A boron-containing ferulic acid compound, derivative or pharmaceutically acceptable salt thereof, and a synthesis method and application thereof

Abstract

The present invention discloses a boron-containing ferulic acid compound, derivative or pharmaceutically acceptable salt thereof, a synthesis method and an application. The boron-containing ferulic acid compound, derivative or pharmaceutically acceptable salt thereof is a compound, derivative and pharmaceutically acceptable salt thereof obtained by combining ferulic acid and its derivatives or substituents with B<supgt;10< / supgt> having neutron absorption ability. It is applied in BNCT boron neutron therapy to provide a high boron content and improve the specificity of the boron carrier for tumor cells.

Description

Technical Field

[0001] The present invention relates to a boron carrier technology, and particularly to a boron-containing ferulic acid compound, derivative or pharmaceutically acceptable salt thereof, and a synthesis method and application thereof. Background Art

[0002] Boron neutron capture therapy (BNCT) is a binary targeted therapy for tumor treatment. The principle is to inject a B 10 compound (boron carrier, also called boron drug) with specific affinity for tumors into the human body. After local irradiation with a neutron beam, the B-10 aggregated in the tumor tissue undergoes a nuclear reaction with thermal neutrons. The range of the 7Li particles released by the reaction of using the generated 7Li and α particles to destroy the tumor tissue is about 4-5 μm, and the range of α particles is about 9-10 μm. The diameter of tumor cells is generally less than 10 μm. Therefore, the radiation killing range of this method is only limited to within tumor cells, and the damage to surrounding normal tissues is very small. Thus, patients undergoing treatment do not require special protection.

[0003] So far, the boron carriers used in boron neutron capture therapy (BNCT) have evolved through three generations. The first-generation boron carriers are inorganic substances such as boric acid and borax. The high toxicity of boric acid and borax results in general tumor treatment effects and more serious toxic side effects. Organic substances 4-dihydroxyboronyl-L-phenylalanine (BPA) and sodium undecahydrododecaborane mercaptide (BSH) have stood out among boron carriers due to their selective targeting characteristics and have become the second-generation organic boron carriers. BPA can specifically bind to certain proteins and selectively accumulate in proteins related to tumor growth during the rapid proliferation of tumor cells; BSH has evolved with the development of carborane chemistry. The carborane structure generally contains more than 10 boron atoms. As a boron carrier, it will effectively increase the boron concentration in tumors; however, limited by defects at the molecular property level, the tumor specificity of BPA and BSH is insufficient. With the development of boron carriers, the third-generation boron carriers are mainly amino acid-based, nucleotide-based, boron-containing nanoparticles, etc.; the third-generation amino acid-based boron carriers have strong metabolic stability and exhibit high tumor specificity. However, limited by the limited specificity of their binding to tumor cells, they are currently mostly used for the treatment of brain tumors, and there are still limitations for the treatment of tumors in other organs. Summary of the Invention

[0004] In order to overcome the deficiencies of the prior art, one of the objectives of the present invention is to provide a boron-containing ferulic acid compound, derivative or pharmaceutically acceptable salt thereof. It combines ferulic acid with specific properties with B 10 and is applied to BNCT boron neutron therapy to provide a high boron content and improve the specificity of the boron carrier for tumor cells.

[0005] The second object of the present invention is to provide a method for synthesizing a boron-containing ferulic acid compound, derivative or pharmaceutically acceptable salt thereof as described above.

[0006] The third object of the present invention is to provide the use of a boron-containing ferulic acid compound, derivative or pharmaceutically acceptable salt thereof in the preparation of a cancer adjuvant therapy drug.

[0007] The first object of the present invention is achieved by the following technical solution: A boron-containing ferulic acid compound, derivative or pharmaceutically acceptable salt thereof, characterized in that it is a compound obtained by combining ferulic acid and its derivatives or substituents with B having neutron absorption ability, and its derivatives and pharmaceutically acceptable salts thereof have or contain the following chemical structural formula (I) or a substituent and derivative having the structural formula (II): 10 Combined to obtain a compound and its derivatives and pharmaceutically acceptable salts thereof, having or containing the following chemical structural formula (I) or a substituent and derivative having the structural formula (II):

[0008]

[0009] Further, the substitution position of the substituent of the structural formula (I) occurs on the benzene ring; in the structural formula (II), R 1 ~R 2 is a short-chain hydrocarbon group of C 1 -C 10 an alkoxy group of C 1 -C 10 or a boron-carrying group in a hetero-chain group containing a nitrogen atom or / and an oxygen atom of C 1 -C 10 ; the R 1 ~R 2 are the same or different groups.

[0010] Further, the derivative is a derivative formed by one or more alicyclic structures, benzene ring structures, piperazine ring structures, amino functional groups or oxa heterocycles in the ferulic acid molecule; the pharmaceutically acceptable salt is a salt obtained by adding an acid or adding a base to the boron-containing ferulic acid and its derivatives, and the acid is hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, citric acid, oxalic acid, malonic acid, salicylic acid, malic acid, fumaric acid, succinic acid, ascorbic acid, maleic acid, tartaric acid, methanesulfonic acid or hydroxyethanesulfonic acid: the base is sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, ammonia, triethylamine or triethanolamine.

[0011] The first object of the present invention is achieved by the following technical solution: A method for synthesizing a boron-containing ferulic acid compound, derivative or pharmaceutically acceptable salt thereof as described above, comprising the following steps:

[0012] (1) Protect the double bond: Using ferulic acid as the raw material, it undergoes an addition reaction with excessive HCl under a small amount of catalyst to obtain crude ferulic acid chloro-substitute (a);

[0013] (2) Protect the carboxyl group: In the apparatus containing the ferulic acid chloro-substitute with chemical structural formula (a), excessive MeOH and excessive trimethylchlorosilane TMSCl are added and reacted fully to form a compound with chemical structural formula (b);

[0014] (3) Electrophilic substitution reaction: Take the compound with chemical structural formula (b), add a small amount of Fe catalyst, introduce excessive chlorine gas, and react fully to form ferulic acid chloride with chemical structural formula (c);

[0015] (4) Amine substitution reaction: Excessive ammonia and a small amount of Cu 2 O are introduced into the ferulic acid chloride with chemical structural formula (c) to conduct an amine substitution reaction to form a ferulic acid amine-substitute with chemical structural formula (d);

[0016] (5) Diazotization reaction: Excessive sodium nitrite and excessive hydrochloric acid are added to the ferulic acid amine-substitute with chemical structural formula (d) to conduct a diazotization reaction to form a compound with chemical structural formula (e);

[0017] (6) Iodine substitution reaction: Excessive KI is added to the compound with chemical structural formula (e), and a substitution reaction is conducted under heating conditions, and -N 2 Cl is substituted by -I to obtain a ferulic acid iodine-substitute with chemical structural formula (f);

[0018] (7) Grignard reagent reaction: The ferulic acid iodine-substitute with chemical structural formula (f) is added to a dry apparatus, reacted with excessive metallic magnesium in excessive anhydrous ether, and heated under reflux to form a compound with chemical structural formula (g);

[0019] (8) Synthesis reaction for introducing boron-10: Excessive B 10 (OCH 2 CH 3 ) 3 is added to the above compound with chemical structural formula (g), and the reaction is conducted at room temperature to introduce boron-10 into the above compound to obtain a compound with chemical structural formula (h);

[0020] (9) Remove the double bond protection: Excessive MeOH is added to the compound with chemical structural formula (h), and the reaction is conducted under alkaline conditions to obtain a compound with chemical structural formula (i);

[0021] (10) Removal of carboxyl protection: The compound of the above chemical structural formula (i) undergoes hydrolysis of the ester group in an acidic aqueous solution to generate a carboxyl group, obtaining boron-containing ferulic acid with the chemical structural formula (II).

[0022] Further, in step (1), the catalyst is selected from one of ferric chloride, phosphoric acid, aluminum trichloride, and titanium tetrachloride, and the reaction condition is a constant temperature of 25 °C; in step (2), the reaction condition is to react for 20 h at room temperature; in step (3), the reaction condition is to react at 0 - 5 °C for 12 - 48 hours; in step (4), the reaction condition is to react at 200 °C and 60 atm for 2 hours.

[0023] Further, in step (5), the reaction condition is to carry out a diazotization reaction at 0 - 5 °C for 2 hours; in step (6), the reaction condition is to stand still in the dark for 3 hours; in step (7), the reaction condition is to heat to 90 °C and react fully for 30 min; in step (8), the reaction condition is to react at room temperature for 2 hours.

[0024] Further, in step (9), the reaction condition is to react under alkaline conditions with a pH of 8 for 1 hour; in step (10), the reaction condition is to react under acidic conditions with a pH of 3 for 1 hour.

[0025] The third object of the present invention is achieved by the following technical solution: Use of a boron-containing ferulic acid compound, derivative, or a pharmaceutically acceptable salt thereof in the preparation of a cancer adjuvant therapy drug.

[0026] Specifically, the cancer adjuvant therapy drug includes adjuvant therapy drugs for kidney cancer, liver cancer, lung cancer, gastric cancer, colon cancer, breast cancer, and melanoma-related diseases.

[0027] The present invention also provides a boron carrier, which contains the boron-containing ferulic acid compound, derivative, or a pharmaceutically acceptable salt thereof as described above. This boron carrier is used for boron neutron capture therapy to treat related diseases such as endoderm, mesoderm, and ectoderm cancers.

[0028] The present invention also provides a pharmaceutical composition, which contains the boron-containing ferulic acid compound, derivative, or a pharmaceutically acceptable salt thereof as described above, and a carrier permitted in medicine and pharmacy.

[0029] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0030] 1. The boron-containing ferulic acid, derivative, or a pharmaceutically acceptable salt thereof described in the present application is a product synthesized from ferulic acid and boron-10 with strong neutron absorption ability, which can provide a boron carrier with a high content, has a high affinity for tumor cells, good stability, and low biological toxicity.

[0031] 2. The boron-containing ferulic acid compound, derivative or pharmaceutically acceptable salt thereof described in the present application binds to protein kinase CK2 in vivo. CK2 is a pleiotropic protein kinase that regulates multiple survival pathways and exerts a global anti-apoptotic function. It is highly expressed in various cancers including renal cancer and liver cancer. After the boronated ferulic acid binds to CK2, it targets tumor cells, achieving the effect of treating cancer by BNCT, and has good application prospects in boron neutron therapy.

[0032] 3. The boron-containing ferulic acid compound, derivative or pharmaceutically acceptable salt thereof described in the present application is used as a boron carrier (boron drug), expanding the selection range of boron drugs used in BNCT treatment.

[0033] 4. The boron-containing ferulic acid compound, derivative or pharmaceutically acceptable salt thereof described in the present application as a boron carrier has good curative effects mainly on endodermal cancers such as renal cancer, liver cancer, lung cancer, gastric cancer, and colon cancer, and also has effects on mesodermal and ectodermal cancers such as breast cancer and melanoma. This patent positions boronated ferulic acid as an endodermal boron drug represented by the treatment of renal cancer and liver cancer. Description of the Drawings

[0034] Figure 1 CNMR spectrum of the boron-containing ferulic acid product (II) in Example 1 of the present invention;

[0035] Figure 2 For the boron-10 reagent B 10 (OCH 2 CH 3 ) 3 CNMR spectrum. Detailed Embodiments

[0036] Next, in combination with the specific embodiments, the present invention will be further described. It should be noted that, on the premise of non-conflict, any combination of the following-described embodiments or technical features can form a new embodiment. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the scope protected by the present application.

[0037] The term "including" and other equivalent descriptive methods involved in the specification and claims of the present application are all intended to cover non-exclusive inclusion, that is, it includes both the content clearly described in the specification and claims and may also include steps or units that are inherent in the product, method or structure but not described in the specification and claims.

[0038] An embodiment of the present application provides a boron-containing ferulic acid compound, derivative or a pharmaceutically acceptable salt thereof, which is characterized in that ferulic acid having the chemical structural formula (Ⅰ) is used as a raw material, and boron-10 is introduced into its structure to obtain a compound having or containing the chemical structural formula (Ⅱ):

[0039]

[0040]

[0041] B 10 is a stable isotope of boron. In BNCT therapy, atoms with strong neutron absorption ability need to be found, and B 10 has a very strong thermal neutron absorption ability for the thermal neutrons emitted by the BNCT device. In particular, boron drugs in BNCT (such as BPA, BSH, etc.) must contain highly enriched B 10 , and generally the enrichment degree is required to exceed 99%. In the present application, the boron-containing ferulic acid is a new boron carrier obtained by introducing boron-10 into the structure of ferulic acid.

[0042] The present application also provides a synthesis process for a boron-containing ferulic acid, derivative or a pharmaceutically acceptable salt thereof. Through this synthesis process, B 10 with very strong neutron absorption ability is introduced into the structure of ferulic acid to obtain a new type of boron carrier / boron drug.

[0043] The synthesis process includes the following steps

[0044] (1) Protecting the double bond: In order to prevent the carbon-carbon double bond in ferulic acid from undergoing an addition reaction in the presence of chlorine, where the double bond C═C is destroyed and cannot be restored, double bond protection is required before the electrophilic substitution reaction.

[0045] That is, ferulic acid and HCl undergo an addition reaction under the conditions of a catalyst such as ferric chloride to obtain a crude ferulic acid chloro-substitute (a). It should be noted that the carbon-chlorine bond product has high toxicity and corrosiveness, and operations should be carried out with gloves in a fume hood. During the operation, the reaction conditions should be strictly controlled and personnel safety measures should be taken.

[0046] Specifically, the operation is as follows: 5 g of ferulic acid and excessive HCl undergo an addition reaction under the conditions of a small amount of ferric chloride catalyst and a constant temperature of 25 °C to obtain a crude ferulic acid chloro-substitute (a).

[0047]

[0048] (2) Protecting the carboxyl group: Generally, carboxylic acids do not react spontaneously with chlorine gas. However, under appropriate conditions, they can undergo chlorination reactions through the action of catalysts or photoaccelerators to form the corresponding acyl chlorides. Among them, common catalysts include phosphoric acid, aluminum trichloride, titanium tetrachloride, etc. In addition, the reaction between carboxylic acids and chlorine gas can also be achieved under the action of ultraviolet light or high-energy radiation. Since chlorine elements need to be introduced into the benzene ring of ferulic acid in subsequent reactions (the reaction raw material is Cl 2 ), a catalyst is required. In order to prevent the carboxyl group in ferulic acid from undergoing chlorination reactions in the presence of chlorine gas and destroying the carboxyl structure of this compound, it is necessary to protect the carboxyl group before this electrophilic substitution reaction.

[0049] That is, in the instrument containing the ferulic acid chloro-substitute with chemical structural formula (a), add MeOH and trimethylchlorosilane TMSCl, and react fully to form a compound with chemical structural formula (b). The raw materials for each subsequent step are derived from the product of the previous step and do not require weighing.

[0050] The specific operation is as follows:

[0051] In the instrument containing the product with the ferulic acid chloro-substitute with chemical structural formula (a), add an excess of MeOH and an excess of trimethylchlorosilane TMSCl, and react at room temperature for 20 h. After full reaction, a compound with chemical structural formula (b) is formed.

[0052]

[0053] (3) Electrophilic substitution reaction: In order to introduce chlorine elements into the benzene ring of ferulic acid, in the instrument containing the compound with chemical structural formula (b), add a polar inert solvent such as anhydrous acetaldehyde or tetrahydrofuran, and one of the catalysts such as transition metals and their compounds like iron and copper, ferrous oxide carbon tetrachloride, sodium phenoxide, etc. Pass chlorine gas into the above reaction system through a gas flowmeter, and the entire reaction system reacts at 0 - 5 °C for 12 - 48 hours. After the reaction is completed, the product is dried to obtain a compound containing chemical structural formula (c). -OH and -OCH 3 both have electron-withdrawing inductive effects and electron-donating conjugative effects. However, the electron-donating ability of the conjugative effect is stronger than the electron-withdrawing ability of the inductive effect. Therefore, the reaction mechanisms of passing chlorine gas through both are the same, and substitution occurs at their ortho and para positions.

[0054] The specific operation is as follows:

[0055] Take the product containing the compound with chemical structural formula (b), add a small amount of Fe catalyst, pass in an excess of chlorine gas, and react fully at 0 - 5 °C for 12 - 48 hours to form the ferulic acid chloride with chemical structural formula (c).

[0056]

[0057] (4) Amine substitution reaction: Excess ammonia and a small amount of Cu 2 O are introduced into ferulic acid chloride with chemical structural formula (c), and the amine substitution reaction is carried out at 200 °C and 60 atm to produce a ferulic acid amine-substituted product with chemical structural formula (d).

[0058] Specific operation is as follows:

[0059] Take the above product containing the compound with chemical structural formula (c), add excess ammonia and a small amount of Cu 2 O, and react fully for 2 hours at 200 °C and 60 atm to produce a ferulic acid amine-substituted product with chemical structural formula (d).

[0060]

[0061] (5) Diazotization reaction: Sodium nitrite and hydrochloric acid are added to the ferulic acid amine-substituted product with chemical structural formula (d), and the diazotization reaction is carried out at 0 - 5 °C to produce a compound with chemical structural formula (e).

[0062] Specific operation is as follows:

[0063] Take the above product containing the compound with chemical structural formula (d), add excess sodium nitrite and excess hydrochloric acid, and carry out the diazotization reaction at 0 - 5 °C. React fully for 2 hours to produce a compound with chemical structural formula (e). The "excess" referred to in the present invention means that it is ensured that the reaction will not be incomplete due to insufficient raw materials. In this step, in actual operation, hydrochloric acid can be added until the pH value is less than 3. In the art, usually adding 37% hydrochloric acid with the same volume as the substrate can reach the excess condition, and there is no need to specifically measure or weigh. Sodium nitrite can be taken as the excess standard when it is just saturated.

[0064]

[0065] (6) Iodine substitution reaction: KI is added to the compound with chemical structural formula (e), and the substitution reaction is carried out under heating conditions, and -N 2 Cl is replaced by -I to obtain a ferulic acid iodine-substituted product with chemical structural formula (f).

[0066] Specific operation is as follows:

[0067] Take the above product containing the compound with chemical structural formula (e), add excess KI, and let it stand in the dark for 3 hours to fully carry out the substitution reaction (the explanation of "excess" here is the same as above. Here, KI can be replaced by an ethanol solution of potassium iodide to increase the reaction efficiency) to produce a ferulic acid iodine-substituted product with chemical structural formula (f).

[0068]

[0069] (7) Grignard reaction: Add the iodo-substituted ferulic acid with the chemical structural formula (f) into a dry instrument, react it with metallic magnesium in anhydrous ether, and heat under reflux to generate the compound with the chemical structural formula (g).

[0070] The specific operation is as follows:

[0071] Take the above product containing the iodo-substituted ferulic acid with the chemical structural formula (f), add an excessive amount of metallic magnesium and an excessive amount of anhydrous ether, and carry out the Grignard reaction under the condition of heating to 90 °C, and react fully for 30 min to generate the iodo-substituted ferulic acid with the chemical structural formula (g).

[0072]

[0073] (8) Synthesis reaction for introducing boron-10: Add B 10 (OCH 2 CH 3 ) 3 to the above compound with the chemical structural formula (g), and react at room temperature for 2 hours to introduce boron-10 into the above compound, obtaining the compound with the chemical structural formula (h).

[0074] The specific operation is as follows:

[0075] Add an excessive amount of B 10 (OCH 2 CH 3 ) 3 to the instrument containing the compound with the chemical structural formula (g) of the above product, and react at room temperature for 2 h to generate the compound with the chemical structural formula (h).

[0076]

[0077] (9) Removal of double bond protection: Since the elimination of the chlorine atom occurs in an alcoholic solution under alkaline conditions, in order not to introduce impurities, add MeOH to the above compound with the chemical structural formula (h), and react under the alkaline condition with a pH of 8 to obtain the compound with the chemical structural formula (i), which restores the carbon-carbon double bond in ferulic acid.

[0078] The specific operation is as follows: Take the above product containing the compound with the chemical structural formula (h), add an excessive amount of MeOH, and react under the alkaline condition with a pH of 8 for 1 hour to generate the compound with the chemical structural formula (i).

[0079]

[0080] (10) Removal of carboxyl protection: The compound of the above chemical structural formula (i) undergoes hydrolysis of the ester group in an acidic aqueous solution to form a carboxyl group, obtaining boron-containing ferulic acid with the chemical structural formula (Ⅱ).

[0081] Specific operation: Take the above product containing the compound of chemical structural formula (i), add an excessive amount of acid, and react under acidic conditions with a pH of 3 for 1 hour to form a compound with the chemical structural formula (Ⅱ).

[0082]

[0083]

[0084] The following are specific examples of this application. In the following examples, unless otherwise specifically limited in this application, the raw materials, reagents, instruments and equipment used can all be obtained by purchase.

[0085] Example 1

[0086] This example provides a boron-containing ferulic acid compound, and the synthesis method of this compound includes the following steps:

[0087] (1) Protecting the double bond: React 5 g of ferulic acid with the following structural formula (Ⅰ) and excessive HCl under the condition of a small amount of catalyst ferric chloride and constant temperature at 25 °C to obtain a crude ferulic acid chloro-substitute (a).

[0088]

[0089] (2) Protecting the carboxyl group: Add excessive MeOH and excessive trimethylchlorosilane TMSCl to the instrument containing the ferulic acid chloro-substitute (a) with the chemical structural formula (a) in the above product, and react at room temperature for 20 h. After full reaction, a compound with the chemical structural formula (b) is formed.

[0090]

[0091] (3) Electrophilic substitution reaction: Take the product containing the compound with the chemical structural formula (b) above, add a small amount of Fe catalyst, introduce excessive chlorine gas, and react fully at 0 - 5 °C for 12 - 48 hours to form ferulic acid chloride with the chemical structural formula (c).

[0092]

[0093] (4) Amine substitution reaction: Take the product containing the compound with the chemical structural formula (c) above, add excessive ammonia and a small amount of Cu 2 O and react fully at 200 °C and 60 atm for 2 hours to form a ferulic acid amine-substitute with the chemical structural formula (d).

[0094]

[0095] (5) Diazotization reaction: Take the product containing the compound of chemical structural formula (d), add an excessive amount of sodium nitrite and a sufficient amount of hydrochloric acid, and carry out the diazotization reaction at 0 - 5 °C for 2 hours to fully react to generate a compound with chemical structural formula (e).

[0096]

[0097] (6) Iodine substitution reaction: Take the product containing the compound of chemical structural formula (e), add an excessive amount of KI, and let it stand in the dark for 3 hours to fully carry out the substitution reaction to generate an iodine-substituted ferulic acid with chemical structural formula (f).

[0098]

[0099] (7) Grignard reagent reaction: Take the iodine-substituted ferulic acid containing the compound of chemical structural formula (f) in the above product, add an excessive amount of metallic magnesium and a sufficient amount of anhydrous ether, and carry out the Grignard reagent reaction under the condition of heating to 90 °C for 30 min to fully react to generate an iodine-substituted ferulic acid with chemical structural formula (g).

[0100]

[0101] (8) Synthesis reaction for introducing boron-10: In the apparatus containing the compound of chemical structural formula (g) in the above product, add an excessive amount of B 10 (OCH 2 CH 3 ) 3 , and react at room temperature for 2 h to generate a compound with chemical structural formula (h).

[0102]

[0103] (9) Removal of double bond protection: Take the product containing the compound of chemical structural formula (h), add an excessive amount of MeOH, and react under the alkaline condition with a pH of 8 for 1 hour to generate a compound with chemical structural formula (i).

[0104]

[0105] (10) Removal of carboxyl protection: Take the product containing the compound of chemical structural formula (i), add a sufficient amount of acid, and react under the acidic condition with a pH of 3 for 1 hour to finally obtain the boron-containing ferulic acid product (II) with the following structure:

[0106]

[0107] Such as Figure 1As shown, the CNMR diagram data of the boron-containing ferulic acid product (II) of the present invention Figure 2 is for the boron-10 reagent (B 10 (OCH 2 CH 3 ) 3 CNMR diagram. Combining Figure 1-2 , it can be seen that if the boron-containing reagent is not thoroughly distilled to remove impurities, a peak will appear near 57, and as Figure 1 shown, the peak at 57 is not obvious, proving that the boron-containing reagent B 10 (OCH 2 CH 3 ) 3 has been removed completely.

[0108] Test Examples

[0109] Test Example 1 Biological toxicity experiment of the compound prepared in Example 1 of the present invention

[0110] Prepare 10 Kunming mice (the source of the mice is Guangdong Biomedical Animal Experiment Center), and their weights are measured as 21.6 g, 22.4 g, 21 g, 22.1 g, 22.5 g, 20.3 g, 22.7 g, 23.0 g, 21.8 g, 21.5 g respectively. Inject the boron-containing ferulic acid of the above Example 1 into the tails of 10 Kunming mice at 100 mg / kg, 150 mg / kg, 170 mg / kg, 200 mg / kg, 300 mg / kg, 100 mg / kg, 200 mg / kg, 300 mg / kg, 320 mg / kg, 330 mg / kg respectively. According to the above doses and the weights of Kunming mice, the calculated amounts of boron-containing ferulic acid to be added are 0.00216 g, 0.00336 g, 0.00357 g, 0.00442 g, 0.00675 g, 0.00203 g, 0.00454 g, 0.0069 g, 0.006976 g, 0.007095 g respectively. Dissolve the above masses of boron-containing ferulic acid in 2 mL of dimethyl sulfoxide to prepare boron-containing ferulic acid solutions. After injection and oral administration respectively, they are marked as groups 1-5 and 6-10, and observed for 3 days.

[0111] Result observation: In the 1st and 2nd groups of injections, no abnormalities were found in Kunming mice, so it is speculated that the toxicity has little harm to Kunming mice. In the 3rd group, Kunming mice became slow in movement 15 minutes after gavage, specifically manifested as slow limb movements. After 15 minutes, they returned to normal. On the first and second days, the vitality of Kunming mice decreased and returned to normal on the third day. In the 4th group, the vitality of Kunming mice decreased 5 minutes after gavage, refused to eat and did not move for the next 3 days, and died after the third day. In the 5th group, the vitality of Kunming mice decreased immediately after gavage, refused to eat and did not move on the same day until they died.

[0112] In the 6th and 7th groups given oral administration, no abnormalities were observed in the Kunming mice. Therefore, it is speculated that the toxicity has less harm to Kunming mice. In the 8th group, the Kunming mice became sluggish 15 minutes after gavage, specifically manifested as slow limb movements. After 15 minutes, they returned to normal. On the first and second days, the vitality of the Kunming mice decreased and returned to normal on the third day. In the 9th group, the vitality of the Kunming mice decreased 5 minutes after gavage. Subsequently, they refused to eat and did not move for the next 3 days. They started eating on the third day, but their vitality was still very low. In the 10th group, the vitality of the Kunming mice decreased 3 minutes after gavage. Subsequently, they refused to eat and did not move for the next 3 days until they died.

[0113] Conclusion: The mice can survive when the boron-containing ferulic acid of the present invention is intravenously injected at a dose of 170 mg / kg and orally administered at a dose of 320 mg / kg. The mice cannot survive when the intravenous injection dose is higher than 170 mg / kg and the oral administration dose is higher than 320 mg / kg.

[0114] Experimental Example 2 In vitro anti-tumor experiment of the compound prepared in Example 1 of the present invention

[0115] Inject 100 mg / kg of boronated ferulic acid into the tail vein of renal cancer mice. After 3 days, cut the tail and take 20 μl of blood and mix it in 1 ml of physiological saline (containing 4 mg of EDTA·2Na / 100 ml or heparin, 1 mg of heparin anticoagulates 2 - 5 ml of blood). After centrifuging at 4000 r / min for 3 min, combine the CH 3 OH extraction solution, then evaporate CH 3 OH, take the residue, add 0.8 mL of CH 3 OH to dissolve it and then perform vortex mixing and centrifugation. The obtained upper clear liquid is used as a sample. In a polytetrafluoroethylene beaker, add an EDTA solution. The absorbance is measured under the same conditions as the determination of the boron standard solution. Subsequently, the mice are dissected, and 0.2 g of the blood, heart, and kidney tissues of the mice are taken, minced, added with 1.8 ml of physiological saline, and homogenized in an ice bath using a glass homogenizer to prepare a 10% tissue homogenate. The tissue homogenate is centrifuged at 4000 r / min for 10 min at 4°C, and the supernatant is used for determination.

[0116] Inject 100 mg / kg of boronated ferulic acid into the tail vein of liver cancer mice. After 3 days, cut the tail and take 20 μl of blood and mix it in 1 ml of physiological saline (containing 4 mg of EDTA·2Na / 100 ml or heparin, 1 mg of heparin anticoagulates 2 - 5 ml of blood). After centrifuging at 4000 r / min for 3 min, combine the CH 3 OH extraction solution, then evaporate CH 3 OH, take the residue, add 0.8 mL of CH 3After the OH was dissolved, it was vortex-mixed and centrifuged. The obtained upper clear liquid was used as a sample. In a polytetrafluoroethylene beaker, an EDTA solution was added, and the absorbance was measured under the same conditions as the determination of the boron standard solution. Subsequently, the mouse was dissected, and 0.2 g of the mouse's blood, heart, and liver tissue were taken, minced, 1.8 ml of normal saline was added, and it was homogenized in an ice bath with a glass homogenizer to prepare a 10% tissue homogenate. The tissue homogenate was centrifuged at 4000 r / min for 10 min at 4 °C, and the supernatant was used for determination.

[0117] 100 mg / kg of ferulic acid borate was injected into the tail vein of lung cancer mice. Three days later, 20 μl of blood was taken from the tail and mixed in 1 ml of normal saline (containing 4 mg of EDTA·2Na / 100 ml or heparin, 1 mg of heparin anticoagulates 2 - 5 ml), centrifuged at 4000 r / min for 3 min, and then the CH 3 OH extract was combined, and then the CH 3 OH was evaporated to dryness. The residue was taken, 0.8 mL of CH 3 After the OH was dissolved, it was vortex-mixed and centrifuged. The obtained upper clear liquid was used as a sample. In a polytetrafluoroethylene beaker, an EDTA solution was added, and the absorbance was measured under the same conditions as the determination of the boron standard solution. Subsequently, the mouse was dissected, and 0.2 g of the mouse's blood, heart, and lung tissue were taken, minced, 1.8 ml of normal saline was added, and it was homogenized in an ice bath with a glass homogenizer to prepare a 10% tissue homogenate. The tissue homogenate was centrifuged at 4000 r / min for 10 min at 4 °C, and the supernatant was used for determination.

[0118] The spectrophotometric method was used to determine the concentration of ferulic acid borate in Kunming mice. The specific operation was as follows: A certain gradient concentration of boron standard solution (20 μg / mL) was respectively pipetted into 7 polyethylene beakers. In each beaker, an EDTA solution was added to mask interfering ions such as iron, aluminum, and copper. It was adjusted to pH 5.0 with sodium hydroxide or hydrochloric acid standard solution, an ammonium acetate buffer solution with pH 5.2 and a methyleneimine H acid color reagent solution were added, transferred to a volumetric flask, diluted to the mark with water, mixed well, and left to stand in the dark at room temperature for 3 h. Using a 1 cm absorption cell, the absorbance was measured successively at a wavelength of 415 nm with water as the reference to plot a working curve.

[0119] 1.0 mL of the supernatant of the blood, heart, kidney tissue, liver tissue, and lung tissue of the above-mentioned diseased Kunming mice was taken to measure the absorbance. The corresponding boron amount was found according to the plotted standard curve, and then the boron content in the sample was calculated. At the same time, a blank test was carried out. The methyleneimine H acid color reagent must be prepared and used immediately.

[0120] Results showed that the measured concentrations of boron-containing ferulic acid in the plasma, heart, kidney, liver, and lung tissues of diseased Kunming mice were 8.93 mg / kg, 14.96 mg / kg, 40.00 mg / kg, 40.03 mg / kg, and 35.34 mg / kg, respectively. From the concentration results, it can be seen that the drug concentration in tumor cells was greater than 35, and the concentration was more than twice that of normal tissues and blood, which proved the effectiveness of the drug. No obvious adverse reactions were observed in the mice after injection of the drug, indicating that the drug had little toxicity to Kunming mice.

[0121] Control Example

[0122] Inject 130 mg / kg of the organic boron carrier BPA into the tail vein of renal cancer mice. After 3 days, cut the tail and collect 20 μl of blood, mix it in 1 ml of physiological saline (containing 4 mg of EDTA·2Na / 100 ml or heparin, 1 mg of heparin anticoagulates 2 - 5 ml), centrifuge at 4000 r / min for 3 min, then combine the CH3OH extraction solution, evaporate the CH3OH to dryness, take the residue, add 0.8 mL of CH3OH to dissolve it, perform vortex mixing, centrifuge, and use the obtained upper clear liquid as the sample. In a polytetrafluoroethylene beaker, add an EDTA solution, and measure the absorbance under the same conditions as the determination of the boron standard solution. Subsequently, dissect the mice, take 0.2 g of the blood, heart, and kidney tissues of the mice, cut them into pieces, add 1.8 ml of physiological saline, homogenize them in an ice bath using a glass homogenizer to prepare a 10% tissue homogenate. Centrifuge the tissue homogenate at 4000 r / min for 10 min at 4°C, and use the supernatant for determination.

[0123] Inject 130 mg / kg of the organic boron carrier BPA into the tail vein of liver cancer mice. After 3 days, cut the tail and collect 20 μl of blood, mix it in 1 ml of physiological saline (containing 4 mg of EDTA·2Na / 100 ml or heparin, 1 mg of heparin anticoagulates 2 - 5 ml), centrifuge at 4000 r / min for 3 min and then combine the CH 3 OH extraction solution, then evaporate the CH 3 OH, take the residue, add 0.8 mL of CH 3 OH to dissolve it, perform vortex mixing, centrifuge, and use the obtained upper clear liquid as the sample. In a polytetrafluoroethylene beaker, add an EDTA solution, and measure the absorbance under the same conditions as the determination of the boron standard solution. Subsequently, dissect the mice, take 0.2 g of the blood, heart, and liver tissues of the mice, cut them into pieces, add 1.8 ml of physiological saline, homogenize them in an ice bath using a glass homogenizer to prepare a 10% tissue homogenate. Centrifuge the tissue homogenate at 4000 r / min for 10 min at 4°C, and use the supernatant for determination.

[0124] Inject 130 mg / kg of the organic boron carrier BPA into the tail vein of lung cancer mice. After 3 days, cut the tail and take 20 μl of blood, mix it in 1 ml of physiological saline (containing 4 mg of EDTA·2Na / 100 ml or heparin, 1 mg of heparin can anticoagulate 2 - 5 ml), centrifuge at 4000 r / min for 3 min, and then combine the CH 3 OH extraction solution, and then evaporate the CH 3 OH. Take the residue, add 0.8 mL of CH 3 OH, dissolve it and perform vortex mixing, then centrifuge. The obtained upper clear liquid is used as a sample. In a polytetrafluoroethylene beaker, add an EDTA solution, and measure the absorbance under the same conditions as the determination of the boron standard solution. Subsequently, dissect the mice, take 0.2 g of the blood, heart, and lung tissue of the mice, cut them into pieces, add 1.8 ml of physiological saline, and homogenize them in an ice bath with a glass homogenizer to prepare a 10% tissue homogenate. The tissue homogenate is centrifuged at 4000 r / min at 4°C for 10 min, and the supernatant is used for determination.

[0125] Use spectrophotometry to determine the concentration of the organic boron carrier BPA in Kunming mice. The specific operation is as follows: Respectively pipette a certain gradient concentration of boron standard solution (20 μg / mL) into 7 polyethylene beakers. Add an EDTA solution to mask interfering ions such as iron, aluminum, and copper in each beaker, adjust to pH 5.0 with sodium hydroxide or hydrochloric acid standard solution, add an ammonium acetate buffer solution with pH 5.2 and a methyleneimine H acid color reagent solution, transfer it to a volumetric flask, dilute it to the mark with water, mix well, and place it in the dark at room temperature for 3 h. Use a 1 cm absorption cell, and measure the absorbance at a wavelength of 415 nm with water as the reference in sequence to draw a working curve.

[0126] Take 1.0 mL of the supernatant of the blood, heart, kidney tissue, liver tissue, and lung tissue of the above-mentioned Kunming mice and measure the absorbance. According to the drawn standard curve, find out the corresponding boron amount, and then calculate the boron content in the sample. At the same time, perform a blank test. The methyleneimine H acid color reagent must be prepared and used immediately.

[0127] The results show that the measured content of the organic boron carrier BPA in the plasma of Kunming mice is 8.94 μg / g, in the heart is 15.35 μg / g, in the kidney tissue is 36.67 μg / g, in the liver tissue is 39.28 μg / g, and in the lung tissue is 18.16 μg / g. The data shows that the enrichment degree of the present invention's BPA in tumor tissues is higher compared to the control. Therefore, the treatment effect is better when using BNCT therapy with thermal neutron irradiation. Thus, compared with the existing organic boron carrier BPA, the boron-containing ferulic acid of the present invention has a broader application prospect in drug development.

[0128] Comparative Example 2 In vitro anti-tumor experiment of the existing third-generation amino acid-based boron carrier

[0129] The third-generation amino acid-based boron carrier, diethyl borate B 10 (OCH 2 CH 3 ) 2 Synthesized independently by Zhongke Gaoneng (Guangzhou) Medical Technology Development Co., Ltd.

[0130] Inject 170 mg / kg of the third-generation amino acid-based boron carrier into the tail vein of renal cancer mice. After 3 days, cut the tail. Take 20 μl of blood and mix it in 1 ml of normal saline (containing 4 mg of EDTA·2Na / 100 ml or heparin, 1 mg of heparin anticoagulates 2 - 5 ml of blood). Centrifuge at 4000 r / min for 3 min, then combine the CH3OH extraction solution. Then evaporate the CH3OH to dryness, take the residue, add 0.8 mL of CH3OH to dissolve it, then perform vortex mixing and centrifugation. The obtained upper clear liquid is used as the sample. In a polytetrafluoroethylene beaker, add the EDTA solution and measure the absorbance under the same conditions as the determination of the boron standard solution. Subsequently, dissect the mice, take 0.2 g of the kidney tissue of the mice, cut it into pieces, add 1.8 ml of normal saline, and homogenize it in an ice bath with a glass homogenizer to make a 10% tissue homogenate. Centrifuge the tissue homogenate at 4000 r / min for 10 min at 4 °C, and the supernatant is used for determination.

[0131] Inject 100 mg / kg of ferulic acid boride into the tail vein of liver cancer mice. After 3 days, cut the tail and take 20 μl of blood and mix it in 1 ml of normal saline (containing 4 mg of EDTA·2Na / 100 ml or heparin, 1 mg of heparin anticoagulates 2 - 5 ml of blood). Centrifuge at 4000 r / min for 3 min, then combine the CH 3 OH extraction solution, then evaporate the CH 3 OH to dryness, take the residue, add 0.8 mL of CH 3 OH to dissolve it, then perform vortex mixing and centrifugation. The obtained upper clear liquid is used as the sample. In a polytetrafluoroethylene beaker, add the EDTA solution and measure the absorbance under the same conditions as the determination of the boron standard solution. Subsequently, dissect the mice, take 0.2 g of the blood, heart, and liver tissue of the mice, cut them into pieces, add 1.8 ml of normal saline, and homogenize them in an ice bath with a glass homogenizer to make a 10% tissue homogenate. Centrifuge the tissue homogenate at 4000 r / min for 10 min at 4 °C, and the supernatant is used for determination.

[0132] Inject 100 mg / kg of ferulic acid boride into the tail vein of lung cancer mice. After 3 days, cut the tail and take 20 μl of blood and mix it in 1 ml of normal saline (containing 4 mg of EDTA·2Na / 100 ml or heparin, 1 mg of heparin anticoagulates 2 - 5 ml of blood). Centrifuge at 4000 r / min for 3 min, then combine the CH 3 OH extraction solution, then evaporate the CH 3 OH to dryness, take the residue, add 0.8 mL of CH3 After the OH was dissolved, vortex mixing was carried out and then centrifugation was performed. The obtained upper clear liquid was used as a sample. In a polytetrafluoroethylene beaker, an EDTA solution was added, and the absorbance was measured under the same conditions as in the determination of the boron standard solution. Subsequently, the mouse was dissected, and 0.2 g of the mouse's blood, heart, and lung tissue were taken, cut into pieces, 1.8 ml of normal saline was added, and homogenization was carried out in an ice bath using a glass homogenizer to prepare a 10% tissue homogenate. The tissue homogenate was centrifuged at 4000 r / min for 10 min at 4 °C, and the supernatant was used for the determination.

[0133] The spectrophotometric method was used to determine the third-generation amino acid-based boron carrier in Kunming mice. The specific operation was as follows: A certain gradient concentration of boron standard solution (20 μg / mL) was respectively pipetted into 7 polyethylene beakers. In each beaker, an EDTA solution was added to mask interfering ions such as iron, aluminum, and copper. The pH was adjusted to 5.0 with sodium hydroxide or hydrochloric acid standard solution. An ammonium acetate buffer solution with a pH of 5.2 and a methyleneimine H acid color reagent solution were added, transferred to a volumetric flask, diluted to the mark with water, mixed well, and left to stand in the dark at room temperature for 3 h. Using a 1-cm absorption cell, the absorbance was measured successively at a wavelength of 415 nm with water as the reference to plot the working curve.

[0134] 1.0 mL of the supernatant of the blood, heart, and kidney tissue of the above-mentioned diseased Kunming mice was taken to measure the absorbance. The corresponding boron amount was determined according to the plotted standard curve, and then the boron content in the sample was calculated. At the same time, a blank test was carried out. The methyleneimine H acid color reagent must be prepared and used immediately.

[0135] The results showed that the measured content of the third-generation amino acid-based boron carrier in the plasma of Kunming mice was 8.93 μg / g, in the heart was 14.96 μg / g, in the kidney tissue was 40.00 μg / g, in the liver tissue was 40.03 μg / g, and in the lung tissue was 35.34 μg / g. Thus, it can be seen that compared with the existing third-generation amino acid-based boron carriers, the boron-containing ferulic acid of the present invention has a broader application prospect in drug development.

[0136] The above-mentioned embodiments are only the preferred embodiments of the present invention and cannot be used to limit the scope of protection of the present invention. Any non-substantial changes and substitutions made by those skilled in the art based on the present invention belong to the scope of protection required by the present invention.

Claims

1. A boron-containing ferulic acid compound, characterized in that, it has the following structural formula compound:

2. A method for synthesizing the boron-containing ferulic acid compound according to claim 1, characterized in that, it includes the following steps: (1) Protecting the double bond: Using ferulic acid (Ⅰ) as the raw material, reacting it with excessive HCl and a small amount of catalyst at room temperature to carry out an addition reaction to obtain a crude ferulic acid chloro-substituted product (a); (2) Protecting the carboxyl group: Adding excessive MeOH and 2eq trimethylchlorosilane TMSCl to the instrument containing the ferulic acid chloro-substituted product with the chemical structural formula (a), and fully reacting at room temperature for 20h to generate a compound with the chemical structural formula (b); (3) Electrophilic substitution reaction: Taking the compound containing the chemical structural formula (b), adding a small amount of Fe catalyst, introducing excessive chlorine gas, and fully reacting at 0-5°C to generate a ferulic acid chloride with the chemical structural formula (c); (4) Amine substitution reaction: Excess ammonia and a small amount of Cu 2 O are introduced into ferulic acid chloride with chemical structural formula (c), and the amine substitution reaction is carried out at 200 °C and 60 atm to produce a ferulic acid amine substitution product with chemical structural formula (d); (5) Diazotization reaction: Adding excessive sodium nitrite and excessive hydrochloric acid to the ferulic acid amine-substituted product with the chemical structural formula (d), and carrying out a diazotization reaction at 0-5°C to generate a compound with the chemical structural formula (e); (6) Iodine substitution reaction: Excessive KI is added to the compound with chemical structural formula (e), and the substitution reaction is carried out under heating conditions, and -N 2 Cl is substituted by -I to obtain the iodine-substituted ferulic acid with chemical structural formula (f); (7) Grignard reagent reaction: Adding the ferulic acid iodine-substituted product with the chemical structural formula (f) to a dry instrument, reacting with excessive metallic magnesium in excessive anhydrous ether, and heating under reflux to generate a compound with the chemical structural formula (g); (8) Synthesis reaction for introducing boron-10: Add an excessive amount of B to the compound of the above chemical structural formula (g). 10 (OCH 2 CH 3 ) 3 , react under normal temperature conditions to introduce boron-10 into the above compound, obtaining a compound with chemical structural formula (h); (9) Removing the double bond protection: Adding excessive MeOH to the compound with the above chemical structural formula (h), and reacting under alkaline conditions to obtain a compound with the chemical structural formula (i); (10) Removing the carboxyl group protection: The compound with the chemical structural formula (i) undergoes hydrolysis of the ester group in an acidic aqueous solution to generate a carboxyl group, and a boron-containing ferulic acid with the chemical structural formula (Ⅱ) is obtained 3. The method for synthesizing the boron-containing ferulic acid compound according to claim 2, characterized in that, in step (1), the catalyst is selected from one of ferric chloride, phosphoric acid, aluminum trichloride, and titanium tetrachloride; in step (3), the full reaction is for 12-48 hours; in step (4), the reaction is for 2 hours.

4. The method for synthesizing the boron-containing ferulic acid compound according to claim 2, characterized in that, in step (5), the reaction is for 2 hours; in step (6), the reaction condition is to stand in the dark for 3 hours; in step (7), the reaction condition is to heat to 90°C and fully react for 30 min; in step (8), the reaction is for 2 hours.

5. The method for synthesizing the boron-containing ferulic acid compound according to claim 2, characterized in that, in step (9), the reaction condition is to react under alkaline conditions with a pH of 8 for 1 hour; in step (10), the reaction condition is to react under acidic conditions with a pH of 3 for 1 hour.

6. Application of the boron-containing ferulic acid compound according to any one of claims 1-5 and its pharmaceutically acceptable salts in the preparation of adjuvant therapeutic drugs for renal cancer, liver cancer, lung cancer, gastric cancer, colon cancer, breast cancer, and melanoma.

7. A boron carrier, characterized in that, it contains the boron-containing ferulic acid compound according to any one of claims 1-5 and its pharmaceutically acceptable salts.

8. A pharmaceutical composition, It is characterized in that it contains a boron-containing ferulic acid compound as described in any one of claims 1-5 and its pharmaceutically acceptable salt, as well as a carrier permitted in medicine and pharmacy.

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