What Is Recombinant Human Growth Hormone?

Recombinant human growth hormone (r-hGH) is produced by recombinant DNA technology. Its chemical 2-octane structure includes the amino acid sequence, and the amino acid composition is identified to be identical to the pituitary growth hormone. Recombinant human growth hormone (r-hGH), with molecular formula C990H1529N2630299S7, is a polypeptide chain consisting of 191 amino acids. Disulfide bonds are connected between the 53 position and 165 and 182 and 189, and the relative molecular mass is 22125. ^[1]

Chinese name: Recombinant human growth hormone
Foreign name: recombinant human growthhormone

CAS number: 82030-87-3
Molecular formula: C990H1529N2630299S7
Molecular weight: 22125

Introduction to recombinant human growth hormone compounds

Basic information on recombinant human growth hormone

Chinese name: Recombinant human growth hormone

English name: Somatonorm

English alias: RhghPowderAndInjectionVial; R-Hgh; SOMATONORM; Methionyl human growth hormone; NL-Methionylgrowth hormone (human); NL-Methionyl-somatonorm; SoMatotropin (huMan), NL-Methionyl- (9CI)

CAS number: 82030-87-3

Molecular formula: C ₉₉₀ H ₁₅₂₉ N ₂₆₃ 0 ₂₉₉ S ₇

Molecular weight: 22125

Structural formula: ^[2]

Recombinant Human Growth Hormone Pharmacopoeia Standard

Recombinant human growth hormone source (name), content (potency)

This product is a protein composed of 191 amino acid residues produced by recombinant technology. An appropriate amount of excipient or stabilizer can be added. The amount of recombinant human growth hormone (C990H1528N262O300S7) per 1 mg of protein should be not less than 0.91 mg. (Equivalent to 3.0 units of anhydrous human growth hormone per 1mg.) ^[3]

Recombinant human growth hormone traits

This product is white lyophilized powder. ^[3]

Identification of recombinant human growth hormone

(1) Take an appropriate amount of this product, add 0.05mol / L trimethylolaminomethane buffer solution (adjust the pH value to 7.5 with 1mol / L hydrochloric acid solution) to dissolve and make a solution containing 2mg of recombinant human growth hormone per 1ml. As a test solution; another appropriate amount of a recombinant human growth hormone reference was prepared in the same way as a reference solution. According to the chromatographic condition test under the relevant protein check item, the retention time of the main peak of the test solution should be consistent with the retention time of the main peak of the reference solution. ^[3]

(2) Take the recombinant human growth hormone reference substance, add the buffer solution under identification (1) and dissolve it to make a solution containing 2mg per 1ml. Take 300l of this solution and trypsin solution (take the appropriate amount of trypsin treated with TPCK, Add 20 l of the buffer solution under identification (1) to make 2ml solution per 1 ml) and 300 l of the buffer solution under identification (1), mix well, place in a 37 ° C water bath for 4 hours, and immediately set to -20 Stop the reaction at and use it as the reference solution. Take this product and prepare it according to the reference solution method as the test solution; take the test solution without the trypsin solution as the blank solution; test according to high performance liquid chromatography (Appendix D of the Pharmacopoeia Part II of the 2010 edition) test , Using octylsilane bonded silica as a filler (5 to 10 m); using 0.1% trifluoroacetic acid solution as mobile phase A; 90% acetonitrile solution containing 0.1% trifluoroacetic acid as mobile phase B; flow rate is 1.0 per minute ml; column temperature was 35 ° C; detection wavelength was 214nm. Gradient elution was performed according to the following table. Take the blank solution, the reference solution and the test solution 100 l each, and inject them into the liquid chromatograph, record the chromatogram. After subtracting the blank solution chromatographic peak, the peptide map of the test solution should be consistent with the peptide map of the reference solution. ^[3]

Time (minutes)	Mobile phase A (%)	Mobile phase B (%)
0	100	0
20	80	20
45	75	25
70	50	50
75	20	80

(3) In the chromatogram recorded under the content determination item, the retention time of the main peak of the test solution should be consistent with the retention time of the main peak of the reference solution. ^[3]

(4) Take this product, add water to dissolve and make a solution containing 1mg per 1ml, take 90l of this solution, add 10l of amphoteric electrolyte and 2l of methyl red test solution, mix well, and use it as the test solution; Long hormone reference, prepared in the same way, as a reference solution. Take 10l each of the reference solution and the test solution, add them to the sample well, and test according to the isoelectric focusing electrophoresis method (2010 edition Pharmacopoeia Part II Appendix F sixth method) test, the main band location of the test solution should be the same as the reference solution The main band position is the same. ^[3]

Recombinant human growth hormone test

1 total protein ^[3]

Take an appropriate amount of this product, accurately weigh, add potassium dihydrogen phosphate buffer solution (take 1.70 g of potassium dihydrogen phosphate, dissolve in 400 ml of water, adjust the pH to 7.0 with 0.1 mol / L sodium hydroxide solution, and dilute to 500 ml with water) to dissolve It was quantitatively diluted to a solution with an absorbance of 0.5 to 1.0 at the maximum absorption wavelength (about 280 nm). As a test solution, it was measured according to the ultraviolet-visible spectrophotometry (Appendix IVA of Pharmacopoeia Part II of the 2010 edition), and the maximum absorption wavelength was recorded ( About 280nm) and the absorbance at the wavelength of 320nm, calculate the total protein content in the test solution according to the following formula, in mg.

V AMAX-A320 /0.82

Where V is the volume of the test solution.

2 related proteins ^[3]

Take an appropriate amount of this product, add the buffer under identification (1) to dissolve it and make a solution containing 2mg of recombinant human growth hormone per 1ml as the test solution. Measured according to high performance liquid chromatography (Appendix D, Part Two of the Pharmacopoeia, 2010 edition), using butylsilane bonded silica as a filler (5-10 m); to identify the buffer solution under item (1)-n-propanol (71 : 29) is the mobile phase. Adjust the proportion of n-propanol in the mobile phase so that the retention time of the main peak of recombinant human growth hormone is 30 to 36 minutes; the flow rate is 0.5 ml per minute; the column temperature is 45 ° C; the detection wavelength is 220nm. Take the system suitability test solution (take the recombinant human growth hormone reference substance, add the buffer under identification (1) to make a solution containing 2mg per 1ml, filter and sterilize, and leave it at room temperature for 24 hours) 20l, and inject liquid chromatography Instrument, the separation between the main peak of the recombinant human growth hormone and the deaminated recombinant human growth hormone peak should be not less than 1.0, and the tailing factor of the recombinant human growth hormone peak should be 0.9 to 1.8. Take 20l of the test solution, inject it into the liquid chromatograph, record the chromatogram, and calculate it according to the peak area normalization method. The total relevant protein content should not be greater than 6.0%.

3 polymer proteins ^[3]

Take an appropriate amount of this product and check it according to the method of content determination. Except for other peak areas with retention times greater than the main peak, calculate according to the peak area normalization method. The sum of all peak areas with retention times less than the main peak must not be greater than 4.0%.

4 sterile ^[3]

Take this product and inspect it according to law (Appendix H Membrane Filtration Method in Part Two of the 2010 Pharmacopoeia), and it should meet the requirements.

5 moisture ^[3]

Take this product and measure it according to the moisture determination method (the first method of Appendix M of the Pharmacopoeia of the 2010 edition), the moisture content must not exceed 10.0%.

6Bacterial endotoxin ^[3]

Take this product and check it according to law (Appendix E of Part Two of the Pharmacopoeia of 2010 Edition). The amount of endotoxin in 1mg of recombinant human growth hormone should be less than 5.0EU.

7 bacterial protein residues ^[3]

Take an appropriate amount of this product and check it according to law (Appendix C in Part III of the 2010 Pharmacopoeia). The bacterial protein residue in each 1mg of recombinant human growth hormone must not exceed 10ng.

8Exogenous DNA residue ^[3]

Take an appropriate amount of this product and check it according to law (Appendix B of Part III of the 2010 Pharmacopoeia). The host DNA in each dose of recombinant human growth hormone must not exceed 10ng.

9 biological activity ^[3]

Take this product and check it according to the growth hormone bioassay method (Appendix P in Part II of the Pharmacopoeia, 2010 Edition). The growth hormone in each 1mg protein should not be less than 2.5 units (at least once a year).

Determination of recombinant human growth hormone

It was determined by molecular exclusion chromatography (Appendix V H, Part Two of the Pharmacopoeia, 2010 Edition).

1 Chromatographic conditions and system suitability test ^[3]

A hydrophilic modified silica gel suitable for separating globular proteins with a molecular weight of 5000 to 60,000 is used as a filler; isopropyl alcohol-0.063mol / L phosphate buffer solution (anhydrous disodium phosphate 5.18g, sodium dihydrogen phosphate 3.65g, Add 950ml of water, adjust the pH to 7.0 with phosphoric acid, and make 1000ml with water) (3:97) as the mobile phase; flow rate is 0.6ml per minute; detection wavelength is 214nm. Take human growth hormone monomer and dimer mixture reference, and use 0.025mol / L phosphate buffer (pH 7.0) [take 0.063mol / L phosphate buffer (1 2.5)] to make about 1ml Take 1.0l of the solution, take 20l and inject it into the liquid chromatograph. The resolution of the recombinant human growth hormone monomer peak and the dimer peak should meet the requirements.

2 determination method ^[3]

Take this product, weigh it accurately, dissolve it with 0.025mol / L phosphate buffer solution (pH 7.0) and quantitatively dilute it to make a solution containing about 1.0mg per 1ml. Precisely take 20l into the chromatograph and record the chromatogram; Take the recombinant human growth hormone reference substance and measure it in the same way. Calculate the peak area according to the external standard method.

Effect of recombinant human growth hormone

First, adjust the endocrine system to restore the level close to puberty, balance emotional fluctuations, make breasts re-grow, increase lift, increase flexibility and delay menopause.

Second, to improve the quality of oocytes and ovarian reactivity, increase endometrial receptivity.

Third, it can activate and maintain the normal work of the immune system, improve immunity and disease resistance, and reduce the body's illness.

4. Regenerate the growth of the underlying cells of the skin that has stopped dividing, and the active electrons adsorb water molecules to increase the skin's moisture content, make the facial deep cracks and wrinkles lighter, the complexion brighter, whiter, delicate and elastic, and restore youthful skin texture.

5. Promote muscle growth and prevent muscle atrophy, thereby consuming body fat, especially the central fat of the human body (fat in the intestine, liver, abdominal cavity and subcutaneous fat in the abdomen and waist), so that the arrangement of muscle and fat is restored Youth status.

6. It is the most effective substance for resuscitating sexual function, supporting normal glandular function, increasing the endocrine index of sexual desire, and opening microvessels.

7. The changes in baldness and white hair, through the pituitary gland secrete chemical information, regenerate hair follicle cells, fully increase nutrients, increase enzymes (causes of white hair = enzyme reduction), and directly stimulate dying hair follicles to resume growth.

8. Regulate the central nervous system of the human body, promote the metabolism of brain cells, enhance memory, improve sleep and relieve insomnia, stress and anxiety. It has adjuvant treatment effect on Alzheimer's disease and Alzheimer's disease.

Nine, promote the regeneration of heart, liver, kidney, islet and other viscera cells, restore liver, kidney, islet function and vitality, for hepatitis B virus carriers; size Sanyang; alanine aminotransferase is high; blood glucose is high; urine protein is high , Kidney disease, diabetes have adjuvant effects.

10. It can promote wound healing, regenerate burned skin, reduce local edema and scar formation.

11. Promote the absorption and storage of calcium by bone cells and prevent osteoporosis.

Twelve, can enhance physical strength, make people energetic, work long hours without fatigue.

Thirteen, it can improve people's temper, make people confident and emotional, and make people look younger.

Pharmacological effects of recombinant human growth hormone

Growth-stimulating growth hormone has various physiological effects such as promoting tissue growth and regulating metabolism. Genotropin is exactly the same as the natural pituitary HGH (191 amino acids), while Smatonorm is a 192 amino acid single-chain polypeptide with the same amino acid sequence as the natural pituitary HGH, with only multiple N-termini. One methionine. Recombinant human growth hormone can promote bone growth, increase the number and volume of muscle cells, promote myocardial growth, increase myocardial contractility, promote protein synthesis in the body, increase nitrogen storage in the body, increase fat oxidation and sugar Xenobiotics, and increase the conversion rate of nutrients, regulate the immune system to enhance immune capacity. ^[4]

Pharmacokinetics of recombinant human growth hormone

Recombinant human growth hormone is absorbed faster by subcutaneous or intramuscular injection. The peak time of plasma drug concentration after intramuscular injection is 3 hours, and the plasma half-life of intravenous injection is 20-30 minutes. There is no accumulation effect after long-term administration. ^[4]

Recombinant human growth hormone indication

It is used for dwarfism and dwarf disease caused by insufficient or no secretion of pituitary growth hormone. ^[4]

Contraindications to recombinant human growth hormone

Malignant tumors, diabetics and pregnant women are prohibited. ^[4]

Recombinant human growth hormone considerations

1. For patients with brain tumors caused by a deficiency of growth hormone deficiency (pituitary dwarfism), be used with caution in patients with heart or kidneys.

2. Before using recombinant human growth hormone, an accurate diagnosis should be made. To this end, a detailed examination of pituitary function is required, including special stimulation tests.

3. Long-term continuous use of recombinant human growth hormone to induce antibody response may reduce the efficacy of recombinant human growth hormone. Drugs should be discontinued for appropriate treatment, and the radioimmunoassay method should be used to regularly detect antibody concentrations.

4. Recombinant human growth hormone should be prepared immediately and used immediately, and stored in the refrigerator after dissolution, and used up within 24 hours.

5. For neonates need to be formulated with benzyl alcohol-free water for injection. ^[4]

Recombinant human growth hormone adverse reactions

Allergies, general itching, and redness at the injection site are common. Occasionally vomiting, flatulence, abdominal pain and other gastrointestinal reactions, edema, headache, subcutaneous fat atrophy at the injection site, microscopic examination of hematuria and so on. Elevated ALT and AST, shoulder pain, and periodic limb paralysis are sometimes seen. ^[4]

Dosage of recombinant human growth hormone

Intramuscular and subcutaneous injections: The dosage of recombinant human growth hormone administered varies widely from patient to patient. The general dose is 0.5 to 0.7 U / kg per week or 12 U / m per week based on body surface area, divided into 2 to 4 intramuscular injections or 6 to 7 subcutaneous injections, and the latter dose is usually used. When using, dissolve the powder with 1 ~ 2ml water for injection. It should be rotated slowly and slowly, and the liquid medicine should not be shaken to prevent the active ingredient from being denatured. The pH value of the prepared injection is about 7.8. The injection site should be selected correctly. If severe pain occurs during the injection or blood is drawn through the needle, the site should be changed to avoid repeated injections in the same site. For burns 0.3 0.6U / kg. ^[4]

Recombinant human growth hormone dosage forms and specifications

4U (additional aminocaproic acid, anhydrous sodium phosphate, etc.), with 1 water for injection.

2. Jian Gaoling, injection (powder): 4U.

3. Quick step, injection (powder): 4U. ^[4]

Interaction of recombinant human growth hormone with other drugs

1. Recombinant human growth hormone combined with glucocorticoids, its growth promoting effect can be suppressed.

2. Anabolic steroids, androgens, estrogen, or thyroxine, when used with recombinant human growth hormone, are at risk of accelerating closure of the epiphysis. ^[4]

Recombinant human growth hormone poisoning and measures

Human growth hormones (growth hormones, growth factors) are secreted by the pituitary somatotropin cells, which can promote material metabolism and growth and development; promote the growth of skeletal muscle and cardiomyocytes, strengthen muscle strength, increase bone length; increase fat oxidation; promote protein Synthesis; regulating immune function. It is mainly used in children for growth hormone deficiency. ^[5]

Clinical manifestations of recombinant human growth hormone

1. Fat atrophy at the injection site.

2. Increased blood glucose may occur during medication.

3. Excessive dosage may cause hypothyroidism. ^[5]

Recombinant human growth hormone laboratory test

Blood and stomach tests can prove poison. ^[5]

Recombinant human growth hormone diagnosis

The main points of diagnosis of human growth hormone poisoning are:

1. A clear history of medication.

2. Clinical manifestations of local atrophy, hyperglycemia, hypothyroidism, etc.

3. Drug analysis Blood or gastric juice and poisons. ^[5]

Recombinant human growth hormone measures

The main points of treatment for human growth hormone poisoning are:

1. Clinically pay attention to changing the injection site every time to avoid fat atrophy at the injection site.

2. When there is an increase in blood sugar or hypothyroidism, adjust the dose in time. ^[5]

Recombinant human growth hormone expert review

Twenty cases of primary pituitary dwarfism, two cases of simple GH deficiency type IA, and one case of intrauterine growth arrest were challenged with classic drugs and blood was taken to measure GH during night sleep to confirm GH deficiency. All cases were treated with recombinant hGH. Twenty patients were treated with 191 peptide Genotropin and 3 patients were treated with Somatonorm with 192 peptide. The course of treatment was 12 months (1 patient in the latter group was treated for 6 months). Results After one year, the height increase in the Genotropin group was (13.3 ± 1.8) cm, 2 cases in the Somatonorm group were 16.0 and 16.6 cm, and 1 case was 7.9 cm after 6 months of treatment. 47.8%, 30.4%, and 30.4% of patients experienced a decrease in serum T4, mild hepatomegaly, and transient hematuria during the course of treatment. Recombinant hGH was used to treat idiopathic growth hormone deficiency dwarfism. Somatonorm was administered at a dose of 0.17U / kg, intramuscularly 3 times a week or 0.1U / kg, and subcutaneously once daily. After 12 months of treatment, the height growth rate increased from 1.9 ± 1.0cm per year to 9.2 ± 1.2cm per year. (P is less than 0.05). The results showed that 7 subcutaneous injections at a dose of 0.7U / kg per week were better than 3 intramuscular injections at 0.15U / kg per week. Nine patients had lower T4 after administration, and had no clinical symptoms of hypothyroidism. Two patients had anti-hGH antibodies, but did not hinder growth. No other side effects. It has obvious curative effect and few adverse reactions. ^[6]