阅读说明：本技术 乌苯美司醋酸盐单晶及其制备方法和应用 (Ubenimex acetate monocrystal and preparation method and application thereof ) 是由 吴子强 刘东华 于玉根 陈其阳 袁文 雷光华 于 2020-06-03 设计创作，主要内容包括：本发明提供了乌苯美司醋酸盐单晶及其制备方法,该单晶体纯度高,能够用于进行X射线单晶衍射确定乌苯美司原料药的绝对构型,且溶解性好,能够用于制备乌苯美司醋酸盐药物组合物,具有临床使用价值。本发明提供的乌苯美司醋酸盐单晶的制备方法析晶时间短,能够制备得到稳定均匀的乌苯美司醋酸盐单晶。(The ubenimex acetate single crystal and the preparation method thereof provided by the invention have the advantages that the purity of the single crystal is high, the single crystal can be used for determining the absolute configuration of ubenimex raw material medicines by X-ray single crystal diffraction, the solubility is good, the single crystal can be used for preparing ubenimex acetate pharmaceutical compositions, and the clinical use value is high. The preparation method of the ubenimex acetate single crystal provided by the invention is short in crystallization time, and can be used for preparing the stable and uniform ubenimex acetate single crystal.)

1. The ubenimex acetate monocrystal is characterized in that the monocrystal is an orthorhombic system and a crystal axis of the monocrystal The included angle between the crystal planes α/° 90, β/° 90, γ/° 90, the unit lattice area V2024.63 (6), the number of molecules Z in the unit lattice 4, and the density d 1.209.

2. The preparation method of ubenimex acetate monocrystal according to claim 1, wherein ubenimex is dissolved in glacial acetic acid at room temperature, a low-polarity solvent is slowly added, the ubenimex is heated to 50-80 ℃, the ubenimex is uniformly stirred until the solution is clear, the ubenimex is filtered while the ubenimex is hot, the filtrate is sealed and is kept stand at room temperature for 0.5-5 hours, and a colorless blocky monocrystal is obtained.

3. The preparation method according to claim 2, wherein the low-polarity solvent is one of dichloromethane, n-hexane, acetone, pentanone, ethyl acetate, butyl acetate and isopropyl acetate or a mixed solvent of ketones and esters thereof.

4. The method according to claim 3, wherein the low polarity solvent is acetone, ethyl acetate or a mixture thereof.

5. The method according to claim 3, wherein the low polarity solvent is ethyl acetate.

6. The preparation method according to claim 2, wherein the weight volume ratio of ubenimex to glacial acetic acid is 1: 2-10, and the volume ratio of glacial acetic acid to the low-polarity solvent is 1: 1-10; when the lower-grade solvent is a mixed solvent of ketones and esters, the volume ratio of the ketones to the esters is 1: 1-10.

7. The preparation method according to claim 2, wherein 0.5g of ubenimex is dissolved in 5ml of glacial acetic acid under stirring at room temperature, then 25ml of ethyl acetate is slowly added, the mixture is heated in a water bath to 50 ℃ and stirred uniformly, the mixture is filtered while hot, the filtrate is sealed, the mixture is kept standing at room temperature for 0.5 hour, and colorless blocky crystals are obtained after filtration.

8. Pharmaceutical composition containing a single crystal of ubenimex acetate according to claim 1, characterized in that said composition is a capsule or tablet.

Technical Field

The invention relates to the field of drug synthesis, in particular to a single crystal of ubenimex acetate and a preparation method and application thereof.

Background

Ubenimex is a compound isolated from a culture solution of streptomyces, competitively inhibits aminopeptidase b and leucine peptidase b, enhances the function of T cells, enhances the killing activity of NK cells, and stimulates the regeneration and differentiation of bone marrow cells by increasing the synthesis of colony stimulating factors. Ubenimex has the function of enhancing immunity, is a clinically unique CD13 antagonist, can comprehensively block the occurrence and development of various tumor cells, thereby inhibiting tumor metastasis and recurrence, inhibiting tumor neovascularization and tumor cell proliferation, promoting tumor cell apoptosis, and can be applied to patients with leukemia, multiple myeloma, myelodysplastic syndrome and hematopoietic stem cell transplantation in combination with chemotherapy and radiotherapy, and other solid tumors; and patients with other solid tumors such as lung cancer, gastric cancer, nasopharyngeal carcinoma, liver cancer, esophageal cancer, intestinal cancer, breast cancer, melanoma, bladder cancer, penile cancer, etc., and the immune function is low due to various factors, and the life quality of the patients can be effectively improved.

Acetate of ubenimex has molecular formula C₁₆H₂₄N₇O₄HOAC, structural formula:

the structural formula of ubenimex contains three chiral centers, and when the absolute configuration of ubenimex bulk drug prepared by chemical synthesis is determined during structure confirmation, a single crystal X-ray diffraction method is preferred, but firstly, a single crystal of a compound to be tested needs to be prepared for testing. In the prior art, US4786754 discloses alpha and beta crystal forms of ubenimex, US4786754 discloses a gamma crystal form, CN108892625A discloses a preparation method of a ubenimex gamma crystal form, CN 106631880B discloses an ubenimex crystal form and a preparation method thereof, and CN 103910648B discloses a crystal form of ubenimex hydrochloride, but the preparation method of the crystal form disclosed by the technology and the adopted crystal form detection method are known to be an X-ray powder diffraction method, the crystals of ubenimex or ubenimex hydrochloride prepared by the ubenimex or the ubenimex hydrochloride are not single crystals, and the crystal form sample is a polycrystal. Therefore, when preparing ubenimex bulk drug, in order to ensure that the prepared ubenimex has the correct spatial configuration, it is necessary to research that the absolute configuration of the prepared ubenimex single crystal is determined by X-ray single crystal diffraction.

Disclosure of Invention

The invention provides a single crystal of ubenimex acetate and a preparation method thereof, which are prepared by dissolving ubenimex in glacial acetic acid and then adding a low-polarity organic solvent for slow crystallization, wherein the required crystallization time is short, and the prepared single crystal has high purity, uniform distribution of internal particles and good stability.

The ubenimex acetate monocrystal provided by the invention is characterized in that the monocrystal is an orthorhombic system and a crystal axis of the monocrystal The included angle between the crystal planes α/° 90, β/° 90, γ/° 90, the unit lattice area V2024.63 (6), the number of molecules Z in the unit lattice 4, and the density d 1.209.

The ubenimex acetate single crystal provided by the invention is discovered under the condition that a single crystal cannot be prepared by adopting the prior art related to ubenimex crystal forms. When the ubenimex single crystal is prepared by a slow volatilization method after heating dissolution, the ubenimex single crystal cannot be prepared by dissolving water, adding methanol/ethanol/acetone to dilute the solution into solutions with different concentrations, standing the solution for crystallization, or adding different organic solvents (ethanol, ethyl acetate, tetrahydrofuran, isopropanol, acetone and the like) to dilute the solution into solutions with different concentrations after the methanol is dissolved, standing the solution for crystallization. The method comprises the steps of culturing a single crystal after salifying ubenimex, dissolving ubenimex in common acids such as hydrochloric acid, citric acid, tartaric acid and acetic acid, and adding an organic solvent for crystallization. The results showed that the dissolution with hydrochloric acid did not yield single crystals, citric acid and tartaric acid yielded only very small amounts of single crystals, and the dissolution in acetic acid alone allowed the crystallization culture to yield single crystals with uniform particle size distribution and high purity.

The preparation method of the ubenimex acetate monocrystal comprises the steps of dissolving ubenimex in glacial acetic acid at room temperature, slowly adding a low-polarity solvent, heating to 50-80 ℃, uniformly stirring until the solution is clear, filtering while the solution is hot, sealing the filtrate, standing at room temperature for 0.5-5 hours, and obtaining a colorless blocky monocrystal.

The lower solvent is selected from one or a mixed solvent of ketones and esters of dichloromethane, n-hexane, acetone, pentanone, ethyl acetate, butyl acetate and isopropyl acetate, preferably acetone, ethyl acetate or a mixture of acetone and ethyl acetate, and most preferably ethyl acetate.

The volume ratio of the weight (g) of the ubenimex to the glacial acetic acid (ml) is 1: 2-10, and the volume ratio of the glacial acetic acid to the low-polarity solvent is 1: 1-10; when the lower-grade solvent is a mixed solvent of ketones and esters, the volume ratio of the ketones to the esters is 1: 1-10.

The technological parameters and results of dissolving 0.5g of ubenimex in 5mL of glacial acetic acid and preparing single crystals by crystallization with different low-polarity solvents are shown in the following table:

the low-polarity solvent can be used for preparing single crystals. Under a microscope, when ethyl acetate is used as a crystallization solvent, the obtained single crystal is most uniform, so the preferable preparation method of the ubenimex acetate single crystal comprises the following steps: dissolving ubenimex 0.5g in glacial acetic acid 5ml under stirring at room temperature, slowly adding ethyl acetate 25ml, heating in water bath to 50 deg.C, stirring, filtering while hot, sealing the filtrate, standing at room temperature for 0.5 hr, and filtering to obtain colorless blocky crystal.

The ubenimex acetate monocrystal can be used for preparing a pharmaceutical composition, and the pharmaceutical composition is an oral preparation and can be tablets or capsules. The pharmaceutical composition is prepared from ubenimex acetate monocrystal, a filling agent, a disintegrating agent, a lubricating agent and other pharmaceutically acceptable auxiliary materials.

In the pharmaceutical composition prepared from the ubenimex acetate monocrystal, the ubenimex acetate content is 45-180 mg, preferably 119.5 mg; the filler is lactose, microcrystalline cellulose or mannitol, preferably lactose and microcrystalline cellulose, wherein the weight ratio of the ubenimex acetate to the filler is 1: 1-30; the disintegrating agent is selected from at least one of sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, pregelatinized starch, cross-linked sodium carboxymethyl cellulose and low-substituted hydroxypropyl cellulose, preferably sodium carboxymethyl starch, and the ratio of the disintegrating agent to ubenimex acetate is 1-3: 1; the lubricant is preferably magnesium stearate and/or aerosil, and the content of the lubricant is 1-2% of the total weight of ubenimex acetate and the filler.

A binder is also required to be added in the preparation by adopting a wet granulation method, and the binder is selected from hypromellose, polyvinylpyrrolidone and sodium carboxymethyl cellulose, and preferably sodium carboxymethyl starch.

The ubenimex acetate pharmaceutical composition provided by the invention has good dissolution rate and high bioavailability, and is suitable for enhancing immune function, adjuvant treatment of anticancer chemotherapy and radiotherapy, senile immune function deficiency and the like.

The ubenimex acetate single crystal is prepared by researching the absolute configuration of ubenimex to prepare the ubenimex single crystal sample, the single crystal is high in purity, can be used for determining the absolute configuration of ubenimex raw material medicines through X-ray single crystal diffraction, is good in solubility, can be used for preparing ubenimex acetate pharmaceutical compositions, and has clinical use value. The preparation method of the ubenimex acetate single crystal provided by the invention is short in crystallization time, and can be used for preparing the stable and uniform ubenimex acetate single crystal.

The invention is further described with reference to the drawings and the detailed description.

Drawings

FIG. 1 shows the X-ray single-crystal diffraction space structure diagram of ubenimex acetate single crystal

Detailed Description