Simple method for detecting content of non-deuterated substances in deuterated drug deurimidevir hydrobromide based on infrared spectrum

Abstract

Compared with the existing nuclear magnetic resonance spectroscopy, the simple method for detecting the content of the non-deuterated substances in the deuterated drug deuteremidevir hydrobromide based on the infrared spectrum has the advantages that (1) an expensive, high-quality and advanced analytical instrument is not needed, and an infrared spectrometer conventionally equipped by a pharmaceutical production enterprise is adopted; (2) sample preparation is not needed, the operation is simpler and more convenient, and the method is environment-friendly; and (3) the test time of the sample does not exceed 1 minute, so that the medicine can be quickly delivered and released in a production enterprise.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical analysis, and particularly to a simple method for detecting the content of non-deuterated substances in deuterated drug remdesivir hydrobromide based on infrared spectroscopy. Background Art

[0002] Remdesivir hydrobromide, also known as VV116, with the chemical name of (2R,3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl-5-deuterio)-2-cyano-5-[(2-methylpropanoyl)oxy]methyltetrahydrofuran-3,4-diyl bis(2-methylpropanoate) hydrobromide, is a viral RNA-dependent RNA polymerase inhibitor, which has been approved by the National Medical Products Administration for marketing and is used for the treatment of mild to moderate novel coronavirus infection in adults.

[0003] Remdesivir hydrobromide is a deuterated drug formed by replacing hydrogen (H) at specific metabolic sites in the compound structure with deuterium (D). Through this deuteration modification, its oxidation and ring-opening degradation in vivo can be inhibited, and its bioavailability and antiviral activity can be improved. According to the literature (Cell Research, 2021, 31(11): 1212-1214) and patent CN114516875A reports, its deuteration synthesis strategy is an isotope exchange method of deuterating the synthesis intermediate through iodine-deuterium exchange reaction under the action of a catalyst and deuterium gas. The reaction conditions are harsh, the deuteration efficiency is limited, and the corresponding non-deuterated substances will inevitably be generated during the reaction process. Therefore, the content of non-deuterated substances in remdesivir hydrobromide is a key quality attribute affecting its effectiveness and safety.

[0004] Deuterated drugs and their corresponding non-deuterated substances have almost the same physical and chemical properties. When using analytical techniques such as chromatography to determine the content or check for impurities of deuterated drugs, deuterated drugs and the corresponding non-deuterated substances cannot be distinguished. Therefore, developing a quantitative analysis method for non-deuterated substances in remdesivir hydrobromide is of great significance for its production quality control.

[0005] Patent CN119310125A discloses a method for determining the content of non-deuterated substances in remdesivir hydrobromide by using hydrogen nuclear magnetic spectroscopy. This method has high accuracy and is easy to operate. However, nuclear magnetic resonance spectrometers are very expensive and have high professional technical requirements for operators. The quality control laboratories of pharmaceutical production enterprises usually do not have the detection conditions, so the practical application of this method in pharmaceutical production enterprises is limited. In addition, hydrogen nuclear magnetic spectroscopy also requires expensive deuterated organic solvents, which further increases the detection cost and is harmful to the environment. The test time of this method is also relatively long, and each test takes more than 30 minutes.

[0006] Therefore, to ensure the safety and effectiveness of deuterated remdesivir hydrobromide, there is an urgent need to develop a non-deuterated substance determination method with low detection cost, short time, and greater simplicity and practicality for the release inspection of deuterated remdesivir hydrobromide in the quality control laboratories of pharmaceutical manufacturing enterprises. Summary of the Invention

[0007] The technical problem to be solved by the present invention is to provide a simple and practical method for detecting the content of non-deuterated substances in deuterated remdesivir hydrobromide. This method can achieve the rapid release of deuterated remdesivir hydrobromide by using the analytical instruments commonly equipped in the quality control laboratories of pharmaceutical manufacturing enterprises.

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

[0009] A simple method for detecting the content of non-deuterated substances in deuterated drug deuterated remdesivir hydrobromide based on infrared spectroscopy, comprising the following steps:

[0010] (1) By mixing the reference substance of deuterated remdesivir hydrobromide and the reference substance of non-deuterated substance, a series of deuterated remdesivir hydrobromide samples with different non-deuterated substance contents are prepared;

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

[0012] (3) Use an infrared spectrometer to collect the infrared spectral data of the calibration set and the validation set;

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

[0014] (5) Select specific infrared wave numbers as characteristic variables and establish a calibration model using partial least squares regression;

[0015] (6) Validate the established calibration model using cross-validation or the validation set;

[0016] (7) Import the infrared spectrum of the test sample with unknown non-deuterated substance content into the calibration model, and the content of non-deuterated substance in the test sample can be calculated;

[0017] The deuterated remdesivir hydrobromide and the corresponding non-deuterated substance have the following chemical structures:

[0018]

[0019] Furthermore, a method for detecting the content of non-deuterated substances in deuterated drug deuterated remdesivir hydrobromide comprises the following steps:

[0020] S1. By mixing the reference substance of deuterated remdesivir hydrobromide and the reference substance of non-deuterated substance, a series of deuterated remdesivir hydrobromide samples with non-deuterated substance contents of 0.8% - 5.0% are prepared.

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

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

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

[0024] S32. The infrared spectrum acquisition range is 4000 - 550 cm -1 , and the resolution is 4 cm -1 , and the number of scans is 16 - 64 times, specifically it can be 32 times.

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

[0026] S5. Select specific infrared wavenumbers as characteristic variables and establish a calibration model using partial least squares regression.

[0027] S51. The selected infrared wavenumbers are 1600 - 550 cm -1 , specifically it can be 1300 - 1040 cm -1 , 900 - 690 cm -1, preferably 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, 1300.3 cm -1 。

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

[0029] S7. Import the infrared spectrum of the sample to be measured with unknown non-deuterated content into the calibration model, and the content of non-deuterated substances in the sample to be measured can be calculated.

[0030] The calibration model established by using the spectral preprocessing method and characteristic infrared wave numbers screened by the present invention can eliminate irrelevant information and redundant or non-linear variables, making the deviation between the calculated value and the actual value of non-deuterated substances small. Among them, the determination coefficients (R 2 ) of the calibration set and the validation set are both greater than 0.99; the root mean square error (RMSE) is both less than 0.1%. Therefore, the quantitative accuracy of the infrared spectroscopy is greatly increased, making it successfully applied to the detection of the content of non-deuterated substances in the deuterated drug deoxydremetrivir hydrobromide.

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

[0032] The beneficial effects of the present invention are as follows: The present invention provides a simple and practical method for detecting the content of non-deuterated substances in deuterated drug deuterated remdesivir 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, and uses an infrared spectrometer commonly equipped in pharmaceutical production enterprises; (2) It does not require sample preparation, is more convenient to operate, and is environmentally friendly; (3) The sample testing time does not exceed 1 minute, enabling the rapid release of this drug from the production enterprise. Description of the Drawings

[0033] Figure 1 It is the test spectrum of the deuterated remdesivir hydrobromide sample after maximum-minimum normalization preprocessing and selection of the modeling wavenumber (red area) in Example 1;

[0034] Figure 2 It is the test spectrum of the deuterated remdesivir hydrobromide sample after maximum-minimum normalization preprocessing and selection of the modeling wavenumber (red dots) in Example 2. Detailed Embodiments

[0035] The following examples illustrate the present invention, but do not limit the present invention. In the art, simple substitutions or improvements made by those skilled in the art to the present invention fall within the scope of the technical solutions protected by the present invention.

[0036] Example 1:

[0037] Since the contents of existing actual samples are all within 0.8% - 2%, the sample range we used for modeling is within 0.8% - 5.0%.

[0038] By mixing the deuterated remdesivir hydrobromide reference substance and the non-deuterated substance reference substance, 21 deuterated remdesivir hydrobromide samples with non-deuterated substance 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%, 0.98% were prepared. Among them, the first 14 samples were used as the calibration set, and the last 7 samples were used as the validation set samples.

[0039] Use an infrared spectrometer to collect the infrared spectral data of the calibration set and validation set samples. The collection mode is attenuated total reflection, the collection range is 4000 - 550 cm -1 , the resolution is 4 cm -1 , the number of scans is 32 times, and each sample is collected 3 times repeatedly.

[0040] The 63 collected infrared spectra were preprocessed spectroscopically according to the maximum-minimum normalization method, and 1300 - 1040 cm-1 and 900 - 690 cm -1 were selected as the modeling bands, and a calibration model was established by partial least squares regression. As a result, the absolute coefficient of the calibration set of the calibration model was 0.99619, the root mean square error was 0.07747%, and the relative errors between the calculated values and the actual values were all less than 10%.

[0041] The validation set was used to verify the accuracy of the established calibration model. As a result, the absolute coefficient was 0.99673, the root mean square error was 0.07879%, and the relative errors between the calculated values and the actual values were all less than 10%.

[0042] The infrared spectrum of the sample to be tested with unknown non-deuterated content was imported into this calibration model, and the content of non-deuterated substances in the sample to be tested was calculated.

[0043] Example 2:

[0044] 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, 1300.3 cm -1 A total of 81 wavenumbers were selected as the modeling wavenumbers, and a calibration model was established by partial least squares regression. Other method parameters were the same as those in Example 1.

[0045] The absolute coefficient of the calibration set of the result correction model is 0.99844, the root mean square error is 0.04960%, and the relative error between the calculated value and the actual value is less than 5%.

[0046] The accuracy of the established calibration model was cross-validated by cross-validation. The result absolute coefficient was 0.99698, the root mean square error was 0.06900%, and the relative error between the calculated value and the actual value was less than 7%. The accuracy of the established calibration model was verified using a validation set. The result absolute coefficient was 0.99763, the root mean square error was 0.06713%, and the relative error between the calculated value and the actual value was less than 9%.

[0047] The infrared spectrum of the sample to be tested with unknown non-deuterated content was imported into this calibration model, and the content of non-deuterated substances in the sample to be tested was calculated.

[0048] The above are only the preferred embodiments of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the creative concept of the present invention, several modifications and improvements can still be made, and these all belong to the protection scope of the present invention.

Claims

1. A simple method for detecting the content of non-deuterated substances in deuterated drug remdesivir hydrobromide based on infrared spectroscopy, characterized in that, It includes the following steps: (1) Prepare a series of deuterated remdesivir hydrobromide samples with different non-deuterated compound contents by mixing deuterated remdesivir hydrobromide reference substance and non-deuterated compound reference substance; (2) Divide these samples into a calibration set and a validation set; (3) Collect the infrared spectral data of the calibration set and the validation set using an infrared spectrometer; (4) Perform spectral preprocessing on the collected infrared spectral data; (5) Select specific infrared wave numbers as characteristic variables and establish a calibration model using partial least squares regression; (6) Validate the established calibration model using cross-validation or the validation set; (7) Import the infrared spectrum of the sample to be tested with unknown non-deuterated compound content into the calibration model, and the content of the non-deuterated compound in the sample to be tested can be calculated; The chemical structures of the deuterated remdesivir hydrobromide and the corresponding non-deuterated compound are shown as follows:

2. The method according to claim 1, wherein In the step (1), the sample is a deuterated remdesivir hydrobromide sample containing 0.8% - 5.0% of non-deuterated compound.

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

1.

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

1.

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

6. The method according to claim 5, wherein The number of scans is 32 times.

7. The method according to claim 1, characterized in that, In the step (4), maximum-minimum normalization method is used for preprocessing.

8. The method according to claim 1, characterized in that The selected infrared wave number in the step (5) is 1600-550 cm -1 .

9. The method according to claim 8, characterized in that The infrared wave numbers selected in the step (5) are 1300 - 1040 cm -1 , 900 - 690 cm -1 .

10. The method according to claim 9, characterized in that, The selected infrared wave numbers 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, 1300.3 cm -1 .

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