Preparation method and application of thyroxine-releasing hormone

Abstract

The invention provides a preparation method and application of a thyroxine release hormone, and relates to the technical field of polypeptide medicine preparation.The preparation method of the thyroxine release hormone comprises the steps that in the step of preparing Z-pGlu-His-Pro-NH2, acetonitrile is used for dissolving a concentrated product, crystals are separated out after low-temperature standing, and the Z-pGlu-His-Pro-NH2 is obtained; the conventional mode of recrystallization after two-time diethyl ether precipitation in the prior art is avoided, and the process period is short. In addition, a small amount of acetonitrile is used for dissolution, and reagents such as diethyl ether with a low flash point are not used. The method can be carried out in general areas and has no special requirements on plants. No safety risk exists, and industrialization can be achieved.

Description

technical field

[0001] The invention relates to the technical field of polypeptide drug preparation, in particular to a preparation method and application of thyrotropin-releasing hormone. Background technique

[0002] Thyrotropin releasing hormone (thyrotropin releasing hormone), a peptide hormone, its chemical structure is pGLU-His-Pro-NH 2 . It is synthesized and secreted by some nerve cells in the paraventricular nucleus of the hypothalamus. Delivered to the adenohypophysis via the pituitary portal system. Consists of pyroglutamic acid, histidine and prolinamide. Thyrotropin-releasing hormone can promote the pituitary gland to secrete thyroid-stimulating hormone (TSH), and affect the secretion of thyroid through TSH. In addition, thyrotropin-releasing hormone also has the physiological function of promoting the secretion of lactogen, oxytocin and roller pressin, as well as affecting the functions of the brain, such as anti-depression, changing sleep wakefulness and b...

Examples

preparation example Construction

[0079] In the step of preparing Z-pGlu-His-OH provided by the present invention, a large amount of purified water is used in the later stage to fully remove Cl - And His impurities, so that the next step to form Z-pGlu-His-Pro-NH 2 The reaction rate is greatly improved, and the target peptide content is increased. The reaction solution obtained before concentration under reduced pressure was filtered to remove the main impurity DCU. After a small amount of acetonitrile is dissolved, the concentration and purity of the target peptide in the acetonitrile solution are relatively high, which meets the basic requirements of the crystallization process.

[0080] In some preferred embodiments, hydrogenolysis of Z-pGlu-His-Pro-NH 2 The steps include:

[0081] To Z-pGlu-His-Pro-NH 2 H in the solution 2 , was detected by TLC to complete the reaction, filtered, and concentrated to dryness under reduced pressure to obtain the thyrotropin-releasing hormone.

[0082] In order to furth...

Embodiment 1

[0101] Example 1 Parameter optimization and screening for the preparation of Z-pGlu-ONB

[0102] An intermediate product was obtained which was dissolved in ethyl acetate.

[0103] Experiment 1: 4% KHCO 3 solution and 5% NaCl solution were washed three times each, Na 2 SO 4 Dry for 8 to 24 hours, filter, wash with ethyl acetate, and recrystallize petroleum ether.

[0104] Experiment 2: 4% KHCO 3 solution and 10% NaCl solution were washed three times each, Na 2 SO 4 Dry for 8 to 24 hours, filter, wash with ethyl acetate, and recrystallize petroleum ether.

[0105] Experiment 3: 8% KHCO 3 solution and 5% NaCl solution were washed three times each, Na 2 SO 4 Dry for 8 to 24 hours, filter, wash with ethyl acetate, and recrystallize petroleum ether.

[0106] Experiment 4: 8% KHCO 3 solution and 10% NaCl solution were washed three times each, Na 2 SO 4 Dry for 8 to 24 hours, filter, wash with ethyl acetate, and recrystallize petroleum ether.

[0107] Experiment 5: ...

Embodiment 2

[0114] Embodiment 2 docking dipeptide procedure carries out monitoring reaction

[0115]

[0116] Experiment 8: Weigh 19.43 grams of L-His, dissolve in 325ml of water, and add KHCO 3 14.54 g was stirred to dissolve.

[0117] Take 46 grams of the product Z-pGlu-ONB obtained in Experiment 7, dissolve it in 380ml of acetonitrile, add it to the above-mentioned L-His-OK aqueous solution, and stir at 30°C for 24 hours. After the Z-pGlu-ONB point disappears under TLC monitoring, at 40°C, Under the condition of -0.09MPa~-0.1MPa, concentrate under reduced pressure to a small volume, adjust the pH to about 6 with 2mol / L HCl under ice bath cooling, place at 2~10℃ for 8~24 hours, filter, and wash the product with ice-purified water to no Cl - and His, set P 2 o 5 After drying in a desiccator, 48.4 g of the product was obtained, with a yield of 73%.

[0118] Experiment 9: Weigh 10 grams of L-His, dissolve in 162ml of water, and add KHCO 3 7.2 g was stirred to dissolve.

[0119]...

technical field

[0001] The invention relates to the technical field of polypeptide drug preparation, in particular to a preparation method and application of thyrotropin-releasing hormone. Background technique

[0002] Thyrotropin releasing hormone (thyrotropin releasing hormone), a peptide hormone, its chemical structure is pGLU-His-Pro-NH 2 . It is synthesized and secreted by some nerve cells in the paraventricular nucleus of the hypothalamus. Delivered to the adenohypophysis via the pituitary portal system. Consists of pyroglutamic acid, histidine and prolinamide. Thyrotropin-releasing hormone can promote the pituitary gland to secrete thyroid-stimulating hormone (TSH), and affect the secretion of thyroid through TSH. In addition, thyrotropin-releasing hormone also has the physiological function of promoting the secretion of lactogen, oxytocin and roller pressin, as well as affecting the functions of the brain, such as anti-depression, changing sleep wakefulness and b...

Examples

preparation example Construction

[0079] In the step of preparing Z-pGlu-His-OH provided by the present invention, a large amount of purified water is used in the later stage to fully remove Cl - And His impurities, so that the next step to form Z-pGlu-His-Pro-NH 2 The reaction rate is greatly improved, and the target peptide content is increased. The reaction solution obtained before concentration under reduced pressure was filtered to remove the main impurity DCU. After a small amount of acetonitrile is dissolved, the concentration and purity of the target peptide in the acetonitrile solution are relatively high, which meets the basic requirements of the crystallization process.

[0080] In some preferred embodiments, hydrogenolysis of Z-pGlu-His-Pro-NH 2 The steps include:

[0081] To Z-pGlu-His-Pro-NH 2 H in the solution 2 , was detected by TLC to complete the reaction, filtered, and concentrated to dryness under reduced pressure to obtain the thyrotropin-releasing hormone.

[0082] In order to furth...

Embodiment 1

[0101] Example 1 Parameter optimization and screening for the preparation of Z-pGlu-ONB

[0102] An intermediate product was obtained which was dissolved in ethyl acetate.

[0103] Experiment 1: 4% KHCO 3 solution and 5% NaCl solution were washed three times each, Na 2 SO 4 Dry for 8 to 24 hours, filter, wash with ethyl acetate, and recrystallize petroleum ether.

[0104] Experiment 2: 4% KHCO 3 solution and 10% NaCl solution were washed three times each, Na 2 SO 4 Dry for 8 to 24 hours, filter, wash with ethyl acetate, and recrystallize petroleum ether.

[0105] Experiment 3: 8% KHCO 3 solution and 5% NaCl solution were washed three times each, Na 2 SO 4 Dry for 8 to 24 hours, filter, wash with ethyl acetate, and recrystallize petroleum ether.

[0106] Experiment 4: 8% KHCO 3 solution and 10% NaCl solution were washed three times each, Na 2 SO 4 Dry for 8 to 24 hours, filter, wash with ethyl acetate, and recrystallize petroleum ether.

[0107] Experiment 5: ...

Embodiment 2

[0114] Embodiment 2 docking dipeptide procedure carries out monitoring reaction

[0115]

[0116] Experiment 8: Weigh 19.43 grams of L-His, dissolve in 325ml of water, and add KHCO 3 14.54 g was stirred to dissolve.

[0117] Take 46 grams of the product Z-pGlu-ONB obtained in Experiment 7, dissolve it in 380ml of acetonitrile, add it to the above-mentioned L-His-OK aqueous solution, and stir at 30°C for 24 hours. After the Z-pGlu-ONB point disappears under TLC monitoring, at 40°C, Under the condition of -0.09MPa~-0.1MPa, concentrate under reduced pressure to a small volume, adjust the pH to about 6 with 2mol / L HCl under ice bath cooling, place at 2~10℃ for 8~24 hours, filter, and wash the product with ice-purified water to no Cl - and His, set P 2 o 5 After drying in a desiccator, 48.4 g of the product was obtained, with a yield of 73%.

[0118] Experiment 9: Weigh 10 grams of L-His, dissolve in 162ml of water, and add KHCO 3 7.2 g was stirred to dissolve.

[0119]...

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