A simple method for detecting the content of non-deuterated substances in deuterated drug deuterated remdesivir hydrobromide based on infrared spectrum

Abstract

The present application provides a simple method for detecting the content of non-deuterated substances in deuterated drug deuterated remdesivir hydrobromide based on infrared spectroscopy, which has many advantages compared with the existing nuclear magnetic resonance spectroscopy method: (1) no need for expensive, high-precision and high-tech analysis instruments, but infrared spectrometer commonly equipped by pharmaceutical production enterprises; (2) no need for sample preparation, more simple operation, and environment-friendly; (3) sample testing time is not more than 1 minute, realizing the rapid release of the drug in the production enterprise.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical analysis, specifically to a simple method for detecting the content of non-deuterated compounds in the deuterated drug deuterated remidevir hydrobromide based on infrared spectroscopy. Background Technology

[0002] Deuterium remdesivir hydrobromide, also known as VV116, with the chemical name (2R,3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazine-7-yl-5-deuterium)-2-cyano-5-[(isobutyryloxy)methyl]tetrahydrofuran-3,4-diylbis(2-methylpropionate) hydrobromide, is a viral RNA-dependent RNA polymerase inhibitor. It has been approved for marketing by the National Medical Products Administration for the treatment of mild to moderate COVID-19 infection in adults.

[0003] Deuterated remdesivir hydrobromide is a deuterated drug formed by replacing hydrogen (H) at specific metabolic sites in the compound structure with deuterium (D). This deuteration modification inhibits its oxidation and ring-opening degradation in vivo, improving bioavailability and antiviral activity. According to literature (Cell Research, 2021, 31(11):1212-1214) and patent CN114516875A, its deuteration synthesis strategy involves isotope exchange via an iodine-deuterium exchange reaction to deuterate the synthetic intermediate under the action of a catalyst and deuterium gas. This reaction condition is harsh, resulting in limited deuteration efficiency, and inevitably, undeuterated non-deuterated compounds are generated during the reaction. Therefore, the content of non-deuterated compounds in deuterated remdesivir hydrobromide is a key quality attribute affecting its efficacy and safety.

[0004] Deuterated drugs and their corresponding non-deuterated compounds have almost identical physicochemical properties. When using analytical techniques such as chromatography to determine the content or check for impurities in deuterated drugs, it is impossible to distinguish between the deuterated drugs and their corresponding non-deuterated compounds. Therefore, developing a quantitative analytical method for non-deuterated compounds in deuterated remidevir hydrobromide is of great significance for its production quality control.

[0005] Patent CN119310125A discloses a method for determining the non-deuterated content in deuterated remidevir hydrobromide using proton nuclear magnetic resonance (NMR) spectroscopy. This method boasts high accuracy and ease of operation. However, NMR spectrometers are extremely expensive and require highly skilled operators. Pharmaceutical manufacturers' quality control laboratories typically lack the necessary equipment, thus limiting the practical application of this method in pharmaceutical companies. Furthermore, proton NMR spectroscopy necessitates expensive deuterated organic solvents, further increasing testing costs and posing environmental hazards. The method also has a long testing time, requiring over 30 minutes per test.

[0006] Therefore, to ensure the safety and efficacy of deuterated remdesivir hydrobromide, there is an urgent need to develop a low-cost, short-time, and simpler and more practical non-deuterated compound determination method for the release of deuterated remdesivir hydrobromide in the quality control laboratories of pharmaceutical manufacturers. Summary of the Invention

[0007] The technical problem this invention aims to solve is to provide a simple and practical method for detecting the content of non-deuterated compounds in deuterated remdesivir hydrobromide. This method allows for the rapid release of deuterated remdesivir hydrobromide from the factory using analytical instruments routinely equipped in the quality control laboratories of pharmaceutical manufacturing companies.

[0008] The technical solution of the present invention to solve the above-mentioned technical problems is as follows:

[0009] A simple method for detecting the content of non-deuterated compounds in the deuterated drug deuterated remidevir hydrobromide based on infrared spectroscopy includes the following steps:

[0010] (1) A series of deuterated remdesivir hydrobromide samples with different non-deuterated content were prepared by mixing deuterated remdesivir hydrobromide reference standard and non-deuterated reference standard.

[0011] (2) Divide these samples into a calibration set and a validation set;

[0012] (3) Infrared spectral data of the calibration set and the validation set were collected using an infrared spectrometer;

[0013] (4) Perform spectral preprocessing on the collected infrared spectral data;

[0014] (5) Select a specific infrared wavenumber as a characteristic variable and use partial least squares regression to establish a calibration model;

[0015] (6) Use cross-validation or validation sets to validate the established calibration model;

[0016] (7) By importing the infrared spectrum of the sample to be tested with unknown non-deuterated content into the calibration model, the content of non-deuterated content in the sample to be tested can be calculated.

[0017] The chemical structures of the aforementioned deuterated remidevir hydrobromide and its corresponding non-deuterated derivatives are shown below:

[0018]

[0019] Furthermore, a method for detecting the content of non-deuterated compounds in the deuterated drug deuterated remidevir hydrobromide includes the following steps:

[0020] S1. A series of deuterated remdesivir hydrobromide samples containing 0.8% to 5.0% of the non-deuterated compound were prepared by mixing deuterated remdesivir hydrobromide reference standard and non-deuterated reference standard.

[0021] S2. Divide these samples into a calibration set and a validation set in a ratio of 4:1 to 1:1, specifically 2:1.

[0022] S3. Use an infrared spectrometer to collect infrared spectral data for the calibration set and the validation set.

[0023] S31. The infrared spectrum acquisition mode is attenuated total internal reflection;

[0024] S32, infrared spectrum acquisition range is 4000~550cm -1 The resolution is 4cm. -1 The number of scans ranges from 16 to 64, but can be as many as 32.

[0025] S4. Perform spectral preprocessing on the collected infrared spectral data, specifically using the maximum-minimum normalization method.

[0026] S5. Select a specific infrared wavenumber as a feature variable and use partial least squares regression to establish a calibration model.

[0027] S51, The selected infrared wavenumber is 1600–550 cm⁻¹ -1 Specifically, it can be 1300-1040cm -1 900~690cm -1Preferably, the values ​​are 692.3, 696.7, 716.9, 717.4, 717.9, 718.4, 718.8, 719.3, 719.8, 720.3, 720.8, 767.5, 768.0, 768.5, 769.0, 770.4, 771.9, 772.4, 772.8, 773.8, 774.8, 7 75.2,794.5,795.5,796.0,797.4,797.9,798.4,798.9,799.8,802.2,802.7,803.7,804.2,804.7,806.1,821.0,821.5,822.5,823.0,823.5,824.4,834.1, 835.5,836.0,836.5,837.4,1068.9,1172.5,1173.5,1174.0,1174.4,1174.9,1175.4,1222.2,1222.6,1223.1,1223.6,1224.1,1225.1,1226.0,1226.5,12 27.0,1227.5,1228.4,1228.9,1229.9,1230.8,1231.3,1231.8,1279.5,1281. 5,1281.9,1283.4,1295.4,1295.9,1296.9,1297.4,1298.3,1299.8,1300.3cm -1 .

[0028] S6. Use cross-validation or validation sets to validate the established calibration model.

[0029] S7. By importing the infrared spectrum of the sample to be tested with unknown non-deuterated content into the calibration model, the content of non-deuterated content in the sample can be calculated.

[0030] The spectral preprocessing method and the calibration model established using characteristic infrared wavenumbers selected in this invention can eliminate irrelevant information and redundant or nonlinear variables, resulting in a smaller deviation between the calculated and actual values ​​of non-deuterated compounds. Specifically, the coefficients of determination (R²) of the calibration set and the validation set... 2 All values ​​were greater than 0.99; the root mean square error (RMSE) was less than 0.1%. Therefore, this significantly increased the quantitative accuracy of infrared spectroscopy, enabling its successful application in the detection of non-deuterated compounds in the deuterated drug deuterated remidevir hydrobromide.

[0031] In this invention, if there is a conflict between the Chinese name and the structural formula of a compound, the structural formula shall prevail.

[0032] The beneficial effects of this invention are as follows: This invention provides a simple and practical method for detecting the content of non-deuterated compounds in the deuterated drug deuterated remidevir hydrobromide. Compared with the existing nuclear magnetic resonance spectroscopy method, this method has many advantages: (1) It does not require expensive and sophisticated analytical instruments, but uses an infrared spectrometer that is routinely equipped by pharmaceutical manufacturers; (2) It does not require sample preparation, making the operation simpler and more environmentally friendly; (3) The sample testing time is less than 1 minute, enabling the rapid release of the drug from the manufacturer. Attached Figure Description

[0033] Figure 1 The spectrum of the deuterium remidevir hydrobromide sample in Example 1 after maximum-minimum normalization preprocessing and selection of modeling wavenumber (red area);

[0034] Figure 2 The image shows the test spectrum of the deuterium remidevir hydrobromide sample after maximum-minimum normalization preprocessing and selection of modeling wavenumber (red dot) in Example 2. Detailed Implementation

[0035] The present invention is illustrated below with reference to examples, but is not intended to limit the invention. Any simple substitutions or modifications made to the present invention by those skilled in the art are within the scope of the technical solutions protected by this invention.

[0036] Example 1:

[0037] Since the actual content of existing samples is between 0.8% and 2%, the sample range we use for modeling is between 0.8% and 5.0%.

[0038] Twenty-one samples of deuterated remdesivir hydrobromide with non-deuterated compound contents of 4.96%, 4.67%, 4.24%, 3.8%, 3.6%, 3.17%, 2.99%, 2.71%, 2.41%, 2.07%, 1.78%, 1.51%, 0.87%, 1.09%, 4.85%, 4.26%, 3.11%, 2.72%, 2.08%, 1.05%, and 0.98% were prepared by mixing deuterated remdesivir hydrobromide reference standards and non-deuterated compound reference standards. The first 14 samples were used as the calibration set, and the last 7 samples were used as the validation set.

[0039] Infrared spectral data of the calibration and validation sets were acquired using an infrared spectrometer. The acquisition mode was attenuated total reflectance, and the acquisition range was 4000–550 cm⁻¹. -1 The resolution is 4cm. -1 The number of scans was 32, and each sample was collected 3 times.

[0040] The 63 infrared spectra acquired were preprocessed using the maximum-minimum normalization method, and the spectral ranges of 1300–1040 cm⁻¹ and 900–690 cm⁻¹ were selected. -1 To model the band, a calibration model was established using partial least squares regression. The results showed that the absolute coefficients of the calibration set were 0.99619, the root mean square error was 0.07747%, and the relative errors between the calculated and actual values ​​were both less than 10%.

[0041] The accuracy of the established calibration model was verified using a validation set. The results showed an absolute coefficient of 0.99673, a root mean square error of 0.07879%, and relative errors between the calculated and actual values ​​of less than 10%.

[0042] The infrared spectrum of the sample to be tested with unknown non-deuterated content is imported into the calibration model to calculate the content of non-deuterated substances in the sample.

[0043] Example 2:

[0044] Select 692.3, 696.7, 716.9, 717.4, 717.9, 718.4, 718.8, 719.3, 719.8, 720.3, 720.8, 767.5, 768.0, 768.5, 769.0, 770.4, 771.9, 772.4, 772.8, 773.8, 774.8, 77 5.2,794.5,795.5,796.0,797.4,797.9,798.4,798.9,799.8,802.2,802.7,803.7,804.2,804.7,806.1,821.0,821.5,822.5,823.0,823.5,824.4,834.1,8 35.5,836.0,836.5,837.4,1068.9,1172.5,1173.5,1174.0,1174.4,1174.9,1175.4,1222.2,1222.6,1223.1,1223.6,1224.1,1225.1,1226.0,1226.5,122 7.0,1227.5,1228.4,1228.9,1229.9,1230.8,1231.3,1231.8,1279.5,1281. 5,1281.9,1283.4,1295.4,1295.9,1296.9,1297.4,1298.3,1299.8,1300.3cm -1 A total of 81 wavenumbers were used as modeling wavenumbers. A calibration model was established using partial least squares regression. Other methods and parameters were the same as in Example 1.

[0045] The absolute coefficients of the calibration set of the calibration model were 0.99844, the root mean square error was 0.04960%, and the relative errors between the calculated and actual values ​​were both less than 5%.

[0046] Cross-validation was used to verify the accuracy of the established calibration model. The results showed an absolute coefficient of 0.99698, a root mean square error of 0.06900%, and relative errors between calculated and actual values ​​of less than 7%. A validation set was also used to verify the accuracy of the established calibration model. The results showed an absolute coefficient of 0.99763, a root mean square error of 0.06713%, and relative errors between calculated and actual values ​​of less than 9%.

[0047] The infrared spectrum of the sample to be tested with unknown non-deuterated content is imported into the calibration model to calculate the content of non-deuterated substances in the sample.

[0048] The above description is only a preferred embodiment of the present invention. It should be noted that those skilled in the art can make several modifications and improvements without departing from the inventive concept of the present invention, and these all fall within the protection scope of the present invention.

Claims

1. A simple method for detecting the content of non-deuterated compounds in the deuterated drug deuterated remidevir hydrobromide based on infrared spectroscopy, characterized in that, Includes the following steps: (1) A series of deuterated remdesivir hydrobromide samples with different non-deuterated content were prepared by mixing deuterated remdesivir hydrobromide reference standard and non-deuterated reference standard; (2) Divide these samples into a calibration set and a validation set; (3) Infrared spectral data of the calibration set and the validation set were collected using an infrared spectrometer; (4) Perform spectral preprocessing on the collected infrared spectral data; the maximum-minimum normalization method is used for preprocessing in step (4); (5) Select specific infrared wavenumbers as feature variables and establish a calibration model using partial least squares regression; the infrared wavenumbers selected in step (5) are 692.3, 696.7, 716.9, 717.4, 717.9, 718.4, 718.8, 719.3, 719.8, 720.3, 720.8, 767.5, 768.0, 768.5, 769.0, 770.4, 771.9, 772.4, 772.8, 773.8, 774.8, 775.2, 794.5, 795.5, 796.0, 797.4, 797.9, 798.4, 798.9, 799.8,802.2, 802.7, 803.7, 804.2, 804.7, 806.1, 821.0, 821.5, 822.5, 823.0, 823.5,824.4, 834.1, 835.5, 836.0, 836.5, 837.4, 1068.9, 1172.5, 1173.5, 1174.0,1174.4, 1174.9, 1175.4, 1222.2, 1222.6, 1223.1, 1223.6, 1224.1, 1225.1,1226.0, 1226.5, 1227.0, 1227.5, 1228.4, 1228.9, 1229.9, 1230.8, 1231.3,1231.8, 1279.5, 1281.5, 1281.9, 1283.4, 1295.4, 1295.9, 1296.9, 1297.4,1298.3, 1299.8, 1300.3cm -1 ; (6) Validate the established calibration model using cross-validation or a validation set; (7) By importing the infrared spectrum of the sample to be tested with unknown non-deuterated content into the calibration model, the content of non-deuterated content in the sample to be tested can be calculated. The chemical structures of the aforementioned deuterated remidevir hydrobromide and its corresponding non-deuterated derivatives are shown below: 。 2. The method according to claim 1, characterized in that, The sample in step (1) is a deuterated remidevir hydrobromide sample containing 0.8% to 5.0% of non-deuterated compounds.

3. The method according to claim 1, characterized in that, In step (2), the samples are divided into a calibration set and a validation set in a ratio of 4:1 to 1:

1.

4. The method according to claim 3, characterized in that, In step (2), the samples are divided into a calibration set and a validation set in a 2:1 ratio.

5. The method according to claim 1, characterized in that, In step (3), the infrared spectrum acquisition mode is attenuated total internal reflection, and the infrared spectrum acquisition range is 4000~550 cm⁻¹. -1 The resolution is 4 cm. -1 The number of scans is 16 to 64.

6. The method according to claim 5, characterized in that, The number of scans was 32.

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