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Polyethylene glycol-modified calcitonin

A kind of polyethylene glycol and calcitonin technology, applied in the field of polyethylene glycol modified calcitonin

Inactive Publication Date: 2010-01-06
SHANGHAI INST OF PHARMA IND +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Structural modifications of calcitonin partially solve all the defects of calcitonin itself, but some introduce new problems

Method used

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  • Polyethylene glycol-modified calcitonin
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  • Polyethylene glycol-modified calcitonin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Synthesis of Salmon Calcitonin Modified by Methoxypolyethylene Glycol Succinimidyl Propionate 5000 (mPEG-SPA-5000)

[0045] Choice of reaction temperature: Take 2ml of 1.0mg / ml salmon calcitonin solution, add 2ml of phosphate buffer to make the pH of the solution 7.0, then add 15mg of mPEG-SPA-5000 solid, dissolve, mix well, each take 0.8 ml was placed in 4 test tubes with stoppers, and then placed at 4°C, 10°C, 25°C and 37°C for 30 minutes respectively, and then 5 mg of glycine was added to terminate the reaction. The modification rate of salmon calcitonin modified with single polyethylene glycol (one molecule of polyethylene glycol chain connected to one salmon calcitonin molecule, hereinafter referred to as polyethylene glycol modified salmon calcitonin) was compared to determine the modification conditions. The results show that the PEG-modified salmon calcitonin can be obtained at these temperatures, and the modification rate is the highest at 25°C. See Table 1 for...

Embodiment 2

[0055] Purification and Identification of Polyethylene Glycol Modified Salmon Calcitonin

[0056] Take 10ml of 1.0mg / ml salmon calcitonin solution, add 10ml of phosphate buffer to make the pH of the solution 7.0, then add 44mg of mPEG-SPA-5000 solid, dissolve, mix well, react at 25°C for 30min, add 25 mg glycine terminated the reaction.

[0057] Take the above-mentioned reaction solution, use an ultrafiltration membrane with a cut-off molecular weight of 1000, replace it with 0.01mol / L, pH4.0 acetic acid-sodium acetate buffer solution, concentrate to 5ml, and put it on the column for separation. The chromatographic conditions are as follows:

[0058] Chromatography medium: SOURCE 30S

[0059] Column volume: 5ml

[0060] Flow rate: 3.0ml / min

[0061] Column equilibration: Equilibrate 5 times column volume with 0.01mol / L, pH4.0 acetic acid-sodium acetate (initial buffer)

[0062] Sample volume: 5ml

[0063] Elution: first use 3 times the column volume of the initial buffer t...

Embodiment 3

[0077] Salmon calcitonin and polyethylene glycol modified salmon calcitonin (prepared by the method of Example 2) reduce the experimental comparison of rat blood calcium

[0078] With reference to the potency determination method of calcitonin in the two appendix XII O of Chinese Pharmacopoeia in 2005, 90 Wistar female rats with a body weight of 200 ± 15g were healthy and qualified, fasted for 16 hours before the test, drank distilled water freely, and were randomly divided into 3 groups, 30 rats in each of blank, salmon calcitonin and polyethylene glycol modified salmon calcitonin groups. Abdominal subcutaneous injection, blank group injection of normal saline, salmon calcitonin and polyethylene glycol modified salmon calcitonin doses were 0.05 μg / kg (salmon calcitonin in polyethylene glycol modified salmon calcitonin Calculated according to the amount of calcitonin), the administration volume is 0.4ml / 100g. Before the administration and 1, 2, 4, 8, and 12 hours after the ad...

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Abstract

The invention relates to polyethylene glycol-modified calcitonin and preparation method thereof, wherein amino groups on the calcitonin are covalently linked with a long chain of polyethylene glycol derivatives through methylene groups or carbonyl groups. The invention also comprises uses thereof for regulating calcium-phosphorus metabolism and treating various bone-related diseases.

Description

technical field [0001] The present invention relates to a chemically modified calcitonin and its preparation method and application, in particular to a polyethylene glycol modified calcitonin and its preparation method, and also includes its role in regulating calcium-phosphorus metabolism and treating various Uses for bone-related diseases. Background technique [0002] Calcitonin (CT) is a 32-peptide hormone secreted by parafollicular cells (C cells) in the thyroid gland. It was first discovered and proved by Hirsh that it has the function of regulating calcium-phosphorus metabolism. At present, calcitonin has shown good curative effect in the treatment of osteoporosis, Paget's disease, hypercalcemia, hyperparathyroidism, osteitis deformans and bone healing. In addition, CN101035556 discloses the use of calcitonin in bone growth; CN1856321 discloses the use of calcitonin in the treatment and / or prevention of osteoarthritis; CN101180070 discloses the use of calcitonin in t...

Claims

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Application Information

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IPC IPC(8): A61K47/48A61K38/23A61P3/00A61P3/02A61P25/04A61P5/20A61P19/08A61P29/00A61P19/02A61K47/60
Inventor 冯军张喜全薛春佳朱裕辉张来芳赵伟胡博新
Owner SHANGHAI INST OF PHARMA IND
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