizing, making, using, selling, promising to sell, producing, importing, exporting, or reacting to generate the Gly-CPTR falls within the scope of protection of the present invention.

[0079] In some embodiments, the camptothecin derivative CPTR has the structure of the following formula I:

[0080] Formula I:

[0081]

[0082] In Formula I, R 1 Selected from: H, NO 2 , CH 2 CH=CH 2 , CH 2 NMe 2 ; R 2 Selected from: H or CH 2 CH 3 ;

[0083] According to the above R 1 and R 2 Based on the general principles of chemistry, other atoms, molecules, and groups with similar structures or properties can be selected to replace the above H, NO 2 , CH 2 CH=CH 2 , CH 2 NMe 2 , CH 2 CH 3 And a camptothecin derivative CPTR with similar structure or properties was obtained.

[0084] Preferably, the saccharide group is selected from the group consisting of glucosyl, galactosyl, fucosyl, lactosyl, xylosyl, mannosyl, cellobiosyl, maltobiosyl or maltotriosyl.

[0085] According to the teachings and inspiration of the specific sugar groups listed above and based on the general principles of chemistry, those skilled in the art can select other sugar molecules or groups with similar structures or properties to replace the above sugar groups for preparing the Gly-CPTR of the present invention.

[0086] In a more preferred embodiment, the glycosylated camptothecin compound Gly-CPTR has a structure shown in the following formula II:

[0087] Formula II:

[0088]

[0089] In Formula II, R 1 Selected from: H, NO 2 , CH 2 CH=CH 2 , CH 2 NMe 2 ; R 2 Selected from: H or CH 2 CH 3 .

[0090] According to the above R 1 and R 2 Based on the general principles of chemistry, other atoms, molecules, and groups with similar structures or properties can be selected to replace the above H, NO 2 , CH 2 CH=CH 2 , CH 2 NMe 2 , CH 2 CH 3 And a camptothecin derivative CPTR with similar structure or properties was obtained.

[0091] In a specific embodiment, the glycosylated camptothecin compound Gly-CPTR is selected from the compound shown in the following structural formula:

[0092]

[0093] According to the teachings and inspiration of the compounds shown in the above structural formulas and based on the general principles of the chemical field, those skilled in the art can select glycosyl molecules or groups other than "glucosyl, galactosyl, fucosyl, lactosyl, xylosyl, mannosyl, cellobiosyl, maltobiosyl or maltotriosyl" to combine with the CPTR shown in the above formula II, react, prepare, synthesize, and produce other Gly-CPTRs with similar structures or properties connected by ether bonds.

[0094] In some embodiments, the glycosylated camptothecin compound Gly-CPTR is a prodrug of a drug for treating colorectal cancer;

[0095] Preferably, the drug for treating colorectal cancer uses a camptothecin derivative as a pharmacologically active ingredient;

[0096] Preferably, the camptothecin derivative CPTR is selected from: 7-ethyl-10-hydroxycamptothecin, 10-hydroxycamptothecin, 9-methylene-N,N-dimethyl-10-hydroxycamptothecin, 9-nitro-10-hydroxycamptothecin, 9-allyl-10-hydroxycamptothecin;

[0097] According to the teachings and inspirations of the above-mentioned specific camptothecin derivatives CPTR and based on the general principles of the chemical field, those skilled in the art can select other camptothecin derivatives with similar structures or properties to replace the above-mentioned camptothecin derivatives.

[0098] Preferably, the prodrug further comprises a pharmaceutically acceptable excipient.

[0099] Group 2 Example: Pharmaceutical Use of the Compound Gly-CPTR of the Present Invention

[0100] This group of embodiments provides the use of Gly-CPTR in which a glycosyl molecule is connected to a camptothecin drug via an ether bond in the preparation of an anti-colorectal cancer drug or prodrug.

[0101] In some embodiments, Gly-CPTR has a structure as shown in Formula II below:

[0102] Formula II:

[0103] In Formula II, R 1 Selected from: H, NO 2 , CH 2 CH=CH 2 , CH 2 NMe 2 ; R 2 Selected from: H or CH 2 CH 3 ;

[0104] According to the above R 1 and R 2 Based on the general principles of chemistry, other atoms, molecules, and groups with similar structures or properties can be selected to replace the above H, NO 2 , CH 2 CH=CH 2 , CH 2 NMe 2 , CH 2 CH 3 And a camptothecin derivative CPTR with similar structure or properties was obtained.

[0105] Preferably, the glycosyl is selected from the group consisting of: glucosyl, galactosyl, fucosyl, lactosyl, mannosyl, xylosyl, mannosyl, cellobiosyl-, maltobiosyl, maltotriosyl;

[0106] Based on the teachings and inspiration of the specific sugar groups listed above and the general principles of chemistry, those skilled in the art can select other sugar molecules or groups with similar structures or properties to replace the above sugar groups for the preparation of the anti-colorectal cancer drugs or prodrugs of the present invention.

[0107] Preferably, the anti-colorectal cancer drug or prodrug is selected from the compounds shown in the following structural formula:

[0108]

[0109] According to the teachings and inspiration of the compounds shown in the above structural formulas and based on the general principles of the chemical field, those skilled in the art can select sugar molecules or groups other than "glucose, galactosyl, fucosyl, lactosyl, xylosyl, mannosyl, cellobiosyl, maltobiosyl or maltotriosyl" to combine with the CPTR shown in the above formula II, react, prepare, synthesize, and produce other drugs or prodrugs with similar structures or properties connected by ether bonds.

[0110] The anti-colorectal cancer drug uses the camptothecin derivative CPTR as a pharmacologically active ingredient;

[0111] Preferably, the camptothecin derivative CPTR is selected from: 7-ethyl-10-hydroxycamptothecin, 10-hydroxycamptothecin, 9-methylene-N,N-dimethyl-10-hydroxycamptothecin, 9-nitro-10-hydroxycamptothecin, 9-allyl-10-hydroxycamptothecin;

[0112] According to the teachings and inspiration of the above-mentioned specific camptothecin derivative CPTRs and based on the general principles of the chemical field, those skilled in the art can select other camptothecin derivative CPTRs with similar structures or properties to replace the above-mentioned H camptothecin derivative CPTR and obtain camptothecin derivative CPTRs with similar structures or properties.

[0113] Preferably, the drug or prodrug further comprises a pharmaceutically acceptable excipient;

[0114] In a specific embodiment, the pharmaceutically acceptable excipients are selected from: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, and release retardants.

[0115] According to the content of the present invention, for different needs in actual production applications, combined with conventional technical means in the field of drug preparation (for example, "Encyclopedia of Preparation Technology", "Drug Preparation Technology", etc.), those skilled in the art can select and formulate the above-mentioned excipients, and prepare the Gly-CPTR of the present invention into different dosage forms, such as powders, tablets, injections, oral liquids, capsules, granules, sprays, gels, ointments, etc.

[0116] Preferably, the anti-colorectal cancer drug or prodrug is an oral drug.

[0117] Based on the contents described in the present invention and in combination with conventional technical operations in the pharmaceutical field, those skilled in the art can prepare the drugs or prodrugs of the present invention into other dosage forms, such as oral liquids, powders, tablets, capsules, granules, decoctions, pills, sprays, inhalants, atomizers, injections, etc.

[0118] Those skilled in the art can make routine selections or adjustments to pharmaceutical excipients based on actual production needs, combined with conventional technical means or basic common sense of pharmaceutical production processes (e.g., "Encyclopedia of Pharmaceutical Technology", "Pharmaceutical Preparation Technology", etc.), and then prepare drugs with different dosage forms, different storage conditions, and different shelf lives. This is possible and easy to do without technical barriers for those skilled in the art.

[0119] Group 3 Examples: Anti-colorectal cancer drugs or prodrugs of the present invention

[0120] This group of embodiments provides an anti-colorectal cancer drug or prodrug. All embodiments in this group have the following common features: the anti-colorectal cancer drug or prodrug includes an active ingredient; the active ingredient includes: a glycosylated camptothecin compound Gly-CPTR provided in any one of the first group of embodiments.

[0121] In a further embodiment, the anti-colorectal cancer drug or prodrug further comprises: a pharmaceutically acceptable excipient.

[0122] In a specific embodiment, the pharmaceutically acceptable excipients are selected from: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, and release retardants.

[0123] According to the content of the present invention, for different needs in actual production applications, combined with conventional technical means in the field of drug preparation (for example, "Encyclopedia of Preparation Technology", "Drug Preparation Technology", etc.), those skilled in the art can select and formulate the above-mentioned excipients, and prepare the Gly-CPTR of the present invention into different dosage forms, such as powders, tablets, oral liquids, capsules, granules, sprays, gels, ointments, decoctions, pills, sprays, inhalants, atomizers, injections, etc.

[0124] Group 4 Example: Method for preparing the anti-colorectal cancer drug or prodrug of the present invention

[0125] This group of embodiments provides a method for preparing an anti-colorectal cancer drug or prodrug. All embodiments in this group have the following common features: the drug or prodrug is prepared by using a fully acyl halide sugar and CPTR through a Koenigs-Knorr glycosylation reaction.

[0126] In a specific embodiment, the peracyl halide sugar is selected from the group consisting of peracetyl bromide sugar, peracetyl chloride sugar, peracetyl iodide sugar, perbenzoyl bromide sugar, perbenzoyl chloride sugar, and perbenzoyl iodide sugar;

[0127] According to the teachings and inspiration of the above-mentioned specific peracyl halide sugars and based on the general principles of the chemical field, those skilled in the art can select other peracyl halide sugars other than "peracetyl bromide sugars, peracetyl chloride sugars, peracetyl iodide sugars, perbenzoyl bromide sugars, perbenzoyl chloride sugars, and perbenzoyl iodide sugars" to replace the above-mentioned specific peracyl halide sugars to participate in the reaction.

[0128] Preferably, the peracetyl brominated sugar is selected from the compounds of the following structural formula:

[0129]

[0130] According to the teachings and inspiration of the specific peracyl halide sugars shown in the above structural formulas and based on the general principles of the chemical field, those skilled in the art can select peracyl halide sugars with similar structures and properties, other than the specific peracyl halide sugars shown in the above structural formulas, and having another structural formula to replace the specific peracyl halide sugars shown in the above structural formulas to participate in the reaction.

[0131] In some embodiments, the Koenigs-Knorr glycosidation reaction refers to: a catalyst is added after the peracyl halide sugar and CPTR are dissolved in an organic solvent;

[0132] Preferably, the catalyst is selected from: silver oxide or anhydrous potassium carbonate or silver trifluoromethanesulfonate;

[0133] Preferably, the organic solvent is selected from: dry acetonitrile, dichloromethane, tetrahydrofuran, N,N-dimethylformamide, acetone, N,N-dimethyl sulfoxide;

[0134] Preferably, the reaction temperature is 25-45°C;

[0135] Preferably, the reaction time is 3 hours;

[0136] Preferably, the usage ratio of the full acyl halide sugar, CPTR, and catalyst is: 1.0-2.0eq: 1.0-2.0eq: 1.0-2.0eq;

[0137] Preferably, after the reaction is completed, the liquid obtained by filtration is concentrated to obtain a crude product;

[0138] Preferably, the crude product is purified by acidic resin and silica gel column in sequence to obtain the final product;

[0139] Preferably, the acid resin purification refers to the crude product being dissolved in a mixed solution of methanol and dichloromethane and reacting at room temperature for 2 hours before adding the acid resin;

[0140] Preferably, the methanol-dichloromethane mixed solution refers to a solution obtained by mixing methanol and dichloromethane in a volume ratio of 1:1 to 1:4;

[0141] Preferably, the pH of the solution dissolved in methanol-dichloromethane is adjusted to 9-11;

[0142] Preferably, the pH adjustment refers to adding 1M sodium methoxide dropwise for adjustment;

[0143] Preferably, the acidic resin refers to acidic resin Resin IR120;

[0144] Preferably, the acidic resin is added and stirred until the pH of the system is 7.0;

[0145] Preferably, the silica gel column purification refers to: after the acid resin purification, the product is filtered and the filtrate obtained is passed through a silica gel column, and the mobile phase is ethyl acetate: methanol: water = 7:2:0 to 7:2:1.

[0146] Experimental Example 1: Glycosylated camptothecin drug Gly-CPTR of the present invention

[0147] The synthetic route of Gly-CPTR is as follows:

[0148]

[0149] Synthesis process:

[0150] Full acyl halide sugar (1.0-2.0eq) and CPTR (100mg) are dissolved in dry acetonitrile (or dichloromethane, tetrahydrofuran, N,N-dimethylformamide, acetone, N,N-dimethylformamide), silver oxide or anhydrous potassium carbonate or silver trifluoromethanesulfonate (1.0-2.0eq) are added, and the reaction is carried out at 25 degrees Celsius to 45 degrees Celsius for 3 hours. The solid is filtered off and the liquid is concentrated. The obtained crude product is dissolved in a methanol-dichloromethane (1 / 1-1 / 4) mixed solution, and 1M sodium methoxide is added dropwise to the system pH 9-11. After reacting at room temperature for 2 hours, acidic resin Resin IR120 is added and stirred to the system pH 7.0, the resin is filtered off, and the filtrate is concentrated and passed through a silica gel column with a mobile phase of ethyl acetate: methanol: water = 7:2:0-7:2:1 to obtain a white powder product.

[0151] Test result data:

[0152] Glucose-10-hydroxycamptothecin (1): total yield 60%. 1 H NMR(500MHz,d6-DMSO)δ8.10(d,J=9.2Hz,1H),7.73(s,1H),7.78(s,1H),7.57(d,J=9.0Hz,1H),7.27(s ,1H),6.51(s,1H),5.43(d,J=5.8Hz,1H),5.42(s,2H),5.30(s,2H),5.17(s,1H),5.15(d,J=6.5Hz,1H), 5.11(d,J=5.1Hz,1H),4.68(s,J=5.45Hz,1H),3.76(dd,J=9.3,5.3Hz,1H),3.50(d,J=7.3Hz,2H),3.36– 3.30(m,5H),3.26–3.14(m,1H),1.87(dt,J=14.4,6.9Hz,2H),0.87(t,J=7.3Hz,3H).HR-ESIMS(m / z)for C 26 H 27 N 2 O 10 [M+H] + ,calcd:527.16657,found:527.16663.

[0153] Glucose-7-ethyl-10-hydroxycamptothecin (2): total yield 65%. 1 H NMR(500MHz,d6-DMSO)δ8.11(d,J=9.2Hz,1H),7.73(s,1H),7.58(d,J=9.0Hz,1H),7.28(s,1H),6.51(s,1 H),5.44(d,J=5.9Hz,1H),5.43(s,2H),5.31(s,2H),5.17(s,1H),5.15(d,J=6.5Hz,1H),5.11(d,J=5.1Hz, 1H),4.68(s,J=5.45Hz,1H),3.76(dd,J=9.3,5.3Hz,1H),3.50(d,J=7.3Hz,2H),3.36–3.30(m,5H),3.26–3 .14(m,3H),1.87(dt,J=14.4,6.9Hz,2H),1.31(t,J=7.4Hz,4H),0.88(t,J=7.2Hz,3H).HR-ESIMS(m / z)for C 28 H 30 N2 O 10 [M+H] + ,calcd:555.19787,found:555.19853.

[0154] Glucose-9-methylene-N,N-dimethyl-10-hydroxycamptothecin (3): total yield 54%. 1 H NMR(500MHz,d6-DMSO)δ8.10(d,J=9.2Hz,1H),7.73(s,1H),7.78(s,1H),7.36(s,1H),6.52(s,1H),5.44(d, J=5.9Hz,1H),5.43(s,2H),5.30(s,2H),5.17(s,1H),5.15(d,J=6.5Hz,1H),5.11(d,J=5.1Hz,1H),4.67(s, 2H),4.68(d,J=5.45Hz,1H),3.76(dd,J=9.3,5.3Hz,1H),3.50(d,J=7.3Hz,2H),3.36–3.30(m,5H),3.26–3. 14(m,1H),2.20(s,3H),2.21(s,3H),1.86(dt,J=14.3,6.8Hz,2H),0.88(t,J=7.2Hz,3H).HR-ESIMS(m / z)for C 29 H 34 N 3 O 10 [M+H] + ,calcd:584.22442,found:584.22451.

[0155] Galactose-7-ethyl-10-hydroxycamptothecin (4): total yield 72%. 1H NMR (500MHz, d6-DMSO) δ8.11(d,J=9.2Hz,1H),7.75(d,J=2.4Hz,1H),7.58(dd,J=9.2,2.5Hz,1H),7.29( s,1H),6.51(s,1H),5.43(s,2H),5.35–5.25(m,3H),5.09(d,J=7.7Hz,1H),4.94(s,1H),4.75(t,J=4.9H z,1H),4.58(d,J=4.2Hz,1H),3.82–3.66(m,3H),3.58(dd,J=10.4,5.4Hz,2H),3.50(d,J=10.2Hz,2H),3 .18(dd,J=14.8,7.0Hz,2H),1.87(dt,J=14.2,6.9Hz,2H),1.32(t,J=7.6Hz,3H),0.88(t,J=7.3Hz,3H). 13 C NMR(125MHz,d6-DMSO)δ173.02,157.33,156.66,150.62,150.53,146.72,145.22,144.71,131.75,128.79,128.15,123.00,118.8 7,107.15,101.68,96.62,76.40,73.82,72.87,70.70,68.73,65.73,60.98,50.01,30.71,22.57,14.04,8.23.HR-ESIMS(m / z)for C 28 H 30 N 2 O 10 [M+H] + ,calcd:555.19787,found:555.19692.

[0156] Xylose-7-ethyl-10-hydroxycamptothecin (5): total yield 75%. 1H NMR(500MHz,d6-DMSO)δ8.13(d,J=9.2Hz,1H),7.66(d,J=2.6Hz,1H),7.60(dd,J=9.2,2.6Hz,1H),7. 29(s,1H),6.51(s,1H),5.47(d,J=4.6Hz,1H),5.43(s,2H),5.32(s,2H),5.22(d,J=7.1Hz,1H),5.19 (d,J=4.1Hz,1H),5.13(d,J=3.7Hz,1H),3.81(d,J=5.8Hz,1H),3.48–3.39(m,2H),3.18(ddd,J=48.7 ,13.6,7.5Hz,3H),1.86(ddd,J=21.4,14.1,7.0Hz,2H),1.31(t,J=7.6Hz,3H),0.88(t,J=7.3Hz,3H). 13 C NMR(125MHz,d6-DMSO)δ173.02,157.33,156.29,150.77,150.52,146.68,145.25,144.61,131.93,128.87,128.07,122.55,11 8.94,107.46,101.18,96.66,76.88,73.55,72.87,69.86,66.17,65.73,50.02,30.70,22.74,14.01,8.23.HR-ESIMS(m / z)for C 27 H 28 N 2 NaO 9 [M+Na] + ,calcd:547.16925,found:547.16876.

[0157] Fucose-7-ethyl-10-hydroxycamptothecin (6): total yield 79%. 1H NMR(500MHz,d6-DMSO)δ8.11(d,J=9.2Hz,1H),7.67(d,J=2.5Hz,1H),7.58(dd,J=9.2,2.6Hz,1H),7.29(s,1H),6.51(s ,1H),5.43(s,2H),5.27(d,J=5.0Hz,1H),5.16(d,J=7.7Hz,1H),4.89(d,J=5.7Hz,1H),4.62(d,J=4.7Hz,1H),3.93(q,J =6.6Hz,1H),3.76–3.59(m,1H),3.54(t,J=3.8Hz,1H),3.53–3.47(m,1H),3.25(dd,J=13.6,7.6Hz,1H),3.09(dd,J=13. 6,7.5Hz,1H),1.87(ddt,J=21.3,14.0,7.2Hz,2H),1.32(t,J=7.6Hz,3H),1.19(d,J=6.4Hz,3H),0.88(t,J=7.3Hz,3H). 13 C NMR (125MHz, d6-DMSO) δ171.94,156.24,155.35,149.54,149.44,145.65,144.07,143.37,130.75,127.75,126.99,121.74,117. 81,105.92,99.89,95.53,72.84,71.79,70.33,69.82,69.22,64.65,48.94,29.62,21.71,16.03,12.89,7.14.HR-ESIMS(m / z)for C 28 H 31 N 2 O 9 [M+H] + ,calcd:539.20296,found:539.20357.

[0158] Mannosyl-7-ethyl-10-hydroxycamptothecin (7): total yield 54%. 1H NMR (500MHz, d6-DMSO) δ8.11(d,J=9.1Hz,1H),7.76(d,J=2.5Hz,1H),7.59(dd,J=9.2,2.5Hz,1H),7.28( s,1H),6.51(s,1H),5.43(s,2H),5.35–5.25(m,3H),5.01(d,J=1.3Hz,1H),4.94(s,1H),4.76(t,J=4.9H z,1H),4.60(d,J=4.2Hz,1H),3.82–3.63(m,3H),3.58(dd,J=10.4,5.4Hz,2H),3.50(d,J=10.2Hz,2H),3 .18(dd,J=14.8,7.0Hz,2H), 1.86(dt,J=14.1,7.0Hz,2H), 1.32(t,J=7.6Hz,3H), 0.88(t,J=7.3Hz,3H). 13 C NMR(125MHz,d6-DMSO)δ173.03,157.32,156.69,150.61,150.52,146.71,145.20,144.71,131.75,128.79,128.15,123.00,118.8 6,107.15,101.68,96.62,76.40,73.82,72.86,70.70,68.73,65.73,60.98,50.01,30.71,22.57,14.02,8.22.HR-ESIMS(m / z)for C 28 H 30 N 2 O 10 [M+H] + ,calcd:555.19787,found:555.19702.

[0159] Lactose-7-ethyl-10-hydroxycamptothecin (8): total yield 54%. 1H NMR(500MHz,d6-DMSO)δ8.12(d,J=9.1Hz,1H),7.74(d,J=2.4Hz,1H),7.58(dd,J=9.2,2.4Hz,1H),7.28(s,1H),6.51(s,1H),5.60(d,J=5.1Hz,1H),5. 43(s,2H),5.31(s,2H),5.27(d,J=8.5Hz,1H),5.24(d,J=5.1Hz,1H),5.05( d,J=4.9Hz,1H),5.01(d,J=5.5Hz,1H),4.83(d,J=2.2Hz,1H),4.74(t,J=5. 8Hz,1H),4.64(t,J=5.5Hz,1H),4.31(d,J=7.9Hz,1H),3.83(dd,J=10.0,5 .3Hz,1H),3.72(ddd,J=17.9,11.2,5.2Hz,3H),3.55(dd,J=8.7,2.1Hz,1H) ,3.45(ddd,J=11.5,8.6,5.9Hz,3H),3.27–3.12(m,4H),3.12–2.98(m,2H), 1.86(dt,J=14.2,6.7Hz,2H), 1.31(t,J=7.6Hz,3H), 0.87(t,J=7.2Hz,3H). 13 CNMR(125MHz,d6-DMSO)δ171.93,156.21,155.39,149.64,149.43,145.61,1 44.25,143.73,130.75,127.78,127.06,121.78,117.83,106.03,102.64,99 .34,95.56,79.57,76.21,75.91,74.68,74.46,72.72,72.31,72.01,69.47, 64.65,60.46,59.45,48.94,29.62,21.63,12.95,7.14.HR-ESIMS(m / z)forC 34 H 41 N 2 O 15 [M+H] + ,calcd:717.25069,found:717.2504.

[0160] Lactose-9-nitro-10-hydroxycamptothecin (9): total yield 56%. 1H NMR(500MHz,d6-DMSO)δ8.48(s,1H),8.12(d,J=9.1Hz,1H),7.99(d,J=9.2Hz,1H),7.54(d,J=9.2Hz,1H),6.51(s,1H),5.60(d,J=5.1Hz,1 H),5.43(s,2H),5.31(s,2H),5.27(d,J=8.5Hz,1H),5.24(d,J=5.1Hz,1H),5.05(d,J=4.9Hz,1H),5.01(d,J=5.5Hz,1H),4.83(d,J=2.2Hz, 1H),4.74(t,J=5.8Hz,1H),4.64(t,J=5.5Hz,1H),4.31(d,J=7.9Hz,1H),3.83(dd,J=10.0,5.3Hz,1H),3.72(ddd,J=17.9,11.2,5.2Hz,3H ),3.55(dd,J=8.7,2.1Hz,1H),3.45(ddd,J=11.5,8.6,5.9Hz,3H),3.27–3.12(m,4H),1.86(dt,J=14.2,6.7Hz,2H),0.87(t,J=7.2Hz,3H). 13 C NMR(125MHz,d6-DMSO)δ171.93,156.21,150.39,149.64,149.43,145.61,144.25,130.75,130.5,128.92,127.78,127.06,118.78,117.83,106 .03,99.34,95.56,79.57,76.21,75.91,74.68,74.46,72.72,72.31,72.01,69.47,64.65,60.46,59.45,48.94,29.62,7.14.HR-ESIMS(m / z)for C 32 H 36 N 3 O 17 [M+H] + ,calcd:734.20447,found:734.2038.

[0161] Cellobiose-7-ethyl-10-hydroxycamptothecin (10): total yield 52%. 1H NMR(500MHz,d6-DMSO)δ8.12(d,J=9.2Hz,1H),7.73(d,J=2.5Hz,1H),7.59(dd,J=9.2,2.5Hz,1H),7.29(s,1H),6.51(s,1H),5.60(d,J=5.1Hz,1H),5. 43(s,2H),5.32(s,2H),5.27(d,J=8.6Hz,1H),5.25(d,J=5.2Hz,1H),5.05( d,J=4.9Hz,1H),5.01(d,J=5.5Hz,1H),4.84(d,J=2.1Hz,1H),4.75(t,J=5. 8Hz,1H),4.63(t,J=5.4Hz,1H),4.31(d,J=7.9Hz,1H),3.83(dd,J=10.0,5 .3Hz,1H),3.71(ddd,J=17.8,11.1,5.3Hz,3H),3.54(dd,J=8.7,2.1Hz,1H) ,3.44(ddd,J=11.5,8.6,5.9Hz,3H),3.27–3.13(m,4H),3.12–2.98(m,2H), 1.87(dt,J=14.2,6.8Hz,2H), 1.31(t,J=7.6Hz,3H), 0.88(t,J=7.3Hz,3H). 13 C NMR(125MHz,d6-DMSO)δ171.94,156.24,155.38,149.65,149.44,145.62,1 44.20,143.74,130.75,127.78,127.06,121.79,117.83,106.03,102.64,99 .34,95.56,79.57,76.21,75.91,74.68,74.46,72.72,72.31,71.79,69.47, 64.65,60.46,59.45,48.94,29.62,21.63,12.95,7.15.HR-ESIMS(m / z)forC 34 H 41 N 2 O 15 [M+H] + ,calcd:717.25069,found:717.2503.

[0162] Maltobiose-7-ethyl-10-hydroxycamptothecin (11): total yield 50%. 1H NMR(500MHz,d6-DMSO)δ8.12(d,J=9.2Hz,1H),7.73(d,J=2.5Hz,1H),7.58(dd,J=9.2,2.5Hz,1H),7.29(s,1H),6.51(s,1H),5.71(d ,J=2.8Hz,1H),5.58(d,J=5.2Hz,1H),5.53(d,J=6.0Hz,1H),5.43(s,2H),5.32(s,1H),5.25(d,J=7.7Hz,1H),5.09(d,J=3.5Hz,1H), 5.02–4.88(m,4H),4.69(t,J=5.6Hz,1H),4.56(t,J=5.3Hz,2H),3.83–3.76(m,2H),3.65(ddd,J=17.6,16.9,10.5Hz,6H),3.59–3.3 8(m,10H),3.30–3.24(m,2H),3.24–3.15(m,2H),3.15–2.96(m,3H),1.94–1.76(m,3H),1.32(t,J=7.5Hz,3H),0.88(t,J=7.3Hz,3H). 13 C NMR(125MHz,d6-DMSO)δ171.94,156.24,155.40,149.62,149.44,145.62,1 44.20,143.78,130.72,127.77,127.07,121.87,117.82,105.92,100.20,99 .51,95.56,78.75,75.73,74.95,72.95,72.69,72.16,71.84,71.79,69.25 ,64.65,60.18,59.72,48.93,29.62,21.62,12.98,7.15.HR-ESIMS(m / z)for C 34 H 41 N 2 O 15 [M+H] + ,calcd:717.25069,found:717.2499.

[0163] Maltobiose-9-allyl-10-hydroxycamptothecin (12): total yield 48%. 1H NMR(500MHz,d6-DMSO)δ7.98(d,J=9.2Hz,1H),7.58(d,J=9.2Hz,1H),7.28(s,1H),6.51(s,1H),5.71(d,J=2.8Hz,1H),6.02 –5.93(m,1H),5.58(d,J=5.2Hz,1H),5.53(d,J=6.0Hz,1H),5.43(s,2H),5.32(s,1H),5.25(d,J=7.7Hz,1H),5.09(d,J=3.5H z,1H),5.02–4.88(m,6H),4.69(t,J=5.6Hz,1H),4.56(t,J=5.3Hz,2H),3.83–3.76(m,4H),3.65(ddd,J=17.6,16.9,10.5Hz, 6H),3.59–3.38(m,10H),3.30–3.24(m,2H),3.24–3.15(m,2H),3.15–2.96(m,1H),1.94–1.76(m,3H),0.88(t,J=7.3Hz,3H). 13 C NMR(125MHz,d6-DMSO)δ171.94,158.24,155.40,149.62,149.44,145.62,144 .20,143.78,134.82,128.72,127.77,127.07,122.87,117.82,116.81,112.42 ,105.92,99.51,95.56,78.75,75.73,74.95,72.95,72.69,72.16,71.84,71.7 9,69.25,64.65,60.18,59.72,48.93,32.15,29.62,7.15.HR-ESIMS(m / z)forC 35 H 41 N 2 O 15 [M+H] + ,calcd:729.25069,found:729.2512.

[0164] Maltotriose-7-ethyl-10-hydroxycamptothecin (13): total yield 45%. 1H NMR(500MHz,d6-DMSO)δ8.12(d,J=9.2Hz,1H),7.73(d,J=2.2Hz,1H),7.58(d d,J=9.2,2.3Hz,1H),7.29(s,1H),6.51(s,1H),5.71(d,J=3.1Hz,1H),5.62( d,J=6.4Hz,1H),5.59(d,J=5.2Hz,1H),5.51(d,J=5.9Hz,2H),5.43(s,2H),5 .31(s,2H),5.23(d,J=7.7Hz,1H),5.10(d,J=3.3Hz,1H),5.02(d,J=3.5Hz,1H ),4.91(d,J=5.3Hz,2H),4.73(t,J=5.7Hz,1H),4.62–4.47(m,2H),3.84–3.7 5(m,1H),3.75–3.54(m,10H),3.55–3.44(m,4H),3.44–3.29(m,9H),3.29–3. 22(m,2H),3.22–3.12(m,2H),3.08(dd,J=11.9,6.0Hz,1H),1.87(dt,J=14.3 ,6.9Hz,2H),1.32(t,J=7.5Hz,3H),0.88(t,J=7.3Hz,4H).HR-ESIMS(m / z)for C 34 H 41 N 2 O 15 [M+H] + ,calcd:879.3035,found:879.3035.

[0165] Experimental Example 2: In vitro anti-HCT-116 cell activity of Gly-CPTR

[0166] HCT-116 cells were cultured at 5 × 10 3 ~1×10 4 / well density was inoculated in a 96-well plate, the culture medium was Mcyoy's 5A, and the culture medium was gently aspirated after culturing at 37°C for 24 hours. Gly-CPTR was dissolved into a mother solution with DMSO, and diluted with the above culture medium to different concentration gradients. 100 μL of Gly-CPTR drug with different concentration gradients was added to each well (final drug concentration 0.1-10000 nM, final DMSO concentration ≤0.1%). The blank group was added with culture medium containing 0.1% DMSO. After continuing to culture at 37°C for 72 hours, 10 μL CCK8 reagent was added to each well and incubated at 37°C for 90 minutes. The OD value was measured at 450 nm by a Multiskan Sky full-wavelength microplate reader, and the EC was calculated. 50The above experiment was repeated three times with three parallel wells each time. Compound 14 was used as a control, wherein compound 14 was synthesized according to the method of Organic & Biomolecular Chemistry 17.29 (2019): 6970-6974. The structural formula of compound 14 is shown in Figure 2 .

[0167] Table 1. Cytotoxicity of Gly-CPTR to HCT116 cells

[0168]

[0169] The results of biological activity detection show that Gly-CPTR still retains some anticancer cell activity in vitro, indicating that the Gly-CPTR designed and synthesized by the present invention has higher drug stability than CPTR and has good drug-making ability and value.

[0170] Experimental Example 3: Drug release of Gly-CPTR in different tissue fluids

[0171] Male SD rats (200-220 g) were killed with isoflurane and then incised in the midline. The contents of the stomach, small intestine, and large intestine were collected separately. The stomach contents were diluted with pH 2.2 acetate buffer, and the other tissue contents were diluted with pH 6.8 phosphate buffer. The tissue contents were homogenized after 10 min of oscillation in a mixer and stored at -80°C for later use. 500 μL of 100 μM Gly-CPTR was added to 500 μL buffer corresponding to 20 μL tissue content homogenate, and 100 μL of reaction solution was taken after incubation at 37°C for 0.5 s, and high temperature treatment was performed for 1 min. The mixture was centrifuged at 15000 rpm for 5 min, and 60 μL of supernatant was taken for HPLC analysis to calculate the original drug release efficiency of Gly-CPTR in different tissue fluids (drug release efficiency = CPTR / (CPTR+Gly-CPTR)X100%), which is the ratio of the amount of CPTR substance released in the tissue fluid to the total amount of CPTR and original drug Gly-CPTR in the tissue fluid.

[0172] The results of biological activity test showed that Gly-CPTR was basically not degraded in the gastric and small intestinal fluids, and most of the Gly-CPTR could completely release the drug in the colonic fluid within half an hour, indicating that the Gly-CPTR designed and synthesized by the present invention had very good targeted drug release efficacy (Table 2).

[0173] Table 2. Release efficiency of Gly-CPTR in different tissue fluids (%)

[0174]

[0175]

[0176] Experimental Example 4: Antitumor activity of Gly-CPTR in vivo

[0177] 6-8 week old SPF male nude mice (Changzhou Cavens Laboratory Animal Co., Ltd., Animal Certificate No.: SCXK (Su) 2021-0010) were fasted for 12 hours and then anesthetized. 0.1 mL HCT116 cell suspension (cell density of 2 × 10 7 / mL) was injected into the colon serosa of nude mice and fed conventionally for 30 days. After the colon cancer mouse model was successfully established, the nude mice were randomly divided into groups, with 6 mice in each group. The control group was gavaged with 0.5% sodium carboxymethyl cellulose, the experimental group was gavaged with Gly-CPTR (0.0255mmol / Kg, dissolved in 0.5% sodium carboxymethyl cellulose), and the positive group was gavaged with irinotecan hydrochloride (0.0255mmol / Kg, dissolved in 0.5% sodium carboxymethyl cellulose). All the drugs were given once every two days, with a gavage volume of 0.1mL / Kg each time, and a total of 7 doses were given. The body weight of nude mice was weighed every two days. After 14 days, the nude mice were killed by cervical dislocation and dissected, the tumor tissue was removed, the size of the tumor tissue was measured, and the tumor weight was weighed. Graphpad Prism 5 (Version 5.01) was used for data analysis and drawing ( Figure 3 ).

[0178] The results of biological activity tests show that compared with the representative drug SN38 without glycosylation, most of the compounds of the present invention have obvious anti-tumor effects, and the tumor inhibition effect is more obvious than that of the control compound 14.

[0179] Experimental Example 5: Acute toxicity of Gly-CPTR in mice

[0180] Balb / C mice (male, about 20 g) aged 6 to 8 weeks were randomly divided into three groups, with 6 mice in each group. Different drugs were given by gavage every day. The blank control group was given 0.5% sodium carboxymethyl cellulose, and the experimental group was given irinotecan hydrochloride and Gly-CPTR (0.074 mmol / Kg, all drugs were dissolved in 0.5% sodium carboxymethyl cellulose). The volume of each administration was 0.1 mL / Kg. The drugs were given continuously for 6 days. The body weight of the mice was recorded every day and the mice were observed for diarrhea and other conditions. After 6 days, the mice were anesthetized and killed, and blood was collected from the heart. The blood biochemical indexes of the mice were tested on the veterinary fully automatic blood cell analyzer BC-5000Vet (Shenzhen Mindray Bio-Medical Electronics Co., Ltd.).

[0181] Irinotecan is a camptothecin drug used clinically and is a prodrug of SN38. It is mainly used for chemotherapy of metastatic colorectal cancer and is administered by drip. The main side effects are delayed diarrhea and neutropenia. The results of biological activity test showed that under the condition of oral administration, the body weight of irinotecan decreased significantly, while the body weight of mice in the Gly-CPTR group prepared by the present invention was similar to that of the blank group. The body weight of some compounds decreased slightly in the middle, but increased to the same level as the blank group on the sixth day, while the body weight of the irinotecan group showed a continuous downward trend ( Figure 4 ). Therefore, the Gly-CPTR drug prepared by the present invention has stronger safety.

Claims

1. A glycosylated camptothecin compound Gly-CPTR, characterized in that: Selected from the compounds shown in the following structural formula: 。 2. A glycosylated camptothecin compound Gly-CPTR according to claim 1, characterized in that: The glycosylated camptothecin compound Gly-CPTR is a drug for treating colorectal cancer.

3. A glycosylated camptothecin compound Gly-CPTR according to claim 2, characterized in that: The drug for treating colorectal cancer uses a glycosylated camptothecin compound Gly-CPTR as a pharmacologically active ingredient.

4. A glycosylated camptothecin compound Gly-CPTR according to claim 2 or 3, characterized in that: The drug also includes pharmaceutically acceptable excipients.

5. Use of Gly-CPTR in the preparation of an anti-colorectal cancer drug, characterized in that: Gly-CPTR is selected from the compounds shown in the following structural formula: 。 6. The use of Gly-CPTR in the preparation of anti-colorectal cancer drugs according to claim 5, characterized in that: The anti-colorectal cancer drug uses Gly-CPTR as a pharmacologically active ingredient; And / or, the drug further comprises a pharmaceutically acceptable excipient; And / or, the anti-colorectal cancer drug is an oral drug.

7. An anti-colorectal cancer drug, comprising a pharmacologically active ingredient; characterized in that: The pharmacologically active ingredient comprises: a glycosylated camptothecin compound Gly-CPTR as described in any one of claims 1-4.

8. An anti-colorectal cancer drug according to claim 7, characterized in that: Also includes: Pharmaceutically acceptable excipients.

9. A method for preparing a glycosylated camptothecin compound Gly-CPTR according to any one of claims 1 to 4, characterized in that: It is prepared by Koenigs-Knorr glycosidation reaction of all-acyl halide sugars and CPTR.

10. The method for preparing a glycosylated camptothecin compound Gly-CPTR according to claim 9, characterized in that: The Koenigs-Knorr glycosidation reaction refers to: the full acyl halide sugar and CPTR are dissolved in an organic solvent and then a catalyst is added to react; And / or, the catalyst is selected from: silver oxide or anhydrous potassium carbonate or silver trifluoromethanesulfonate; and / or, the organic solvent is selected from: dry acetonitrile, dichloromethane, tetrahydrofuran, N,N-dimethylformamide, acetone, N,N-dimethyl sulfoxide; And / or, the reaction temperature is 25-45°C; And / or, the reaction time is 3 hours; And / or, the usage ratio of the full acyl halide sugar, CPTR, and catalyst is: 1.0-2.0eq: 1.0-2.0eq: 1.0-2.0eq; and / or, after the reaction is completed, the liquid obtained by filtration is concentrated to obtain a crude product; and / or, the crude product is purified by acidic resin and silica gel column in sequence to obtain the final product; And / or, acid resin purification refers to the crude product being dissolved in a mixed solution of methanol-dichloromethane and reacting at room temperature for 2 hours before adding an acid resin; And / or, the methanol-dichloromethane mixed solution refers to a solution obtained by mixing methanol and dichloromethane in a volume ratio of 1:1 to 1:4; and / or, dissolving in a mixed solution of methanol and dichloromethane and adjusting the pH to 9-11; And / or, adjusting pH means adding 1M sodium methoxide dropwise for adjustment; and / or, the acidic resin refers to acidic resin Resin IR120; and / or, adding an acidic resin and stirring until the pH of the system reaches 7.0; And / or, silica gel column purification means: the product is purified by acidic resin and the filtrate is filtered through a silica gel column, and the mobile phase is ethyl acetate: Methanol:water = 7:2:0~7:2:1.

Images