Preparation method and application of catechol derivative, and administration and preparation device of catechol derivative

Abstract

The invention relates to the technical field of biological material synthesis, and particularly discloses a preparation method and application of a catechol derivative and a drug delivery and preparation device.The preparation method comprises the steps that a sodium sulfite-boric acid-argon protection system is adopted, and dopa or carbidopa derivatives are prepared in an organic phase-water phase mixed solution; or dopa is taken as a raw material, SOCl2 and absolute methanol are subjected to a methyl esterification reaction, then the reaction product and methyl trifluoroacetate are subjected to an exchange reaction, and one-pot synthesis of Tfa-DOPA-OMe is realized; or preparing acetonide-protected tanshinol derivatives based on dopa derivatives or tanshinol and derivatives thereof; or the acetonide-protected tanshinol and the ester thereof are used for preparing natural catechol compounds such as Shimobastic acid C and the like, and the acetonide-protected tanshinol and the ester thereof are subjected to coupling reaction with organic acid, amino acid and derivatives thereof, oligopeptide, polypeptide, alcohol or saccharide to prepare non-natural catechol compounds; the purposes of completely and thoroughly removing acetonide protection of catechol and not destroying ester bonds contained in a molecular structure are achieved through a high-selectivity deprotection condition.

Description

Technical Field

[0001] The invention relates to the technical field of biomaterial synthesis, and in particular to a preparation method, application, administration and preparation device of a class of catechol derivatives. Background Art

[0002] Catechol (catechol) is an organic compound. Catechol exists as a functional group (catechol group) in natural products such as caffeic acid, dopamine, levodopa (L-DOPA), danshensu (DSS), rosmarinic acid (RMA), salvianolic acid (including but not limited to A to H, J to Y), lithospermic acid, salicylic acid, yunnaneicacids, shimobashiric acids (B, C) and globoidnanA and their derivatives. They often have certain biological activities, and some have even become drugs or health products. For example, levodopa is the most effective and important drug for the treatment of Parkinson's disease; danshensu is a drug for the treatment of chest tightness and angina pectoris caused by coronary heart disease; salvianolic acid is used to treat acute myocardial infarction, cerebral thrombosis and other diseases; rosmarinic acid is used as an antioxidant and anti-ultraviolet additive.

[0003] Both L-DOPA and Danshensu are catechol derivatives and are natural products of the phenylpropionic acid class (C6+C3); CarbiDOPA is a synthetic L-DOPA derivative, also known as methyldopa hydrazine, and is a DDC inhibitor of dopa decarboxylase (DDCI). They are very similar in structure, with the only difference being the substituents on α-C.

[0004] The catechol groups in catechol derivatives are easily oxidized into quinones, which can undergo irreversible cross-linking reactions with themselves or other reagents, and even produce substances with toxic side effects, which seriously affect the purity and activity of catechol derivatives. Therefore, exploring suitable catechol protecting groups or protection measures is the key to the successful preparation of natural products or derivatives containing catechol.

[0005] Moreover, as early as 1974, patents US 3803120 and US4035507 proposed the use of DOPA-containing oligopeptides as anti-Parkinson's disease prodrugs, and pointed out that these oligopeptides have good water solubility; however, the synthesis method of DOPA-containing oligopeptides is relatively complicated, requiring multiple steps of reaction, and there are problems such as complicated reaction process and high cost.

[0006] At the same time, in terms of drug administration, currently developed methods include oral administration, gastrostomy administration of DuoDOPA enteric gel, subcutaneous injection, continuous intravenous injection, continuous oral administration, continuous transdermal administration, and pulmonary administration. However, oral administration will have a weaning period over time, and gastrointestinal dysfunction and food effects affect the absorption of levodopa. In the gastrostomy administration of DuoDOPA enteric gel, Parkinson's patients need to go through a gastrostomy - implanting a tube to the duodenum for administration, which has the problem that the suspension preparation is easily caused by long-term placement. The drug can cause bidopa to precipitate and aggregate; the byproducts are ammonia and hydrazine, which makes the drug easy to deteriorate; the shelf life is short, and it needs to be stored and transported at -20°C, which is cumbersome to use and expensive to treat; in addition, the gastrostomy catheter administration method is prone to catheter dislocation, displacement, occlusion, kinking, connector damage and accidental removal, and is also prone to stoma and other infections; the subcutaneous injection dosage is relatively small, and drug absorption is greatly affected by the skin location and thickness, and is also significantly affected by individual differences and gender differences; continuous intravenous administration requires patients to actively adhere to subcutaneous injections, This results in poor patient compliance and the risk of infection during the injection process. In the continuous administration method with dental braces, the levodopa / carbidopa compound is placed in a medicine box on one side of the braces and absorbed by swallowing after soaking in saliva. The fluctuation range of blood drug concentration in patients using this method is significantly lower than that of intermittent oral administration. However, the size of the dental braces is large, and patients who wear them for a long time have a more serious foreign body sensation in their mouths, which affects their daily lives. In addition, the compound preparation lacks the necessary flavoring agents, is bland, and patient compliance is not high. Transdermal continuous administration uses a micropump to deliver a continuous and stable subcutaneous dose for 24 hours. The administration of levodopa can provide a stable and effective level of levodopa, and its variability is lower than that of standard oral levodopa, but the minimally invasive method will still cause wounds, and it is mainly aimed at people with advanced Parkinson's disease, and the applicable population is small; in pulmonary administration, levodopa, as an inhaled drug, does not rely on digestive tract absorption. It quickly enters the blood through lung absorption and is administered using a passive breath-driven drug delivery system. However, since there are many factors that affect the lung deposition rate of drug particles, the reproducibility is relatively poor. In addition, this method of administration is prone to cause sneezing, coughing, and breath holding and other adverse reactions.

[0007] Danshensu derives a large number of natural products, such as salvianolic acid, by forming ester bonds with its α-hydroxyl group; Danshensu and its derivatives are widely used in the pharmaceutical industry. The Pharmacopoeia of the People's Republic of China (2020 edition) records 179 prescriptions containing Danshen ingredients; in 2022, the in-hospital sales of drugs containing Danshen ingredients exceeded 7.34 billion yuan. At present, in the process of artificially synthesizing danshensu derivatives (such as salvianolic acid AD), because the catechol protection measures (such as methyl ether) used are too strong, when deprotecting catechol after the carbon skeleton is constructed, harsh reaction conditions are required to achieve effective deprotection; this causes a large number of ester bonds in danshensu derivatives to break, resulting in low yield of the target product, many impurities, difficult separation and purification, and poor economic benefits. Summary of the invention

[0008] In view of the above problems, the present invention is committed to providing a preparation method and a preparation device of a catechol derivative, and an application and an application device of the catechol derivative.

[0009] The first object of the present invention is to provide a method for preparing a class of catechol derivatives, which comprises adopting sodium sulfite-inert gas protection, adding or not adding boric acid, which can effectively prevent catechol from being oxidized, and preparing levodopa or carbidopa derivatives in an aqueous phase or an aqueous-organic phase mixture; by adjusting the protection sequence of the amino group and the carboxyl group of L-DOPA, a one-pot synthesis of Tfa-DOPA-OMe is achieved; and based on levodopa derivatives, danshensu or its derivatives, acetonide-protected danshensu derivatives are prepared; acetonide-protected danshensu and its esters are used to prepare natural catechol compounds, and non-natural catechol compounds are prepared by coupling reaction with organic acids, amino acids and their derivatives, oligopeptides and polypeptides, alcohols or sugars.

[0010] The second object of the present invention is to provide the use of the above-mentioned catechol derivatives, including solid preparations or liquid preparations containing DOPA prepared based on the catechol derivatives.

[0011] The third object of the present invention is to provide an oral continuous or intermittent drug delivery device, which can be used for the delivery of the above-mentioned liquid preparations. A micro peristaltic pump is used to pump out an appropriate amount of medicine from the cup body into a quantitative pool on time. The drug delivery device has extremely high compliance and a very good drug control effect. It can control the blood drug concentration to be neither too high nor too low within a certain period of time, and has small fluctuations. It is suitable for patients in the middle and late stages of Parkinson's disease, has a wide coverage, and is extremely feasibility and practical.

[0012] A fourth object of the present invention is to provide a preparation apparatus for acetal / ketone protection of a catechol group.

[0013] The first technical scheme adopted by the present invention is: a method for preparing a class of catechol derivatives, wherein the method for preparing a class of catechol derivatives includes ① the preparation of levodopa or carbidopa derivatives, ② one-pot synthesis of Tfa-DOPA-Ome, ③ the preparation of acetone-protected tanshinone derivatives, and ④ the preparation of natural catechol compounds and non-natural catechol compounds.

[0014] ① Preparation of levodopa or carbidopa derivatives;

[0015] By adopting a sodium sulfite-boric acid-argon system for protection, levodopa or carbidopa derivatives are prepared in an organic phase-aqueous phase mixture with high yield and high purity; further, for levodopa or carbidopa derivatives protected by a protective group that is unstable to acid, hydrochloric acid or trifluoroacetic acid (TFA) can be used for deprotection to obtain a dipeptide of levodopa or carbidopa; and for levodopa or carbidopa derivatives protected by a protective group that is stable to acid, an acetonide cyclization reaction can be further performed to protect the phenolic hydroxyl group of catechol.

[0016] Wherein, the preparation method of levodopa or carbidopa derivatives comprises the following steps:

[0017] S1: adding a reducing agent, a boric acid compound and a mixed solvent into a reaction device, and bubbling an inert gas for 10-30 minutes to remove oxygen in the reaction device;

[0018] S2: Add compound I and a base into the reaction device (compound I and a base are added as solid powder or compound I and a base are prepared with a mixed solvent into a solution and then dripped into the reaction device), stir and continue to pass an inert gas for 10-30 minutes;

[0019] S3: Add the reactant II in batches as solid powder or make it into a solution and drip it into the reaction device, stir and continue to pass inert gas for 3-24 hours, and then acidify to obtain compound III; the reaction formula is as follows:

[0020]

[0021] The molar ratio of the compound I to the reactant II is 1:1.3 to 1:0.3, preferably 1:0.8.

[0022] Step S3 also includes: after the reaction is completed (i.e., stirring and continuing to pass inert gas for 3-24 hours), washing multiple times with an organic solvent (such as ethyl acetate) at a pH of 6.5-11; then, extracting with an organic extractant (such as ethyl acetate) under a strong acid condition of pH 1-5, taking an organic phase; then washing the organic phase with acid, brine, and pure water respectively; then taking the organic layer, drying with anhydrous magnesium sulfate, filtering, and rotary evaporation to remove the solvent to obtain a crude product; then recrystallizing to obtain a high-purity compound III. The organic solvent and organic extractant include but are not limited to ethyl acetate (EA), tetrahydrofuran (THF), dichloromethane (DCM), toluene (TLN), petroleum ether (PE), n-hexane (HXN), methyl tert-butyl ether, etc.; preferably EA.

[0023] Wherein, the mixed solvent includes but is not limited to a combination of two or more of water, tetrahydrofuran (THF), acetone, dioxane (DIOX), dimethylformamide (DMF), etc., preferably a mixed solvent formed by THF and water. The reducing agent includes but is not limited to sodium thiosulfate, sodium sulfite, sodium pyrosulfite, vitamin C, isoascorbic acid, etc., preferably sodium sulfite and sodium thiosulfate. Boric acid compounds include but are not limited to sodium borate, potassium borate, boric acid, phenylboric acid, etc.; preferably sodium borate. In the case where boric acid causes adverse side reactions, boric acid can be omitted, and a reducing agent and an inert gas can be used in combination, or only a reducing agent can be added. Inert gases include but are not limited to argon (Ar) and nitrogen (N2), preferably Ar.

[0024] Bases include but are not limited to triethylamine (Et3N), sodium carbonate, sodium bicarbonate, potassium carbonate, sodium phosphate, sodium hydroxide, etc.; preferably sodium carbonate. Compound I includes but is not limited to levodopa (L-DOPA), methyldopa, carbidopa (CarbiDOPA), dopamine, etc., and R1 in Compound I includes but is not limited to H, Me (methyl), COOH (carboxyl), CH2COOH and other substituents; R2 includes but is not limited to H, Me, Et, F, Cl, Br, OH and other substituents; R3 includes but is not limited to NH2, NHNH2 and other substituents.

[0025] Reactant II (R4-X) is an acyl halide, anhydride, N-hydroxysuccinimide type activated ester (-OSu), 1-hydroxybenzotriazole (HOBt) type activated ester, pentafluorophenol type activated ester (PFP) or other active compounds that can react with amine groups; wherein R4 is an amino protecting group, an amino acid and its derivatives or an organic acyl group; the amino protecting group includes but is not limited to fluorenylmethoxycarbonyl (Fmoe-), benzyloxycarbonyl (Cbz-), tert-butyloxycarbonyl (Boc-), phthaloyl (Phth-), etc., and the amino acid and its derivatives include but are not limited to Fm oc-AA (amino acid)-, Boc-AA-, Cbz-AA-, Phth-AA-, Boc-AA-AA- and other aminoacyl or oligopeptide acyl (i.e., N-modified amino protecting groups), organic acyl includes but is not limited to acetyl, propionyl, butyryl, valeryl, fatty acyl (C6-C30) and other organic acyl, or a combination of two or more of the above amino protecting groups, amino acids and their derivatives, organic acyl or fatty acids, including but not limited to fatty acyl aminoacyl, such as fatty dipeptide acyl, etc.; X is an activated form, including but not limited to acyl halide, acid anhydride, activated ester or other activated forms. As some examples of R4-X, the reactant II is fluorenylmethoxycarbonyl chloride (Fmoc-Cl), phthaloyl chloride, fluorenylmethoxycarbonyl succinimide (Fmoc-OSu), (Boc)20, Fmoc-AA-OSu, Fmoc-AA-OBt, Boc-AA-OSu, Boc-AA-PFP, Boc-AA-AA-OSu, C 11 H 23 CO-Asp(OtBu)-OSu, N-ethoxycarbonylphthalimide (Phth-CE), OSu or OBt esters of hyaluronic acid, activated esters of carboxymethyl cellulose, and the like.

[0026] Acidification is performed by adding one or more of hydrochloric acid, citric acid, phosphoric acid, sulfuric acid or trifluoroacetic acid.

[0027] R5 in compound III is a group formed by condensation reaction of reactant II (R4-X) and R3. As examples of R1 / R2 / R3 / R4, compound III includes but is not limited to Fmoc-DOPA-OH, Cbz-DOPA-OH, Fmoc-Val-DOPA-OH, Boc-Met-DOPA-OH, Cbz-Aps(Bzl)-DOPA-OH, Boc-Ser(tBu)-DOPA-OH, C12H25CO-Asp(OtBu)-DOPA-OH, Fmoc-CarbiDOPA-OH, and Boc-Met-CarbiDOPA-OH.

[0028] The present invention adopts three measures to avoid oxidative crosslinking of catechol groups under alkaline conditions: a. the reducing agent reacts with residual oxygen to consume dissolved oxygen, thereby avoiding oxidation of catechol groups by oxygen, b. the inert gas keeps the reaction system in an anaerobic condition, and c. the catechol groups and boric acid form an oxidation-resistant complex in situ. In addition, the present invention avoids the occurrence of a large amount of Lossen rearrangement byproducts or oligopeptide byproducts by controlling the molar ratio of compound I to reactant II.

[0029] At the same time, the present invention utilizes compound III to be dissolved in a mixed solvent in the form of a salt under high pH (6.5-11) conditions, so that low-polar impurities can be washed away easily using an organic extractant, and compound III is only slightly washed away or not washed away at all; the pH of the mixed solution is adjusted to 1-5, compound III exists in the form of a neutral molecule, and is extracted into an organic phase by an organic extractant, while unreacted compound I is protonated and does not enter the organic phase; finally, the organic phase is washed successively with an acid solution, brine, and pure water to further remove impurities with high polarity, thereby obtaining compound III (i.e., a catechol derivative) with high purity; and the purity of compound III can be further improved by recrystallization in a suitable solvent to meet the purity requirements of pharmacy.

[0030] Preferably, for levodopa or carbidopa derivatives protected by acid-labile protecting groups (such as Boc), hydrochloric acid or TFA can be used for deprotection to obtain levodopa or carbidopa dipeptides, that is, when compound III is a Boc-modified catechol derivative (such as Boc-AA-DOPA-OH and Boc-AA-CarbiDOPA-OH), the Boc- in the catechol derivative is further converted to H- (that is, the amino protecting group is removed and the catechol derivative is deprotected to obtain an amino acid-modified catechol derivative), for example, Boc-AA-DOPA-OH and Boc-AA-CarbiDOPA-OH are converted to H-AA-DOPA-OH and H-AA-CarbiDOPA-OH.

[0031] Preferably, for levodopa or carbidopa derivatives protected by acid-stable protecting groups (e.g., Fmoc-, Phth-, 2-C1-Cbz-), an acetonide cyclization reaction can be further performed to protect the phenolic hydroxyl group of catechol to form a useful reagent (e.g., Fmoc-DOPA(Acetonide)-OH, Phth-DOPA(Acetonide)-OH, 2-C1-Cbz-DOPA(Acetonide)-OH), i.e., the catechol group in the levodopa or carbidopa derivative is further acetal / ketone protected; the levodopa or carbidopa derivative can also be deprotected (i.e., the amino protecting group is removed) to obtain an amino acid-modified catechol derivative / dipeptide; wherein, the acetal / ketone protection of the catechol group in the catechol derivative comprises the following steps:

[0032] S10: In a reaction device (e.g., a three-necked flask), a certain amount of compound IV (compound IV includes N-modified (e.g., N-Fmoc, N-Phth, N-Tfa, etc., fatty acids, lipopeptides, etc.) catechol derivatives in compound III, but does not include Boc-modified catechol derivatives) and a cosolvent are added, and heated to form a solution;

[0033] S20: adding a certain amount of carrier to the reaction device, introducing inert gas, heating to 70-120° C., reflux for about 20 minutes, and using the absorbent in the absorption device to absorb water and by-products;

[0034] S30: adding reactant V (the molar ratio of compound IV to reactant V is controlled in the range of 1:1 to 1:10, preferably 1:2) and a catalyst, and continuing to reflux for about 2-5 hours until most of compound IV is consumed; adding a small amount of reactant V again (adding 0.5-3 molar equivalents of reactant V, preferably 1 molar equivalent), and after distilling out part of the volatile substances (about 20 minutes), stopping the reaction to obtain compound VI; the reaction formula is as follows:

[0035]

[0036] The step S30 further includes: after stopping the reaction, when the temperature drops to near room temperature, slowly adding an appropriate amount of pyridine (2 equivalents) using a syringe, removing volatiles by rotary evaporation, extracting the residue with EA, washing with 0.01N0.5mol / L citric acid solution, and washing with pure water; taking the organic phase, adding magnesium sulfate to dry, filtering, and rotary evaporating to dryness; the obtained crude product is recrystallized in TLN / n-hexane (HXN) to obtain a high-purity compound VI.

[0037] Wherein, the compound IV includes but is not limited to compound III, R1 includes but is not limited to Me, COOH, CH2COOH and other substituents; R2 includes but is not limited to H, Me, Et, F, Cl, Br, OH and other substituents; R5 is a combination of amino or hydrazine and its corresponding protecting groups, including but not limited to -NHFmoc, -NHNHFmoc, -NHCbz, -NPhth, -NHTfa, -NTFa-NHTfa, C 15 H 31 CO-NH-, C 12 H 25 CO-NH-, C 12 H 25 CO-Val-NH-, Fmoc-Val-NH-, etc.

[0038] The co-solvent is a solvent with high solubility for compound IV, including but not limited to THF, acetone, methyl ethyl ketone, ether, etc., preferably acetone. The carrier includes but is not limited to benzene, TLN, xylene, etc., preferably benzene; the carrier is used to remove water and by-products from the reaction system by azeotropic distillation. The inert gas includes but is not limited to Ar, N2, preferably Ar.

[0039] The absorption device includes but is not limited to a Soxhlet extractor, a constant pressure dropping funnel and an improved constant pressure dropping funnel disclosed in the present invention. The absorbent includes but is not limited to calcium chloride, calcium oxide, magnesium sulfate, sodium sulfate, phosphorus pentoxide, molecular sieves, etc., preferably calcium chloride.

[0040] The catalyst is an acid, including but not limited to p-toluenesulfonic acid (TsOH), camphorsulfonic acid, trifluoromethanesulfonic acid, TFA, HCl, lactic acid, hydrogen ion exchange resin, etc.; according to the different reaction activities of catechol and the accompanying side reactions, the catalyst with appropriate concentration and equivalent is selected; preferably 2.5% equivalent TsOH.

[0041] The reactant V includes but is not limited to aldehydes, ketones, hemiacetals and hemiketals of aldehydes and ketones, etc.; aldehydes include formaldehyde, acetaldehyde, benzaldehyde, etc.; ketones include acetone, butanone, cyclopentanone, cyclohexanone, benzophenone, diphenylethylenedione, hemiacetals and hemiketals of aldehydes and ketones include 2,2-dimethoxypropane (DMP) and 2,2-dimethoxy-2-phenylacetophenone; preferably DMP, cyclohexanone and benzophenone.

[0042] In the compound VI, R6 is a structure formed by acetalization or ketalization of the reactant V and the catechol group, including but not limited to formal, acetal, benzaldehyde, acetone, cyclopentanone, cyclohexanone, benzophenone, methoxymethylene (MOM), ethoxymethylene (EOM), etc., preferably acetone. The compound VI includes but is not limited to Fmoc-DOPA (Acetonide) -OH, Fmoc-DOPA (Cyclohexanonide) -OH, Fmoc-DOPA (benzophenone) -OH, Phth-DOPA (Acetonide) -OH, Cbz-DOPA (Acetonide) -OH, Tfa-DOPA (Acetonide) -OH, Fmoc-Val-DOPA (Acetonide) -OH, Fmoc-(2-Cl-Cbz)CarbiDOPA (Acetonide) -OH, C 12 H 25 CO-Val-DOPA(Acetonide)-OH, etc.

[0043] The present invention adopts a method of adding the reactant V in batches, which not only avoids the generation of more by-products when the reactant V is excessive, but also avoids the problem of too low conversion rate of compound IV when the reactant V is insufficient. The first addition of reactant V: the molar amount of compound IV to the molar amount of reactant V is controlled in the range of 1:1 to 1:10, preferably 1:2; the second addition of reactant V: 0.5-3 molar equivalents of reactant V are added before the reaction is almost finished, preferably 1 molar equivalent.

[0044] The present invention adopts a more effective by-product removal device (absorption device of the present invention), improves conversion efficiency, increases product purity, and reduces the difficulty of product separation and purification; in addition, the present invention successfully amplifies the ketal reaction by multiple times using absorption device A, thereby realizing mass production. Moreover, the present invention adopts a TLN / HXN solvent system for recrystallization, better separates the target product from its methyl ester, reduces the impurity content, and improves the purity of the target product VI.

[0045] Preferably, the method further comprises converting the byproduct containing dopa methyl ester into a target carboxylic acid;

[0046] When the catechol group in the catechol derivative is protected by acetal / ketone, a by-product containing DOPA methyl ester is generated. The present invention provides a method for converting the by-product containing DOPA methyl ester into a target carboxylic acid, and a method for preparing acetal / ketone protected carboxylic acid substances of DOPA analogs. The method can selectively decompose the methyl ester compound VII and convert it into compound VIII (including but not limited to compound VI).

[0047] The conversion of the dopa methyl ester-containing byproduct into the target carboxylic acid comprises:

[0048] 1) Adding a certain amount of methyl ester compound VII and an organic solvent into a reaction device to obtain a methyl ester compound VII solution;

[0049] 2) dissolving a certain amount of lithium salt (LiX) in a small amount of water to obtain a lithium salt solution, and adding the lithium salt solution to the above-mentioned methyl ester compound VII solution; adjusting the pH of the reaction system to 7-8 with hydrochloric acid, and then adding a certain amount of alkali, and stirring for 3-24 hours;

[0050] 3) Concentrate the organic phase by rotary evaporation, add water, adjust the pH to 3-5 with HCl solution, extract with EA, wash the organic phase with acid water and pure water respectively; take the organic phase, add anhydrous magnesium sulfate to dry, filter, and rotary evaporate to remove volatiles, and recrystallize in TLN / HXN to obtain high-purity compound VIII; the reaction formula is as follows:

[0051]

[0052] Wherein, the lithium salt includes but is not limited to lithium halide, lithium trifluoromethanesulfonate, lithium perchlorate, etc.; preferably lithium bromide. The organic solvent includes but is not limited to THF, acetonitrile, acetone, etc.; preferably acetonitrile. The base is a common organic base, including but not limited to Et3N, diisopropylethylamine (DIEA), etc.; preferably DIEA.

[0053] The definitions of R2, R5, and R6 in the compound VII and compound VIII are the same as those of R2, R5, and R6 in the compound VI; the compound VII includes but is not limited to Fmoc-DOPA(Acetonide)-OMe, Phth-DOPA(Acetonide)-OMe, Cbz-DOPA(Acetonide)-OMe, Tfa-DOPA(Acetonide)-OMe, etc. The compound VIII includes but is not limited to Fmoc-DOPA(Acetonide)-OH, Phth-DOPA(Acetonide)-OH, Cbz-DOPA(Acetonide)-OH, Tfa-DOPA(Acetonide)-OH, etc.

[0054] ② One-pot synthesis of Tfa-DOPA-OMe;

[0055] One-pot synthesis of Tfa-DOPA-OMe uses levodopa as raw material, adopts SOCl2 / MeOH methylation to replace the CH3I / KHCO3 / DMF esterification reaction of the existing synthesis route, and then realizes a one-pot synthesis method through an ester amide exchange reaction with methyl trifluoroacetate, which is simple and efficient to protect the carboxyl group and the amino group; the specific steps include:

[0056] L-DOPA and anhydrous methanol (MeOH) were added to the reaction apparatus, argon was introduced, SOCl2 was slowly added dropwise under ice bath conditions, and stirring was continued overnight after the ice bath was removed; heating and stirring were performed, argon bubbling was turned on, the temperature was lowered, triethylamine and methyl trifluoroacetate were slowly added, and stirring was performed until the reaction was complete; after rotary evaporation and concentration, the pH of the solution was adjusted to 3-4, extraction was performed, and the obtained organic phase was washed with water, dried over anhydrous magnesium sulfate, rotary evaporated, and vacuum dried to obtain Tfa-DOPA-OMe.

[0057] The reaction mechanism of the one-pot synthesis of Tfa-DOPA-OMe is as follows: first, L-DOPA is converted into methyl ester hydrochloride under the action of SOCl2 / MeOH; although methanol is a poor solvent for L-DOPA, it is a good solvent for the intermediate product methyl ester hydrochloride, so it can promote the completion of this step of the reaction; after the methylation is completed, the excess SOCl2 can be removed from the reaction system by heating to reflux in an oil bath and bubbling with argon; there are many schemes for the trifluoroacetylation of the amino group, and the ester-amide exchange reaction is more effective than the direct acylation with trifluoroacetic anhydride, as the former has mild reaction conditions and almost no racemization; in addition, since the reaction activity of methyl trifluoroacetate (Tfa-OMe) is slightly higher than that of ethyl ester, it is more suitable; among them, the amount of triethylamine is critical. If the amount is too small, the reaction will be relatively slow, and if the amount is too large, it will lead to waste and decomposition of the methyl ester. After screening, the most suitable molar ratio of triethylamine to L-DOPA is 4.

[0058] Furthermore, based on Tfa-DOPA-OMe, an acetonide-protected L-DOPA intermediate (Tfa-DOPA(Acetonide)-OMe) can be successfully prepared; and when the amino and carboxyl protections are completely removed and then coupled with the N-protecting group or only the methyl ester is selectively removed, other acetonide-protected L-DOPA reagents can be prepared, such as Boc-DOPA(Acetonide)-OH, Fmoc-DOPA(Acetonide)-OH, Phth-DOPA(Acetonide)-OH, Tfa-DOPA(Acetonide)-OH, etc.

[0059] Although the present invention claims an advantageous method in which the two-step reaction is carried out in one reactor, it is also claimed that according to this improvement, the one-pot process is simply changed to carrying out the two-step reaction claimed by the present invention in two vessels.

[0060] ③ Preparation of acetone-protected danshensu derivatives;

[0061] The preparation of acetone-protected danshensu derivatives based on dopa derivatives specifically includes:

[0062] The acetonide-protected L-dopa derivative is deaminated and reacted with nitrous acid in water or a mixture of water and an organic solvent under acid catalysis, and then hydrolyzed to generate an acetonide-protected L-danshensu derivative. Correspondingly, an acetonide-protected D-danshensu derivative is prepared using a dextrodopa derivative.

[0063] The preparation of acetone-protected danshensu derivatives based on danshensu or its derivatives specifically includes:

[0064] First, the carboxyl group of danshensu is protected as an ester or amide to improve its solubility in organic solvents, and the alcoholic hydroxyl group of danshensu is protected as an ester or ether, preferably an ester, to avoid the Oxa-Pictet-Spengler side reaction, so as to obtain a danshensu intermediate; for example, methyl rosmarinic acid, the α-hydroxyl group of the danshensu structure is protected as an ester by caffeic acid, and the carboxyl group is protected as a methyl ester; then, the danshensu intermediate is reacted with an acetonide cyclization reagent to achieve the acetonide protection of the catechol group of danshensu; finally, through selective hydrolysis, alcoholysis or aminolysis, an acetonide-protected danshensu derivative is obtained.

[0065] ④ Preparation of natural catechol compounds and non-natural catechol compounds;

[0066] Using acetone-protected danshensu and its esters as raw materials, natural catechol compounds such as rosmarinic acid, lithospermic acid, salvianolic acid, salicylic acid, Yunnaneic acids, SBA-C and globoidnanA are prepared through ester bond formation (including Mitsunobu configuration inversion ester bond formation reaction), amide bond formation or ether bond formation reaction, and non-natural catechol compounds formed with organic acids, amino acids and their derivatives, oligopeptides and polypeptides, alcohols, sugars, etc. are prepared.

[0067] Among them, in the structure of the reactant that undergoes ester, amide or ether bond formation reaction with acetonide-protected danshensu, if there is a catechol group, the catechol group is also protected by acetonide; if there is no catechol group but an isolated phenolic hydroxyl group is contained, the phenolic hydroxyl group should be appropriately protected, preferably with a protecting group in an acid-labile orthogonal system, including but not limited to MOM, EOM, Boc, and tBu. This ensures that when the acetonide protection is removed, other protecting groups that need to be removed are also removed, thereby reducing the difficulty of synthesis.

[0068] Finally, using medium concentration of TFA, HCl, HBr or TsOH as catalyst, under mild conditions, the acetone protection is removed to obtain the target catechol compound; the deprotection conditions do not destroy the ester bonds, glycosidic bonds, amide bonds, hydrazide bonds, ether bonds, and disulfide bonds that are desired to be retained but unstable in the structure of the intermediate product and the target product; preferably, a mixed solution of 60% TFA and DCM is added with 10 equivalents of water and 10 equivalents of triisopropylsilane (TIS); the yield of the target product is close to 100%, and the ester bond formed by the methyl ester and the α-hydroxyl of Danshensu is not destroyed. For example, the synthesis of the natural product SBA-C.

[0069] The second technical solution adopted by the present invention is: the application of a class of catechol derivatives, including the preparation of solid preparations and liquid preparations containing dopa oligopeptides based on the catechol derivatives.

[0070] (1) Dopa oligopeptide solid preparation (DOSF);

[0071] The solid preparation containing dopa oligopeptides includes the catechol derivative API-1. The category of API-1 is a derivative providing L-DOPA. API-1 includes but is not limited to one or more of L-DOPA, dopa esters, fatty acyl dopa, L-DOPA-containing oligopeptides (preferably dipeptides, tripeptides), L-DOPA-containing lipo-oligopeptides (preferably lipodipeptides), etc.

[0072] The solid preparation containing DOPA oligopeptide also includes other active ingredients and / or excipients (FE); other active ingredients include API-2 and / or API-3, the category of API-2 is DDCI (DOPA decarboxylase inhibitor), API-2 includes one or more of benserazide hydrochloride, CarbiDOPA, oligopeptides containing CarbiDOPA (preferably dipeptides, tripeptides), ester oligopeptides containing CarbiDOPA (preferably lipid dipeptides), etc.; the category of API-3 is other active pharmaceutical ingredients, and API-3 includes other anti-Parkinson's disease drugs, such as COMT inhibitors, MAO-B inhibitors, etc.

[0073] Excipients (FE) include ion regulators (FE-1), pH regulators (FE-2), thickeners / physical gelling agents (FE-3), antioxidants (FE-4), flavoring agents (FE-5), preservatives and antibiotics (FE-6), etc.; ion regulators include NaCl, pH regulators include citric acid salts, phosphate salts, acetic acid salts, phthalic acid salts, etc., thickeners / physical gelling agents include carboxymethyl cellulose salts, hyaluronic acid salts, polyacrylic acid salts, etc., antioxidants include isoascorbic acid, sodium sulfite, sodium bisulfite, sodium metabisulfite, sodium thiosulfate, vitamin C, vitamin E, etc., flavoring agents include aspartame, sucrose, stevioside, agar, sodium bicarbonate, pineapple essence, etc., preservatives and antibiotics include benzoic acid, methyl / ethyl parahydroxybenzoate, chlorhexidine acetate, etc.

[0074] Since the above-mentioned solid preparation containing DOPA oligopeptide does not contain water, the rate of degradation to produce ammonia and hydrazine during production, packaging, transportation and storage is very slow, and the rate of oxidation of the catechol group therein is also very slow, thus meeting the requirement for a longer shelf life of the drug; when used by patients, if it is used to replace liquid preparations such as Duopa, Sirio, ABBV-951, etc., a pre-prepared solvent (Pre-Prepared menstrua, PPM) can be added, and after shaking for 5-30 minutes, a clear, uniform, transparent liquid can be formed for use.

[0075] The solvent (PPM) is, for example, a pharmaceutical solvent or a prefabricated solution; the pharmaceutical solvent (PPM-1) is, for example, water for injection, physiological saline for injection or glucose solution for injection; the prefabricated solution is, for example, PPM-2, PPM-3 or PPM-4, PPM-2 includes, for example, water, 0.1-10% (mass percentage) carboxymethyl cellulose salt, preservative and antibiotic, or includes, for example, water, 0.1-10% (mass percentage) carboxymethyl cellulose salt, alkali, preservative and antibiotic, or 0.1-10% (mass percentage) carboxymethyl cellulose salt aqueous solution; PPM-3 includes, for example, water, 0.1-10% (mass percentage) hyaluronate, alkali, preservative and antibiotic, or includes, for example, 0.1-10% hyaluronate aqueous solution; PPM-4 includes, for example, water, 0.1-10% (mass percentage) polyacrylate, alkali, preservative and antibiotic.

[0076] (2) Liquid preparations containing dopa oligopeptides;

[0077] The liquid preparation containing DOPA oligopeptide includes DOPA-containing dipeptide and carbidopa dipeptide and flavor corrector, thereby forming an oral liquid preparation with pleasant taste (high patient compliance); the drug exists in the form of solution and is not easy to deteriorate or lose effectiveness in a short period of time.

[0078] Flavoring agents include sweeteners and / or aromatics.

[0079] Sweeteners are divided into two categories: natural sweeteners and synthetic sweeteners. Natural sweeteners include sucrose, monosaccharide syrup, stevioside, etc.; synthetic sweeteners include saccharin sodium, cyclamate, aspartame, etc.; other flavor enhancers (also called flavoring agents) that do not affect the efficacy of the medicine can also be added to prepare oral solutions with different special flavors, such as orange flavor, apple flavor, etc.; effervescent agents and syrups that interfere with the taste can also be added.

[0080] The third technical solution adopted by the present invention is: to provide an oral continuous or intermittent drug delivery device, including a cup body and a cup cover, which are detachably connected; the cup body is a container for holding liquid preparations; the cup cover is provided with a switch button, a battery slot, a micro peristaltic pump, a delivery tube, a quantitative tank and an output tube, the switch button is used to turn on or off the micro peristaltic pump, the battery slot is used to install batteries to power the micro peristaltic pump, the micro peristaltic pump is used to pump the liquid medicine in the cup body into the quantitative tank on time and in quantity through the delivery tube, the delivery tube is installed on the micro peristaltic pump, one end of the delivery tube is located at the bottom of the cup body, and the other end is located at the bottom of the quantitative tank, the output tube passes through the cup cover and extends into the quantitative tank for the patient to obtain the liquid medicine.

[0081] The fourth technical solution adopted by the present invention is: a preparation device for acetalization and ketone protection of catechol groups, comprising a reactor, a condensing device, an absorption tower, a gas pipeline and a liquid pipeline; the reactor is connected to the condensing device through a gas pipeline, the reactor is connected to the absorption tower through a liquid pipeline, the absorption tower is connected to the condensing device through a liquid pipeline, and a valve is provided on the liquid pipeline.

[0082] Among them, the reactor includes but is not limited to flasks and reactors; the gas pipeline includes but is not limited to glass tubes, plastic tubes, ceramic tubes, metal tubes, etc., preferably glass tubes with vacuum jackets; the condensation equipment includes but is not limited to glass or metal condensers, cold traps, etc., preferably glass condensers; the absorption tower includes but is not limited to glass tubes, ceramic tubes, metal tubes, etc., preferably stainless steel tubes: the absorbent includes but is not limited to one or a combination of any two or more of calcium chloride, molecular sieves, calcium oxide, caustic soda, magnesium sulfate, sodium sulfate, etc., preferably calcium chloride; the liquid pipeline includes but is not limited to metal tubes, plastic tubes, rubber tubes, glass tubes, etc., preferably polytetrafluoroethylene plastic tubes.

[0083] Preferably, in industrial production equipment, the condensing device and the absorption tower are connected in parallel, and in laboratory small-scale production equipment, the condensing device and the absorption tower are connected in a vertical manner with the upper and lower ends connected.

[0084] Preferably, the present invention also discloses an improved constant pressure dropping funnel, wherein the reactor is connected to the lower interface of the improved constant pressure dropping funnel through a liquid pipeline, and the condensation equipment is vertically connected to the upper interface of the improved constant pressure dropping funnel through a liquid pipeline.

[0085] The improved constant-pressure dropping funnel includes a dropping funnel body, a constant-pressure tube, a vacuum sleeve and a condensation sleeve. A regulating valve is arranged in the middle of the dropping funnel body, and the regulating valve divides the dropping funnel body into an upper part of the dropping funnel and a lower part of the dropping funnel. The constant-pressure tube connects the upper part of the dropping funnel and the lower part of the dropping funnel. An upper interface is arranged at the upper part of the dropping funnel, and a lower interface is arranged at the lower part of the dropping funnel. A dropping nozzle is arranged in the lower interface, and the dropping nozzle is connected with the upper part of the dropping funnel. A sand core plate is arranged at the lower end of the upper body of the dropping funnel. The vacuum sleeve is wrapped around the outside of the constant-pressure tube, and the condensation sleeve is arranged on the upper part of the dropping funnel.

[0086] The improved Soxhlet extractor intermediate tube comprises a body, a constant pressure tube (with or without a vacuum sleeve), a siphon tube and a finger-shaped condenser tube socket or an outer condenser sleeve. The siphon tube is used to replace the regulating valve of the dropping funnel, which is convenient to use.

[0087] Beneficial effects of the above technical solution:

[0088] (1) In the preparation method of a class of catechol derivatives disclosed in the present invention, the preparation of levodopa or carbidopa derivatives utilizes inert gas / reducing agent / borate for combined protection, which effectively prevents the catechol derivatives from being oxidized. The method is synthesized in two steps and has the advantages of simple operation, high yield and purity of the product, etc.

[0089] (2) Compared with the prior art (CN110294789A), the synthesis scheme using propylene protection requires multiple steps, is too complicated, and has high cost. The present invention optimizes the method for synthesizing AA-L-DOPA type dipeptides, uses inert gas / reducing agent / borate combined protection, and develops a two-step synthesis method that is simple, has high yield, and high purity.

[0090] (3) The present invention extends to the synthesis of other N-modified L-DOPA complexes (catechol derivatives), including various amino protecting groups, such as N-Fmoc, N-Phth, N-Tfa, fatty acids, lipid dipeptides, etc.; it mainly solves the following problems: 1) It solves the problem that the product purity is not high and high-purity products need to be passed through a liquid column to be obtained, and an inert gas / reducing agent / borate combined protection is required; 2) It reduces the ratio of N-modified groups to L-DOPA, avoiding the Lossen rearrangement caused by -OSu activated ester and the generation of dipeptide / tripeptide impurities caused by acyl chloride.

[0091] (4) In the present invention, the DMP (reactant V)-TsOH (catalyst) system is used to protect the catechol group by acetonide; at the same time, the reactant V is added in batches, which not only avoids the generation of more by-products when the reactant V is excessive, but also avoids the problem of too low conversion rate of compound IV when the reactant V is insufficient. The first addition of reactant V: the molar amount of compound IV to the molar amount of reactant V is controlled in the range of 1:1 to 1:10, preferably 1:2; the second addition of reactant V: 0.5-3 molar equivalents of reactant V are added before the reaction is almost finished, preferably 1 molar equivalent.

[0092] (5) The present invention provides a simpler and more efficient method for preparing DOPA-containing oligopeptides (catechol derivatives). The prepared L-DOPA derivatives have high purity and are suitable for use as prodrugs in preparations or in scientific research. The present invention provides a more efficient and economical method for preparing DOPA-containing oligopeptide-type prodrugs, provides a synthetic method for preparing carbidopa oligopeptide-type prodrugs with good water solubility, and provides intermediates used in preparing DOPA-containing oligopeptides, especially acetone-protected intermediates.

[0093] (6) Compared with the prior art, the one-pot synthesis of Tfa-DOPA-OMe disclosed in the present invention greatly simplifies the reaction process by adjusting the protection order of the amino group and the carboxyl group of L-DOPA, first protecting the carboxyl group and then protecting the amino group, avoiding the use of high-boiling-point DMF as a solvent and not using the expensive and environmentally harmful methyl iodide reagent. That is, the present invention does not use the non-volatile DMF and the expensive reagent CH3I, is environmentally friendly, and the two-step reaction steps are combined into a one-pot reaction, which has the advantages of simplicity, high efficiency, reagent saving, and low cost.

[0094] (7) The present invention prepares a solid preparation containing DOPA oligopeptides based on catechol derivatives, which can be used to replace liquid preparations such as Duopa, Sirio, and ABBV-951. When used as a substitute for Duopa preparations, it mainly solves the following problems: 1) The aqueous liquid suspension preparation is unstable and easily decomposes to produce harmful components such as ammonia and hydrazine. Compared with Duopa preparations, the present invention prepares a solid preparation, and when used, water is injected into the preparation by a syringe or the like, mixed, and a solution is formed, thereby meeting the requirement for a longer shelf life of the drug; 2) L-DOPA and carbidopa in the active ingredients of Duopa are in the The solubility in the aqueous phase pH (5.8-6.8 range) is relatively low, 5 g / L and 5 mg / L respectively. In the present invention, levodopa is prepared into oligopeptides, dipeptides or tripeptides to improve the solubility (about 10 times, which can meet the demand), and carbidopa is also converted into a prodrug, and the solubility is further improved (also meeting the use requirements), but the present invention still uses soluble benserazide hydrochloride as a substitute for carbidopa; 3) The synthesis method for preparing the dipeptide or tripeptide of N-terminal levodopa is optimized, so that the synthesis cost of this type of prodrug is greatly reduced, and the cost can be borne by patients.

[0095] As an alternative drug to Sirio, Madopar and Sinime developed by Italian Cathy Pharmaceuticals, levodopa has poor water solubility, so the levodopa powder contained in Madopar and Sinime is prone to drug accumulation and dose dumping in the stomach. Sirio developed by Italian Cathy Pharmaceuticals uses dopa methyl ester (a levodopa prodrug) instead of levodopa, avoiding the above problems. However, dopa methyl ester is an organic amine and does not have an isoelectric point, so it can only exist in the form of hydrochloride, which is particularly hygroscopic and difficult to handle. Therefore, the present invention synthesizes dopa dipeptide / tripeptide type prodrugs, which are similar to amino acids in structure and have an isoelectric point, so they can form neutral inner salts, greatly reducing absorbability.

[0096] As an alternative drug to ABBV-951 preparation; ABBV-951 uses FoslevoDopa and foscarbidopa as prodrugs, and the preparation method is complicated and the cost is very high; compared with the ABBV-951 preparation, the present invention uses dopa oligopeptide prodrugs and carbidopa dipeptide prodrugs as substitutes to reduce costs.

[0097] (8) The "milk tea cup" type drug delivery device of the present invention can pump out an appropriate amount of medicine from the cup body into the quantitative pool on time, so that the patient can drink it once every 5 to 10 minutes, so that the amount of medicine each time the patient drinks is adapted to the interval time; the medicine is convenient to drink, tastes good, is non-invasive and painless, and can basically stabilize the blood drug concentration of Parkinson's patients without surgical fistula, braces, or skin perforation. The patient compliance is very high, and a good therapeutic effect can be achieved.

[0098] (9) The temperature of the reflux liquid in the inner cylinder of the Soxhlet extractor is relatively high, which is not conducive to the absorption of methanol. The inner cylinder is relatively small, and the absorbent dosage is insufficient, resulting in large fluctuations in the yield and difficulty in scaling up the reaction volume. In view of the above technical problems, the present invention designs an experimental instrument device A for mass production. The experimental device A-1 (including its absorbent holding part device A-1-1) is designed to ensure that the absorbent absorbs methanol under low temperature conditions, and gradient absorption is introduced, which has a good effect, slightly increases the yield, and is stable and highly repeatable. That is, the present invention adopts a more effective by-product removal device (the absorption device of the present invention) when performing acetal / ketone protection on the catechol group, thereby improving the conversion efficiency, increasing the product purity, and reducing the difficulty of product separation and purification. In addition, the present invention uses the absorption device A to successfully scale up the ketal reaction by multiple times, thereby achieving mass production.

[0099] (10) In the prior art CN111153885A, Fmoc-DOPA(Acetonide)-OH and the impurity Fmoc-DOPA(Acetonide)-OMe are mixed together and are difficult to remove. In view of the above technical problems, the present invention explores a better recrystallization method, using toluene and petroleum ether (n-hexane) for recrystallization, with a purity of more than 98.5%. That is, the present invention uses a TLN / HXN solvent system for recrystallization, which better separates the target product from its methyl ester, reduces the impurity content, and improves the purity of the target product VI. Moreover, the by-product Fmoc-DOPA(Acetonide)-OMe in CN111153885A affects the yield, and it is seriously wasted if thrown away. The present invention discloses a method for efficiently converting the by-product Fmoc-DOPA(Acetonide)-OMe to Fmoc-DOPA(Acetonide)-OH, which has a good effect and a total yield of more than 90%. BRIEF DESCRIPTION OF THE DRAWINGS

[0100] Figure 1 A side view of a device for oral continuous or intermittent drug delivery provided by one embodiment of the present invention;

[0101] Figure 2 A top view of a cup cover in a device for oral continuous or intermittent drug delivery provided by one embodiment of the present invention;

[0102] Figure 3 A schematic diagram of the structure of a preparation device A provided in one embodiment of the present invention;

[0103] Figure 4 A schematic diagram of the structure of a preparation device A-1 provided in one embodiment of the present invention;

[0104] Figure 5 A schematic structural diagram of an improved constant pressure dropping funnel A-1-1 provided in one embodiment of the present invention;

[0105] Figure 6 A schematic structural diagram of an improved Soxhlet extractor intermediate tube A-1-2 provided in one embodiment of the present invention;

[0106] Figure 7 A schematic structural diagram of an improved Soxhlet extractor intermediate tube A-1-3 provided in one embodiment of the present invention;

[0107] Among them, 100-cup cover, 101-switch button, 102-battery slot, 103-micro peristaltic pump, 104-transmission tube, 105-output tube, 106-speed adjustment button, 107-flow rate display screen, 108-quantitative pool; 200-cup body, 201-cup body shell; 1-dropping funnel body, 11-regulating valve, 12-dropping funnel upper part, 13-dropping funnel lower part, 14-first upper interface, 15-first lower interface; 2-first constant pressure tube; 3-vacuum jacket; 4-condensation jacket, 41-coolant inlet connector, 42-coolant outlet connector; 5-sand core plate; 2.1- The second lower interface, 2.2-the second constant pressure tube, 2.3-the closed bottom of the first glass tube, 2.4-the frosted bottom of the first glass tube, 2.5-the Soxhlet extractor body, 2.6-the first siphon, 2.7-the side ground mouth, 2.8-the second upper interface, 2.9-the vacuum jacket of the middle tube of the Soxhlet extractor; 301-the third upper interface, 302-the closed cooling jacket, 303-the coolant outlet, 304-the coolant inlet, 305-the frosted bottom of the second glass tube, 306-the second siphon, 307-the third lower interface, 308-the third constant pressure tube, 309-the closed bottom of the second glass tube, 310-the tube body. DETAILED DESCRIPTION

[0108] The present invention is further described below through specific embodiments. It should be pointed out that, for ordinary technicians in this field, several modifications and improvements can be made without departing from the principle of the present invention, and these should also be regarded as belonging to the protection scope of the present invention.

[0109] The contents not described in detail in the specification of the present invention belong to the common knowledge of the professionals in this field.

[0110] Example 1: Preparation of Levodopa or Carbidopa Derivatives

[0111] 1-1: Synthesis of Fmoc-DOPA-OH

[0112]

[0113] Method A: In a 500mL three-necked flask, add 9.53g (25mmol) Na2B407.10H2O, 1.75g ​​Na2S203 and 100mL water, stir and notify for 30min; add 9.86g (50mmol) L-DOPA, 3.98g (37.5mmol) sodium carbonate, 75mL water and 75mL THF; add a total of 13.48g (40mmol, 0.8eq) Fmoc-OSu in batches and stir overnight; then, concentrate the reaction solution by rotary evaporation, wash the residual liquid with EA for several times, and retain the aqueous phase; adjust the pH of the aqueous phase to 2 with 2N hydrochloric acid, and extract 3 times with EA; combine the organic phases and wash them with 0.1N hydrochloric acid, saturated salt and water for several times. Next, the organic phase was dried over MgSO4, filtered, and rotary evaporated to obtain a crude product, which was finally recrystallized from EA / HXN to obtain 16.3 g of white powder, Fmoc-DOPA-OH, with a purity of >98%, a yield of 97% (based on Fmoc-OSu), and an HPLC retention time (HPLC RT) of 14.7 min.

[0114] High performance liquid chromatography (HPLC) analysis conditions: Waters e2695 HPLC instrument, detection wavelength 280 / 215 nm, Symmetry C18 (4.6×250 mm, 5 μm) reverse phase chromatographic column; mobile phase A (89.8% water, 10% acetonitrile, 0.1% TFA) and B (89.9% acetonitrile, 10% water, 0.1% TFA), linear gradient elution from 100% A to 100% B for 20 min, followed by isocratic elution with 100% B for 20 min; flow rate 1 mL / min.

[0115] Method B uses Fmoc-Cl instead of Fmoc-OSu, with a purity of >98% and a yield of about 75%.

[0116] The purity of Fmoc-DOPA-OH prepared by the two methods is very high. Lossen rearrangement byproducts caused by Fmoc-OSu and dipeptide byproducts caused by Fmoc-Cl are not detected, and liquid column chromatography purification is not required.

[0117] 1-2: Synthesis of Phth-DOPA-OH

[0118] In the reaction kettle, add 71.5g (187mmol) Na2B407.10H2O, 1500mL water and 15g Na2SO3, stir mechanically, and inform for 30min. Then, add 73.9g L-DOPA (375mmol), 30g Na2CO3 and 1500mL THF. Add 73.5g (335mmol) Phth-CE in batches; stir and react at room temperature for about 3-16h. Concentrate the reaction solution by rotary evaporation, add HCl to adjust the pH to 1-2, extract with EA for several times, combine the organic phases, wash with 1mM hydrochloric acid, saturated brine and water in turn, dry with MgSO4, filter, and concentrate by rotary evaporation to obtain a yellow crude product. Recrystallize with a mixture of DCM and PE (5:1) to obtain a white solid with a yield of 75%, a purity of 100%, and an HPLC RT of 13.2min.

[0119] 1-3: Synthesis of Boc-Met-DOPA-OH

[0120] In a 500mL three-necked flask, add 9.53g (25mmol) Na2B407·10H2O, 1.75g ​​Na2S203 and 175mL water, and stir for 30min; add 9.85g (50mmol) L-DOPA, 3.0g (26.3mmol) Na2CO3, and 75mL THF; then, add 13.85g (40mmol) Boc-Met-OSu in batches, and stir to react overnight; concentrate the reaction solution by rotary evaporation, adjust the pH to 7-8, and wash with EA several times; adjust the pH of the aqueous phase to 3 with 1M citric acid solution, and extract with EA 3 times; wash the organic phase with 1mM HCl, saturated brine and water in sequence; take the organic phase, dry it over MgSO4, filter it, and spin dry it. The obtained solid is recrystallized from EA / HXN to obtain 10.9g of white powder with a yield of 64% and an HPLC RT of 11.50min.

[0121] 1-4: Synthesis of Boc-Met-CarbiDOPA-OH

[0122] Using reaction conditions similar to those of Example 1-3, 12.2 g (50 mmol) of L-CarbiDOPA and 13.85 g (40 mmol) of Boc-Met-OSu were used as reactants. The crude product was recrystallized from EA / HXN to obtain a white powder of Boc-Met-CarbiDOPA-OH, 16.4 g, with a yield of 90%, and an HPLC RT of 11.6 min.

[0123] 1-5: Synthesis of Fmoc-NH-NH-CarbiDOPA-OH

[0124] In a 1000mL three-necked flask, add 19.06g (50mmol) Na2B407·10H2O, 2.0g sodium sulfite, 150mL water, and pass Ar for 30min; then add 21.32g (100mmol) carbidopa, 7.96g (75mmol) Na2CO3, 75mL water and 150mL THF. Add a total of 26.96g (80mmol) Fmoc-OSu in batches and stir overnight. Using a similar treatment method as 1-3, 32.27g of white solid powder was obtained, with a yield of 90%, and HPLC RT was 14.99min. HRMS (ESI): C25H24N206Calcd.for [M+H] + 449.1707, found 449.1706.

[0125] 1-6: Synthesis of H-Met-DOPA-OH

[0126] In a 500 mL flask, add 10 g Boc-Met-DOPA-OH, 250 mL 2M HCl / DIOX solution, and 25 mL TIS, and react under magnetic stirring for 4 h; remove HCl and solvent by rotary evaporation, add a small amount of methanol to dissolve, and then use cold PE to precipitate. After centrifugation and vacuum drying, 9.3 g of white foamy solid is obtained.

[0127] 1-7: Synthesis of H-Met-CarbiDOPA-OH

[0128] Take 0.1g Boc-Met-CarbiDOPA-OH, add 20mL TFA / TIS / H2O / DCM (25:5:5:65) deprotection solution, and react under magnetic stirring for 2-4 hours; rotary evaporate to dryness, add a small amount of methanol to dissolve, and precipitate with glacial ether; after vacuum drying the obtained solid, add 10mL water to dissolve, and then freeze-dry; obtain 0.066g white solid H-Met-CarbiDOPA-OH, with a yield of 64.0% and an HPLC RT of 12.1.min.

[0129] 1-8: Coupling reaction of hyaluronic acid and levodopa methyl ester

[0130] Take 1.9 grams of hyaluronic acid (1.5 MDa, 5 mmol structural unit), dissolve in 200 mL of water, add 1-ethyl-(3-dimethylaminopropyl) carbodiimide (EDC) hydrochloride (0.96 g, 5 mmol) and HOSu (1.15 g, 10 mmol), seal with a rubber stopper, and stir at room temperature for 24 hours. Add Na2SO3 (0.5 g, 4 mmol) and H-DOPA-OMe hydrochloride (1.23 g, 5 mmol), adjust the pH to 6.8 with 1 M NaOH solution, seal and stir at room temperature for 24-48 hours. Use a semipermeable membrane (molecular weight cutoff 2000 Da) for 2-3 days, freeze-dry, and obtain a white solid. Due to the protective effect of the antioxidant Na2SO3, the solution remains colorless during the entire reaction process.

[0131] 1-9: Coupling reaction of hyaluronic acid and carbidopa methyl ester

[0132] A method similar to that of Example 1-8 was used, except that carbidopa methyl ester hydrochloride was used instead of H-DOPA-OMe hydrochloride; due to the protective effect of the antioxidant Na2SO3, the solution exhibited a light yellow color during the entire reaction process.

[0133] 1-10: Synthesis of other Boc-AA-DOPA-OH and Boc-AA-CarbiDOPA-OH

[0134] Boc-AA-DOPA-OH and Boc-AA-CarbiDOPA-OH were prepared using Boc-AA-OSu as a reactant in a manner similar to that of Examples 1-3 and 1-4. The corresponding characterization data are summarized as follows.

[0135] Boc-Met-DOPA-OH: Yield 64%, HRMS Calcd.for [MH] - 427.1544, found 427.1544. 11H NMR (400 MHz, MEOD) δ 6.66 (dd, J = 7.1, 4.9 Hz, 3H), 6.53 (dd, J = 8.0, 1.8 Hz, 1H), 4.59 (t, J = 6.3 Hz, 1H), 4.22 - 4.03 (m, 1H), 3.03 (dd, J = 13.9, 5.1 Hz, 1H), 2.90 (dd, J = 13.9, 7.5 Hz, 2H), 2.10 - 2.03 (m, 3H), 2.02 - 1.90 (m, 1H), 1.80 (td, J = 14.0, 8.4 Hz, 1H), 1.43 (s, 9H). 13C NMR (101 MHz, MEOD) δ 174.48, 174.31, 157.70, 146.11, 145.22, 129.29, 121.78, 117.36, 116.28, 80.81, 55.04, 54.97, 37.68, 32.85, 31.02, 28.67, 15.17.

[0136] Boc-Val-DOPA-OH: Yield 82%, HRMS calcd. for [M - H]- 395.1824, found 395.1825. 1 1H NMR (400 MHz, MEOD) δ 6.66 (d, J = 8.1 Hz, 2H), 6.54 (dd, J = 8.1, 1.8 Hz, 1H), 4.60 (dd, J = 7.7, 5.6 Hz, 1H), 3.86 (d, J = 6.8 Hz, 1H), 3.02 (dd, J = 13.9, 5.3 Hz, 1H), 2.86 (dd, J = 13.9, 8.0 Hz, 1H), 2.04 1.93 (m, 1H), 1.43 (s, 9H), 0.88 (dd, J = 9.2, 6.8 Hz, 6H). 13C NMR (101 MHz, MEOD) δ 174.63, 174.15, 157.89, 146.14, 145.20, 129.52, 121.74, 117.32, 116.27, 80.64, 61.51, 55.12, 37.95, 32.04, 28.70, 19.71, 18.40.

[0137] Boc-Ala-DOPA-OH: Yield 57%, HRMS Calcd for [M-H]-367.1511, found 367.1509. 1H NMR (400 MHz, MEOD) δ 6.71 - 6.50 (m, 3H), 4.62 - 4.50 (m, 1H), 4.14 - 3.93 (m, 1H), 3.02 (dd, J = 13.7, 5.0 Hz, 1H), 2.90 (dd, J = 13.7, 6.9 Hz, 1H), 1.56 - 1.09 (m, 12H). 13C NMR (101 MHz, MEOD) δ 175.45, 174.46, 157.55, 146.08, 145.21, 129.28, 121.82, 117.44, 116.26, 80.71, 54.94, 51.55, 37.73, 28.67, 18.30。

[0138] Boc-β-Ala-DOPA-OH: Yield 42%, HRMS Calcd for [M+H] + 369.1656, found 369.1657. 1 1H NMR (400 MHz, MEOD) δ 6.72 - 6.51 (m, 3H), 4.64 - 4.54 (m, 1H), 3.31 - 3.18 (m, 1H), 3.04 (dd, J = 14.0, 5.1 Hz, 1H), 2.81 (dd, J = 14.0, 8.8 Hz, 1H), 2.43 - 2.30 (m, 2H), 1.42 (s, 9H). 13C NMR (101 MHz, MEOD) δ 174.95, 173.70, 158.26, 146.15, 145.16, 129.75, 121.61, 117.22, 116.24, 80.19, 55.28, 37.81, 36.99, 28.71。

[0139] Boc-Phe-DOPA-OH: Yield 63%, HRMS Calcd. for [M-H] - 443.1824, found 443.1829. 1H NMR (400 MHz, MEOD) δ 7.41 - 7.03 (m, 5H), 6.68 (d, J = 8.1 Hz, 2H), 6.54 (d, J = 7.9 Hz, 1H), 4.60 (t, J = 6.3 Hz, 1H), 4.30 (dd, J = 9.6, 4.7 Hz, 1H), 3.13 - 2.97 (m, 2H), 2.89 (dd, J = 13.9, 7.4 Hz, 1H), 2.73 (dd, J = 13.8, 9.9 Hz, 1H), 1.58 - 1.02 (m, 9H). 13C NMR (101 MHz, MEOD) δ 174.40, 174.08, 157.54, 146.14, 145.26, 138.63, 130.29, 129.33, 127.59, 121.83, 117.43, 116.30, 80.71, 57.35, 55.07, 39.13, 37.81, 28.61.

[0140] Boc-Leu-DOPA-OH: Yield 78%, HRMS Calcd for [M+Na] + 433.1945, found 433.1942. 1 1H NMR (400 MHz, MEOD) δ 6.71 - 6.47 (m, 3H), 4.63 - 4.50 (m, 1H), 4.14 - 3.92 (m, 1H), 3.03 (dd, J = 13.9, 5.3 Hz, 1H), 2.89 (dd, J = 13.9, 7.4 Hz, 1H), 1.70 - 1.32 (m, 12H), 1.03 - 0.85 (m, 6H). 13C NMR (101 MHz, MEOD) δ 175.32, 174.43, 157.75, 146.07, 145.21, 129.27, 121.81, 117.44, 116.24, 80.67, 54.83, 54.57, 42.07, 37.76, 28.68, 25.84, 23.40, 21.84。

[0141] Boc-Ile-DOPA-OH: Yield 88%, HRMS Calcd for [M+Na] + 433.1945, found 433.1941. 11H NMR (400 MHz, MEOD) δ 6.69 - 6.52 (m, 3H), 4.71 - 4.51 (m, 1H), 3.90 (d, J = 7.2 Hz, 1H), 3.02 (dd, J = 13.9, 5.2 Hz, 1H), 2.86 (dd, J = 13.9, 8.1 Hz, 1H), 1.73 (d, J = 6.5 Hz, 1H), 1.43 (s, 10H), 1.19 - 1.00 (m, 1H), 0.97 - 0.74 (m, 6H). 13C NMR (101 MHz, MEOD) δ 174.51, 174.23, 157.82, 146.13, 145.20, 129.46, 121.73, 117.29, 116.25, 80.64, 60.66, 54.98, 38.29, 37.92, 28.70, 25.63, 15.85, 11.45。

[0142] Boc-Gly-DOPA-OH: Yield 66%, HRMS Calcd for [M - H] - 353.1354, found 353.1354. 1 1H NMR (400 MHz, MEOD) δ 6.75 - 6.46 (m, 3H), 4.66 - 4.53 (m, 1H), 3.81 - 3.60 (m, 2H), 3.02 (dd, J = 13.9, 5.1 Hz, 1H), 2.89 (dd, J = 13.9, 7.3 Hz, 1H), 1.43 (s, 9H). 13C NMR (101 MHz, MEOD) δ 174.58, 172.11, 158.33, 146.18, 145.27, 129.29, 121.73, 117.35, 116.31, 80.81, 55.02, 44.44, 37.77, 28.66 (3C).

[0143] Boc-Glu(OtBu)-DOPA-OH: Yield 72%, HRMS Calcd for [M + Na] + 505.2160, found 505.2150. 1H NMMR (400 MHz, MEOD) δ 6.71 - 6.51 (m, 3H), 4.58 (t, J = 6.4 Hz, 1H), 4.13 - 3.91 (m, 1H), 3.03 (dd, J = 13.9, 5.1 Hz, 1H), 2.89 (dd, J = 13.8, 7.4 Hz, 1H), 2.28 (t, J = 7.4 Hz, 2H), 2.01 - 1.73 (m, 2H), 1.44 (s, 18H). 13C NMR (101 MHz, MEOD) δ 174.38, 174.16, 174.06, 157.66, 146.13, 145.25, 129.28, 121.78, 117.34, 116.29, 81.78, 80.78, 54.98, 37.72, 32.62, 28.67, 28.55, 28.33。

[0144] Boc-Asp(OtBu)-DOPA-OH: Yield 76% HRMS Calcd for [M+Na] + 491.2001, found 491.2000. 1H NMR (400 MHz, MEOD) δ 6.74 - 6.48 (m, 3H), 4.62 - 4.52 (m, 1H), 4.50 - 4.32 (m, 1H), 3.01 (dd, J = 13.9, 5.3 Hz, 1H), 2.90 (dd, J = 13.9, 6.9 Hz, 1H), 2.71 (dd, J = 16.0, 5.2 Hz, 1H), 2.48 (dd, J = 16.0, 8.7 Hz, 1H), 1.44 (s, 18H). 13 C NMR (101 MHz, MEOD) δ 174.22, 173.20, 171.43, 157.54, 146.16, 145.30, 129.10, 121.82, 117.39, 116.30, 82.38, 80.97, 55.04, 52.61, 38.71, 37.66, 28.66, 28.26。

[0145] Boc-Ala-CarbiDOPA-OH: Yield 52%, HRMS Calcd for [M-H] -396.1776, found 396.1778. 1H NMR (400 MHz, MEOD) δ 6.68 (dd, J = 9.1, 5.0 Hz, 2H), 6.55 (dd, J = 8.1, 2.0 Hz, 1H), 4.02 (q, J = 7.2 Hz, 1H), 2.87 (dd, J = 35.9, 13.6 Hz, 2H), 1.43 (s, 9H), 1.29 (t, J = 3.6 Hz, 6H). 13C NMR (101 MHz, MEOD) δ 177.81, 175.12, 157.51, 146.05, 145.45, 128.23, 122.61, 118.36, 116.18, 80.73, 66.66, 51.06, 44.11, 28.66, 21.27, 18.00.

[0146] Boc-Val-CarbiDOPA-OH: Yield 78%, HRMS Calcd for [M+H]+ 426.2235, found 426.2236. 1 1H NMR (400 MHz, MeOD) δ 6.72 - 6.52 (m, 3H), 3.83 (d, J = 6.7 Hz, 1H), 2.87 (d, J = 3.8 Hz, 2H), 2.10 - 2.00 (m, 1H), 1.43 (s, 9H), 1.28 (s, 3H), 1.03 - 0.83 (m, 6H). 13C NMR (101 MHz, MeOD) δ 177.61, 173.53, 157.95, 146.03, 145.44, 128.26, 122.68, 118.41, 116.16, 80.72, 66.69, 60.84, 44.00, 31.45, 28.68, 21.22, 19.74, 18.44.

[0147] Boc-Met-CarbiDOPA-OH: Yield 90%, HRMS Calcd for [M-H]-456.1810, found 456.1806. 1H NMR (400 MHz, MeOD) δ 6.71-6.51 (m, 3H), 4.19-4.11 (m, 1H), 3.04-2.74 (m, 2H), 2.61-2.47 (m, 2H), 2.07 (s, 3H), 2.04-1.97 (m, 1H), 1.91-1.79 (m, 1H), 1.43 (s, 9H), 1.28 (s, 3H). 13C NMR (101 MHz, MeOD) δ 177.83, 173.95, 157.71, 146.04, 145.44, 128.26, 122.64, 118.38, 116.18, 80.86, 66.56, 54.36, 32.35, 31.19, 28.66, 21.38, 15.17.

[0148] Boc-Leu-CarbiDOPA-OH: Yield 92%, HRMS Calcd for [M-H]-438.2246, found 438.2243. 1H NMR (400 MHz, MeOD) δ 6.72-6.49 (m, 3H), 4.03 (t, J = 7.6 Hz, 1H), 2.97-2.75 (m, 2H), 1.77-1.61 (m, 1H), 1.51 (t, J = 7.1 Hz, 2H), 1.43 (s, 9H), 1.28 (s, 3H), 0.97-0.88 (m, 6H). 13C NMR (101 MHz, MeOD) δ 177.91, 174.82, 157.70, 146.02, 145.42, 128.27, 122.62, 118.36, 116.17, 80.72, 66.57, 53.86, 44.09, 41.70, 28.67(3C), 25.89, 23.34, 21.75, 21.35.

[0149] Boc-Ile-CarbiDOPA-OH: Yield 81%, HRMS Calcd for [M+Na]+462.2211, found 462.2212. 1 H NMR (400 MHz, MeOD) δ 6.71-6.53 (m, 3H), 3.88 (d, J = 7.0 Hz, 1H), 2.87 (d, J = 5.5 Hz, 2H), 1.89-1.73 (m, 1H), 1.49 (s, 10H), 1.18 (s, 4H), 1.01-0.81 (m, 6H). 1313C NMR (101 MHz, MeOD) δ 177.78, 173.50, 157.87, 145.98, 145.38, 128.31, 122.67, 118.39, 116.14, 80.69, 66.56, 59.86, 44.05, 37.76, 28.67, 25.82, 21.30, 16.01, 11.52.

[0150] Boc-Gly-CarbiDOPA-OH: Yield 67%, HRMS Calcd for [M-H] - 382.1620, found: 382.1619. 1 1H NMR (400 MHz, MeOD) δ 6.72 - 6.52 (m, 3H), 3.78 - 3.61 (m, 2H), 3.02 - 2.70 (m, 2H), 1.43 (s, 9H), 1.29 (s, 3H). 13C NMR (101 MHz, MeOD) δ 177.99, 171.68, 158.29,

[0151] 146.01, 145.41, 128.34, 122.63, 118.37, 116.16, 80.86, 66.50, 44.08, 43.84, 28.64, 21.34.

[0152] Boc-Phe-CarbiDOPA-OH: Yield 59%, HRMS Calcd. for [M + H] + 474.2235, found 474.2235. 1 1H NMR (400 MHz, MEOD) δ 7.28 - 7.18 (m, 5H), 6.69 - 6.63 (m, 2H), 6.53 (dd, J = 8.1, 2.0 Hz, 1H), 4.27 (dd, J = 9.1, 6.1 Hz, 1H), 3.07 (dd, J = 13.8, 6.1 Hz, 1H), 2.91 - 2.72 (m, 3H), 1.36 (s, 9H), 1.13 (s, 3H). 13 13C NMR (101 MHz, MEOD) δ 177.69, 173.42, 157.53,

[0153] 146.01, 145.41, 138.45, 130.25, 129.49, 128.30, 127.79, 122.63, 118.38, 116.15, 80.78, 66.48, 56.68, 44.07, 38.80, 28.61, 21.15.

[0154] Boc-β-Ala-CarbiDOPA-OH: Yield 42%, HRMS Calcd. for [M+H] + 398.1922, found 398.1923. 1H NMR (400 MHz, MEOD) δ 6.68 (dd, J = 10.5, 5.0 Hz, 2H), 6.55 (dd, J = 8.1, 2.1 Hz, 1H), 3.31 - 3.27 (m, 2H), 2.95 - 2.78 (m, 2H), 2.42 - 2.27 (m, 2H), 1.41 (s, 9H), 1.27 (s, 3H). 13C NMR (101 MHz, MEOD) δ 177.64, 172.87, 158.26, 145.96, 145.33, 128.48, 122.71,

[0155] 118.44, 116.10, 80.20, 66.44, 44.04, 37.98, 35.39, 28.71, 20.97.

[0156] Boc-Glu(OtBu)-CarbiDOPA-OH: Yield 84%, HRMS Calcd for [N+Na]+ 534.2422, found 534.2421. 1H NMR (400 MHz, MeOD) δ 6.73 - 6.52 (m, 3H), 4.09 - 3.95 (m, 1H), 2.98 - 2.75 (m, 2H), 2.32 (t, J = 7.4 Hz, 2H), 2.07 - 1.94 (m, 1H), 1.87 - 1.73 (m, 1H), 1.45 (t, J = 8.7 Hz, 18H), 1.28 (s, 3H). 13C NMR (101 MHz, MeOD) δ 177.81, 173.86, 173.72, 157.64, 146.01, 145.41, 128.27, 122.64, 118.37, 116.17, 81.85, 80.82, 66.50, 54.62, 44.07, 32.69, 28.66, 28.32, 21.32。

[0157] Boc-Asp(OtBu)-CarbiDOPA-OH: Yield 79%, HRMS Calcd for [M+Na] +520.2266, found: 520.2266.1HNMR (400MHz, MeOD) δ6.73-6.51 (m, 3H), 4.45-4.33 (m, 1H), 2.91 (d, J =13.6Hz, 1H), 2.82 (d, J = 13.6Hz, 1H), 2.74 (dd, J = 16.3, 5.8Hz, 1H), 2.53 (dd, J = 16.2, 8.1Hz, 1H), 1 .43 (s, 18H), 1.28 (s, 3H).13CNMR (101MHz, MeOD) δ177.89, 172.74, 171.26, 157.49, 146.02, 145.41 , 128.25, 122.61, 118.35, 116.16, 82.41, 80.98, 66.50, 51.88, 44.00, 38.32, 28.65, 28.27, 21.38.

[0158] 1-11 Synthesis of other H-AA-DOPA-OH and H-AA-CarbiDOPA-OH

[0159] Other H-AA-DOPA-OH and H-AA-CarbiDOPA-OH dipeptides were prepared using a Boc deprotection method similar to that of Examples 1-6 and 1-7. The corresponding characterization data are summarized below.

[0160] H-Met-DOPA-OH: yield 93%, HRMS: m / z Calcd for C 14 H 20 N2O5S, 327.1020, found: 327.1017[MH] - . 1 H NMR (400MHz, MeOD) δ6.70 (m, 2H), 6.59 (m, 1H), 4.61 (m, 1H), 3.96 (m, 1H), 3.11 (dd, J=14.1, 4.7Hz, 1H), 2.86 (dd, J=14.1, 9.4Hz, 1H), 2.61 (m, 2H), 2.2 2-1.98 (m, 5H).13CNMR (101MHz, MeOD) δ174.24, 169.70, 146.21, 145.25, 1 29.58, 121.56, 117.19, 116.34, 55.68, 53.51, 37.33, 32.23, 29.44, 15.03.

[0161] H-Met-DOPA-OH: yield 93%, HRMS Calcd for [MH] -327.1020, found 327.1017. 1 HNMR(400 MHz, MeOD) δ 6.70 (m, 2H), 6.59 (m, 1H), 4.61 (m, 1H), 3.96 (m, 1H), 3.11 (dd, J = 14.1, 4.7 Hz, 1H), 2.86 (dd, J = 14.1, 9.4 Hz, 1H), 2.61 (m, 2H), 2.22 - 1.98 (m, 5H). 13C NMR(101 MHz, MeOD) δ 174.24, 169.70, 146.21, 145.25, 129.58, 121.56, 117.19, 116.34, 55.68, 53.51, 37.33, 32.23, 29.44, 15.03.

[0162] H-Phe-DOPA-OH: Yield 87%, HRMS Calcd for [M + H] + 345.1145, found 345.1142. 1 HNMR(400 MHz, MeOD) δ 7.42 - 7.23 (m, 5H), 6.80 - 6.64 (m, 2H), 6.64 - 6.48 (m, 1H), 4.63 (m, 1H), 4.19 - 4.08 (m, 1H), 3.34 - 3.26 (m, 1H), 3.16 - 2.98 (m, 2H), 2.87 (m, 1H). 13 13C NMR(101 MHz, MeOD) δ 174.20, 169.61, 146.20, 145.27, 135.42, 130.61, 130.08, 129.57, 128.82, 121.63, 117.28, 116.33, 55.73, 55.44, 38.46, 37.69.

[0163] H-Ala-DOPA-OH: Yield 85%, HRMS Calcd for [M + H] + 269.1132, found 269.1131. 1HNMR(400MHz,MeOD)δ6.72-6.64(m,2H),6.57(dd,J8.1,2.0Hz,1H),4.56(dd,J9.1,4.8Hz,1H),3.90(q,J N7.0Hz,1H),3.10(dd,J)14.1,4.8Hz,1H),2.84(dd,J D14.1,9.2Hz,1H),1.48(d,J7.0Hz,3H).13CNMR(101 MHz,MeOD)δ175.15,170.90,146.15,145.16,129.93,121.64,117.28,116.29,56.16,50.09,37.69,17.58.

[0164] H-Val-DOPA-OH:Volume 98%,HRMS Calcd for[M+H] + 297.1445,found 1 HNMR(400MHz,MeOD)δ6.78-6.55(m,3H),4.68-4.57(m,1H),3.83 -3.73(m,1H),3.09(dd,J)14.1,5.0Hz,1H),2.87(dd,J14.1,9 Hz,1H),2.42-2.10(m,1H),1.12-0.99(m,6H).13CNMR(101MHz,MeOD)δ173.09,168.16,14 4.84,143.89,128.33,120.24,115.91,114.97,58.08,54.44,36.23,30.21,17.52,1

[0165] H-Leu-DOPA-OH:dispersion 82%,HRMS Calcd for[M+H] + 311.1601,found 311.1603. 1HNMR (400 MHz, MeOD) δ 6.74 - 6.54 (m, 3H), 4.64 - 4.54 (m, 1H), 3.93 - 3.82 (m, 1H), 3.10 (dd, J = 14.1, 5.0 Hz, 1H), 2.88 (dd, J = 14.1, 9.3 Hz, 1H), 1.78 - 1.61 (m, 3H), 1.01 - 0.95 (m, 6H). 13C NMR (101 MHz, MeOD) δ 174.38, 170.69, 146.20, 145.24, 129.73, 121.58, 117.26, 116.33, 55.77, 52.85, 41.68, 37.46, 25.19, 23.18, 21.98。

[0166] H-Ile-DOPA-OH: Yield 84%, HRMS Calcd for [M+H] + 311.1601, found 311.1600. 1 HNMR (400 MHz, MeOD) δ 6.81 - 6.35 (m, 3H), 4.70 - 4.50 (m, 1H), 3.79 - 3.71 (m, 1H), 3.09 (dd, J = 14.1, 5.0 Hz, 1H), 2.87 (dd, J = 14.1, 9.1 Hz, 1H), 2.02 - 1.92 (m, 1H), 1.63 - 1.50 (m, 1H), 1.22 - 1.15 (m, 1H), 1.09 - 0.91 (m, 6H). 13C NMR (101 MHz, MeOD) δ 174.64, 169.47, 146.04, 145.09, 129.78, 121.67, 117.27, 116.32, 58.90, 55.81, 38.08, 37.52, 24.97, 15.10, 11.71。

[0167] H-Gly-DOPA-OH: Yield 86%, HRMS Calcd for [M-H] - 253.0830, found 253.0831. 1HNMR (400 MHz, D2O) δ 6.85 - 6.62 (m, 3H), 4.67 - 4.60 (m, 1H), 3.77 (s, 1H), 3.73 (s, 1H), 3.08 (dd, J = 14.1, 5.5 Hz, 1H), 2.87 (dd, J = 14.1, 8.7 Hz, 1H). 13C NMR (101 MHz, MeOD) δ 175.01, 167.15, 146.15, 145.20, 129.68, 121.68, 117.31, 116.32, 55.82, 41.40, 37.93。

[0168] H-Asp-DOPA-OH: Yield 72%, HRMs Calcd for [M+H] + 313.1030, found 313.1027. 1 HNMR(400MHz, MeOD) δ6.81 - 6.47(m, 3H), 4.61(dd, J = 9.3, 4.7Hz, 1H), 4.19(dd, J = 9.2, 3.9Hz, 1H), 3.77 - 3.65(m, 1H), 3.14 - 2.97(m, 2H), 2.86 - 2.79(m, 1H).13C NMR(101MHz, MeOD) δ174.22, 172.87, 169.23, 146.20, 145.26, 129.53, 121.60, 117.20, 116.33, 55.74, 50.80, 37.44.35.98。

[0169] H-Met-CarbiDOPA-OH: Yield 64%, HRMS Calcd for [M+H]+ 358.1431, found358.1431. 1 H NMR(400MHz, MeOD) δ6.78 - 6.54(m, 6H), 4.48 - 4.40(m, 1H), 3.93(m, 1H), 2.97 - 2.88(m, 2H), 2.78 - 2.66(m, 2H), 2.64 - 2.48(m, 4H), 2.20 - 1.96(m, 10H), 1.23(s, 6H) 13 C NMR(101MHz, MeOD) δ181.62, 181.02, 173.90, 169.77, 145.90, 145.83, 145.14, 145.10, 129.48, 129.42, 122.97, 122.88, 118.64, 118.54, 116.17, 116.05, 67.39, 67.30, 52.80, 51.41, 44.96, 44.44, 32.25, 31.16, 30.20, 30.16, 22.33, 21.58, 15.28, 15.15。

[0170] H-Ala-CarbiDOPA-OH: Yield 66%, HRMS Calcd for [M+Na] + 320.1217, found320.1219. 11H NMR (400 MHz, MeOD) δ 6.74 - 6.67 (m, 4H), 6.60 - 6.50 (m, 2H), 4.23 (q, J = 6.9 Hz, 1H), 3.97 (q, J = 6.9 Hz, 1H), 2.96 - 2.67 (m, 4H), 1.56 - 1.41 (m, 6H), 1.31 - 1.16 (m, 6H). 13C NMR (101 MHz, MeOD) δ 179.47, 178.47, 175.25, 170.76, 145.89, 145.82, 145.27, 128.56, 128.47, 123.06, 122.75, 118.78, 118.45, 116.18, 116.00, 66.87, 66.27, 64.25, 54.79, 44.40, 44.18, 21.05, 20.17, 17.93, 16.68.

[0171] H-Val-CarbiDOPA-OH: Yield 67%, HRMS Calcd for [M+H] + 326.1710, found 326.1708. 1 1H NMR (400 MHz, MeOD) δ 6.75 - 6.65 (m, 4H), 6.60 - 6.55 (m, 2H), 4.17 (d, J = 4.5 Hz, 1H), 3.68 (d, J = 5.8 Hz, 2H), 2.89 (d, J = 13.3 Hz, 1H), 2.76 (d, J = 10.4 Hz, 1H), 2.46 - 2.28 (m, 1H), 2.25 - 2.05 (m, 2H), 1.26 (s, 6H), 1.13 - 0.90 (m, 12H). 13 13C NMR (101 MHz, MeOD) δ 180.99, 179.77, 173.81, 168.94, 145.90, 145.83, 145.22, 145.15, 129.05, 129.00, 123.05, 122.79, 118.84, 118.48, 116.13, 115.95, 67.03, 66.87, 58.76, 56.66, 44.53, 44.42, 31.44, 30.28, 21.93, 20.59, 19.46, 18.87, 18.12, 16.70.

[0172] H-Phe-CarbiDOPA-OH: Yield 71%, HRMS Calcd for [M+H] + 374.1710, found 374.1712. 1H NMR (400MHz, MeOD) δ7.44-7.02(m, 10H), 6.79-6.48(m, 6H), 4.56-4.46(m, 1H), 4.07-3.96( m, 1H), 3.31-3.09 (m, 2H), 3.04-2.78 (m, 4H), 2.76-2.56 (m, 2H), 1.24 (s, 3H), 0.96 (s, 3H). 13 C NMR (101MHz, MeOD) δ181.69, 180.85, 173.60, 169.68, 145.94, 145.85, 145.16, 145.11, 136.32, 136.01, 130.59, 130.48, 130.08, 130.00, 129. 47, 129.42, 128.67, 128.56, 123.02, 122.80, 118.73, 118.47, 116.06, 67.39, 67.18, 55.07, 53.14, 44.85, 44.49, 38.93, 37.86, 22.01, 21.37.

[0173] 1-12: Testing the solubility of the synthesized L-DOPA dipeptide and CarbiDOPA dipeptide

[0174] (1) Draw a standard straight line between absorbance and concentration: Take 50.2 mg of L-DOPA and dissolve it in 50 mL of 11 mmol / L citric acid-sodium citrate (pH 6.4) buffer solution. After multiple dilutions, measure the absorbance of the sample at 280 nm and draw a standard straight line between L-DOPA concentration and absorbance. The R value of the standard straight line is greater than 0.99.

[0175] (2) The effectiveness of the method was evaluated by measuring the solubility of L-DOPA: 100 mg of L-DOPA was added to 5 mL of 11 mmol / L citric acid-sodium citrate buffer solution (pH 6.4), vortexed, allowed to stand, and the supernatant was collected. The solubility was measured to be between 2.4 and 2.6 mmol / 100 mL, which is close to the value in the relevant literature (5 g / L or 2.5 mmol / 100 mL).

[0176] (3) Preparation of L-DOPA dipeptide or CarbiDOPA dipeptide test sample: Weigh about 0.1 mmol of L-DOPA dipeptide or CarbiDOPA dipeptide (in the form of HCl or TFA salt) solid, add it to 200-1200 μl of 2 M sodium citrate solution, vortex and centrifuge, take 100 μl of the supernatant, dilute it to 3 mL with water, measure its pH, and record it as the initial pH.

[0177] Re-prepare 100ul of the above supernatant, add 11mmol / L citric acid-sodium citrate buffer solution (pH 6.4) to dilute to an appropriate multiple, and calculate the solubility of the sample in the citric acid-sodium citrate solution at the initial pH value by measuring its absorbance; the actual solubility should be greater than the calculated value. The specific solubility of L-DOPA dipeptide or CarbiDOPA dipeptide at different pH values ​​is shown in Table 2, and the unit of solubility is mmol / 100mL.

[0178] Table 1 Solubility of L-DOPA dipeptide or CarbiDOPA dipeptide at different pH values

[0179]

[0180] As shown in Table 1, the solubility of L-DOPA dipeptide (H-AA-DOPA-OH) measured between pH 6.1 and 6.5 is greater than that of L-DOPA, and L-DOPA dipeptides all have good water solubility; among them, H-Gly-DOPA-OH has the best solubility, which is more than 10 times that of L-DOPA at pH 6.3; this is consistent with the conclusions in the early US3803120 and US4035507. At pH 6.1, the measured L-DOPA dipeptides can provide an L-DOPA drug loading of more than 10mmol / 100mL; at pH 6.3, L-DOPA modified with amino acids with smaller side chains and less non-polarity has an advantage and can also provide an L-DOPA drug loading of 10mmol / 100mL.

[0181] The measured CarbiDOPA dipeptide has better solubility than CarbiDOPA; under the measured pH conditions, CarbiDOPA dipeptide can provide a soluble drug loading of about 0.7mmol / 100mL. Therefore, the solid powder preparations made of L-DOPA dipeptide and CabiDOPA dipeptide can form a clear solution after adding water or an aqueous solution containing excipients.

[0182] Example 2

[0183] 2-1: One-pot synthesis of Tfa-DOPA-OMe

[0184] In a 500mL two-necked flask, add L-DOPA (19.7g, 100mmol) and 150mL of anhydrous methanol. Under ice bath conditions, use a dropping funnel to slowly add SOCl2 (14.5mL, 200mmol); remove the ice bath and continue stirring for 3h. Replace the dropping funnel with a condenser, heat the reaction solution in an oil bath to reflux and notify bubbling for 1h. Stop heating and cool to room temperature, slowly add 30.2mL (300mmol) of methyl trifluoroacetate and 56.0mL (400mmol) of Et3N, and continue stirring for 6-12h. After rotary evaporation and concentration, add 100mL of saturated sodium chloride solution, adjust the pH to 3 with 1M HCl, extract with EA, wash the obtained organic phase with water, dry it with MgSO4, rotary evaporation, and vacuum drying to obtain 28.8g of white solid Tfa-DOPA-OMe with a yield of 94%. Then, using the experimental method of US8227628, Tfa-DOPA-OMe was successfully converted into Tfa-DOPA(Acetonide)-OMe.

[0185] 2-2: Synthesis of Tfa-DOPA(Acetonide)-OH

[0186] Weigh 6.9 g (20 mmol) of Tfa-DOPA (Acetonide) -OMe, add 100 mL of acetonitrile, 2 mL of water, 17.4 g (200 mmol) of LiBr and 5.1 mL (30 mmol) of DIEA, and stir the reaction at room temperature for about 24 h; add 20 mL of 1 M citric acid solution, remove acetonitrile by rotary evaporation, and then extract with 100 mL of EA; wash with water, dry on MgSO4, filter, rotary evaporate, and vacuum dry to obtain an off-white solid, which is recrystallized in DCM to obtain 4.8 g of white needle crystals with a yield of 73%. 1 13C NMR (101MHz, CDCl3) δ: 175.0, 157.0 (q, JC-F=38.0Hz), 148.0, 147.2, 127.1, 12 2.0, 118.5, 115.6 (q, JC-F=287.5Hz), 109.3, 108.6, 53.5, 36.8, 25.9 (2C).HRMS Calcd For[M+H] + 334.0897, found 334.0867.

[0187] 2-3: Synthesis of Fmoc-DOPA(Acetonide)-OH

[0188] In a 500mL three-necked flask, add 8.4g (20mmol) Fmoc-DOPA-OH, 45mL acetone, and 300mL benzene; connect the device A-1-1 (containing calcium chloride) to the middle port of the three-necked flask, and then connect a reflux condenser to it, pass argon, reflux for 20min, add 5mL (40.66mmol) DMP, 86mg (0.5mmol) TsOH, react for 2-3h, add 2mL (16.3mmol) DMP, and continue to react for 0.5h. Cool to room temperature, add 1.7mL (20.5mmol) pyridine, and evaporate to dryness; add 100mL EA to extract the product, wash with 0.5M citric acid and water in turn, dry with MgSO4, filter and evaporate to obtain a crude product.

[0189] HPLC analysis showed that Fmoc-DOPA(Acetonide)-OH accounted for 93% of the crude product, and Fmoc-DOPA(Acetonide)-OMe accounted for 7%; recrystallization in TLN / HXN gave 7.2 g of a white solid, the purity of Fmoc-DOPA(Acetonide)-OH in the product was >98%, and the yield was 78% (the mother liquor of the recrystallization was subjected to conversion reaction to give 1.8 g of Fmoc-DOPA(Acetonide)-OH, and the total yield was >90%).

[0190] 2-4: Synthesis of Boc-DOPA(Acetonide)-OH.

[0191] Method A: 3.47 g (10 mmol) of Tfa-DOPA (Acetonide) -OMe was dissolved in 60 mL of THF, and 720 mg (30 mmol) of LiOH was dissolved in 20 mL of water. The mixture was stirred at room temperature for about 2.5 h. The pH was adjusted to 7-8 with 1 M HCl, and 2.12 g (20 mmol) of Na2CO3 and 2.62 g (12 mmol) of di-tert-butyl carbonate (Boc2O) were added. The mixture was stirred at room temperature for 3 h. The pH was adjusted to 3-4 with 1 M HCl, and the mixture was extracted with EA. The organic phase was washed with 0.001 M hydrochloric acid and water, dried with MgSO4, and rotary evaporated to obtain a crude product. The product was recrystallized in EA / HXN to obtain 2.86 g of a white solid with a yield of 85%.

[0192] Method B: 4.6g (10mmol) Fmoc-DOPA (Acetonide) -OH was dissolved in 50mL THF, 0.96g (40mmol) LiOH was dissolved in 50mL water, mixed with 50mL PE, and stirred at room temperature for 3h. The aqueous phase was separated by a separatory funnel, washed with EA and PE once in turn, filtered to obtain a clear aqueous solution. Then, 4.37g (20mmol) Boc2O was dissolved in 50mL THF, added to the above-mentioned clear aqueous solution, and stirred for 5h. 1M citric acid solution was used to adjust the pH to 3-4, EA was added to extract 3 times, the organic phases were combined, washed with water, dried with MgSO4, filtered and rotary evaporated, and dried in a vacuum drying oven to obtain a white foamy solid with a yield of about 81%.

[0193] 1 H NMR (400MHz, CDCl3) δ: 10.00 (br, 1H), 6.73-6.49 (m, 3H), 5.00 (d, J=8.0Hz, 1H), 4.63- 4.20 (m, 1H), 3.11-3.06 (m, 1H), 3.02-2.75 (m, 1H), 1.65 (s, 6H), (1.42, 1.31) (s, 9H). 13 C NMR (101MHz, CDCl3)δ: (176.6, 176.0) (1C), (156.6, 155.5) (1C), 147.7 (1C), 146.7 (1C), (129.6, 128.8) (1C), (122.3, 122.0) (1C) ), 118.0(1C), 109.6(1C), 108.3(1C), (81.5, 80.4)(1C), (56.3, 54.5)(1C), (39.2, 37.6)(1C), (28.4, 27.5)(3C), 26.0(2C).HRMS Calcdfor[MH] - 336.1453, Found 336.1450.

[0194] Example 2-5: Synthesis of Phth-DOPA (Acetonide) -OH

[0195] Method A: N-ethoxycarbonylphthalamide (CE-Phth) was used to replace Boc2O as the acylating agent to obtain 2.78 g of white crystal Phth-DOPA(Acetonide)-OH with a yield of 75%.

[0196] Method B adopts the method of CN111153885A, taking 75g (0.229mol) of Phth-DOPA-OH, using 200mL of acetone as a cosolvent, 1.97g of TsOH as a catalyst, and 42.3mL of DMP to prepare through an acetonide cyclization reaction to obtain 53g of solid, with a yield of 70%.

[0197] 1 H NMR (400MHz, CDCl3) δ: 7.79 (dd, J=5.5, 3.0Hz, 2H), 7.68 (dd, J=5.5, 3.0Hz, 2H), 6.5 7-6.49 (m, 3H), 5.16-5.11 (m, 1H), 3.48 (d, J=8.3Hz, 2H), 1.59 (s, 3H), 1.55 (s, 3H); 13 C NMR (101MHz, CDCl3) δ: 174.8, 167.6 (2C), 147.6 (2C), 146.4, 134.3 (2C), 131.7 ,129.5,123.7(2C),121.5,117.9,109.2,108.3,53.5,34.2,25.83,25.78.HRMS Calcd for[M+H] + 368.1129, found 368.1114.

[0198] 2-6: Synthesis of Phth-DOPA(Acetonide)-OtBu

[0199] In a 500mL flask, add Phth-DOPA (Acetonide) -OH (43g, 0.117mol), Boc2O (51g, 2eq.), tBuOH (22.4mL, 2eq), 300mL dry DCM and DMAP (8.54g, 0.6eq); connect the flask mouth with oil seal, stir and react at room temperature for 5h. The organic phase is washed with ammonium chloride solution and water, dried with MgSO4, filtered and rotary evaporated to obtain 25g of white solid, with a yield of 60%. 1 H NMR (400MHz, CDCl3) δ7.73 (ddd, J=43.1, 5.4, 3.1Hz, 4H), 6.53 (dd, J=16.1, 8.6Hz, 3H) , 4.98 (dd, J=9.9, 6.5Hz, 1H), 3.49-3.35 (m, 2H), 1.57 (d, J=14.9Hz, 6H), 1.44 (s, 9H). 13C NMR (101MHz, CDCl3) δ167.98, 167.82, 147.59, 146.25, 134.11, 131.88, 130.31, 123.49, 121.39, 117.77, 82.84, 54.57, 34.53, 28.03, 25.86.HRMS Calcd for[M+H] + 424.1755, found 424.1733.

[0200] 2-7: Synthesis of H-DOPA(Acetonide)-OtBu

[0201] In a 500mL flask, add Phth-DOPA(Acetonide)-OtBu (0.423g, 1mmol), 60mL DCM and 60mL methanol, hydrazine hydrate (2mL, 5mmol), seal with a rubber stopper, and stir at room temperature for 3h. Remove the volatile solvent by rotary evaporation, extract with 60mL DCM, and refrigerate. Filter to remove the white precipitate, and after rotary evaporation, obtain 0.24g of colorless oil, with a yield of 83%. 1 H NMR (400MHz, CDCl3) δ6.59 (m, 3H), 3.66 (s, 1H), 2.93 (dd, 1H), 2.84 (dd, 1H), 1.60 (s, 6H), 1.38 (s, 9H). 13 C NMR (101MHz, CDCl3) δ172.35, 147.63, 146.48, 129.31, 121.98, 117.88, 109.51, 108.11, 82.12, 55.76, 39.47, 27.94, 25.83.HRMS Calcd for [M+H] + 294.1700, found 294.1686.

[0202] 2-8: Synthesis of Boc-DOPA(Acetonide)-Asp-Phe-OMe

[0203] In a 250mL flask, add Boc-DOPA (Acetonide) -OH (3.37g, 10mmol), HOSu (1.73g, 15mmol), 150mL dry DCM and DCC (2.26g, 11mmol), stir and react for 12h, filter and keep the filtrate. Take another flask, add aspartame (2.94g, 10mmol), 50mL DMF and 3.23mL DIEA (20mmol), slowly add the above filtrate dropwise, and continue to react for 12-16h. Add 0.1M citric acid solution to adjust pH to 3, extract with EA; wash the organic phase with water, dry with MgSO4, filter and rotary evaporate to obtain a crude product; use EA / HXN (0.5% acetic acid) as the mobile phase, separate by silica gel column chromatography, and obtain 3.1g of white powder, with a yield of 50%, HPLC RT 17.5min.

[0204]

[0205] 1 H NMR (400MHz, CDCl3) δ7.44 (d, J=8.0Hz, 1H), 7.22-7.01 (m, 6H), 6.52 (d, J=7.0Hz, 1H), 4.87-4.54 (m, 2H ), 3.57 (s, 3H), 2.99 (m, 3H), 2.91-2.69 (m, 3H), 2.61 (dd, J=17.1, 6.4Hz, 1H), 1.55 (s, 6H), 1.28 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 13 C NMR (101MHz, CDCl3) δ175.67, 174.39, 172.15, 171.69, 170.26, 155.93, 147.72, 146.52, 135.94, 129.35, 128.68, 12 7.22, 121.88, 117.98, 109.45, 108.22, 80.63, 55.84, 54.03, 52.43, 49.32, 37.99, 37.78, 35.83, 28.30, 25.92.HRMS Calcd for[MH] - 612.2562, found 612.2563.

[0206] Synthesis of 2-9 Tripeptide H-DOPA-Asp-Phe-OMe

[0207] Take a 50mL flask, add Boc-DOPA(Acetonide)-Asp-Phe-OMe (0.2g), deprotection solution (2MHCl DIOX solution 10mL, TIS 0.25mL and water 0.25mL), stir and react for 2h; the mixed solution is rotary evaporated to obtain a crude product, 1mL of methanol is added to dissolve, and precipitated with glacial ether. After centrifugation and drying, a white solid is obtained, with a yield of 54%, HPLC RT 11.6min.

[0208] Example 3: Preparation of Danshensu-type catechol derivatives

[0209]

[0210] 3-1: Oxa-Pictet-Spengler reaction product of D-tanshinone methyl ester (3B-2)

[0211] Salvianolic acid B (content 60%) was converted into salvianolic acid A and danshensu by high-pressure hydrolysis; then it was reacted with SOCl2 / MeOH to generate a mixture of D-danshensu methyl ester and salvianolic acid A methyl ester; this mixture was subjected to acetonide cyclization reaction in an acetone-benzene-TsOH-DMP system to separate a colorless crystal, which was structurally characterized as compound 3B-2.

[0212] 1 H NMR (400MHz, CDC13) δ6.45 (s, 1H), 6.43 (s 2H), 4.42 (dd, J=11.4, 3.2Hz, 1H), 3.79 (s, 3H), 2.94 (dd, J=15.6, 11.4Hz, 1H), 2.81 (dd, J=15.7, 3.2Hz, 1H), 1.64 (s, 6H), 1.55 (s, 3H), 1.49 (s, 3H). 13 C NMR (101MHz, CDC13) δ172.04, 146.47, 146.04, 134.71, 123.80, 117.93, 108.03, 105.10, 76.74, 68.84, 52.33, 32.34, 31.39, 28.43, 25.94.HRMSCalcd for[M+Na] + , 315.1203, found 315.1201.

[0213] 3-2: Synthesis of methyl rosmarinic acid (3B-3)

[0214] Method A: In a 500mL flask, add rosmarinic acid (5g, 13.8mmol) and 100mL anhydrous methanol, cool in an ice bath, and stir magnetically. Slowly add SOCl2 (1.5mL, 20.7mmol), remove the ice bath and return to temperature, and monitor the reaction progress by HPLC. The reaction is complete after 1-1.5h. Remove the solvent by rotary evaporation and dry in vacuo to obtain the crude product 3B-3 of rosmarinic acid methyl ester with a purity of 95%.

[0215] Method B: In a 250 mL flask, rosmarinic acid (5 g, 13.8 mmol), 25 mL anhydrous methanol, 100 mL DMP and 17.25 mL catalyst Me3SiCl were added and stirred for 3 h; the solvent was removed by rotary evaporation, and the residue was dried in a vacuum drying phase with a purity of 95%.

[0216] HPLC RT 11.4min; HRMS Calcd for[M+Na] + 397.0894, found 397.0898.

[0217] 3-3: Synthesis of bis-acetone protected rosmarinic acid methyl ester (3B-4)

[0218] In a 250mL three-necked flask, methyl rosmarinic acid (1.46g 3.6mmol) was dissolved in 10mL acetone, and 150mL benzene was added. Argon was passed through the oil seal, magnetic stirring was performed, and the mixture was heated to reflux for 10-30min; DMP (3.5mL, 8eq.) and TsOH (0.12g, 0.2eq.) were added, and the reaction equilibrium was moved using CaCl2 adsorbent; the mixture was refluxed for 2-5h, and the reaction progress was monitored by TLC. After rotary evaporation and silica gel column chromatography, 1.37g of yellow oil was obtained, with a yield of 83%, and HPLC RT of 22.1min. 1H NMR (400MHz, CDCl3) δ7.60 (d, J=15.9Hz, 1H), 7.00-6.93 (m, 2H), 6.71 (d, J=7.9Hz, 1H), 6.66-6.64 (m, 3H), 6.27 (d, J=15.9Hz, 1H), 5.27 (dd , J=7.7, 4.8Hz, 1H), 3.74 (s, 3H), 3.12 (dd, J=14.1, 4.8Hz, 1H), 3.07 (dd, J=14.1, 7.7Hz, 1H), 1.68 (s, 6H), 1.654 (s, 3H), 1.648 (s, 3H).13C NMR (101MHz, CDCl3) δ170.48, 166.60, 149.93, 148.28, 147.59, 146.57, 146.26, 129.04, 128.16, 124.54, 1 21.93, 119.09, 117.98, 114.39, 109.64, 108.49, 108.15, 106.54, 73.26, 52.43, 37.38, 26.00, 25.96.HRMS Calcd for[M+Na] + , 477.1520, found 477.1535.

[0219] 3-4: Synthesis of bis-acetone protected rosmarinic acid (3B-5)

[0220] Compound 3B-4 (0.26 g, 0.57 mmol), 12 mL THF, and 6 mL water were added to a 50 mL single-necked flask. The mixture was cooled in an ice bath, and LiOH (0.027 g, 1.14 mmol) solution was added dropwise. The reaction was continued by stirring for 2 h. The reaction progress was monitored by TLC (EA / PE / acetic acid = 1:2:0.01, R f =0.43). A saturated ammonium chloride solution was added, EA was extracted, the organic phase was washed with water, dried over MgSO4, filtered and rotary evaporated to obtain a crude product 3B-5, which was then purified by liquid column chromatography (EA / PE) with a yield of 98% and an HPLC RT of 19.7 min. 1 H NMR (400MHz, CDCl3) δ7.61 (d, J=15.9Hz, 1H), 6.96 (d, J=8.7Hz, 2H), 6.67 (dt, J=16.7, 9.3Hz, 4H), 6. 25 (t, J=14.9Hz, 2H), 5.29 (d, J=4.2Hz, 1H), 3.14 (ddd, J=22.8, 14.4, 6.0Hz, 2H), 1.73-1.56 (m, 12H). 13C NMR (101MHz, CDCl3) δ174.54, 166.77, 149.95, 148.26, 147.57, 146.57, 146.52, 129.02, 128.14, 124.63, 122.01, 119.10, 118.00, 114.22, 109.69, 108.48, 108.20, 106.61, 77.16, 73.12, 37.15, 25.95.HRMSCalcd for [MH]-439.1398, found 439.1416.

[0221] 3-5: Synthesis of methyl rosmarinic acid ester (3B-6) protected by monoacetone

[0222] In a 250mL three-necked flask, methyl rosmarinic acid (1.46g 3.6mmol) was dissolved in 10mL acetone, and 150mL benzene was added. Argon was passed through the oil seal, magnetic stirring was performed, and heating was refluxed for 10-30min; DMP (0.88mL, 2eq.) and TsOH (17mg, 2.5%) were added, and a small amount of CaCl2 was used to move the reaction equilibrium; refluxed for 1-1.5h, and the reaction progress was detected by HPLC, and it was found that the peak (RT17.5min) accounted for about 80%. 1 HNMR (400MHz, MEOD) δ7.56 (d, J=15.8Hz, 1H), 7.04 (d, J=2.1Hz, 1H), 6.94 (dd, J=8.1, 2.1Hz, 1H), 6.77 (d, J=8.1Hz, 1H), 6.70-6.63 (m, 3H), 6.25 ( d, J=15.8Hz, 1H), 5.21 (dd, J=7.6, 5.1Hz, 1H), 3.71 (s, 3H), 3.11 (dd, J=14.2, 5.1Hz, 1H), 3.06 (dd, J=14.2, 7.6Hz, 1H), 1.62 (s, 3H), 1.61 (s, 3H). 13 C NMR (101MHz, MEOD) δ171.98, 168.23, 149.87, 148.82, 148.01, 147.83, 146.80, 130.31, 127.46, 12 3.24, 123.00, 118.97, 116.50, 115.24, 114.01, 110.50, 108.89, 74.50, 52.71, 38.09, 25.88.HRMS Calcd for [MH]-413.1242, found 413.1257.

[0223] 3-6: Synthesis of D-danshensu methyl ester (3B-7) protected by acetonide

[0224] In a 50mL single-necked flask, add compound 3B-4 (0.454g, 1mmol), add anhydrous methanol 10mL, cool in an ice bath; add sodium methoxide (0.054g, 1mmol) anhydrous methanol solution 10mL, TLC detection reaction progress, about 1h reaction is complete. Add saturated ammonium chloride solution 30mL, EA extraction; organic phase is washed with water, dried with MgSO4, filtered and rotary evaporated to obtain a crude product, and then purified by liquid column chromatography (EA / PE elution = 1:5) to obtain 0.2g yellow solid, yield 80%, HPLC RT 14.7min. 1 H NMR (400MHz, MEOD) δ6.63 (t, J=1.2Hz 1H), 6.61 (d, J=1.2Hz, 2H), 4.30 (dd, J=7.6, 4.9Hz, 1H), 3.69 (s, 3H), 2.95 (dd, J=13.9, 4.9Hz, 1H), 2.81 (dd, J=13.9, 7.6Hz, 1H), 1.61 (s, 6H); 13 C NMR (101MHz, MEOD) δ175.70, 148.68, 147.51, 131.48, 122.93, 118.71, 110.57, 108.73, 73.17, 52.34, 41.30, 25.91.HRMS Calcd for [MH] - , 251.0925, found 251.0938.

[0225] 3-7: Synthesis of L-danshensu methyl ester (3B-8) protected by acetonide

[0226] In a 100 mL two-necked flask, add H-DOPA (Acetonide) -OMe (1.12 g, 4.4 mmol) and THF 5 mL, cool in a brine-ice bath (-5°C), add acetic acid solution (36%, 3.5 mL); then add sodium nitrite (0.455 g, 6.6 mmol) solution 10 mL. Remove the ice bath and return to the temperature, continue stirring for 1 h, extract with EA, wash the organic phase with water, dry it with anhydrous MgSO4, and the HPLC detection peak (RT 14.7 min) accounts for about 35%; separate by semi-preparative HPLC, and characterize the structure to determine that the peak corresponds to 3B-8. 1H NMR (400 MHz, MEOD) δ 6.63 (s, 1H), 6.61 (s, 2H), 4.30 (m, 1H), 3.69 (s, 3H), 2.95 (m, 1H), 2.81 (m, 1H), 1.61 (s, 6H); 13C NMR (101MHz, MEOD) δ175.70, 148.73, 147.57, 131.52, 122.94, 118.72, 110.58, 108.73, 73.21, 52.31, 41.34, 25.90.HRMS Calcd for [M+H] + , 253.1071, found 253.1075.

[0227] 3-8: Synthesis of Boc-DSS-(Acetonide)-OMe (3B-9)

[0228] In a 10 mL flask, add 3B-7 (0.10 g, 0.4 mmol), add DCC (0.13 g, 0.6 mmol), 10 mL dry DCM, stir to dissolve, add 4-dimethylaminopyridine (DMAP) (10 mg, 0.2 eq.), react at room temperature for 30 min. Add 10 mL of 0.1 M lemon solution, extract 3 times with EA, wash the organic phase with water, dry with MgSO4, filter and rotary evaporate to obtain a crude product, and then purify it with liquid column chromatography (EA / PE=1:5) to obtain a yellow oil with a yield of 98%, HPLC RT 20.4 min. 1 H NMR (400MHz, CDCl3) δ6.66-6.58 (m, 3H), 5.02 (dd, J=8.2, 4.7Hz, 1H), 3.76-3.69 (m, 3H), 3.02 (qd, J=14.4, 6.5Hz, 2H), 1.64 (s, 6H), 1.44 (s, 9H). 13 C NMR (101MHz, CDCl3) 6170.28, 152.87, 147.56, 146.56, 128.78, 121.96, 117.90 ,109.64,108.14,83.11,77.16,77.16,75.51,52.36,37.34,27.74,25.92.HRMS Calcd for[M+Na] + 375.1414, found 375.1421.

[0229] 3-9: Synthesis of Boc-DSS-(Acetonide)-OH (3B-10)

[0230] In a 10 mL flask, add 3B-9 (0.18 g, 0.5 mmol), 10 mL THF, and 5 mL water; slowly add 12 mL LiOH (0.027 g, 1.14 mmol) solution dropwise, and stir at room temperature for 2 h. Add saturated ammonium chloride solution to quench, extract with EA, wash the organic phase with water, dry with MgSO4, filter and rotary evaporate to obtain a crude product, which is then purified by liquid column chromatography (EA / PE) to obtain a yellow solid with a yield of 98% and HPLC RT of 17.6 min. 1 H NMR (400MHz, CDCl3) δ6.68-6.63 (m, 3H), 5.07 (dd, J=8.6, 4.2Hz, 1H), 3.15-2.99 (m, 2H), 1.65 (s, 6H), 1.44 (d, J=10.0Hz, 9H). 13 C NMR (101MHZ, CDCl3) δ175.65, 152.87, 147.59, 146.63, 128.61, 122.05, 117.98, 109.69, 108.22, 83.43, 77.16, 76.84, 37.14, 27.75, 25.95.HRMS Calcd for[MH] - 337.1293, found 337.1273.

[0231] 3-10: Synthesis of Fmoc-Val-D-DSS(Acetonide)-OMe(3B-11)

[0232] In a 50 mL flask, add 3B-7 (0.1 g, 0.4 mmol), Fmoc-Val-OH (0.136 g, 0.4 mmol), 10 mL DCM, DCC (0.1 g, 0.48 mmol) and DMAP (10 mg, 0.2 eq.); stir and react at room temperature for 3 h. Filter to remove the precipitate, wash the organic phase with water, dry it with MgSO4, filter and rotary evaporate to obtain a crude product, which is then purified by liquid column chromatography (EA / PE) to obtain a white solid with a yield of 89% and HPLC RT 23.35 min. 1HNMR (400MHz, CDCl3) δ7.77 (d, J=7.5Hz, 2H), 7.61 (d, J=7.1Hz, 2H), 7.36 (dt, J= 34.6, 7.4Hz, 4H), 6.62 (d, J=8.3Hz, 3H), 5.26 (d, J=7.8Hz, 1H), 4.57-4.27 (m, 3H ), 4.24 (t, J=7.0Hz, 1H), 3.75 (s, 3H), 3.07 (ddd, J=23.8, 14.2, 6.5Hz, 2H), 2.11 (dt, J=34.9, 17.4Hz, 1H), 1.63 (s, 6H), 0.99-0.82 (m, 3H), 0.72 (d, J=6.8Hz, 3H). 13 C NMR (101MHz, CDCl3) δ171.48, 169.59, 156.23, 147.68, 146.64, 144.04, 143.90, 141.41, 128.65, 127.81, 127.17, 125.22, 125.19, 121.83, 120.08, 118.03, 109.39, 108.20, 77.16, 73.93, 67.18, 58.94, 52.93, 47.29, 37.06, 31.33, 26.10, 18.91, 16.88.HRMSCalcd for[M+Na] + , 337.1293, found 337.1273.

[0233] 3-11: Synthesis of acetone-protected caffeic acid dimer (3B-12)

[0234] In a 10 mL flask, add 3B-1 (0.11 g, 0.5 mmol), 1 mL ethanol and 4 mL water to form a suspension, place it in front of a 300 nm-400 nm black light, stir overnight at room temperature, filter to obtain a crude product, and recrystallize in methanol to obtain white crystals with a purity of ≥99%, a yield of 94%, and HPLC RT 15.4 min. 1H NMR (400MHz, DMSO-d6) δ6.83 (s, 2H), 6.79-6.66 (m, 4H), 4.15 (dd, J=10.3, 7.4Hz, 2H), 3.67 (dd, J=10.3, 7.3Hz, 2H), 1.62 (s, 12H).13C NMR (101MHz, DMSO-d6) δ173.08, 146.77, 145.66, 132.77, 120.43, 117.78, 107.98, 107.71 ,46.69,40.81,40.15,39.94,39.73,39.52,39.52,39.31,39.10,38.89,25.62.HRMSCalcd for[MH] - 439.1398, found 439.1394.

[0235] 3-12: Synthesis of tert-butyl rosmarinate (3B-13) protected by bis-acetone

[0236] In a 50mL flask, add 3B-5 (0.09g, 0.2mmol), tert-butyl alcohol (0.86mL) and 9mL dry DCM, stir and mix, then add DCC (0.05g, 0.24mmol), DMAP (0.122g, 0.5mmol), stir and react for 3-4h. Use EA extraction, wash the organic phase with water, dry it with MgSO4, filter and rotary evaporate to obtain the crude product, separate it by semi-preparative HPLC, freeze-dry to obtain a yellow oil, with a yield of 60%, HPLC RT 24.1min. 1 H NMR (400MHz, CDCl3) δ7.75-7.69 (m, 1H), 7.09-7.05 (m, 2H), 6.84-6.75 (m, 4H), 6.42-6.36 (m, 1H), 5.25 (dd, J =7.1, 5.8Hz, 1H), 3.22-3.16 (m, 2H), 1.80-1.75 (m, 12H), 1.54 (s, 9H).13CNMR (101MHz, CDCl3) δ168.97, 166.6 0, 149.81, 148.25, 147.48, 146.43, 145.85, 129.30, 128.18, 124.38, 122.02, 119.02, 118.00, 114.74, 109.80 , 108.53, 107.93, 106.56, 82.32, 77.16, 73.65, 37.34, 28.12, 25.55.HRMSCalcd.for[M+Na]+519.1989, found 519.1983.

[0237] 3-13: Synthesis of bis-acetone protected rosmarinic acid and isoborneol complex (3B-14)

[0238] In a 50mL single-necked flask, add 3B-5 (0.88g, 0.2mmol), isoborneol (62mg, 0.4mmol) and 8mL DCM, stir and mix. Then add DCC (50mg, 0.4mmol), DMAP (48.8mg, 0.4mmol), stir for 3-6h. Use EA extraction, the organic phase is dried over water and MgS04, filtered and rotary evaporated, separated by semi-preparative HPLC, and a white solid powder is obtained, with a yield of 70%, HPLC RT 30.3min. 1 H NMR (400MHz, MeOD) δ8.27-5.68 (m, 6H), 4.88 (s, 10H), 4.60 (s, 1H), 3.06 (dd, J=15.8, 10.1Hz, 2H), 1.56 (dd, J=69.8, 64.3Hz, 12H), 1.19 (ddd, J=81.4, 32.5, 25.5Hz, 3H), 0.98-0.57 (m, 6H). 13 C NMR (101MHz, MeOD) δ171.16, 168.07, 151.36, 149.69, 148.90, 147.98, 147.45, 130.10, 129.46, 125.78, 123.22, 120.26, 119.01, 115. 27, 110.66, 109.38, 108.94, 107.38, 83.52, 74.97, 49.00, 47.84, 46.37, 39.40, 38.23, 34.69, 27.85, 26.00, 25.95, 20.45, 11.71.HRMS Calcd for[M+Na]+ 599.2615, found 599.2626.

[0239] 3-14: Synthesis of the complex of rosmarinic acid and phenylalanine protected by bis-acetone (3B-15)

[0240] In a 10 mL three-necked flask, Ar was passed through for 10 min, 3B-5 (0.88, 0.2 mmol), DCC (50 mg, 0.24 mmol), HOSu (46 mg, 0.4 mmol), and 10 mL of dry DCM were added and stirred to dissolve. Then, H-Phe-OtBu hydrochloride (51.5 mg, 0.2 mmol) and DIEA (0.1 mL, 0.2 mmol) were added and stirred for 4 h. The yield was 98% (HPLC), and the HPLC RT was 23.9 min. 1H NMR (400MHz, Methanol-d4) δ7.57 (d, J=15.8Hz, 1H), 7.25-7.10 (m, 6H), 7.08 -6.99 (m, 2H), 6.75 (d, J=8.0Hz, 1H), 6.63 (s, 1H), 6.59 (s, 2H), 6.31 (d, J=15. 8Hz, 1H), 5.22(dd,J=7.4, 5.5Hz, 1H), 4.53(dd,J=8.2, 6.0Hz, 1H), 3.07(dd,J =13.8, 6.0Hz, 1H), 2.99-2.84(m, 3H), 1.66(s, 6H), 1.58(s, 6H), 1.41(s, 9H). 13 C NMR (101MHz, MEOD) δ171.97, 171.57, 167.51, 151.34, 149.67, 148.83, 147.80, 147.43, 137.97, 130.45, 130.42, 129.51, 129.47, 127.87, 125.80,123.10,120.23,118.89,115.29,110.63,109.35,108.87,107.41,83.16,75.65,55.45,38.83,38.35,28.21,25.94,25.92.HRMS Calcd for[M+H] + 644.2854, found 644.2851.

[0241] 3-15: Coupling Synthesis of Acetone-Protected Methyl Rosmarinic Acid 3B-4

[0242] In a 50mL flask, acetone protected caffeic acid (0.044g, 0.2mmol), acetone protected D-danshensu methyl ester (0.05g, 0.2mmol), 10mL of dry dichloromethane, dicyclohexylcarbodiimide (0.045g, 0.22mmol), DMAP (0.016g, 0.1mmol) were added, and the mixture was reacted for 12h under magnetic stirring. The reaction progress was detected by HPLC, and the peak (RT 22.1min) accounted for about 98%; its retention time was consistent with that of the mixed sample of 3B-4.

[0243] 3-16: Synthesis of acetonide-protected Shimobashiric acid C methyl ester (3B-16)

[0244] In a 50mL flask, 3B-7 (0.75g, 3mmol), 3B-11 (0.66g, 1.5mmol) and 100mL dry DCM, DMAP (0.75g, 4.6mmol), DCC (0.75g, 3.6mmol) were added; the reaction progress was monitored by HPLC and completed in about 4h. The precipitate was filtered off, and the organic phase was washed with water, dried, filtered and rotary evaporated to obtain a crude product, which was then purified by liquid column chromatography (DCM) to obtain 1.27g of a white solid, with a yield of 85%, and HPLC RT 25.1min. 1 H NMR (400MHz, CDCl3) δ6.64-6.41 (m, 12H), 4.91 (dd, J=7.8, 5.3Hz, 1H), 4.76 (t, J=6.1Hz, 1H), 4.31-4.22 (m, 2H), 3.90 -3.83 (m, 2H), 3.59 (d, J=14.4Hz, 6H), 2.88 (ddd, J=22.0, 14.1, 6.6Hz, 2H), 2.76-2.61 (m, 2H), 1.63 (t, J=8.4Hz, 24H). 13 C NMR (101MHz, CDCl3) δ171.52, 170.83, 169.86, 169.60, 147.69, 147.64, 147.62, 147.50, 146.6 6, 146.59, 146.57, 146.56, 131.60, 131.48, 128.67, 128.61, 128.38, 128.33, 121.96, 121.92, 1 20.23, 119.96, 118.11, 117.99, 117.96, 109.65, 109.54, 108.25, 108.20, 108.10, 107.99, 107 .74, 77.16, 73.48, 73.44, 52.23, 47.07, 46.98, 41.77, 41.37, 37.43, 37.06, 25.97, 25.93.HRMS Calcd for[M+Na] + 931.3148, found 931.319.

[0245] 3-17: Synthesis of acetonide-protected Shimobashiric acid C (3B-17)

[0246] In a 100mL flask, add compound 3B-16 (0.054g, 0.58mmol), 24mL THF, 12mL water, cool in an ice bath, slowly add 12mL LiOH (0.086g, 3.6mmol) solution, and continue stirring and reacting for 4h. Add 0.5M citric acid solution to adjust pH to 4, extract with EA, wash the organic phase with water, dry, filter and rotary evaporate to obtain a crude product; then purify with liquid column chromatography (EA / PE) to obtain 0.05g of white solid, with a yield of 95%, HPLC RT 21.3min. 1 H NMR (400MHz, CDCl3) δ6.62-6.45(m, 12H), 4.96-4.72(m, 2H), 4.33-4.18(m, 2H), 3.85(d d, J=17.1, 9.9Hz, 2H), 2.82 (dddd, J=41.4, 34.8, 14.3, 4.7Hz, 4H), 1.70-1.54 (m, 24H). 13 C NMR (101MHz, CDCl3) δ174.34, 173.91, 171.53, 170.81, 147.68, 147.63, 147.49, 146.65, 146.64, 146.60, 131.49, 131.20, 128.50, 128.27, 122.03, 120.19, 119. 96, 118.16, 118.06, 118.02, 109.73, 109.58, 108.29, 108.18, 107.66, 73.08, 73 .02, 47.02, 46.91, 41.72, 41.44, 37.21, 36.83, 26.01, 25.94, 25.86.HRMSCalcd for[MH] - , 879.2870, found 879.2872.

[0247] 3-18: Synthesis of Shimobashiric acid C methyl ester (3B-18)

[0248] In a 10 mL flask, add compound 3B-16 (0.06 g) and 6 mL of deprotection solution (TFA / DCM / H2O / Tis=55:44:0.5:0.5); stir and react for 2 h in an ice bath, concentrate by rotary evaporation, add water and freeze-dry to obtain 0.049 g of a white solid with a yield of nearly 100%, HPLC RT 11.4 min. 1H NMR (400MHz, MEOD) δ6.77-6.37 (m, 12H), 4.93 (s, 8H), 4.77 (d, J=6.2Hz, 1H), 4.56 (t, J=6.2Hz, 1H), 4.25-4.15 (m, 2H), 3.88 (dd, J =10.6, 7.2Hz, 1H), 3.77 (dd, J=10.5, 7.0Hz, 1H), 3.57 (t, J=14.4Hz, 6H), 2.86-2.75 (m, 2H), 2.62 (ddd, J=38.7, 14.0, 6.2Hz, 2H). 13 C NMR (101MHz, MEOD) δ173.21, 172.62, 171.95, 171.47, 146.18, 146.16, 146.03, 1 45.99, 145.50, 145.35, 145.29, 145.28, 131.47, 131.37, 128.34, 128.08, 122.01 ,120.37,119.66,117.54,117.40,116.40,116.36,116.31,116.27,115.96,115 .54, 74.94, 52.65, 52.56, 49.00, 48.32, 48.00, 42.66, 42.08, 37.88, 37.58.HRMS Calcd for[MH] - , 747.1931, found 747.1937.

[0249] 3-19: Synthesis of Shimobashiric acid C (3B-19)

[0250] In a 10 mL flask, add 6 mL of deprotection solution (TFA / DCM / H2O / Tis=55:44:0.5:0.5), cool in an ice bath, then add 3B-17 (60 mg), stir and react for 4 h in an ice bath. Concentrate by rotary evaporation, add water and freeze-dry to obtain 49 mg of white solid, with a yield of nearly 100%, a purity of 99%, and an HPLC RT of 10.6 min. 1 H NMR (400MHz, MEOD) δ6.82-6.25 (m, 12H), 4.98 (s, 10H), 4.72 (dd, J=7.3, 5.2Hz, 1H), 4.57 (t, J=5.9Hz, 1H), 4.25-4. 14 (m, 2H), 3.91 (dd, J=10.5, 7.5Hz, 1H), 3.75 (dd, J=10.2, 6.8Hz, 1H), 2.75 (dddd, J=51.3, 45.1, 14.1, 5.4Hz, 4H). 13C NMR (101MHz, MEOD) δ173.37, 173.29, 172.83, 172.70, 146.15, 146.09, 145.93, 1 45.87, 145.40, 145.26, 145.16, 145.11, 131.62, 131.50, 128.83, 128.53, 122.18 , 122.15, 120.51, 119.34, 117.77, 117.49, 116.46, 116.39, 116.39, 116.24, 115. 98, 115.49, 74.91, 74.81, 49.00, 48.57, 47.88, 42.74, 41.99, 37.80, 37.48.HRMS Calcd for[MH] - , 719.1618.found 719.1618.

[0251] 3-20: Screening of reaction conditions for highly selective deprotection of acetone

[0252] In order to explore highly selective deprotection conditions that can quickly and completely remove the acetone protection without destroying the unstable ester bond in the molecule, the present invention uses 3B-4 and 3B-16 as substrates, TFA, HCl, acetic acid as catalysts, DCM, THF, DIOX, H2O as solvents, and uses HPLC to monitor the effects of various deprotection conditions. Take about 20 mg of 3B-4 and use 2 ml of mixed solution (TFA / DCM / H2O / TIS, 50 / 40 / 5 / 5) for deprotection at room temperature: after 30 minutes of reaction, 3B-4 can no longer be detected, and the product 3B-3 accounts for about 90%; after 1 hour of reaction, 3B-3 accounts for about 87%. Use 2 ml of mixed solution (TFA / DCM / H2O / TIS, 25 / 65 / 5 / 5) for deprotection at room temperature; after 30 minutes of reaction, 3B-4 can no longer be detected, and 3B-3 accounts for 93%; after 1 hour of reaction, 63B-3 accounts for 91%. Deprotection was performed using 2 ml of a mixture (TFA / DCM / H2O / TIS, 50 / 40 / 5 / 5) in an ice bath. After 30 minutes of reaction, 3B-4 was no longer detected and 3B-3 accounted for 94%. It was finally found that the selective deprotection of acetone was best when 50-70% TFADCM solution was used in the presence of a trace amount of water. For example, the deprotection method used in Examples 3-19 and 3-18.

[0253] Example 4: Preparation of dopa oligopeptide solid preparation (DOSF) based on catechol derivatives

[0254] 4-1: DOSF-1 freeze-dried powder;

[0255] Weigh 9.3 g of H-Met-DOPA-OH hydrochloride and dissolve it in 30 mL of water. Under Ar protection, add about 220 mL of 0.1 M NaOH solution (Ar deoxygenation), adjust the pH to 5.6, and freeze-dry to obtain a fluffy white solid, which is DOSF-1 freeze-dried powder.

[0256] 4-2: DOSF-2 freeze-dried powder;

[0257] Weigh 8.4 g of H-Val-DOPA-OH hydrochloride and dissolve it in 30 mL of water. Under Ar protection, add about 220 mL of 0.1 M NaOH solution (Ar deoxygenation), adjust the pH to 5.6, and freeze-dry to obtain a fluffy white solid, which is DOSF-2 freeze-dried powder.

[0258] 4-3: DOSF-3 powder;

[0259] Weigh 0.84 g of DOSF-1 freeze-dried powder, 0.148 g of benserazide hydrochloride, 40 mg of sodium sulfite, 24 mg of L-DOPA, 0.2 g of sodium carboxymethyl cellulose, and 0.1 g of sodium citrate, mix them evenly, grind and mix them evenly to form a solid powder preparation, i.e., DOSF-3 powder.

[0260] When using, take a medicine bottle containing DOSF-3 powder, add 20 mL of solvent PPM-2 (1% sodium carboxymethyl cellulose aqueous solution), and shake for 5-30 minutes to obtain a clear and uniform solution.

[0261] 4-4: DOSF-4 powder;

[0262] Weigh 0.84 g of DOSF-2 freeze-dried powder, 0.148 g of benserazide hydrochloride, 40 mg of sodium sulfite, 24 mg of L-DOPA, 0.2 g of sodium carboxymethyl cellulose, and 0.1 g of sodium citrate, mix them evenly, grind and mix them evenly to form a solid powder preparation, i.e., DOSF-4 powder.

[0263] DOSF-4 powder can be used as a drug to replace Duopa; 30 minutes before use, take a medicine bottle encapsulated with DOSF-4 powder, add 20mL of solvent PPM-2 (1% sodium carboxymethyl cellulose aqueous solution), and shake for 5-30 minutes to obtain a clear and uniform solution; the solution does not contain any insoluble matter, has a certain viscosity, and is thixotropic: it becomes a gel in a static state, and forms a sol when pressure is applied, and the sol can pass through a syringe.

[0264] 4-5: DOSF-5 granules;

[0265] Weigh 0.84 g of DOSF-1 freeze-dried powder, 0.148 g of benserazide hydrochloride, 40 mg of sodium sulfite, 24 mg of L-DOPA, and 0.1 g of sodium citrate and mix well, then add 3 g of sucrose, grind and mix well, add a small amount of water to make a soft material, pass the soft material through a 10-mesh nylon sieve to obtain wet granules, place the wet granules in a vacuum drying oven to dry to obtain dry granules, pass the dry granules through a 10-mesh sieve, and then pass through a No. 5 sieve to remove fine powder, and then encapsulate to obtain DOSF-5 granules.

[0266] 4-6: DOSF-6 tablets;

[0267] 0.84 g of DOSF-1 freeze-dried powder, 0.148 g of benserazide hydrochloride, 40 mg of sodium sulfite, 24 mg of L-DOPA, and 0.1 g of sodium citrate were weighed and mixed, and then 3 g of sucrose and 25 mg of magnesium stearate were added, and the mixture was ground and mixed evenly to prepare granules; the granules were pressed into tablets on a tablet press to prepare DOSF-6 tablets.

[0268] 4-7: Oral solution;

[0269] The powder, granules or tablets of the DOPA-containing dipeptide and carbidopa dipeptide are added with 0.01 g / mL sucrose aqueous solution (or 0.0001 g / mL aspartame aqueous solution) to prepare an oral liquid; the DOPA dipeptide in the oral liquid has good water solubility, is relatively stable, does not settle, and has no particles; and has a good taste, greatly improving the compliance of patients in taking medicine, thereby better controlling symptoms, stabilizing blood drug concentration, and improving the life of patients.

[0270] Example 5: Oral continuous or intermittent drug delivery device

[0271] like Figure 1 As shown, the oral continuous or intermittent drug delivery device is designed as a "milk tea cup", including a cup cover 100 and a cup body 200; the cup body and the cup cover are made of plastic or metal material or a combination of the two, so that the cup is light and easy to carry, not easy to break, relatively stable, easy to change the color and shape of the cup body, and easy to adapt to the storage and use of drugs.

[0272] The cup body shell 201 is a shell made of plastic cloth and metal. The space enclosed by the cup body shell becomes the cup body 200. The cup body 200 contains all the liquid medicine used within a period of time. By controlling the amount of the liquid medicine, it is ensured that the patient has taken it within the corresponding time and the medicine will not deteriorate or settle.

[0273] like Figure 2As shown, the cup cover 100 is provided with a switch button 101, a battery slot 102, a micro peristaltic pump 103, a delivery tube 104 (i.e., an oral liquid delivery tube), and an output tube 105 (i.e., an oral liquid output tube); a quantitative pool 108 is provided at the lower end of the cup cover. The quantitative pool 108 is relatively small, with a volume of about 25 ml. When the cup cover and the cup body are in a connected state, the quantitative pool 108 is located inside the cup body.

[0274] The switch button 101 is used to turn on or off the micro peristaltic pump; the battery slot 102 is used to install batteries, for example, 1.5V-3V batteries can be installed to power the micro peristaltic pump; the transmission tube 104 is installed on the micro peristaltic pump, one end of the transmission tube 104 is located at the bottom of the cup body, and the other end is located at the bottom of the quantitative tank. The micro peristaltic pump 103 is used to pump the liquid medicine in the cup body 200 into the quantitative tank 108 on time and in quantity through the transmission tube, so as to control the amount of medicine taken by the patient each time to be appropriate, and the cut-off threshold can be adjusted. If the patient does not drink for more than 10 minutes, the micro peristaltic pump will no longer pump the liquid medicine into the quantitative tank, which ensures that the amount of liquid medicine will not be too large when the patient takes it next time, causing movement complications (dyskinesia), etc.; the output tube 105 passes through the upper part of the cup cover and extends into the quantitative tank 108, which is used for the patient to absorb the liquid medicine from the quantitative tank, that is, the output tube inserted in the quantitative tank can be used as a straw for the patient to drink directly, and can also be transferred to other auxiliary drinking tubes.

[0275] Furthermore, in one embodiment, a speed regulating button 106 and a flow rate display screen 107 are also provided on the cup cover 100. The speed regulating button 106 is used to adjust the flow rate of the current medicine liquid pumped from the cup body into the quantitative pool, that is, the speed regulating button 106 is used to adjust the amount of medicine liquid delivered per minute according to the patient's needs; the flow rate display screen 107 displays the flow rate of the current medicine liquid pumped from the cup body into the quantitative pool.

[0276] like Figure 1 The cup body and the cup cover are detachably connected. For example, the cup body and the cup cover can be tightly combined by threads, or can be combined together by pressing, magnetic attraction or other suitable methods; rubber rings or silicone rings may or may not be used to help seal.

[0277] The oral continuous or intermittent drug delivery device disclosed in the present invention is similar to a water cup, which can pump out an appropriate amount of medicine from the cup body into a storage quantitative pool on time; the patient only needs to take the tablet Parkinson's drug orally in the early morning or at noon for 2 hours, and start using the oral liquid in Example 12 of the present invention when the blood drug concentration is about to drop. The oral liquid is administered through the oral continuous or intermittent drug delivery device of the present invention. The patient drinks the oral liquid once every 5 to 10 minutes, and the blood drug concentration can be stabilized at 2-5 micrograms / ml until the next drinking of the tablet drug is stopped; the oral liquid in Example 12 and the drug delivery method disclosed in the present invention can greatly reduce the pain of patients taking the medicine while stabilizing the blood drug concentration, and the appropriate sweetness ratio can enhance the patient's pleasure of taking the medicine, and the drug delivery device is light, convenient, and easy to carry. It can also be considered to be made into a cup shape such as a water bag, which can be well integrated into normal life, reduce the sense of shame of Parkinson's patients, and provide patients with a more comfortable and decent life and treatment.

[0278] Example 6: Preparation apparatus for acetal / ketone protection of catechol groups

[0279] 6-1: Preparation device A

[0280] A preparation device A for preparing a dopa oligopeptide intermediate, such as Figure 3 As shown, it includes a reactor, a condensing device, an absorption tower, a gas pipeline and a liquid pipeline; the reactor is connected to the condensing device through a gas pipeline, the reactor is connected to the absorption tower through a liquid pipeline, the absorption tower is connected to the condensing device through a liquid pipeline, and a valve is provided on the liquid pipeline.

[0281] Through azeotropic distillation, the volatile by-products in the reactor are carried by the carrier through the gas pipeline to the condensation equipment, and are effectively condensed by the condensation equipment to form a condensed liquid. The condensed liquid then enters the absorption tower (one or more) filled with absorbent through the liquid pipeline, and the by-products in the condensed liquid are completely removed through the gradient absorption of the absorption tower; finally, the condensed liquid after the by-products are removed returns to the reactor through the liquid pipeline, which maximizes the promotion of the reaction equilibrium to move in the forward direction.

[0282] Furthermore, in one embodiment, the preparation device also has a bubbler, and the bubbler is connected to the condensation device; and an inert gas pipeline is arranged on the reactor.

[0283] Among them, the reactor includes but is not limited to flasks and reactors; the gas pipeline includes but is not limited to glass tubes, plastic tubes, ceramic tubes, metal tubes, etc., preferably glass tubes with vacuum jackets; the condensing equipment includes but is not limited to glass or metal condensers, cold traps, etc., preferably glass condensers; the absorption tower includes but is not limited to glass tubes, ceramic tubes, metal tubes, etc., preferably stainless steel tubes; the absorbent includes but is not limited to one or a combination of any two or more of calcium chloride, molecular sieves, calcium oxide, caustic soda, magnesium sulfate, sodium sulfate, etc., preferably calcium chloride; the liquid pipeline includes but is not limited to metal tubes, plastic tubes, rubber tubes, glass tubes, etc., preferably polytetrafluoroethylene plastic tubes.

[0284] Further, in one embodiment, if Figure 4 As shown, the condensing equipment and the absorption tower are vertically connected to form preparation device A-1.

[0285] 6-2: Improved constant pressure dropping funnel A-1-1

[0286] The present invention also discloses a preparation device for preparing dopa oligopeptide intermediates, comprising a reactor, a condensing device, a liquid pipeline and an improved constant pressure dropping funnel (i.e., a device A-1-1 connecting an absorption tower and a gas pipeline); the reactor is connected to the lower end of the improved constant pressure dropping funnel through a liquid pipeline, and the condensing device is vertically connected to the upper end of the improved constant pressure dropping funnel through a liquid pipeline.

[0287] like Figure 5 As shown, the improved constant pressure dropping funnel A-1-1 includes a dropping funnel body 1, a first constant pressure tube 2, a vacuum sleeve 3 and a condensing sleeve 4, a regulating valve 11 is arranged in the middle of the dropping funnel body 1, the regulating valve 11 divides the dropping funnel body 1 into an upper part 12 of the dropping funnel and a lower part 13 of the dropping funnel, the first constant pressure tube 2 connects the upper part 12 of the dropping funnel and the lower part 13 of the dropping funnel; the upper part 12 of the dropping funnel is provided with a first upper interface 14 (i.e., an inner ground mouth 40 / 38), the lower part of the dropping funnel is provided with a first lower interface 15 (i.e., an outer ground mouth 24 / 29), a dropping nozzle is arranged in the lower interface, and the dropping nozzle is connected with the upper part of the dropping funnel; a sand core plate 5 is arranged at the lower end of the body of the upper part 12 of the dropping funnel, and the sand core plate is, for example, a sand core plate G3; the sand core plate provides support for the solid absorbent added into the improved constant pressure dropping funnel, and can ensure that only condensed liquid, but no solid absorbent, flows into the reactor.

[0288] The vacuum jacket 3 is wrapped around the outside of the first constant pressure tube 2 to ensure the smooth passage of the high-temperature distillation gas; and the inner diameter of the constant pressure tube (i.e., the gas pipeline) is larger than that of a conventional constant pressure dropping funnel, and the inner diameter of the constant pressure tube (i.e., the gas pipeline, the steam riser) is, for example, 12 mm; the outer diameter of the vacuum jacket is, for example, 20 mm.

[0289] The condensation sleeve 4 is set on the upper part of the dropping funnel body 1, and the height of the condensation sleeve is preferably half of the height of the upper part of the dropping funnel body. The condensation sleeve 4 ensures that the absorbent works at a lower temperature, and the absorption efficiency and capacity are improved; the coolant inlet joint 41 and the outlet joint 42 of the condensation sleeve are both 8mm pagoda joints; and by controlling the regulating valve 11, the height of the condensate retained on the sand core plate 5 can be selected to adjust the contact time between the condensate and the absorbent; the upper interface in the device A-1-1 of the present invention is wide, which is convenient for taking and placing the absorbent; the upper interface is connected to the condensation equipment (such as a glass condenser); the lower interface matches the reactor interface, which is the outlet of the distilled gas and the inlet of the condensed liquid returning to the reactor. The improved constant pressure dropping funnel A-1-1 is mainly made of glass, and only the regulating valve part adopts a polytetrafluoroethylene stopcock, and its volume is, for example, 250ml; the improved constant pressure dropping funnel A-1-1 disclosed in the present invention combines the advantages of a constant pressure dropping funnel, a Soxhlet extractor, and a condenser.

[0290] Through azeotropic distillation, the volatile by-products in the reactor are carried by the carrying agent through the liquid pipeline to the improved constant pressure dropping funnel A-1-1. After the volatile by-product gas enters the lower part of the dropping funnel, it enters the upper part of the dropping funnel through the constant pressure tube, and then passes through the upper interface of the upper part of the dropping funnel and enters the condensation equipment through the liquid pipeline. After the condensation equipment effectively condenses, a condensed liquid is formed. The condensed liquid then passes through the liquid pipeline through the upper interface into the upper part of the dropping funnel. The condensed liquid passes through the sand core plate on the upper part of the dropping funnel to completely remove the by-products in the condensed liquid. Finally, the condensed liquid after the by-products are removed flows into the lower part of the dropping funnel, enters the liquid pipeline through the lower interface and returns to the reactor.

[0291] 6-3: Improved Soxhlet extractor intermediate tube A-1-2

[0292] The present invention also discloses a preparation device for preparing dopa oligopeptide intermediates, comprising a reactor, a condensing device, a liquid pipeline and an improved Soxhlet extractor intermediate tube A-1-2; the reactor is connected to the lower end of the improved Soxhlet extractor intermediate tube through a liquid pipeline, and the condensing device is vertically connected to the upper end of the improved Soxhlet extractor intermediate tube through a liquid pipeline.

[0293] like Figure 6As shown, the improved Soxhlet extractor intermediate tube comprises a second lower interface 2.1 (i.e., a Soxhlet extractor lower ground mouth, generally an outer ground mouth connected to a reaction flask), a second constant pressure tube 2.2 (i.e., a gas flow tube), a first glass tube sealed bottom 2.3, a first glass tube frosted bottom 2.4, a Soxhlet extractor body 2.5, a first siphon 2.6, a side ground mouth 2.7 (generally an inner ground mouth, connected to a finger-shaped condenser) and a second upper interface 2.8 (i.e., an upper ground mouth, which is generally an inner ground mouth, connected to a condenser); the first glass tube sealed bottom 2.3 divides the Soxhlet extractor body 2.5 into an upper part of the Soxhlet extractor body and a lower part of the Soxhlet extractor body, and the upper end of the upper part of the Soxhlet extractor body is provided with a second upper interface 2.8, and the second upper interface 2.8 is connected to the first glass tube. A side ground mouth 2.7 is arranged between the closed bottom 2.3 of the tube; the second constant pressure tube 2.2 connects the upper part of the Soxhlet extractor body and the lower part of the Soxhlet extractor body, the upper end of the second constant pressure tube 2.2 is located on the opposite side between the side ground mouth 2.7 and the second upper interface 2.8, and the lower end is located between the first glass tube closed bottom 2.3 and the second lower interface 2.1; the lower end of the upper part of the Soxhlet extractor body is provided with a first glass tube frosted bottom 2.4; the first siphon tube 2.6 connects the upper part of the Soxhlet extractor body and the lower part of the Soxhlet extractor body, the upper end of the first siphon tube 2.6 is located between the first glass tube closed bottom 2.3 and the first glass tube frosted bottom 2.4, and the lower end is located between the first glass tube closed bottom 2.3 and the lower end of the second constant pressure tube 2.2.

[0294] Furthermore, in one embodiment, the improved Soxhlet extractor intermediate tube further comprises a vacuum jacket 2.9, and the vacuum jacket 2.9 is wrapped around the outside of the second constant pressure tube 2.2.

[0295] The Soxhlet extractor is very suitable for reusing the refluxed solvent to extract natural products. If an absorbent (such as CaCl2) is placed in the sample tube, the Soxhlet extractor is also used to absorb and remove volatile by-products produced in the reaction. However, it has four disadvantages: the sample tube is small and can hold less absorbent; the refluxed solvent temperature is high, which reduces the adsorption capacity of the absorbent; the siphon is high and occupies more liquid; all immersion absorption does not form an absorption gradient.

[0296] The present invention transforms the middle tube of the Soxhlet extractor to avoid the above disadvantages. Firstly, a sample tube is not used, and a frosted sheet is added to the bottom of the middle tube, so that the entire middle tube is used to hold the absorbent, thereby increasing the volume. Secondly, a ground opening is added to the side of the middle tube for inserting a finger-shaped condenser tube, which effectively reduces the stability of the absorption area and ensures the absorption efficiency and absorption capacity of the absorbent. Thirdly, the height of the siphon tube is appropriately lowered, the volume of the solvent retained in the middle tube is reduced, and the reaction liquid in the reaction flask is kept in an appropriate amount. Fourthly, by appropriately lowering the height of the siphon tube, the absorbent in the middle tube absorbs the by-products to be removed in a gradient manner, the absorbent above the height of the siphon tube absorbs the reflux liquid containing the by-products with a high concentration in a leaching manner, and the absorbent below the height of the siphon tube absorbs the reflux liquid containing the by-products with a lower concentration in a soaking manner.

[0297] 6-4: Improved Soxhlet extractor intermediate tube A-1-3

[0298] The present invention also discloses a preparation device for acetal / ketone protection of catechol groups, i.e., a preparation device for preparing dopa oligopeptide intermediates, comprising a reactor, a condensing device, a liquid pipeline and an improved Soxhlet extractor intermediate tube A-1-3; the reactor is connected to the lower end of the improved Soxhlet extractor intermediate tube A-1-3 through a liquid pipeline, and the condensing device is vertically connected to the upper end of the improved Soxhlet extractor intermediate tube A-1-3 through a liquid pipeline.

[0299] like Figure 7As shown, the improved Soxhlet extractor intermediate tube A-1-3 comprises: a tube body 310, a third lower interface 307 (i.e., the Soxhlet extractor lower interface, generally with an outer ground mouth, such as 24 / 29, connected to a reaction flask), a third constant pressure tube 308 (i.e., a gas flow tube), a second glass tube sealed bottom 309, a second glass tube frosted bottom 305, a second siphon tube 306, a third upper interface 301 (i.e., an upper interface, generally with an inner ground mouth, such as 40 / 38, connected to a condenser) and a closed cooling jacket 302; the second glass tube sealed bottom 309 divides the tube body 310 of the Soxhlet extractor intermediate tube into The upper part and the lower part of the tube body 310 are connected; the upper end of the upper part of the tube body 310 is provided with a third upper interface 301; the third constant pressure tube 308 connects the upper part and the lower part of the tube body 310; the lower end of the upper part of the tube body 310 is provided with a second glass tube frosted bottom 305; the second siphon tube 306 connects the upper part and the lower part of the tube body 310, the upper end of the second siphon tube 306 is located between the second glass tube closed bottom 309 and the second glass tube frosted bottom 305, and the lower end of the second siphon tube 306 extends out of the third lower interface 307 of the tube body 310, and the third lower interface 307 is located at the lower end of the lower part of the tube body 310. The upper and lower parts of the tube body 310 are connected by gas flow through the third constant pressure tube 308, and the steam from the lower part of the middle tube of the Soxhlet extractor is introduced into the upper part, and the gas is converted into liquid by the condensation device connected to the upper part of the Soxhlet extractor; the upper and lower parts are connected by liquid through the second siphon tube 306, and the upper part of the second glass tube frosted bottom 305 (such as a sand core plate) above the closed bottom 309 is equipped with an adsorbent (such as calcium chloride), and the water and methanol in the condensate are selectively absorbed by the adsorbent here, and then the condensate is filtered through the second glass tube frosted bottom 305, and refluxed to the reactor connected to the third lower interface 307 of the tube body 310 through the second siphon tube 306.

[0300] A glass closed cooling jacket 302 is provided on the outer side of the lower end of the upper part of the tube body 310, and its height accounts for 40-100% of the upper part of the tube body 310; the coolant inlet 304 below the cooling jacket 302 is connected to the liquid outlet of the circulating cooling pump, and the coolant outlet 303 above the cooling jacket is connected to the liquid inlet of the circulating cooling pump; the closed cooling jacket 302 ensures that the absorbent in the intermediate tube body 310 of the Soxhlet extractor works at a lower temperature, thereby improving the absorption efficiency and absorption amount of water and methanol.

[0301] The present invention is described in detail above in combination with specific implementation methods and exemplary examples. The above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments; the above description cannot be understood as limiting the present invention. Those skilled in the art understand that, without departing from the spirit and scope of the present invention, the technical solution of the present invention and its implementation methods may be subjected to a variety of equivalent substitutions, modifications or improvements, all of which fall within the scope of the present invention; the protection scope of the present invention shall be subject to the attached claims.

Claims

1. A method for preparing a catechol derivative, characterized in that: The method comprises adopting a sodium sulfite-boric acid-argon system for protection, preparing a catechol derivative in an organic phase-aqueous phase mixture, wherein the catechol derivative comprises a levodopa or carbidopa derivative; the preparation method comprises the following steps: S1: adding a reducing agent, a boric acid compound and a mixed solvent into a reaction device, and using an inert gas to bubble to remove oxygen in the reaction device; S2: adding compound I and a base into a reaction device, stirring and continuing to pass an inert gas; S3: Add reactant II into the reaction device, stir and continue to pass inert gas, and obtain compound III after acidification; the reaction formula is as follows: Wherein, R1 includes one of H, Me, COOH, and CH2COOH; R2 includes one of H, Me, Et, F, Cl, Br, and OH; R3 includes one of NH2 and NHNH2; The reactant II comprises one of acyl halide, acid anhydride, N-hydroxysuccinimide type activated ester and pentafluorophenol type activated ester; the reactant II is R4-X, X is an active group, R4 is an inactive group; and R5 is a fragment formed by the reaction of R3 and R4-X.

2. The method for preparing a catechol derivative according to claim 1, characterized in that: For the levodopa or carbidopa derivatives protected by acid-labile protecting groups, hydrochloric acid or TFA is used for deprotection to obtain the dipeptide of levodopa or carbidopa, including: Boc-AA-DOPA-OH and Boc-AA-CarbiDOPA-OH were converted to H-AA-DOPA-OH and H-AA-CarbiDOPA-OH.

3. The method for preparing a catechol derivative according to claim 1, characterized in that: For the levodopa or carbidopa derivatives protected by acid-stable protecting groups, the catechol groups in the levodopa or carbidopa derivatives are further protected by acetal / ketone protection, including: S10: adding a certain amount of a catechol derivative of compound IV and a cosolvent into a reaction device, and heating to form a solution; the compound IV is an N-modified catechol derivative in compound III; S20: adding a certain amount of carrier to the reaction device, introducing inert gas, heating to 70-120° C., reflux for about 20 minutes, and using the absorbent in the absorption device to absorb water and by-products; S30: adding reactant V and a catalyst, and continuing to reflux for about 2-5 hours; adding reactant V again, and after distilling off the volatile substances, stopping the reaction to obtain compound VI; the reactant V includes aldehydes, ketones, hemiacetals and hemiketals of aldehydes and ketones; the reaction formula is as follows: Wherein, R5 includes -NHFmoc, -NHNHFmoc, -NHCbz, -NPhth, -NHTfa, -NTFa-NHTfa, C 15 H 31 CO-NH-, C 12 H 25 CO-NH-, C 12 H 25 One of CO-Val-NH- and Fmoc-Val-NH-.

4. The method for preparing a catechol derivative according to claim 3, characterized in that: Also included is the conversion of the dopa-containing methyl ester produced when the catechol group is subjected to acetal / ketone protection into the target carboxylic acid, including: 1) adding a methyl ester compound VII and an organic solvent into a reaction apparatus to obtain a methyl ester compound VII solution; 2) dissolving a certain amount of lithium salt in water to obtain a lithium salt solution, and adding the lithium salt solution to the above-mentioned methyl ester compound VII solution; adjusting the pH of the reaction system to 7-8 with hydrochloric acid, and then adding a certain amount of alkali, and stirring for 3-24 hours; 3) Concentrate the organic phase by rotary evaporation, add water, adjust the pH to 3-5 with HCl solution, extract with EA, wash the organic phase with acid water and pure water respectively; take the organic phase, add anhydrous magnesium sulfate to dry, filter, and rotary evaporate to remove volatiles, and recrystallize in TLN / HXN to obtain compound VIII; the reaction formula is as follows: Wherein, the R6 includes formal, acetaldehyde, benzaldehyde, acetone, cyclopentanone, cyclohexanone, benzophenone, and methoxymethylene.

5. The method for preparing a catechol derivative according to claim 3, characterized in that: It also includes the generation of acetonide-protected L-danshensu derivatives based on acetonide-protected L-dopa derivatives: The acetonide-protected L-dopa derivative is deaminated and reacted with nitrous acid in water or a mixture of water and an organic solvent under acid catalysis, and then hydrolyzed to generate an acetonide-protected L-danshensu derivative.

6. A method for preparing a catechol derivative, characterized in that: Using levodopa as a raw material, SOCl2 and anhydrous methanol are used for methyl esterification reaction, and then exchange reaction is carried out with methyl trifluoroacetate to achieve a one-pot synthesis of Tfa-DOPA-OMe, which specifically includes the following steps: L-DOPA and anhydrous methanol were added to the reaction apparatus, argon was introduced, SOCl2 was slowly added dropwise under ice bath conditions, and stirring was continued overnight after removing the ice bath; heating and stirring were performed, argon bubbling was turned on, the temperature was lowered, alkali and methyl trifluoroacetate were slowly added, and stirring was performed until the reaction was complete to obtain Tfa-DOPA-OMe.

7. A method for preparing an acetone-protected danshensu derivative, characterized in that: include: The carboxyl group of danshensu is protected as ester or amide, and the alcoholic hydroxyl group of danshensu is protected as ester or ether by adopting protection measures, thereby forming a danshensu intermediate; The danshensu intermediate is reacted with an acetonide cyclization reagent to protect the catechol group with acetonide; The acetone-protected danshensu derivatives are obtained through selective hydrolysis, alcoholysis or aminolysis.

8. A method for preparing a catechol compound, characterized in that: include: The acetone-protected danshensu is subjected to an ester bond forming reaction, an amide bond forming reaction or an ether bond forming reaction to obtain a complex containing the acetone-protected danshensu; trifluoroacetic acid or hydrochloric acid is used to selectively remove the acetone protection of catechol in the complex under mild reaction conditions to obtain a catechol compound, wherein the catechol compound includes an ester bond.

9. The method for preparing catechol compounds according to claim 8, characterized in that: include: The acetonide-protected caffeic acid undergoes head-to-tail dimerization under black light irradiation to form an acetonide-protected intermediate, and the intermediate is coupled with the acetonide-protected danshensu methyl ester through an esterification reaction to form acetonide-protected Shimobashiric acid C methyl ester, wherein the structural formula of the Shimobashiric acid C methyl ester is 3B-18; Finally, shimobashiricacid C is synthesized by selectively removing acetone protection, and the structural formula of shimobashiricacid C is 3B-19; 10. Use of the intermediate obtained by the preparation method according to any one of claims 1 to 9 in the synthesis, separation and purification of natural catechol compounds or non-natural catechol compounds, in ultraviolet capture, anti-oxidation, anti-aging, memory improvement, microcirculation improvement, neuroprotection, and in the prevention of Parkinson's disease, cardiovascular and cerebrovascular diseases, Alzheimer's disease, and cancer.

11. Use of a catechol derivative prepared by the preparation method according to any one of claims 1 to 6, 8 and 9 in the preparation of a solid preparation or a liquid preparation containing dopa oligopeptide.

12. Use of a tanshinone derivative protected by the preparation method according to claim 7 in the preparation of solid preparations and liquid preparations containing dopa oligopeptides.

13. An oral continuous or intermittent administration device for the liquid preparation according to any one of claims 11 to 12, characterized in that: It comprises a cup body (200) and a cup cover (100), wherein the cup body (200) and the cup cover (100) are detachably connected; The cup body (200) is a container for containing oral liquid; The cup cover (100) is provided with a switch button (101), a battery slot (102), a micro peristaltic pump (103), a transmission tube (104), an output tube (105) and a quantitative pool (108); the switch button (101) is used to turn on or off the micro peristaltic pump, the battery slot (102) is used to install a battery to supply power to the micro peristaltic pump (103); the transmission tube (104) is installed on the micro peristaltic pump, and one end of the transmission tube (104) is located at the bottom of the cup body (200). The cup cover (100) has one end at its top and the other end at the bottom of the quantitative pool (108); the micro peristaltic pump (103) is used to pump the liquid medicine in the cup body (200) into the quantitative pool (108) through the transmission tube (104); the quantitative pool (108) is arranged at the lower end of the cup cover (100); when the cup cover and the cup body are in a connected state, the quantitative pool (108) is located on the inner side of the cup body (200); the output tube (105) passes through the cup cover (100) and extends into the quantitative pool (108).

14. The oral continuous or intermittent drug delivery device according to claim 13, characterized in that: The cup cover is also provided with a speed regulating button and a flow rate display screen.

15. A preparation device for realizing acetal / ketone protection of catechol groups as claimed in claim 3, characterized in that: It includes reactor, condensing equipment, absorption tower, gas pipeline and liquid pipeline; The reactor is connected to the condensing device via a gas pipeline, the reactor is connected to the absorption tower via a liquid pipeline, and the absorption tower is connected to the condensing device via a liquid pipeline; Among them, the reactor includes one of a flask and a reaction kettle; the gas pipeline includes one of a glass tube, a plastic tube, a ceramic tube, and a metal tube; the condensation equipment includes one of a glass or metal condenser tube and a cold trap; the absorption tower includes one of a glass tube, a ceramic tube, and a metal tube; the liquid pipeline includes one of a metal tube, a plastic tube, a rubber tube, and a glass tube.

16. A preparation device for realizing acetal / ketone protection of catechol groups as claimed in claim 3, characterized in that: It includes a reactor, a condensation device, an improved constant pressure dropping funnel and a liquid pipeline; The reactor is connected to the lower interface of the improved constant pressure dropping funnel through a liquid pipeline, and the condensation device is vertically connected to the upper interface of the improved constant pressure dropping funnel through a liquid pipeline; The improved constant pressure dropping funnel comprises a dropping funnel body (1), a first constant pressure tube (2), a vacuum sleeve (3) and a condensation sleeve (4); a regulating valve (11) is arranged in the middle of the dropping funnel body (1); the regulating valve (11) divides the dropping funnel body (1) into an upper part (12) of the dropping funnel and a lower part (13) of the dropping funnel; the first constant pressure tube (2) connects the upper part (12) of the dropping funnel and the lower part (13) of the dropping funnel; The upper part (12) is provided with a first upper interface (14), the lower part (13) of the dropping funnel is provided with a first lower interface (15), a dripping nozzle is provided in the first lower interface (15), and the dripping nozzle is connected with the upper part of the dropping funnel; a sand core plate (5) is provided at the lower end of the main body of the upper part (12) of the dropping funnel; the vacuum sleeve (3) is wrapped around the outside of the first constant pressure tube (2), and the condensation sleeve (4) is sleeved on the upper part (12) of the dropping funnel.

17. A preparation device for realizing acetal / ketone protection of catechol groups as claimed in claim 3, characterized in that: It comprises a reactor, a condensing device, an improved Soxhlet extractor intermediate tube and a liquid pipeline; the reactor is connected to the lower interface of the improved Soxhlet extractor intermediate tube through the liquid pipeline, and the condensing device is vertically connected to the upper interface of the improved Soxhlet extractor intermediate tube through the liquid pipeline; The improved Soxhlet extractor intermediate tube comprises a second lower interface (2.1), a second constant pressure tube (2.2), a first glass tube sealed bottom (2.3), a first glass tube frosted bottom (2.4), a Soxhlet extractor body (2.5), a first siphon tube (2.6), a side ground mouth (2.7) and a second upper interface (2.8); the first glass tube sealed bottom (2.3) divides the Soxhlet extractor body (2.5) into an upper part of the Soxhlet extractor body and a lower part of the Soxhlet extractor body, the upper end of the upper part of the Soxhlet extractor body is provided with a second upper interface (2.8), and a side ground mouth (2.7) is provided between the second upper interface (2.8) and the first glass tube sealed bottom (2.3); the second constant pressure tube (2.2 ) connects the upper part of the Soxhlet extractor body and the lower part of the Soxhlet extractor body, the upper end of the second constant pressure tube (2.2) is located on the opposite side between the side ground mouth (2.7) and the second upper interface (2.8), and the lower end is located between the first glass tube sealed bottom (2.3) and the second lower interface (2.1); the lower end of the upper part of the Soxhlet extractor body is provided with a first glass tube frosted bottom (2.4); the first siphon tube (2.6) connects the upper part of the Soxhlet extractor body and the lower part of the Soxhlet extractor body, the upper end of the first siphon tube (2.6) is located between the first glass tube sealed bottom (2.3) and the first glass tube frosted bottom (2.4), and the lower end is located between the first glass tube sealed bottom (2.3) and the lower end of the second constant pressure tube (2.2).

18. A preparation device for realizing acetal / ketone protection of catechol groups as claimed in claim 3, characterized in that: It comprises a reactor, a condensing device, an improved Soxhlet extractor intermediate tube and a liquid pipeline; the reactor is connected to the lower interface of the improved Soxhlet extractor intermediate tube through the liquid pipeline, and the condensing device is vertically connected to the upper interface of the improved Soxhlet extractor intermediate tube through the liquid pipeline; The improved Soxhlet extractor intermediate tube comprises a tube body (310), a third lower interface (307), a third constant pressure tube (308), a second glass tube sealed bottom (309), a second glass tube frosted bottom (305), a second siphon tube (306), a third upper interface (301) and a sealed cooling jacket (302); the second glass tube sealed bottom (309) divides the tube body (310) into an upper part and a lower part; the upper end of the upper part of the tube body (310) is provided with a third upper interface (301); the third constant pressure tube (308) connects the upper part and the lower part of the tube body (310); ; A second glass tube frosted bottom (305) is provided at the lower end of the upper part of the tube body (310); the second siphon tube (306) connects the upper part and the lower part of the tube body (310); the upper end of the second siphon tube (306) is located between the second glass tube closed bottom (309) and the second glass tube frosted bottom (305); the lower end of the second siphon tube (306) extends out of the third lower interface (307) of the tube body (310), and the third lower interface (307) is located at the lower end of the lower part of the tube body (310); the closed cooling jacket (302) is provided at the lower end of the outer side of the upper part of the tube body (310).

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