INDEX

SERIAL NO.

CONTENTS

PAGE NO.

1.

IDENTIFICATION

2.

CHEMICAL NATURE

1-2

3.

PHYSIOLOGICAL FUNCTIONS

3-5

4.

SYNTHETIC PATH

5.

THERAPEUTIC APPLICATIONS

5-6

6.

MARKETED PRODUCTS

6-8

7. 8.

RESEARCH REFERENCES

8 10-12

IDENTIFICATION:

Human growth hormone is identified on the basis of molecular weight studies and protein analysis(study of amino acid residues). Alternatively its 3D-structure also serves as an identity.

Assay methods for human growth hormone:

GH is assayed by Anion exchange chromatography (HPIEC) Non-denaturing size exclusion chromatography (SEC):

Anion exchange chromatography: In this method the column(TSK DEAE 5PW-1.07.5cm) is run with 50mM potassium phosphate pH 5.5 containing 10%w/v of acetonitrile Temperature of column must be maintained at 450C. Mobile phase flow rate should be optimized at 0.5ml/min. Gradient elution is performed after loading the column with 83 g of protein where the gradient programme lasts for 25min with mobile phase composition ranging from 50-100mM potassium phosphate pH 5.5 with constant 10%w/v acetonitrile. Detected by UV-DETECTOR AT 230nm.

Non-denaturing size exclusion chromatography: This method is useful in detecting the total monomer content present in the formulation.

Column(TSK 2000 SWXL) is run with a mobile phase of 50mM sodium phosphate pH 7.2 containing 150mM sodium chloride . Column load: 50-75g of protein. Flow rate: 1m.l./min Detection: UV-detection at 214 &280 nm.

CHEMICAL NATURE:
Growth hormone is a peptide hormone composing of a single polypeptide chain of 191 amino acids with two disulphide bonds between amino acids 53and 165 ,282 and 189 with a molecular weight of 22Kd. Essentially the structure of human growth hormone has four helices which are required for its interaction with growth hormone receptors.

AMINO ACID SEQUENCE OF HUMAN GROWTH HORMONE

3D STRUCTURE OF HUMAN GROWTH HORMONE

It occurs as two isoforms in human body namely growth hormone1 and growth hormone-2. Growth hormone-1 is the major isoform in the body and is known as somatotropin

PHYSIOLOGICAL FUNCTIONS:
As the name implies growth hormone is responsible for growth and to emphasize it no individual crosses infant stage without growth. To reflect its physiological functions it is called as ANABOLIC hormone. The physiological effects of growth hormone are divided in to two distinct effects: 1. DIRECT EFFECTS 2. INDIRECT EFFECTS

DIRECT EFFECTS: Here the physiological effects produced are due to the direct action of growth hormone on its target receptors.

e.g., lipolysis-Fat cells possess growth hormone receptors through which growth hormone break down triglyceride and thus lipolysis. MECHANISM OF ACTION: HGH binds to human growth receptor hormone(GHR)

Dimerisation of GHR

Activation of JAK2 tyrosine kinase

Activation of MAP kinases, Insulin receptor substrate, DAG, protein kinase c,calcium

Gene expression

INDIRECT EFFECTS: These effects are mediated via Insulin-like Growth Factor-I (IGF-I), a hormone secreted in response to growth hormone on liver and other tissues. e.g., Bone growthIGF-I stimulates

proliferation of cartilage cells Muscle growth-IGF-I stimulates

differentiation and proliferation of myoblasts. Protein synthesis-IGF-I produced from the

growth hormone acting on the liver and other tissues also stimulates the

uptake of aminoacids and thus promote protein

synthesis in muscle and oter tissues.

Thus the overall physiological functions of growth hormone are: Mineralisation of bone by increasing calcium retention Lipolysis(Fat metabolism) Increases protein synthesis(Protein metabolism)

 Growth of all organs except brain Anti-insulin activity i.e.,reduces the uptake of glucose into liver(Carbohydrate metabolism) Promote gluconeogenesis in liver Stimulation of immune system Increases muscle mass via sarcomere hyperplasia

SYNTHETIC PATHWAY:
Human growth hormone is artificially synthesized using r DNA technology. It has 191 amino acids and is synthesized in two fragments which are later bonded together. A c DNA , the human growth hormone (HGH) was synthesized based on 191 amino acid sequence. This Cdna is cleaved using restriction enzyme HAeIII into two fragments, one coding for 1-23 aminoacids of the hormone and the other coding for 24-191 aminoacids . These dna fragments are cloned separately. These fragments are joined together with ligase. The resulting gene is incorporated into the plasmid of E.coli using HAeIII . The recombinant E.coli are cloned to produce large quantities human growth hormone.

THERAPEUTIC APPLICATIONS:
GROWTH HORMONE DEFECIENCY:

 In both peadiatric and adults. In children it increases growth and promote muscle development In adults, it builds muscle, increase energy and reduce body fat TREATMENT OF DWARFISM IN CHILDREN(Since 1950s) AIDS WASTING-Loss of atleast 10% of body weight due to deterioration of muscle by virus. Cachexia-Muscle wasting due to malnutrition(HGH increases muscle density) TREATMENT SYNDROME. GROWTH PROBLEMS CAUSED BY SHORT BOWEL SYNDROME TREATMENT OF SHORT STATURE CAUSED BY PRADER-WILLI SYNDROME TREATMENT FOR IMPROPER GROWTH IN CHILDREN DUE TO RENAL DISEASE BODY BUILDING ENHANEMENT DRUG(increases lean body mass) WEIGHT LOSS DRUG ANTI AGING DRUG TREATMENT FOR CHILDREN SUFFERING WITH RHEUMATOID ARTHRITIS TREATMENT OF OSTEOPOROSIS TREATMENT OF CHILDREN WITH X-LINKED HYPOPHOSPHATEMIC RICKETS OF SHORT STATURE CAUSED BY TURNERS

MARKETED PRODUCTS:

BRANDNAME(MARKETED MANUFACTURER PRODUCT)

BioTropin Genotropin Humatrope Norditropin Nutropin NutropinAQ Protropin Saizen Serostim Serostim Tev-Tropin

BIOTECH GENERAL PFIZER ELI LILLY NOVO NORDISK GENETECH INC. GENETECH INC. GENETECH INC. SERONO SA SERONO SA SERONO SA

Nutropindepot(sustain release)

GATE

GENENTECH ALKERMES

AND

FORMULATIONS:

umatrope 24 mg cartridge cartridge Norditropin flexpro 15 mg/1.5

Norditropin 15 mg/1.5ml Solution 1.5ml Norditropin NordiFlex Pen 15 mg/1.5ml Solution 1 Syringe = 1.5ml Norditropin 15 mg/1.5 ml Norditropin nordiflx 15 mg/1.5 Genotropin 12 mg Solution Humatrope 12 mg cartridge Nutropin 10 mg vial Nutropin aq 20 mg/2ml pen cart Nutropin aq nuspin 20 pen cart Norditropin NordiFlex Pen 10 mg/1.5ml Solution 1 Syringe = 1.5ml Norditropin nordiflex 30 mg/3 Saizen 8.8 mg Solution Vial Saizen Click.Easy 8.8 mg Solution Vial Saizen 8.8 mg viaL Protropin 10 mg Solution Vial Humatrope 6 mg cartridge Genotropin MiniQuick 0.8 mg (7 Cartridges Per Box) Serostim 8.8 mg vial Nutropin 5 mg vial Saizen 5 mg Solution Vial Genotropin 5.8 mg Cartridge Norditropin 5 mg/1.5ml Solution 1.5ml Cartridge Norditropin NordiFlex Pen 5 mg/1.5ml Solution 1 Syringe = 1.5ml Serostim 6 mg vial Genotropin MiniQuick 0.6 mg (7 Cartridges Per Box) Omnitrope 5.8 mg Solution Vial Serostim 5 mg vial Protropin 5 mg Solution Vial Tev-Tropin 5 mg Solution Vial

Tev-tropin 5 mg vial Genotropin MiniQuick 0.4 mg (7 Cartridges Per Box) Serostim 4 mg vial Genotropin MiniQuick 2 mg (7 Cartridges Per Box) Genotropin miniquick 2 mg Genotropin miniquick 1.8 mg Genotropin MiniQuick 1.6 mg (7 Cartridges Per Box) Genotropin miniquick 1.6 mg Genotropin MiniQuick 0.2 mg (7 Cartridges Per Box) Genotropin MiniQuick 1.4 mg (7 Cartridges Per Box) Genotropin miniquick 1.4 mg Genotropin MiniQuick 1.2 mg (7 Cartridges Per Box) Genotropin miniquick 1.2 mg Genotropin MiniQuick 1 mg (7 Cartridges Per Box) Genotropin miniquick 1 mg Genotropin miniquick 0.8 mg Genotropin miniquick 0.6 mg Genotropin miniquick 0.4 mg

RESEARCH:
Research in mice concluded increased levels of growth hormone decreases the life span. RESEARCH ON STRENGH, EXERCISE AND BODY FAT: 88% increase in muscle strength 81% increase in muscle size 72% improvement in body fat loss 81% improvement in exercise tolerance RESEARCH ON SKIN AND HAIR:

 71% improvement in skin texture 68% increase in skin thickness 71% improvement in skin elasticity 51% improvement in wrinkle disappearance 38% improvement towards new hair growth RESEARCH ON HEALING, FLEXIBILITY AND RESISTANCE: 55% improvement in healing old injuries 61% in other injuries 71% in healing capacity 53% in back flexibility 73% in resistance to common illness RESEARCH ON SEXUAL FUNCTION: 75% improvement in sexual potency 62% increase in the penile erection 57% in frequency of night time urination 38% in menstrual cycle regulation RESEARCH ON ENERGY , EMOTIONS AND MEMORY: 84% improvement in energy levels 67% in emotional stability 78% in attitude towards life 62% improvement in memory

REFERENCES:
1. Matsuzaki F, Irie M, Shizume M. (1971) Growth hormone in human fetal gland and cord blood. J Clin EndocrinolMetab33:908-911.

2. Parks JS. (1989) Molecular biology of growth hormone. Acta Paedatr Scand. Suppl 349:127-135

3. Baumann G. (1991) Growth hormone heterogeneity: genes, isohormones, variants and binding proteins. EndocrineRev12:424-449

4.onnebier WJ, Swaab DE. (1973) The influence of anencephaly upon intrauterine growth of the fetus and the placenta, and upon gestational length. J Obstet Gyneco Br Common 80:577-588

5. Leung DW, Spencer SA, Cachianes G, Hammondss RG, Collins C, Henzel WJ, Barnard R, Waters MJ, Wood WI. (1987) Growth hormone receptor and serum binding protein: purification, cloning and expression. Nature 330:537.

6. Godowski PJ, Leung DW, Meacham LR, Galgani JP, Hellmiss R, Keret R, Rotwein PS, Parks JS, Laron Z, Wood WI. (1989) Characterization of the human growth hormone receptor gene and demonstration of a partial gene deletion in two patients with Laron-type dwarfism. Proc Natl Acad SciUS86:8083-8087.

7. Hill DJ, Freemark M, Strain AJ et al. (1988) Placental lactogen and growth hormone receptors in human fetal tissues: relationship to fetal plasma hPL concentrations and fetal growth. J Clin EndocrinolMetab661283-1290.

8. Werther GA, Haynes K, Waters MJ. (1993) GH receptors are expressed on human fetal mesenchymal tissues-identification of mRNA and GH binding protein. J Clin Endocrinol Metab 76:1638-1646

9. Waters MJ, Kaye PL. (2002) The role of growth hormone in fetal development. GH and IGF Res 12:137-146

10.Barinaga M, Bilezikjian LM, Vale WW, Rosenfeld MG, Evans RM (1985) Independent effects of growth hormone releasing factor on growth hormone release and gene transcription. Nature 314:279-281

11. Grossman A, Savage MO, Lytras N (1984) response to analogues of growth hormone releasing hormone in normal subjects and in growth hormone deficient children and young adults. Clin Endocrinol (Oxf) 21:321-331

12.Gaylinn D, Harrison JK, Zysk JR, Lyons Jr CE, Lynch KR, Thorner MO (1993) Molecular cloning and expression of a human anterior pituitary receptor for growth hormone-releasing hormone. Mol Endocrinol 7:77-84

13. Aguiar-Oliveira MH, Gill MS, Barreto de A E, et al. (1999) Effect of severe growth hormone deficiency due to a mutation in the GH-releasing hormone receptor on insulin-like growth factors (IGFs), IGF-binding proteins and ternary complex formation throughout life. J Clin Endocrinol Metab 84:4118-4126.

14. Krulich L, Dhariwal AP, McCann SM (1968) Stimulatory and inhibitory effects of purified hypothalamic extracts on growth hormone release from rat pituitary in vitro. Endocrinology 83:783-790

15. Brazeau P, Vale W, Burgus R, Ling N, Butcher M, Rivier, Guillemin R. (1973) Hypothalamic polypeptide that inhibits the secretion of immunoreactive pituitary growth hormone. Science 179:77-79.

16. Shen LP, Rutter WJ (1984) Sequence of the human somatostatin I gene. Science 224:168-171

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24. http://www.hghhumangrowthhormone.com/hgh_growth_hormone_research.htm