Salt conversion method of utecartide hydrochloride bulk drug

Abstract

The invention relates to the technical field of medicine preparation, and discloses a salt conversion method of an itecartide hydrochloride bulk drug, which comprises the following steps: S1, preparing about 160mg of purified itecartide sample with qualified purity; and S2, loading the Itecartide sample onto the reversed-phase C18 preparative column through a semi-preparative high performance liquid chromatograph. In order to solve the problem that existing methods such as nanofiltration are tedious in operation, a purified sample with qualified purity firstly passes through a high performance liquid chromatograph, and then the content of chloride ions in the itecatide hydrochloride can be quantitatively controlled. In order to solve the problems that the chlorine ion content is difficult to control and salt type conversion is not thorough, a chromatography filler is selected as a reversed-phase C18 filler, an ammonium chloride solution with a certain concentration is used for completing ion exchange on a product, and then a hydrochloric acid aqueous solution with a certain concentration is used for completing salt conversion work through a high performance liquid chromatograph. And carrying out rotary evaporation and freeze-drying to obtain an entecatide hydrochloride finished product.

Description

Technical Field

[0001] The invention belongs to the technical field of drug preparation, and in particular relates to a salt conversion method of etelkatide hydrochloride raw material medicine. Background Art

[0002] SHPT is a common complication of chronic kidney disease (CKD) that seriously affects the prognosis and quality of life of patients. When renal function is impaired, factors such as decreased active vitamin D, hyperphosphatemia, and hypocalcemia stimulate the parathyroid glands to proliferate and secrete excessive parathyroid hormone (PTH). Increased PTH secretion can further cause calcium-phosphorus metabolism disorders, leading to osteitis fibrosa and mixed bone disease, bone and muscle pain, muscle weakness, and fractures. Severe SHPT patients may experience skin itching, hypercalcemia, heart disease, and Vascular disease, muscle necrosis, mental disorder and even death; on the one hand, etelkatide can increase the sensitivity of extracellular calcium ions by regulating the calcium-sensitive receptors of the parathyroid glands, thereby inhibiting the synthesis of PTH; on the other hand, it can upregulate the transcription and translation of parathyroid calcium-sensitive receptor mRNA and vitamin D receptor mRNA to inhibit the synthesis of PTH; due to its properties, etelkatide has excellent water solubility; the conventional salt conversion method has an early peak in the liquid phase, which is not conducive to the conversion and control of salt; and the original research has extremely strict requirements on the chloride ion content.

[0003] The methods currently mentioned on the market for controlling the chloride ion content of products are: 1. Concentrate the purified product with qualified purity by nanofiltration to 1 to 10 times the volume. After a single nanofiltration, add 5 to 10 times the volume of deionized water after concentration, and then repeat the nanofiltration concentration and dilution for more than three times. This method can control the chloride ion of the product.

[0004] 2. In addition, the difference in solubility between etelkatide trifluoroacetate and hydrochloride in isopropanol was used to convert etelkatide into hydrochloride by adding hydrochloric acid dropwise to the isopropanol solution of etelkatide trifluoroacetate.

[0005] However, the above method has certain problems: 1. Controlling the chloride ion content of the product by nanofiltration has certain defects: Since the chloride ion content of etelkatide hydrochloride is extremely strict, if it is controlled by nanofiltration, the chloride ion content in the product may be too high or too low, and it is difficult to accurately control it. And controlling the chloride ion content by nanofiltration requires repeated concentration-diluting with water and re-concentrating more than three times, which is cumbersome.

[0006] 2. The method of converting etelkatide into hydrochloride by adding hydrochloric acid to an isopropanol solution of etelkatide trifluoroacetate. The amount of hydrochloric acid added is excessive and needs to be repeatedly washed with isopropanol. Moreover, the etelkatide hydrochloride precipitated by this method is a viscous solid in isopropanol and cannot be thoroughly washed in subsequent isopropanol washing, resulting in excessive hydrochloric acid content. Some trifluoroacetates may also be incompletely converted, resulting in residual trifluoroacetic acid. Summary of the invention

[0007] In view of the problems existing in the prior art, the present invention provides a method for converting the salt of etelkatide hydrochloride raw material drug, which can overcome the above problems or at least partially solve the above problems.

[0008] The present invention is achieved by a method for converting the salt of etelkatide hydrochloride raw material, comprising: S1: preparing about 160 mg of purified etelkatide sample with qualified purity; S2: The eterkatide sample is loaded onto a reverse phase C18 preparative column by semi-preparative high performance liquid chromatography; S3, first use 100mmol / L ammonium chloride aqueous solution as A1 phase and acetonitrile as B phase with a gradient of 2%-2% to complete ion exchange; S4, then use 0.1% hydrochloric acid aqueous solution as phase A2, acetonitrile as phase B, and the gradient is 2%-2% to complete the salt conversion of the product; S5, collecting the salt conversion elution sample to obtain etelkatide hydrochloride salt conversion solution; S6. The etelkatide hydrochloride salt solution is concentrated by rotary evaporation and freeze-dried to obtain the etelkatide hydrochloride finished product.

[0009] Preferably, the etekatide sample is trifluoroacetate.

[0010] Compared with the prior art, the present invention has the following beneficial effects: 1. In order to solve the problem of complicated operation of existing methods such as nanofiltration, the present invention only requires the purified sample with qualified purity to be passed through a high performance liquid chromatograph to quantitatively control the chloride ion content of eterkatide hydrochloride.

[0011] 2. In order to solve the problems of difficulty in controlling chloride ion content and incomplete salt conversion, the present invention selects reverse phase C18 filler as the chromatography filler, uses a certain concentration of ammonium chloride solution to complete ion exchange of the product, and then uses a high performance liquid chromatograph and a certain concentration of hydrochloric acid aqueous solution to complete the salt conversion, and obtains the finished product of etelkatide hydrochloride by rotary evaporation and freeze drying. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 It is a schematic diagram of a flow chart provided by an embodiment of the present invention; Figure 2 is a schematic diagram of the elution gradient in S2 provided in an embodiment of the present invention; Figure 3 This is a diagram of the freeze-drying operation steps used in S3 of the embodiment of the present invention for concentrating the salt-converted sample under reduced pressure at 35±2°C. Specific implementation methods In order to further understand the content, features and effects of the present invention, the following embodiments are given as examples and described in detail with reference to the accompanying drawings.

[0014] The structure of the present invention is described in detail below in conjunction with the accompanying drawings.

[0015] like Figures 1 to 3 As shown, a salt conversion method of etelkatide hydrochloride raw material provided in an embodiment of the present invention includes: S1: preparing about 160 mg of etelkatide sample with qualified purity after purification; S2: loading the etelkatide sample onto a reverse phase C18 preparative column through a semi-preparative high performance liquid chromatograph; S3, first using 100 mmol / L ammonium chloride aqueous solution as A1 phase, acetonitrile as B phase, and a gradient of 2%-2% to complete ion exchange; S4, then using 0.1% hydrochloric acid aqueous solution as A2 phase, acetonitrile as B phase, and a gradient of 2%-2% to complete salt conversion of the product; S5, collecting the salt conversion elution sample to obtain the etelkatide hydrochloride salt conversion solution; S6, rotary evaporation and concentration of the etelkatide hydrochloride salt conversion solution, and freeze drying to obtain the etelkatide hydrochloride finished product. The etelkatide sample is trifluoroacetate.

[0016] Working principle of the present invention: When in use, the chromatographic filler is reverse phase C18, 10um, 120A, 21.2*250mm. First, about 160mg of purified and qualified etkadipide sample (trifluoroacetate) is loaded onto the reverse phase C18 preparative column through a Hanbang semi-preparative high performance liquid chromatograph; first, 100mmol / L ammonium chloride aqueous solution is used as the A1 phase, acetonitrile is used as the B phase, and the gradient is 2%-2% to complete the ion exchange. Then, 0.1% hydrochloric acid aqueous solution is used as the A2 phase, acetonitrile is used as the B phase, and the gradient is 2%-2% to complete the salt conversion of the product, and the salt conversion elution sample is collected to obtain the etkadipide hydrochloride salt conversion solution. The etkadipide hydrochloride salt conversion solution is subjected to rotary evaporation concentration and freeze drying to obtain the etkadipide hydrochloride finished product. The salt conversion sample is concentrated under reduced pressure at 35±2°C, and the chloride ion content of the obtained etkadipide hydrochloride product is 14.5%.

[0017] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.

[0018] The above description is only a preferred embodiment of the present invention, and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment as above, it is not used to limit the present invention. Any technician familiar with this patent will not depart from the scope of the technical solution of the present invention.

Claims

1. A salt conversion method for etelkatide hydrochloride raw material, characterized in that: include: S1: preparing about 160 mg of purified etelkatide sample with qualified purity; S2: The etelkatide sample is loaded onto a reverse phase C18 preparative column by semi-preparative high performance liquid chromatography; S3, first use 100mmol / L ammonium chloride aqueous solution as A1 phase and acetonitrile as B phase with a gradient of 2%-2% to complete ion exchange; S4, then use 0.1% hydrochloric acid aqueous solution as phase A2, acetonitrile as phase B, and the gradient is 2%-2% to complete the salt conversion of the product; S5, collecting the salt conversion elution sample to obtain etelkatide hydrochloride salt conversion solution; S6. The etelkatide hydrochloride salt solution is concentrated by rotary evaporation and freeze-dried to obtain the etelkatide hydrochloride finished product.

2. The salt conversion method of etelkatide hydrochloride raw material according to claim 1, characterized in that: The etkatide sample was trifluoroacetate salt.

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