CN116407554A - Drugs for reducing intestinal damage caused by targeted antineoplastic drugs and their application - Google Patents

Abstract

The invention provides a compound for improving the curative effect of a targeted anti-tumor drug and application thereof. Specifically, the compound has a structure shown in a formula I, wherein each group is defined as the specification; the invention also discloses application of the compound in reducing intestinal damage caused by oral targeted antitumor drugs, reducing diarrhea and improving the treatment effect of the targeted antitumor drugs.

Description

Drugs for reducing intestinal damage caused by targeted anti-tumor drugs and their applications

Technical Field

The invention belongs to the field of chemical pharmacy, and in particular relates to the use of a drug for reducing intestinal damage caused by targeted anti-tumor drugs.

Background Art

Many chemotherapy drugs are cytotoxic drugs that can quickly target dividing tumor cells and also have a killing effect on normal cells. They can cause gastrointestinal epithelial cell damage and cause various gastrointestinal symptoms. Clinically, it manifests as chemotherapy-related diarrhea (CID), which has been reported as grade 3–4 in randomized clinical trials, with a frequency of severe diarrhea of 5–47%. In addition to chemotherapy drugs that act on gastrointestinal tumors, chemotherapy drugs for other tumors such as docetaxel, capecitabine, or folinic acid antagonists (such as methotrexate) increase the risk of chemotherapy-induced diarrhea. Small molecule targeted drugs, especially EFGR-TKI, VEGFR-TKI, BCR-ABL TKI, and BTK inhibitors, can cause a high incidence of diarrhea and may cause serious consequences such as dehydration, malnutrition, electrolyte imbalance, and immune decline, which are life-threatening. Many drugs require dose adjustment or interruption of administration, and anti-diarrhea intervention measures are also required.

At present, there are few studies on the pathological mechanism of diarrhea caused by anticancer drugs, and it is still unclear. Diarrhea caused by chemotherapy drugs may be related to drug damage to the small intestinal epithelial tissue, inflammatory response, infection and the use of antibiotics.

Because oral targeted anti-tumor drugs cause intestinal damage in cancer patients, the frequency of diarrhea increases significantly, which seriously affects the treatment process. At present, there is a lack of effective drugs for treating diarrhea caused by tumor drugs in clinical practice, and most drugs are symptomatic treatments. The commonly used drugs are only oral preparations of loperamide and injectable preparations of octreotide. The antidiarrheal effect of loperamide on grade 3-4 diarrhea is not ideal. Continuous high-density use of high doses will lead to drug resistance when patients receive irinotecan. It can also cause side effects such as constipation, intestinal obstruction, affect drug absorption, aggravate intestinal infection, and liver damage, and cannot be used for a long time. Octreotide has gastrointestinal symptoms, neurological symptoms, hepatobiliary diseases (the incidence of cholelithiasis is about 15-30%), and metabolic and nutritional disorders. It is inconvenient for patients to be injected and the patient compliance is poor. Tumor targeted drugs need to be used for a long time until the disease progresses, so this type of auxiliary clinical drug that reduces the toxicity of targeted drugs is limited. It is urgent to develop a new oral drug for the prevention and treatment of diarrhea that is safer, more effective, and easier to use.

Summary of the invention

The purpose of the present invention is to provide a drug for reducing the damage of targeted anti-tumor drugs to the intestine and its application.

In a first aspect of the present invention, there is provided a use of a compound of formula I or a pharmaceutically acceptable salt thereof for preparing a pharmaceutical composition or preparation, wherein the pharmaceutical composition or preparation is used for:

(a) improving the therapeutic effect of targeted anti-tumor drugs; and/or

(b) Reduce the damage of targeted anti-tumor drugs to the intestine and reduce diarrhea;

In the formula,

其中所述的取代为磺酸基或羟基取代； _R1 is selected from the group consisting of hydroxy, C1-C6 alkoxy, halogen, substituted or unsubstituted -O-C1-C6 alkyl,

The substitution is sulfonic acid or hydroxyl substitution;

in,

中

的碳原子为手性碳原子，所述的手性碳原子选自下组：

middle

The carbon atom is a chiral carbon atom, and the chiral carbon atom is selected from the following group:

m is a positive integer from 1 to 6;

X is S or O;

_R5 is H or

R ₄ is a metal ion selected from the group consisting of Na ⁺ , K ⁺ , Li ⁺ or Cs ⁺ ;

_R6 is C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, aryl or heteroaryl;

_R2 and _R3 are each independently H, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, C2-C6 hydroxyalkyl or -(C1-C3 alkylene)-COOH;

n is a positive integer from 6 to 12.

In another preferred embodiment, _R1 is selected from the following group: hydroxyl, methoxy,

In another preferred embodiment, m=1.

In another preferred embodiment, X is S.

In another preferred embodiment, R ₄ is Na ⁺ .

In another preferred embodiment, R ₆ is CH ₃ .

In another preferred embodiment, n is 6, 7 or 8.

In another preferred embodiment, _R1 is selected from the following group: hydroxyl, methoxy,

In another preferred embodiment, R ₂ and R ₃ are each independently H, methyl, hydroxypropyl, hydroxyethyl or carboxymethyl.

In another preferred embodiment, the compound of formula I is selected from the following group: methyl cyclodextrin, carboxymethyl cyclodextrin, hydroxyethyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin or sulfobutyl ether-β-cyclodextrin.

In another preferred embodiment, the core of the compound of formula I is cyclodextrin.

In another preferred embodiment, the compound of formula I is

In another preferred embodiment, the compound of formula I is selected from the following group:

In another preferred embodiment, the pharmaceutical composition further contains additional active ingredients.

In another preferred embodiment, the pharmaceutical composition or preparation contains (a) a compound of formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient and (b) a pharmaceutically acceptable carrier.

In another preferred embodiment, in the pharmaceutical composition, the total amount of component (a) is 0.001-99wt%, preferably 0.1-90wt%, more preferably 1-80wt%, based on the total weight of the composition.

In another preferred embodiment, the pharmaceutical composition or preparation is used to improve the therapeutic effect of targeted anti-tumor drugs.

In another preferred embodiment, the pharmaceutical composition or preparation is used to reduce intestinal damage caused by taking targeted anti-tumor drugs and reduce diarrhea.

In another preferred embodiment, the pharmaceutical composition or preparation is used to reduce intestinal flatulence caused by intestinal damage.

In another preferred embodiment, the pharmaceutical composition or preparation is used to protect the integrity of intestinal villi and reduce the destruction of intestinal villi.

In another preferred embodiment, the pharmaceutical composition or preparation does not affect the absorption of oral targeted anti-tumor drugs.

In another preferred embodiment, the dosage form of the pharmaceutical composition or preparation is selected from the following group: tablets, capsules, powders, pills, granules or sustained-release preparations.

The second aspect of the present invention provides a pharmaceutical composition or preparation, which contains (a) an active ingredient, wherein the active ingredient includes a compound of formula I or a pharmaceutically acceptable salt thereof; and (b) a pharmaceutically acceptable carrier; the pharmaceutical composition or preparation is used for:

(1) Improving the therapeutic effect of targeted anti-tumor drugs; and/or

(2) Reduce the damage of targeted anti-tumor drugs to the intestine and reduce diarrhea.

In another preferred embodiment, the pharmaceutical composition or preparation may further contain other pharmaceutically active ingredients or pharmaceutically acceptable carriers.

In another preferred embodiment, when the active ingredient contains two components, the weight ratio of the two components is 1:20 to 20:1, preferably 1:10 to 10:1, and more preferably 1:5 to 5:1.

In another preferred embodiment, the other active pharmaceutical ingredients include targeted anti-tumor drugs.

In another preferred embodiment, the targeted anti-tumor drug is selected from the following group: gefitinib, erlotinib, osimertinib, dacomitinib, afatinib, nilotinib, lenvatinib, ceritinib, ibrutinib, dasatinib, imatinib, sunitinib, pazopanib, sorafenib or lapatinib.

In another preferred embodiment, when the active ingredient contains two components, the weight ratio of the two components is 1:20 to 20:1, preferably 1:10 to 10:1, and more preferably 1:5 to 5:1.

The third aspect of the present invention provides a medicine kit, comprising:

(1) a first container, and a first pharmaceutical composition in the container, wherein the first pharmaceutical composition comprises a first compound or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier;

(2) an nth container, and an nth pharmaceutical composition in the container, wherein the nth pharmaceutical composition comprises an nth compound or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier; wherein n is any positive integer from 2 to 8;

Wherein, the first compound and the nth compound are both compounds selected from the compounds described in claim 1; or at least one of the first compound selected from the compounds described in claim 1 and the nth compound is a targeted anti-tumor active substance;

and/or (3) optional instructions for use.

The fourth aspect of the present invention provides a method for treating a disease, wherein the disease is as described in claim 1, characterized in that it comprises the step of administering a compound of formula I or a pharmaceutically acceptable salt thereof to a subject in need thereof, wherein the compound of formula I is as defined in claim 1.

In another preferred embodiment, the subject is a mammal.

In another preferred embodiment, the subject is a human.

In a fifth aspect, the present invention provides a use of a combination of a targeted anti-tumor drug and a compound of formula I for preparing a drug for treating tumors.

In another preferred embodiment, the combination of the targeted anti-tumor drug and the compound of formula I comprises:

(a) a first pharmaceutical composition, the first pharmaceutical composition comprising: a targeted anti-tumor active ingredient and a pharmaceutically acceptable carrier; and

(b) a second pharmaceutical composition, comprising: a compound of formula I as an active ingredient and a pharmaceutically acceptable carrier;

In another preferred embodiment, the first pharmaceutical composition and the second pharmaceutical composition are independent of each other or are the same pharmaceutical composition.

In another preferred embodiment, the targeted anti-tumor drug is selected from the following group: gefitinib, erlotinib, osimertinib, or a combination thereof.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here.

DETAILED DESCRIPTION

After extensive and in-depth research, the inventor unexpectedly discovered for the first time that a compound represented by Formula I with a cyclodextrin as a parent core can be used to reduce the intestinal damage caused by targeted anti-tumor drugs. Further experiments show that the compound represented by Formula I can reduce the intestinal damage caused by targeted anti-tumor drugs, reduce diarrhea symptoms, and improve the therapeutic effect of targeted anti-tumor drugs, and has high safety. On this basis, the present invention is completed.

the term:

The term "halogen" refers to F, Cl, Br and I.

The term "C1-C6 alkyl" refers to a straight or branched chain alkyl group comprising 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, neopentyl, tert-pentyl, or the like.

The term "C2-C6 alkenyl" refers to a straight or branched alkenyl group having 2 to 6 carbon atoms and containing one double bond, including but not limited to ethenyl, propenyl, butenyl, isobutenyl, pentenyl, hexenyl and the like.

The term "C2-C6 alkynyl" refers to a straight or branched alkynyl group having 2 to 6 carbon atoms and containing one triple bond, including but not limited to ethynyl, propynyl, butynyl, isobutynyl, pentynyl, hexynyl and the like.

The term "C1-C6 hydroxyalkyl" refers to a straight or branched chain alkyl group having 1 to 6 carbon atoms and containing one hydroxyl group, including but not limited to hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, etc., preferably a C1-C3 hydroxyalkyl group.

The term "C3-C8 cycloalkyl" refers to a cyclic alkyl group having 3 to 8 carbon atoms in the ring, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like.

The term "C1-C6 alkoxy" refers to a straight or branched alkoxy group having 1 to 6 carbon atoms, including but not limited to methoxy, ethoxy, propoxy, isopropoxy and butoxy, etc. Preferably, it is a C1-C4 alkoxy group.

The term "aromatic ring" or "aryl" has the same meaning, preferably "C6-C10 aryl". The term "C6-C10 aryl" refers to an aromatic ring group having 6 to 10 carbon atoms without heteroatoms in the ring, such as phenyl, naphthyl, etc.

The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms, including nitrogen, oxygen and S(O)r (wherein r is an integer of 0, 1, or 2). For example, a 4-8 membered heteroaryl refers to a heteroaromatic system containing 4 to 8 ring atoms, and a 4-10 membered heteroaryl refers to a heteroaromatic system containing 4 to 10 ring atoms, including but not limited to pyrrolyl, furanyl, thienyl, pyrazolyl, thiazolyl, imidazolyl, oxazolyl, isoxazolyl, pyridinyl, pyranyl, pyridazinyl, pyrimidinyl, pyrazinyl, benzimidazolyl, triazolyl, and the like.

Unless otherwise specified as being "substituted or unsubstituted", the groups described in the present invention may be substituted by substituents selected from the following groups: halogen, acyloxy, cyano, amino, nitro, carboxyl, amide, carboxymethyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C2-C6 alkenyl, C2-C6 haloalkenyl, C2-C6 alkynyl, C2-C6 haloalkynyl, hydroxyl, C3-C6 cycloalkyl, C3-C6 halocycloalkyl, hydroxyl C1-C4 alkyl, C5-C7 cycloalkenyl, phenyl, naphthyl, etc.

表示基团的接合位置。

Indicates the bonding position of the group.

Active substances

As used herein, the terms "compound of the present invention" and "active ingredient of the present invention" are used interchangeably to refer to a compound of Formula I.

In the present invention, pharmaceutically acceptable salts of the compounds of formula I are also included. The term "pharmaceutically acceptable salt" refers to a salt suitable for use as a drug formed by the compounds of the present invention and an acid or a base. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by the polymer of the present invention and an acid. Acids suitable for forming salts include but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenesulfonic acid, benzenesulfonic acid, and acidic amino acids such as aspartic acid and glutamic acid.

The compounds of formula I of the present invention can be prepared by methods well known to those skilled in the art, and there is no particular restriction on the reaction parameters of each step. In addition, the typical compounds of the present invention can also be obtained by commercial means.

As used herein, in the compounds of Formula I, if a chiral carbon atom exists, the chiral carbon atom may be in the R configuration, the S configuration, or a mixture of the two.

In the present invention, the active ingredient compound of formula I has the following structure:

Where:

其中所述的取代为磺酸基或羟基取代； _R1 is selected from the following group: hydroxyl, C1-C6 alkoxy, halogen, substituted or unsubstituted -O-C1-C6 alkyl,

The substitution is sulfonic acid or hydroxyl substitution;

in,

中

的碳原子为手性碳原子，所述的手性碳原子选自下组：

middle

The carbon atom is a chiral carbon atom, and the chiral carbon atom is selected from the following group:

X is S or O; m is a positive integer from 1 to 6;

_R5 is H or

R ₄ is a metal ion selected from the group consisting of Na ⁺ , K ⁺ , Li ⁺ or Cs ⁺ ;

_R6 is C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, aryl or heteroaryl;

_R2 and _R3 are each independently H, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, C2-C6 hydroxyalkyl or -(C1-C3 alkylene)-COOH;

n is a positive integer from 6 to 12.

In one embodiment, _R1 is selected from the group consisting of hydroxyl, methoxy,

In another embodiment, m=1.

In another embodiment, R ₄ is Na ⁺ .

In another embodiment, R ₆ is CH ₃ .

In another embodiment, n is 6, 7 or 8.

In another embodiment, _R1 is selected from the group consisting of hydroxyl, methoxy,

In another embodiment, _R2 and _R3 are each independently H, methyl, hydroxypropyl, hydroxyethyl or carboxymethyl.

In another embodiment, the compound of formula I is selected from the group consisting of methyl cyclodextrin, carboxymethyl cyclodextrin, hydroxyethyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin or sulfobutyl ether-β-cyclodextrin.

In another embodiment, the core of the compound of formula I is cyclodextrin.

In another embodiment, the compound of formula I is

In another embodiment, the compound of formula I is selected from the following group:

Pharmaceutical compositions and methods of administration

The present invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and one or more safe and effective amounts of the compounds of the present invention.

Since the compounds of the present invention have excellent intestinal protective activity, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to reduce intestinal damage caused by targeted anti-tumor drugs and reduce diarrhea symptoms.

The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 10-1000 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

)、润湿剂(如十二烷基硫酸钠)、着色剂、调味剂、稳定剂、抗氧化剂、防腐剂、无热原水等。"Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as

), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

The pharmaceutical composition is in the form of injection, capsule, tablet, pill, powder or granule.

There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administrations include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

The compound of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds (such as targeted anti-tumor drugs).

The treatment method of the present invention can be used alone or in combination with other treatment methods or therapeutic drugs.

When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 50 to 1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

The main advantages of the present invention include:

(1) The compounds of formula I of the present invention can be used to improve the therapeutic effect of targeted anti-tumor drugs;

(2) The compounds of formula I of the present invention can be used to reduce intestinal damage caused by taking targeted anti-tumor drugs and reduce diarrhea;

(3) The compounds of formula I of the present invention can be used to reduce intestinal flatulence caused by intestinal damage;

(4) The compounds of formula I of the present invention can be used to protect the integrity of the intestinal villi and reduce the destruction of intestinal villi;

(5) The compound of formula I of the present invention has high biological safety and good therapeutic effect.

The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods for which specific conditions are not specified in the following examples are generally performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or under conditions recommended by the manufacturer. Unless otherwise indicated, percentages and parts are weight percentages and weight parts.

Example 1 Evaluation of the protective efficacy of samples on the gastrointestinal tract

Experimental methods:

390 4dpf wild-type AB strain zebrafish were randomly selected and placed in a 6-well plate, with 30 zebrafish treated in each well (experimental group). Except for the normal control group, all other experimental groups were given gefitinib in water to establish a zebrafish gastrointestinal injury model. "Compound (1)", "Compound (2)", "Compound (3)", "Compound (4)", "Compound (5)", "Compound (6)", "Compound (7)", "Compound (8)", "Compound (9)", "Compound (10)", "Compound (11)" and "Compound (12)" were given in water at a concentration of 83.3 μg/mL, and a normal control group and a model control group were set up at the same time, with a capacity of 3 mL per well. After 2 days of treatment with 28L, each experimental group was observed under a microscope, and the observation indicators included: ① intestinal cavity size, ② intestinal cavity degeneration, the incidence was counted and typical pictures were taken, and the gastrointestinal protective efficacy of the compounds of the present invention was evaluated based on the statistical analysis results of each indicator.

Table 1. Gastrointestinal phenotype (smaller intestinal cavity) and incidence of zebrafish after treatment with the compounds of the present invention (n=30)

Compared with the model control group, *p<0.05, **p<0.01

Table 2. Gastrointestinal phenotype (intestinal degeneration) and incidence of zebrafish after treatment with the compounds of the present invention (n=30)

Compared with the model control group, *p<0.05.

Experimental results: At this experimental concentration, the incidence of intestinal shrinkage caused by the above 12 compounds was lower than that of the model control group. The incidence of intestinal degeneration caused by "Compound 4" and "Compound 9" was higher than that of the model control group, and the mortality rates of "Compound 3", "Compound 4", "Compound 5", "Compound 6", "Compound 8", "Compound 9" and "Compound 10" were all higher than those of the model control group.

Experimental conclusion: Under the experimental conditions, "Compound 1", "Compound 2", "Compound 3", "Compound 5", "Compound 6", "Compound 7", "Compound 8", "Compound 10" and "Compound 11" showed an improving trend in gastrointestinal protective efficacy on zebrafish at a concentration of 83.3 μg/mL under the experimental conditions, among which Compound 11 had obvious gastrointestinal protective efficacy on zebrafish.

Example 2 Sample-induced sensitization risk assessment

Experimental methods:

Wild-type AB strain was selected, and each experimental group consisted of 30 zebrafish, which were incubated in a 28-hour incubator. There were 3 experimental groups in total. Each sample had 1 test dose (500 ng/tail), 1 solvent control group, and 1 normal control group. The samples were treated with normal zebrafish by intravenous administration.

After the sample treatment, the expression level of tryptase in zebrafish of each experimental group was detected by using a specific detection reagent for anaphylactic reaction (mast cell degranulation). The statistical processing results were expressed as mean. The calculation formula of sample sensitization is as follows:

SPSS software was used for statistical analysis, and p < 0.05 indicated a significant difference;

Table 3 Allergenic risk of each sample

Experimental results: Under the dosage condition of 500 ng/tail, the tested compounds of the present invention have a low risk of sensitization. Representatively, as shown in Table 3, the sensitization risks of the tested compounds of the present invention (1), (2), (7) and (11) are all significantly lower than the sensitization risk of the positive control.

Among them, "Compound (2)" and "Compound (1)" have no or substantially no sensitization risk, and "Compound (7)" and "Compound (11)" both have low sensitization risk.

Experimental conclusion: Combining the experimental results of Examples 1 and 2, that is, taking safety and effectiveness into consideration, the present invention selects compound (1) as the preferred compound for subsequent experiments.

Example 3 Gastrointestinal toxicity evaluation of compound 1

Experimental methods:

180 4dpf wild-type AB strain zebrafish were randomly selected and placed in a 6-well plate, with 30 in each well. The samples were dissolved in water (concentrations are shown in Table 4), and a solvent control group (1% DMSO) and a normal control group (zebrafish treated with standard diluted water) were set up at the same time, with a capacity of 3mL per well. After 2 days of incubation of zebrafish for 28 days: (1) Each experimental group was observed under a microscope, and the observation indicators included: ① intestinal cavity size, ② intestinal cavity color, ③ presence of bile in the intestinal cavity, ④ intestinal folds, and ⑤ bleeding conditions. The toxicity incidence was statistically analyzed and typical pictures were taken to evaluate the gastrointestinal toxicity of the samples; (2) 10 zebrafish were randomly selected from each concentration group and photographed and data was collected under a dissecting microscope. The intestinal cavity area of zebrafish was analyzed and statistically analyzed, and the gastrointestinal toxicity of the samples was evaluated based on the statistical analysis results. The calculation formula for the effect of the sample on the intestinal cavity area is as follows:

SPSS26.0 software was used for statistical analysis, and p<0.05 indicated that the difference was statistically significant.

Table 4 Gastrointestinal toxicity of compound (1) to zebrafish (n=30)

Note: "Note: indicates no obvious abnormality

Table 5 Effects of compound (1) on the gastrointestinal tract of zebrafish (n=10)

Compared with the solvent control group, *p<0.05, ***p<0.001.

Compared with gefitinib 12.5 μg/mL concentration, ^# p<0.05, ^### p<0.001.

Experimental results:

(1) Compared with gefitinib at a concentration of 12.5 μg/mL, the incidence of zebrafish intestinal cavity shrinkage was reduced in the concentrations of gefitinib 12.5 μg/mL + "Compound (1)" 0.139 μg/mL, gefitinib 12.5 μg/mL + "Compound (1)" 0.416 μg/mL, and gefitinib 12.5 μg/mL + "Compound (1)" 1.25 μg/mL, and the incidence of zebrafish intestinal cavity darkening was reduced.

(2) Gefitinib at a concentration of 12.5 μg/mL, gefitinib at a concentration of 12.5 μg/mL + "Compound (1)" at a concentration of 0.416 μg/mL, and gefitinib at a concentration of 12.5 μg/mL + "Compound (1)" at a concentration of 1.25 μg/mL induced the zebrafish intestinal cavity to shrink. Among them, gefitinib at a concentration of 12.5 μg/mL + "Compound (1)" at a concentration of 0.139 μg/mL had no effect on the zebrafish intestinal cavity area.

(3) Compared with gefitinib 12.5 μg/mL + "Compound (1)" 0.139 μg/mL, gefitinib 12.5 μg/mL + "Compound (1)" 0.416 μg/mL, and gefitinib 12.5 μg/mL + "Compound (1)" 1.25 μg/mL, the intestinal area of each concentration group was p<0.001, p<0.05, and p<0.05, indicating that compared with gefitinib alone, "Compound (1)" + gefitinib can improve the shrinkage of the zebrafish intestinal cavity.

Experimental conclusion: Under the experimental conditions, compared with the administration of gefitinib alone, the combined administration of "Compound (1)" + gefitinib can significantly improve the shrinkage of the zebrafish intestinal cavity, and the incidence of shrinkage and darkening of the intestinal cavity is reduced.

Example 4 Protective effect of compound 1 on gefitinib-induced intestinal damage in rats and pharmacokinetic detection

Experimental methods: 30 SD rats, half male and half female, weighing 175.9-212.0g, were randomly divided into 5 groups according to sex and weight, namely: normal control group, model control group, low, medium and high dose groups of compound 1 (40mg/kg, 120mg/kg, 180mg/kg), 6 rats in each group. Except for the normal control group, the other animals were given gefitinib (22.5mg/kg) by gavage. After modeling, the rats in the normal control group and model control group were given distilled water by gavage. The rats in the low, medium and high dose groups of compound 1 (40mg/kg, 120mg/kg, 180mg/kg) were given the corresponding drug solution by gavage 3 hours after gefitinib administration. The administration volume was 10mL/kg, once a day, and the administration lasted for 28 days. During the administration period, the defecation of the animals was observed every day, the animals were weighed every week, blood was collected for blood drug concentration detection after the last administration, and the ileum and colon were dissected for histopathological examination.

Experimental results:

4.1 General Observations

The rats in the model control group had obvious diarrhea when they were given D11, but no obvious diarrhea was observed in the groups given compound 1. The fecal occult blood test results of the rats in each group were negative, and there was no occult blood in the feces.

4.2 Effect of compound 1 on body weight of rats with gefitinib-induced intestinal injury

As shown in Table 6, compared with the normal control group, the body weight of the rats in the model control group was not significantly different. Compared with the model control group, the body weight of the rats in the low, medium and high dose groups of compound 1 was not significantly different.

n＝6)Table 6 Effect of compound 1 on body weight of rats with intestinal injury induced by gefitinib (

n＝6)

4.3 Effect of compound 1 on blood concentration of gefitinib-induced intestinal injury in rats

Compared with the model control group, there was no significant difference in the blood concentration of gefitinib in the low, medium and high dose groups of compound 1 within 4 hours of administration. Compound 1 had no significant effect on the absorption of gefitinib and did not affect the anti-tumor effect of gefitinib.

4.4 Effect of compound 1 on intestinal histology in rats with gefitinib-induced intestinal injury

Results of ileum histopathological examination: The ileum mucosa structure of rats in the normal control group was clear, with no obvious abnormalities; the vascular dilation of the lamina propria of the ileum mucosa of rats in the model control group, a small amount of inflammatory cell infiltration in the lumen, an increase in the number of goblet cells, an increase in mucin secretion, and no obvious damage to intestinal columnar epithelial cells. Compared with the model control group, the number of goblet cells in the ileum mucosa of rats in the low, medium, and high dose groups of compound 1 was significantly reduced, and mucin secretion was significantly reduced, and no obvious effect was observed on intestinal columnar epithelial cells or caused no damage. That is, the intestinal columnar epithelial cells of the ileum of rats in the model control group and the low, medium, and high dose groups of compound 1 were normal.

Results of colon histopathological examination: The colon mucosal structure of rats in the normal control group was clear, with no obvious abnormalities; the number of goblet cells in the colon mucosa of rats in the model control group increased, mucin secretion increased, and no obvious damage was observed in the intestinal columnar epithelial cells. Compared with the model control group, the number of goblet cells in the colon mucosa of rats in the medium and high dose groups of compound 1 was significantly reduced, mucin secretion was significantly reduced, and no obvious effect or damage was observed on the intestinal columnar epithelial cells. That is, the intestinal columnar epithelial cells of the colon of rats in the model control group and the low, medium, and high dose groups of compound 1 were normal.

n＝6)Table 7 Effect of compound 1 on the number of intestinal goblet cells in rats (

n＝6)

Note: Compared with the normal control group, ⁺⁺ P<0.01; compared with the model control group, *P<0.05, **P<0.01.

Experimental results: Compound 1 can significantly reduce the diarrhea symptoms induced by gefitinib in rats, reduce the number of goblet cells and mucin secretion in the intestinal tissue of rats, have no obvious effect or damage to columnar epithelial cells, and do not affect the blood concentration of gefitinib.

Experimental conclusion: Compound 1 has a significant protective effect on gefitinib-induced intestinal damage in rats and has no effect on the blood concentration of gefitinib.

Example 5 Protective effects of compound (1) and compound (5) on erlotinib-induced intestinal damage in rats

Experimental methods: 1) 38 male SD rats, weighing 200-220g, were randomly divided into 5 groups, namely: normal control group (3 rats), model control group (7 rats), low-dose and high-dose groups of compound (1) (1 mg/kg, 3 mg/kg), and low-dose and high-dose groups of compound (5) (1 mg/kg, 3 mg/kg). Except for the normal control group, the other animals were given erlotinib (48 mg/kg) by gavage on an empty stomach. After modeling, the rats in the normal control group and model control group were given distilled water by gavage. After erlotinib was given, the rats in each dose group of compound (1) and compound (5) were given the corresponding test solution by gavage at the same time. The administration volume was 10 mL/kg, once a day, and the administration was continued for 28 days. During the administration period, the defecation of the animals was observed every day, and the water content of the feces was calculated. 2) Nine male SD rats, weighing 200-220 g, were randomly divided into three groups: erlotinib group (48 mg/kg), erlotinib + compound (5) low-dose group (7.5 mg/kg), and erlotinib + compound (5) high-dose group (30 mg/kg). Blood samples were collected from the two groups at 0 h before oral administration and 2, 4, 6, 7, 8, 9, 10, 12, and 24 h after administration for blood drug concentration detection.

Table 8 Effects of Compound (1) and Compound (5) on Fecal Scores of Erlotinib-induced Intestinal Damage

Table 8 Effects of Compound (1) and Compound (5) on the Rate of Erlotinib-induced Intestinal Injury Diarrhea

Table 9 Effects of Compound (1) and Compound (5) on Fecal Water Content in Intestinal Damage Induced by Erlotinib

Group 7 days of medication 8 days of medication Drug administration for 9 days 10 days of medication Normal group 48.4% 53.9% 50.7% 52.8% Erlotinib group 72.0% 72.2% 73.1% 77.8% Compound (1) low dose group 60.6% 63.1% 65.8% 61.8% Compound (1) high dose group 69.5% 67.4% 68.8% 67.6% Compound (5) low dose group 53.1% 54.4% 58.6% 61.5% Compound (5) high dose group 63.9% 61.9% 63.7% 62.3%

Experimental conclusion: Compared with the erlotinib group, each dose group of compound (1) and (5) can reduce the fecal score and fecal water content of rats, and with the extension of administration time, compound 1 and compound 5 can reduce the diarrhea rate of rats. Low doses of compound (1) and compound (5) are superior to high doses of compound (1) and compound (5) in reducing fecal score, fecal water content and diarrhea rate of rats.

5.4 Effects of single administration of compound (1) and compound (5) on blood concentrations of erlotinib in rats with intestinal injury

Compared with the model control group, there was no significant difference in the blood concentration of erlotinib in the low-dose and high-dose groups of compound (1) and the low-dose and high-dose groups of compound (5) within 24 hours after administration of erlotinib. Compounds (1) and (5) had no significant effect on the absorption of erlotinib, and therefore did not affect the anti-tumor effect of erlotinib.

Example 6 Protective effect of compound 1 on osimertinib-induced intestinal damage in rats

Experimental methods:

SD rats were randomly divided into 5 groups, with 6-8 rats in each group; blank group, model group (osimertinib 30 mg/kg, 15 mg/kg), model + treatment group (compound (1) 240 mg/kg + osimertinib 30 mg/kg; compound (1) 60 mg/kg + osimertinib 30 mg/kg); the blank group rats were given pure water by oral gavage every day, the osimertinib 30 mg/kg group was given 3 mg/ml osimertinib by oral gavage, the osimertinib 15 mg/kg group was given 1.5 mg/ml osimertinib, and compound (1) 240 mg/kg + osimertinib 30 mg/kg) by oral gavage. The 40 mg/kg group was orally gavaged with 3 mg/ml osimertinib, and the compound (1) 60 mg/kg group was orally gavaged with 3 mg/ml osimertinib; 12 hours later, the blank group rats were orally gavaged with pure water, the osimertinib 30 mg/kg group was orally gavaged with pure water, the osimertinib 15 mg/kg group was orally gavaged with pure water, the compound (1) 240 mg/kg group was orally gavaged with 24 mg/ml compound (1), and the compound (1) 60 mg/kg group was orally gavaged with 6 mg/ml compound (1), and the administration was continued for 7 weeks.

Experimental results:

(1) Effect on rat body weight: After one week of administration of osimertinib, the body weight of rats in the 30 mg/kg osimertinib group increased slowly, which was significantly different from that of other groups; there was a significant difference in body weight between the 2nd, 4th, 5th, 6th and 7th weeks and the blank control group; the body weight of rats in the low-dose osimertinib group at the 4th, 5th, 6th and 7th weeks was significantly different from that of the high-dose group; the body weight of rats treated with compound (1) also increased slowly, but their average body weight was higher than that of untreated rats at the same osimertinib dose, and the mean showed a dose-response relationship.

(2) Effect on rat feces: Rats were anesthetized and then killed. Laparotomy was performed. It was found that the ileum of the model group rats had flatulence, intestinal wall expansion, and abnormal contents of the cecum and colon segments near the cecum compared with the control group. The incidence of intestinal flatulence and loose stools in the cecum and colon in the group treated with compound (1) was significantly reduced.

(3) Ileal pathological results: The blank group had intact mucosa, neatly arranged villi, clear glandular structure, and clear and complete crypt structure; the model group had chaotically arranged villi, short villi, destroyed crypt structure, and missing villi; the compound (1) treatment group had basically intact villi structure, and neatly arranged villi.

(4) Colon pathology results: The blank group had intact mucosa and neatly arranged villi; the model group had shorter villi, damaged crypt structure, and thinner intestinal wall; the group treated with compound (1) had basically intact villi structure and neatly arranged villi.

Experimental conclusion: The intestinal bloating of rats treated with compound (1) was improved compared with that of the model group. From the analysis of pathological results, compound (1) has a certain protective effect on the digestive tract damage caused by osimertinib, can protect the integrity of the intestinal wall villi and reduce the damage to the intestinal villi, and has a certain therapeutic effect on the diarrhea caused by osimertinib.

Example 7 Preparation of granules

Table 10 Formula of compound (1) for preparing granules

prescription Prescription amount g effect Compound (1) 123.38 Active ingredients D-Mannitol 525 Fillers lactose 315 Fillers Purified water 77 Adhesive solution

Weigh compound (1), D-mannitol and lactose monohydrate according to the prescription amount; add compound (1), D-mannitol and lactose monohydrate into the pot of a wet mixing granulator, start stirring and mix for 5 minutes; a. Granulation: start the stirring speed and cutting speed, use a peristaltic pump to add the binder solution in atomized form, and the granulation time is 1 minute; b. Granulation and drying: the obtained wet granules are sieved by a swing granulator, placed in an electric constant temperature drying oven at 70°C for drying (turned once every 5-10 minutes, moisture ≤0.5%); the dried material is sieved by a swing granulator and packaged in an aluminum foil bag.

All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims (10)

1. A use of a compound of formula I or a pharmaceutically acceptable salt thereof, characterized in that it is used to prepare a pharmaceutical composition or preparation, wherein the pharmaceutical composition or preparation is used for: (a) improving the therapeutic effect of targeted anti-tumor drugs; and/or (b) Reduce the damage of targeted anti-tumor drugs to the intestine and reduce diarrhea;

In the formula, _R1 is selected from the following group: hydroxyl, C1-C6 alkoxy, halogen, substituted or unsubstituted -O-C1-C6 alkyl,

The substitution is sulfonic acid or hydroxyl substitution; in,

middle

The carbon atom is a chiral carbon atom, and the chiral carbon atom is selected from the following group:

m is a positive integer from 1 to 6; X is S or O; _R5 is H or

R ₄ is a metal ion selected from the group consisting of Na ⁺ , K ⁺ , Li ⁺ or Cs ⁺ ; _R6 is C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, aryl or heteroaryl; _R2 and _R3 are each independently H, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, C2-C6 hydroxyalkyl or -(C1-C3 alkylene)-COOH; n is a positive integer from 6 to 12. 2. The use according to claim 1, characterized in that the compound of formula I is selected from the following group:

3. The use according to claim 1, characterized in that the pharmaceutical composition or preparation contains (a) a compound of formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient and (b) a pharmaceutically acceptable carrier. 4. The use according to claim 1, characterized in that the pharmaceutical composition or preparation is used to improve the therapeutic effect of targeted anti-tumor drugs. 5. The use according to claim 1, characterized in that the pharmaceutical composition or preparation is used to reduce intestinal damage caused by taking targeted anti-tumor drugs and reduce diarrhea. 6. The use according to claim 1, characterized in that the dosage form of the pharmaceutical composition or preparation is selected from the following group: tablets, capsules, powders, pills, granules or sustained-release preparations. 7. A pharmaceutical composition or preparation, characterized in that the pharmaceutical composition or preparation contains (a) an active ingredient, wherein the active ingredient includes a compound of formula I or a pharmaceutically acceptable salt thereof; and (b) a pharmaceutically acceptable carrier; the pharmaceutical composition or preparation is used for: (1) Improving the therapeutic effect of targeted anti-tumor drugs; and/or (2) Reduce the damage of targeted anti-tumor drugs to the intestine and reduce diarrhea. 8. A medicine box, characterized in that the medicine box comprises: (1) a first container, and a first pharmaceutical composition in the container, wherein the first pharmaceutical composition comprises a first compound or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier; (2) an nth container, and an nth pharmaceutical composition in the container, wherein the nth pharmaceutical composition comprises an nth compound or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier; wherein n is any positive integer from 2 to 8; Wherein, the first compound and the nth compound are both compounds selected from the compounds described in claim 1; or at least one of the first compound selected from the compounds described in claim 1 and the nth compound is a targeted anti-tumor active substance; and/or (3) optional instructions for use. 9. A method for treating a disease as described in claim 1, characterized in that it comprises the step of administering a compound of formula I or a pharmaceutically acceptable salt thereof to a subject in need thereof, wherein the compound of formula I is as defined in claim 1. 10. Use of a combination of a targeted anti-tumor drug and a compound of formula I, characterized in that it is used to prepare a drug for treating tumors.