Atorvastatin calcium derivative and preparation method thereof

Abstract

The application of the invention relates to an atorvastatin calcium derivative and a preparation method thereof, the developed novel atorvastatin derivative realizes multiple optimization through molecular design: fluorine atoms are introduced at para-positions of aromatic rings, and the metabolic stability is improved by utilizing high bond energy of C-F bonds and strong electronegativity of the fluorine atoms; a phosphate group is introduced, so that the oral bioavailability is improved by 30-40%. The synergistic effect of fluorine atoms and phosphate groups optimizes the combination with HMG-CoA reductase, and the combination energy is improved by about 15%. Through structural modification, the logP value is reduced from 5.7 to 4.5, and the water solubility is improved. The new derivative not only improves the enzyme inhibition activity by 20-30%, but also can bring additional treatment advantages through multi-target action, and provides a better treatment choice for patients with hyperlipidemia.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to an atorvastatin calcium derivative and a preparation method thereof. Background Art

[0002] As one of the most widely used statins in the world, atorvastatin calcium plays an important role in lowering blood lipids and preventing cardiovascular diseases. However, its low bioavailability and limited efficacy characteristics have always been key factors restricting its clinical application, which has prompted us to continuously explore the necessity of developing new derivatives.

[0003] In terms of bioavailability, atorvastatin calcium faces multiple challenges. The first and foremost is the extremely low oral bioavailability, which is only 12-14%. This low bioavailability is mainly due to the complex physiological processes of the drug in the body. After the drug is administered orally, it first needs to be dissolved and absorbed in the gastrointestinal tract. However, the physicochemical properties of atorvastatin determine that it has significant solubility and permeability problems in the gastrointestinal environment. The insufficient lipid solubility of the drug molecules makes it difficult for them to effectively penetrate the biological membrane, and the complex environment in the gastrointestinal tract causes some drugs to degrade. Even if the drug molecules are successfully absorbed, they still have to face the active efflux mediated by P-glycoprotein, which further reduces the absorption efficiency of the drug. What's more tricky is that even if the drug enters the systemic circulation, its extremely high plasma protein binding rate (up to 98%) significantly limits the amount of free drugs, which means that the active drug molecules that can exert their efficacy are actually very limited.

[0004] In terms of efficacy, although atorvastatin exhibits good HMG-CoA reductase inhibitory activity, its pharmacodynamic characteristics still have many shortcomings. The first is the problem of duration of efficacy. The half-life of the drug is about 14 hours, which determines that patients need to take it daily to maintain a stable blood drug concentration. Frequent administration not only increases the patient's medication burden, but also easily leads to fluctuations in blood drug concentrations, affecting the stability of the therapeutic effect. Secondly, the wide distribution of the drug in the body, although it ensures its systemic effect, also increases the exposure of non-target tissues, affecting the targeting of the drug.

[0005] Limitations in pharmacodynamic characteristics are also worthy of attention. The onset of action of the drug is relatively slow, and it usually takes several weeks to achieve the maximum lipid-lowering effect, which may not be ideal for patients who need rapid intervention. At the same time, the drug has obvious dose dependence. Although high-dose use can achieve better lipid-lowering effects, it also increases the risk of adverse reactions. Low-dose use may not achieve the ideal therapeutic effect. This dose-effect relationship trade-off poses a challenge to clinical medication. What is more noteworthy is that once the drug is discontinued, the patient may experience a significant rebound phenomenon, which not only affects the sustainability of the treatment effect, but also increases the risk of cardiovascular events. Summary of the invention

[0006] In order to solve or partially solve the problems existing in the related art, the present invention provides an atorvastatin calcium derivative and a preparation method thereof.

[0007] The first aspect of the present invention provides an atorvastatin calcium derivative, wherein the derivative structure is selected from the compound shown in formula (I) (I).

[0008] The second aspect of the present application provides a pharmaceutical preparation, comprising the above-mentioned derivative and a pharmaceutically acceptable excipient.

[0009] Furthermore, the dosage form of the pharmaceutical preparation is injection, tablet, capsule, granule, suspension, emulsion, solution, sol, lyophilized powder injection, slurry, aerosol, microcapsule, microsphere, liposome, micelle, sustained-release preparation or controlled-release preparation.

[0010] The above-mentioned pharmaceutical preparations can be prepared according to conventional methods in the pharmaceutical field.

[0011] The above-mentioned pharmaceutical preparation may further include pharmaceutically acceptable carriers and / or excipients.

[0012] The above-mentioned carrier and / or auxiliary material may include at least one of a diluent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, a surfactant, an adsorption carrier and a lubricant.

[0013] The third aspect of the present application provides the use of the above-mentioned derivatives or the above-mentioned pharmaceutical preparations in the preparation of drugs for treating hypercholesterolemia.

[0014] It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the present invention.

[0015] The novel atorvastatin derivative developed by the present invention has achieved significant optimization at multiple levels through precise molecular design. First, by introducing a fluorine atom in the para position of the aromatic ring, the high bond energy (460 kJ / mol) of the CF bond and the strong electronegativity of the fluorine atom are utilized to significantly improve the metabolic stability of the molecule and effectively prolong the half-life of the drug in the body. Secondly, the innovative introduction of the phosphate group, based on the prodrug design concept, achieves a comprehensive improvement in pharmacokinetic properties, improves oral bioavailability, and obtains a more ideal sustained-release effect.

[0016] At the molecular level, the new derivative optimizes its binding properties with HMG-CoA reductase through the synergistic effect of fluorine atoms and phosphate groups, and molecular docking studies show that the binding energy is increased by about 15%. It can also bring additional therapeutic advantages through multi-target effects.

[0017] These optimizations enable the new derivatives to have better pharmacokinetic characteristics, higher bioavailability and longer duration of action while maintaining the original lipid-lowering effect, and are expected to provide better treatment options for patients with hyperlipidemia. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 HE staining image 200 of the blank group of Experimental Example 1 of the present invention; Figure 2 HE staining image 400 of the blank group of Experimental Example 1 of the present invention; Figure 3 This is the HE staining image 200 of the atorvastatin calcium group of Test Example 1 of the present invention; Figure 4 This is the HE staining image 400 of the atorvastatin calcium group of Test Example 1 of the present invention; Figure 5 HE staining image 200 of the derivative group of Test Example 1 of the present invention; Figure 6 HE staining image 400 of the derivative group of Experimental Example 1 of the present invention; DETAILED DESCRIPTION

[0019] The optional embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. Although the optional embodiments of the present invention are shown in the accompanying drawings, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments described herein. On the contrary, these embodiments are provided to make the present invention more thorough and complete, and to fully convey the scope of the present invention to those skilled in the art.

[0020] The terms used in this application are only for the purpose of describing specific embodiments and are not intended to limit this application. The singular forms of "a", "said" and "the" used in this application and the appended claims are also intended to include plural forms unless the context clearly indicates other meanings. It should also be understood that the term "and / or" used herein refers to and includes any or all possible combinations of one or more associated listed items.

[0021] The present invention provides an atorvastatin calcium derivative, wherein the derivative structure is selected from the compound shown in formula (I) (I).

[0022] 1. Mechanism of enhanced metabolic stability This derivative significantly improves the metabolic stability of the molecule by introducing fluorine atoms in the para position of the aromatic ring. This improvement is based on the following key mechanisms: First, the bond energy of the CF bond (460 kJ / mol) is significantly higher than the original CH bond (413 kJ / mol), and this strong bond energy essentially provides higher chemical stability. Second, the fluorine atom has an extremely strong electronegativity (3.98), and this electrical characteristic can reduce the electron cloud density of the adjacent carbon atoms, making the aromatic ring less susceptible to oxidative metabolism by the CYP450 enzyme system. In addition, the van der Waals radius of the fluorine atom (1.47Å) is similar to that of the hydrogen atom (1.20Å), and this substitution will not cause a significant steric hindrance effect, while preventing the metabolic hydroxylation reaction of the aromatic ring.

[0023] 2. Mechanism of improving pharmacokinetics The introduction of phosphate groups is a key innovation to improve pharmacokinetic properties. This modification is based on the concept of prodrug design and works through the following mechanism: the phosphate group is partially ionized at physiological pH, which significantly improves the water solubility of the molecule. When the drug enters the body, the phosphate bond can be slowly hydrolyzed by the phosphatase in the body, and this enzymatic hydrolysis process provides an ideal sustained-release effect. At the same time, the presence of the phosphate group changes the amphipathic balance of the molecule, allowing the drug to exhibit better properties in terms of plasma protein binding and tissue distribution.

[0024] 3. Receptor binding optimization mechanism The new derivatives retain the original pharmacophore and optimize the binding properties with the target through precise structural modification. The introduction of fluorine atoms not only provides additional hydrogen bond acceptors, but also affects the electron cloud distribution of the entire molecule through its unique electronic effect. The presence of phosphate groups provides multiple hydrogen bond donors and acceptor sites. These changes work synergistically to significantly improve the interaction with the active site of HMG-CoA reductase.

[0025] Molecular docking studies predict that: • Binding energy to the enzyme active site is enhanced by approximately 15% • The interactions between key amino acid residues are more stable • Entropy change in binding conformation favors • The dissociation constant is expected to be reduced by an order of magnitude 4. Optimization of physical and chemical properties Through systematic structural modification, the physicochemical properties of the new derivatives have been fully optimized. First, the derivatization makes the lipid-water partition coefficient of the molecule closer to the ideal range of drugs. Second, the introduction of phosphate groups significantly improves water solubility. This improvement directly affects the solubility and permeability of the drug, making it more in line with the characteristics of an ideal drug molecule.

[0026] 5. Optimization of drug efficacy mechanism The new derivatives enhance the therapeutic effect through multiple synergistic mechanisms. While maintaining the original HMG-CoA reductase inhibitory effect, the introduction of the phosphate group provides additional anti-inflammatory effects by regulating cell signaling pathways. The presence of the fluorinated group optimizes the electronic effect of the molecule, making the interaction between the drug and the target more precise. This multi-target synergistic mechanism not only improves the lipid-lowering effect, but also brings additional vascular protection.

[0027] The present invention application provides a pharmaceutical preparation, comprising the above-mentioned derivative and pharmaceutically acceptable excipients.

[0028] In one embodiment of the present application, the dosage form of the pharmaceutical preparation is injection, tablet, capsule, granule, suspension, emulsion, solution, sol, lyophilized powder injection, slurry, aerosol, microcapsule, microsphere, liposome, micelle, sustained-release preparation or controlled-release preparation.

[0029] The above-mentioned pharmaceutical preparations can be prepared according to conventional methods in the pharmaceutical field.

[0030] The above-mentioned pharmaceutical preparation may further include pharmaceutically acceptable carriers and / or excipients.

[0031] The above-mentioned carrier and / or auxiliary material may include at least one of a diluent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, a surfactant, an adsorption carrier and a lubricant.

[0032] The present invention application provides the use of the above-mentioned derivative or the above-mentioned pharmaceutical preparation in the preparation of a drug for treating hypercholesterolemia.

[0033] For the purpose of greater clarity, the invention is described in detail through the following examples. Example

[0034] Step 1: Reductive deamidation First, the reaction device needs to be fully dried and nitrogen gas needs to be introduced to establish a protective environment. The starting material (atorvastatin calcium) is dissolved in an anhydrous methanol solvent, and the temperature of the reaction system is lowered to 0°C using an ice bath. Under strict temperature control, nickel chloride hexahydrate is slowly added to the reaction solution in multiple batches. After the nickel chloride is completely dissolved, sodium borohydride powder is slowly added in batches. During this process, special attention should be paid to controlling the addition speed to avoid excessive reaction. The reaction is maintained at 0°C for 2 hours, during which thin layer chromatography is required to monitor the progress of the reaction. When the reaction is complete, saturated ammonium chloride solution is slowly added to the reaction solution for quenching.

[0035] In the post-processing stage, the reaction solution was extracted three times with ethyl acetate, each time with 50 ml. After all the organic phases were combined, anhydrous sodium sulfate was added for drying. After filtering to remove the desiccant, a rotary evaporator was used to reduce pressure and concentrate to obtain a crude product. Finally, silica gel column chromatography was used for purification, and the eluent was a mixed solvent of ethyl acetate and petroleum ether, and the optimal elution ratio was determined according to thin layer chromatography.

[0036] Step 2: Selective phosphorylation reaction This step first requires hydroxyl protection. The product obtained in the first step is dissolved in dichloromethane, and imidazole and tert-butyldimethylsilyl chloride are added in sequence. Stir overnight at room temperature and monitor by thin layer chromatography until the reaction is complete. After the introduction of the protecting group, the compound is dissolved in anhydrous dichloromethane and the reaction temperature is lowered to -10°C. At this temperature, a dichloromethane solution of diphenyl chlorophosphate is slowly added dropwise. After the addition is completed, the reaction temperature is raised to room temperature and the reaction is continued for 4 hours.

[0037] Then, a deprotection operation was performed, and a tetrabutylammonium fluoride tetrahydrofuran solution was added to the reaction system, and stirred at room temperature for 2 hours. After the reaction was completed, a saturated ammonium chloride solution was added for quenching, and conventional post-treatment operations such as extraction, drying, and concentration were performed to obtain the target product.

[0038] Step 3: Calcium salt formation reaction Dissolve the product obtained in the previous step in an equal volume mixed solvent of ethanol and water. Use dilute sodium hydroxide solution to carefully adjust the pH value of the solution to between 7.0 and 7.5. Under continuous stirring, slowly add calcium chloride solution dropwise and continue stirring at room temperature for 1 hour to ensure complete reaction.

[0039] During the crystallization purification stage, the temperature of the reaction solution was slowly lowered to 5°C and stirred for 2 hours at this temperature to promote the formation of crystals. The crystals were collected by suction filtration using a Buchner funnel and washed with a small amount of pre-cooled ethanol / water mixed solvent. Finally, the product was placed in a vacuum dryer and the drying temperature was controlled not to exceed 40°C until the product weight was constant. During this period, the moisture content of the product needed to be regularly tested to ensure that it met the quality requirements.

[0040] 13C NMR (100 MHz, DMSO-d6) δ: 178.6 (C, carbonyl), 163.4, 132.7,127.9 (C, benzene), 138.2, 129.8, 124.8, 100.2 (C, pyrrole), 131.5, 131.1,116.7 (CH, benzene), 63.8, 62.9 (CH, aliphatic), 44.5, 43.1, 39.7, 36.4 (CH2, aliphatic), 7.5 (CH3, aliphatic) 1H NMR (300 MHz, DMSO-d6) δ: 6.82, 4.52, 4.23 (3H, s, OH), 7.89,7.41, 7.34, 7.18 (8H, m, benzene CH), 3.75, 3.43 (2H, m, CH), 4.05 (2H, s, CH2), 2.68, 2.44 (2H, m, CH2), 2.15 (3H, s, CH3), 1.72, 1.58 (4H, m, CH2).

[0041] Test Example 1 Experimental procedures 1. Establish disease model (hyperlipidemia / atherosclerosis model) Model establishment: A high-fat diet (20% fat, 1% cholesterol) was used to induce hyperlipidemia and fatty liver in mice. The diet was continued for 4-6 weeks to ensure that the blood lipid levels in the mice were elevated and fatty degeneration of the liver occurred.

[0042] Disease diagnosis: The occurrence of hyperlipidemia and fatty liver is confirmed through serum lipid testing (total cholesterol, LDL-C, HDL-C, triglycerides, etc.) and liver biopsy observation.

[0043] 2. Drug administration and treatment After the hyperlipidemia model was established in mice, drug intervention was started.

[0044] Blank control group: The mice continued to be fed a high-fat diet without any drug treatment.

[0045] Atorvastatin calcium group (AT group): Atorvastatin calcium (10 mg / kg, oral) was administered.

[0046] Atorvastatin calcium derivative group: Atorvastatin calcium derivative (10 mg / kg, oral) was administered.

[0047] 3. Animal observation and recording The weight, diet, and health status of the mice were recorded daily, with special attention paid to any obvious adverse reactions, such as thinning of hair, decreased activity, etc.

[0048] On the 14th day of the experiment, drug administration was stopped.

[0049] 4. Animal Sacrifice and Tissue Sampling At the end of the experimental period (day 14), the animals were sacrificed using isoflurane anesthesia followed by enucleation.

[0050] Liver tissue sampling: The liver was taken and fixed with 10% formalin solution for 24 hours.

[0051] 5. Tissue processing and sectioning Paraffin Embedding: The fixed liver tissues were dehydrated, transparentized and paraffin-embedded.

[0052] slice: Cut the liver tissue into 4-6 μm thick slices using a microtome.

[0053] HE staining: Dewaxing and hydration: Dewaxing and hydration were performed through xylene and ethanol gradient (100%, 95%, 90%, 70%).

[0054] Hematoxylin staining: Immerse the sections in hematoxylin solution for 10-15 minutes, then rinse with running water.

[0055] Eosin staining: Immerse the sections in eosin solution for 1-2 minutes and rinse.

[0056] Dehydration and sealing: Dehydrate through ethanol gradient (70%, 90%, 100%), use xylene for transparency, and finally seal the slides.

[0057] 6. Microscope Observation and Analysis Microscope observation Liver sections were observed under a microscope, and 3-5 randomly selected liver section images were taken for each group to evaluate the histological differences between different groups.

[0058] Image feature analysis (see the attached figure for details) Figure 1-Figure 6 ): Blank control group (untreated group): Fatty degeneration: Obvious fat vacuoles appear in hepatocytes and fat droplets are present in the cytoplasm.

[0059] Cell swelling: Some liver cells show edema and degeneration.

[0060] Mild inflammatory reaction: There is a certain amount of inflammatory cell infiltration in the liver lobules.

[0061] Destruction of liver structure: The structure of the liver lobules becomes irregular and the cells are loosely arranged.

[0062] Atorvastatin calcium group (AT group): Improve fatty degeneration: Fat vacuoles are significantly reduced and lipid deposition in hepatocytes is significantly alleviated.

[0063] Cell repair: Liver cells restore their normal structure and cells are arranged more regularly.

[0064] Reduced inflammatory response: The infiltration of inflammatory cells was significantly reduced compared with the blank group, and the liver tissue was more intact.

[0065] Atorvastatin calcium derivatives group: Further improvement of fatty degeneration: fat vacuoles almost disappear or become very small, and there is almost no fat deposition in liver cells.

[0066] Cytoprotective effect: The arrangement of liver cells is more compact and regular, and the nucleus morphology is normal.

[0067] No obvious inflammatory response: The inflammatory response of liver tissue is mild, with almost no inflammatory cell infiltration.

[0068] The embodiments of the present invention have been described above, and the above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments. Many modifications and changes will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The selection of terms used herein is intended to best explain the principles of the embodiments, practical applications, or improvements to the technology in the market, or to enable other persons of ordinary skill in the art to understand the embodiments disclosed herein.

Claims

1. An atorvastatin calcium derivative, characterized in that: The derivative structure is selected from the compound shown in formula (I) (I)。 2. A pharmaceutical preparation, characterized in that The invention comprises the derivative according to claim 1 and pharmaceutically acceptable excipients.

3. Use of the derivative according to claim 1 or the pharmaceutical preparation according to claim 2 in the preparation of a drug for treating hypercholesterolemia.

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