Octreotide acetate is a chemical drug, the main component is octreotide, and the chemical formula is c49h66n10o10s2. White crystal powder. It is mainly suitable for acromegaly, digestive tract diseases and neuroendocrine diseases requiring long-term administration. This product is only for laboratory use, not for other purposes.
Synthesis of octreotide acetate:
Compound (I) was hydrogenated to remove the benzyl protecting group, and then coupled with (II) in dimethylformamide, Dicyclohexylcarbodiimide (DCC) and HOBt to obtain compound (III). (III) the benzyl protecting group is removed by hydrogenation first, and then in dimethylformamide, DCC and HOBt react with (IV) to obtain (V). (5) And hydrazine to convert to (Ⅵ).
In addition, (VII) and (VIII) are coupled to obtain (IX). (IX) after removing the tert butyl protecting group, it is coupled with (VI) to obtain (x). (10) It reacts with trifluoroacetic acid, benzyl sulfide, boron trifluoroacetic acid and trifluoroacetic acid to remove the protective group, and finally oxidizes and cyclizes with air to obtain octreotide.
Relevant experimenters have conducted animal experiments to verify the effect of the finished product. The experiments are as follows: amale PCK rats (n = 24) were randomly assigned to one of the four groups (n = 6 in each group): subcutaneous injection treatment was once every four weeks, octreotide LAR alone was used, pas-lar alone was used, and octreotide and pas-lar alone were used. From 4 to 16 weeks of age, 8mg / kg octreotide and 8mg / kg PAS were administered together, or vehicle (particulate liquid; cont). The carrier contains copolymer microparticles having polylactic acid co glycolic acid (PLGA). Heart rate (HR), diastolic blood pressure (DBP) and systolic blood pressure (SBP) were measured using a tail cuff sphygmomanometer in 4-week-old and 15 week old conscious rats. Twenty four hour urine volume and food consumption were measured using metabolic cages after 15.5 weeks of age. After measuring the body weight, the animals were anesthetized with sodium pentobarbital at the age of 16 weeks, and the kidneys and livers were rapidly removed, resulting in fatal bleeding. Total wet kidney weight and wet liver weight were measured, and blood samples were collected for measuring serum urea nitrogen (sun), aspartate aminotransferase (AST), alanine aminotransferase (ALT), insulin-like growth factor-1 (IGF-1)), glucose, insulin, glucagon, and cortisol.
The finished product of this product is recorded in the Chinese Pharmacopoeia:
The product is d-phenylalanyl-l-cysteinyl-l-phenylalanyl-d-tryptophan-l-lysinyl-l-threoniyl-n - [2-hydroxy-1 - (hydroxymethyl) propyl] - l-cysteamine ring (2 → 7) - disulfide bond acetate, and the content of octreotide (c49h66n10o10s2) should be 95.0% ~ 102.0% based on anhydrous and acetic acid free substances.
character:
1. The product is white or off white lyophilized powder.
2. The product is soluble in water and glacial acetic acid, but insoluble in ether.
3. Specific rotation: take the product, weigh it accurately, make a solution containing 5mg per 1ml with glacial acetic acid, and measure it according to the law (Appendix VI E). The specific rotation shall be - 66.0 ° to - 76.0 ° based on the calculation of anhydrous and acetic acid free substances.
identification:
(1) take about 1mg of the product, add 1ml of water to dissolve it, and add 1ml of alkaline copper tartrate test solution to show blue purple.
(2) in the chromatogram recorded under the content determination item, the retention time of the main peak of the test solution shall be consistent with that of the reference solution.
inspection:
Acidity: take this product and add water to prepare a solution containing 0.125mg per 1ml. Measure it according to the law (Appendix VI h). The pH value should be 5.0-7.0.
Clarity and color of solution: take this product and add water to make a solution containing 0.125mg per 1ml. The solution should be clear and colorless. If it is turbid, it should not be thicker than No. 1 turbidity standard solution (Appendix IX b); If the color is developed, it shall not be deeper than the Yellow No. 1 standard colorimetric solution (Appendix IX a method I).
Amino acid ratio: take an appropriate amount of the product, add 6 mol / L hydrochloric acid solution, replace the air with nitrogen, melt and seal, hydrolyze at 110 ℃ for 24 hours, cool, unseal, vacuum dry, add 0.1mol/l hydrochloric acid solution to dissolve and prepare about 1mg solution per 1ml. Take 100 µ l of this solution, add 10% dithiodipropionic acid solution (take 1g of dithiodipropionic acid, add 10ml of 0.4mol/l borate buffer (ph10.2) to dissolve it, adjust the pH to 10.4 with 40% sodium hydroxide solution), mix well, take 80 ℃ water bath for 10 minutes, cool it, and serve as the test solution. In addition, take cystine, threonine, phenylalanine, lysine and threonine reference materials, and use 0.1mol/l hydrochloric acid solution to prepare a solution with the concentration corresponding to the test sample as the reference solution. Determine according to the amino acid analysis method (Appendix XX, method 1). The content of each amino acid was calculated by the peak area according to the external standard method.
Take 1 / 3 of the total moles of phenylalanine and lysine as 1 to calculate the relative proportion of each amino acid, and the value shall be within the following range:
Cysteine 1.7-2.3, threonine 0.8-1.2, phenylalanine 1.8-2.2, lysine 0.9-1.1; Threonine is present.
The related substances shall be determined according to the method under content determination, and calculated according to the peak area normalization method, except for solvent peak and acetic acid peak, the area of single impurity peak shall not exceed 1.0%, and the total area of each impurity peak shall not exceed 2.0%.
Acetic acid: take an appropriate amount of the product and weigh it accurately. According to the determination method of acetic acid in synthetic polypeptide, the content of acetic acid should be 5.0% ~ 12.0%.
Moisture: take this product and measure it according to the moisture measurement method, and the moisture content shall not exceed 10.0%.
content determination:
Determined by high performance liquid chromatography.
Chromatographic conditions and system adaptability octadecylsilane bonded silica gel was used as filler; Gradient elution was carried out with 10% tetramethylammonium hydroxide solution water acetonitrile (2:88:10) (adjust pH to 5.4 with 10% phosphoric acid solution) as mobile phase A and 10% tetramethylammonium hydroxide solution water acetonitrile (2:38:60) (adjust pH to 5.4 with 10% phosphoric acid solution) as mobile phase B; The detection wavelength was 210nm; Take an appropriate amount of destriothrin octreotide and octreotide reference substance, dissolve them with water and prepare them into a solution containing about 10% of destriothrin octreotide per 1ml μ G and octreotide 0.1mg, and 20 μ L inject into the liquid chromatograph and record the chromatogram. The theoretical plate number shall not be less than 3000 according to the peak of octreotide acetate; The resolution of the peak of octreotide and octreotide should meet the requirements.
Determination method: take an appropriate amount of the product, accurately weigh it, and prepare a solution containing about 0.125mg per 1ml with water. Accurately measure 20 μ L inject into the liquid chromatograph and record the chromatogram; Take an appropriate amount of octreotide acetate reference substance and determine it by the same method. Calculated by peak area according to external standard method.